Comparative performance of electrochemiluminescence immunoassay and EIA for HIV screening in a multiethnic region of China.
The recent approval of 4th generation HIV tests has forced many laboratories to decide whether to shift from 3rd to these tests. There are limited published studies on the comparative evaluation of these two different assays. We compare the performance of fourth-generation electrochemiluminescence immunoassay (ChIA) and third-generation enzyme linked immunosorbent assay (EIA) for human immunodeficiency virus (HIV) screening and gauge whether the shift from EIA to ChIA could be better in a multiethnic region of China. We identified a large number of routine specimens (345,492) using two different assays from Jan 2008 to Aug 2011 in a teaching hospital with high sample throughput. Of the 344,596 specimens with interpretable HIV test results, 526(0.23%) of 228,761 using EIA and 303(0.26%) of 115,835 using ChIA were HIV-1 positive. The false-positive rate of EIA was lower than that of ChIA [0.03% vs. 0.08%, odds ratio 0.33 (95% confidence interval 0.24, 0.45)]. The positive predictive value (PPV) of EIA (89.6%) was significantly higher than that of ChIA (76.1%) (<0.001), reflecting the difference between the two assays. The clinical sensitivities of two assays in this study were 99.64% for EIA and 99.88% for ChIA. Caution is needed before shifting from 3rd to 4th generation HIV tests. Since none of these tests are perfect, different geographic and ethnic area probably require different considerations with regard to HIV testing methods, taking into account the local conditions.


Duration of motherhood has incremental effects on mothers' neural processing of infant vocal cues: a neuroimaging study of women.
The transition to motherhood, and the resultant experience of caregiving, may change the way women respond to affective, infant signals in their environments. Nonhuman animal studies have robustly demonstrated that mothers process both infant and other salient signals differently from nonmothers. Here, we investigated how women with and without young infants respond to vocalisations from infants and adults (both crying and neutral). We examined mothers with infants ranging in age (1-14 months) to examine the effects of duration of maternal experience. Using functional magnetic resonance imaging, we found that mothers showed greater activity than nonmothers to vocalisations from adults or infants in a range of cortical regions implicated in the processing of affective auditory cues. This main effect of maternal status suggests a general difference in vocalisation processing across infant and adult sounds. We found that a longer duration of motherhood, and therefore more experience with an infant, was associated with greater infant-specific activity in key parental brain regions, including the orbitofrontal cortex and amygdala. We suggest that these incremental differences in neural activity in the maternal brain reflect the building of parental capacity over time. This is consistent with conceptualizations of caregiving as a dynamic, learning process in humans.


Effect of Irradiation and Detection of Long-Lived Polyenyl Radicals in Highly Crystalline Ultra-High Molar Mass Polyethylene (UHMMPE) Fibers.
To improve properties such as thermal conductivity, low temperature thermal strain, and creep resistance of ultra-high molar mass polyethylene (UHMMPE) fibers, several researchers have previously undertaken efforts to crosslink these fibers using radiation. Ionizing radiation is commonly used to crosslink bulk UHMMPE in other applications, such as artificial joints. However, UHMMPE fibers differ from bulk UHMMPE in that they have a higher crystallinity (approximately 85% to 90%) and are very highly oriented during manufacturing in which the fibers are stretched 50 to 100 times their original length. Thus, the amorphous fraction of the UHMMPE fibers is also highly ordered. Several experiments were conducted to crosslink the UHMMPE fibers using both low dose rate (gamma) and high dose rate (electron beam) irradiation, all in the absence of oxygen. In all cases, the tensile strength of the fiber was greatly reduced by the irradiation. The oxidation index was also measured for the irradiated samples, and oxidation was not found to play a major role in the reduction of tensile strength in the fibers after irradiation. While this work did not achieve the desired result of improving the mechanical properties of the UHMMPE fiber, a significant result was found. The electron paramagnetic resonance (EPR) spectrum of the UHMMPE fibers was measured shortly after irradiation, and a mixture of allyl and alkyl radicals were detected. The irradiated samples were stored in dark ambient conditions for at least six years, then reexamined using EPR for free radical characterization. Surprisingly, the gamma-irradiated samples showed clear evidence of long-lived polyenyl radicals present in the material. Free radicals are very reactive species that will typically migrate to the surface of the crystalline domain and decay in a relatively short time through various reactions in the amorphous regions. It is hypothesized herein that due to the high crystallinity and large anisotropy of the highly drawn UHMMPE fiber, the polyenyl radicals were trapped in the crystal phase and were unable to migrate and decay. An experiment was performed to test this hypothesis, by which samples of the irradiated fibers were heated to temperatures above first the alpha relaxation and then melting point of polyethylene, and EPR measurements were taken. Results showed that the polyenyl radical signal persisted below the Tm, but was rapidly eliminated upon melting of the crystals. These experiments support the hypothesis that the long-lived polyenyl radicals are trapped in the crystalline region of the polyethylene fibers.


Parenteral nutrition increases susceptibility of ileum to invasion by E coli.
Parenteral nutrition (PN), with the lack of enteral feeding, compromises mucosal immune function and increases the risk of infections. We developed an ex vivo intestinal segment culture (EVISC) model to study the ex vivo effects of PN on susceptibility of the ileum to invasion by extra-intestinal pathogenic Escherichia coli (ExPEC) and on ileal secretion of antimicrobial secretory phospholipase A2 (sPLA2) in response to the pathogen. Study 1: Using mouse (n = 7) ileal tissue, we examined the effects of ileal region (proximal versus distal) and varying ExPEC inoculum concentrations on ex vivo susceptibility to ExPEC invasion and sPLA2 secretion. Study 2: Ten mice were randomized to oral chow or intravenous PN feeding for 5 d (n = 5/group). Using the EVISC model, we compared the susceptibility of ileal tissue to invasion by ExPEC and sPLA2 secretion in response to the pathogen. Study 1: The proximal ileum was more susceptible to invasion (P < 0.0001) and secreted lower amounts of sPLA2 (P = 0.0002) than the distal ileum. Study 2: Ileal tissue from PN-fed animals was more susceptible (approximately 4-fold, P = 0.018) to invasion than those from chow-fed animals. Ileal tissue from PN-fed animals secreted less sPLA2 (P < 0.02) than those from chow-fed animals. The data illustrate EVISC as a reproducible model for studying host-pathogen interactions and the effects of diet on susceptibility to infections. Specifically, the findings support our hypothesis that PN with the lack of enteral feeding decreases mucosal responsiveness to pathogen exposure and provides a plausible mechanism by which PN is associated with increased risk of infectious complication.


Seasonal variations in the progression of myopia in children enrolled in the correction of myopia evaluation trial.
To investigate monthly and seasonal variations in the progression of myopia in children enrolled in the Correction of Myopia Evaluation Trial (COMET). An ethnically diverse cohort of 469 myopic 6- to <12 year-old children was randomized to single vision or progressive addition lenses and followed for 3 years with 98.5% retention. Progression of myopia was measured semiannually by noncycloplegic autorefraction (Nidek ARK 700A) and annually by cycloplegic autorefraction, with the former measurements used in these analyses. The semiannual progression rate was calculated as (change in spherical equivalent refraction between two consecutive semiannual visits/number of days between the two visits) times 182.5. Months were categorized as the midpoint between two visit dates. Seasons were classified as winter (October through March) or summer (April through September). The seasonal difference was tested using a linear mixed model adjusting for demographic variables (age, sex, ethnicity), baseline refraction, and treatment group. Data from 358 children (mean [± SD] age = 9.84 ± 1.27 years; mean myopia = -2.54 ± 0.84 diopters [D]) met the criteria for these analyses. Myopia progression varied systematically by month; it was slower in April through September than in the other months. Mean progression in winter was -0.35 ± 0.34 D and in summer was -0.14 ± 0.32 D, a statistically significant difference (0.21 D, P < 0.0001). The same seasonal pattern was found by age, sex, ethnicity (except in the small sample of Asians), lens type, and clinical center. The slower progression of myopia found in summer is likely related to children's spending more time outdoors and fewer hours in school. The data have clinical implications regarding the time of year and the frequency with which myopic children have eye examinations and the need for precise timing of visits in clinical trials testing new myopia treatments. (ClinicalTrials.gov number, NCT00000113.).


Timing of peak plantar pressure during the stance phase of walking. A study of patients with diabetes mellitus and transmetatarsal amputation.
High plantar pressures contribute to skin breakdown in patients with diabetes mellitus and peripheral neuropathy. The primary purpose of this study was to determine the point during the stance phase of walking that corresponds with forefoot peak plantar pressures. Results indicate that peak plantar pressures occurred at 80% +/- 5% of the stance phase of gait in subjects with diabetes and transmetatarsal amputation, as well as in control subjects. Improved methods of footwear design or walking strategies proposed to patients should focus on the demands of the foot during the late stance phase of walking in order to increase available weightbearing area or to decrease forces, which will minimize plantar pressures and reduce trauma to the neuropathic foot.


Evidence for the Efficacy of an Opioid-Sparing Effect of Intravenous Acetaminophen in the Surgery Patient: A Systematic Review.
Intravenous (IV) acetaminophen is used in multimodal analgesia to reduce the amount and duration of opioid use in the postoperative setting. A systematic review of published randomized controlled trials was conducted to define the opioid-sparing effect of IV acetaminophen in different types of surgeries. Eligible studies included prospective, randomized, double-blind trials of IV acetaminophen compared with either a placebo- or active-treatment group in adult (age ≥18 years) patients undergoing surgery. Trials had to be published in English in a peer-reviewed journal. A total of 44 treatment cohorts included in 37 studies were included in the systematic analysis. Compared with active- or placebo-control treatments, IV acetaminophen produced a statistically significant opioid-sparing effect in 14 of 44 cohorts (32%). An opioid-sparing effect was more common in placebo-controlled comparisons. Of the 28 placebo treatment comparisons, IV acetaminophen produced an opioid-sparing effect in 13 (46%). IV acetaminophen produced an opioid-sparing effect in only 6% (one out of 16) of the active-control groups. Among the 16 active-control groups, opioid consumption was significantly greater with IV acetaminophen than the active comparator in seven cohorts and not significantly different than the active comparator in eight cohorts. The results of this systematic analysis demonstrate that IV acetaminophen is not effective in reducing opioid consumption compared with other adjuvant analgesic agents in the postoperative patient. In patients where other adjuvant analgesic agents are contraindicated, IV acetaminophen may be an option.


Comparative evaluation of marginal adaptation and microleakage of low-shrinking composites after thermocycling and mechanical loading.
The aim of this study was to evaluate and correlate marginal adaptation and microleakage of different low-shrinking composites. Standardized class V cavities (n = 20/group) with occlusal margin in enamel and gingival margin in dentin were restored with low-shrinking silorane-based (Filtek Silorane) or methacrylate-based (Clearfil Majesty Posterior, Grandio, Reflexions XLS, Tetric EvoCeram, Premise, Ceram-X Duo, Aelite LS Posterior) composites and a conventional composite (Filtek Z250). All teeth were stored in water (24 h), thermocycled (5000×), and mechanically loaded (60,000×). Marginal adaptation of epoxy resin replicas was analyzed using scanning electron microscope. Microleakage of the restoration was assessed by dye penetration on sectioned specimens under stereomicroscopy. Data were statistically analyzed with Mann-Whitney U test, with a significance of P < 0.05. Pearson's correlation test was used to evaluate the correlation between results of margin analysis and microleakage. No statistical difference in marginal gap formation was determined between Filtek Silorane and Z250. The lowest microleakage score at dentin margins was recorded for Filtek Silorane, which was not significantly different from that of all other groups. No similar ranking between the results of microleakage at enamel and dentin margins was observed for the materials tested. Marginal adaptation was not correlated to microleakage, except for Filtek Silorane, Grandio, and Filtek Z250. Compared to the conventional composite Filtek Z250, Filtek Silorane demonstrated no improvements with reduced marginal gap formation. Low-shrinking properties of composites appear to have no ability of sealing restoration margins and preventing leakage.


Spectroscopic probes of the quasi-liquid layer on ice.
Raman spectra of the water OH-stretch region were acquired at air-ice and air-water interfaces at a glancing angle, which allowed observation of surface characteristics. The shapes of the OH-stretch bands indicate that the environment at the air-ice interface is different from that at the air-water interface and from that seen in bulk water. Water spectra measured at the surface of dodecane under low relative humidity indicate that this method is sensitive to fewer than 50 monolayers of water. Changes in the local environment of the surfacial water molecules may be induced by the presence of different solute species, giving rise to changes in the shape of the band. Dissolved sodium chloride disrupts hydrogen bonding in liquid water and has the same effect at the air-ice interface. However, when either HCl or HNO(3) is adsorbed from the gas phase onto an ice surface, the opposite effect is seen: Their presence appears to increase the extent of hydrogen bonding at the ice surface. At the same time, shifts in the laser-induced fluorescence spectra of acridine, a fluorescent pH-probe present at the air-ice interface, indicate that dissociation of acids occurs there. These observations suggest that the formation of hydronium ions at the air-ice interface enhances the hydrogen bonding of surfacial water molecules.


Sex partnerships, health, and social risks of young men leaving jail: analyzing data from a randomized controlled trial.
Young men involved in the criminal justice system face disproportionately high rates of sexual risk behavior, drug, use, and violence. Little is known about how their involvement in sex partnerships might mitigate their unique health and social risks. This study explores whether sex partner experience protects against harmful sexual behaviors, drug problems, violence, and recidivism in 16-18-year-old Black and Latino men leaving a US jail. Data were drawn from the Returning Educated African-American and Latino Men to Enriched Neighborhoods (REAL MEN) study conducted between 2003-2007, which tracked 552 adolescents during their time in a New York City jail and 397 of them one year after their release. Logistic regression was used to examine the relationship between sex partner experience and sex behavior, drug use, violence, and recidivism. This study indicates that young men who have long-term sex partners prior to incarceration are less likely to be inconsistent condom users (OR = 0.50, p ≤ 0.01), have sex while high on drugs/alcohol (OR = 0.14, p ≤ 0.001), use marijuana daily (OR = 0.45, p ≤ 0.001), and carry weapons during illegal activity (OR = 0.58, p ≤ 0.05), especially compared with peers who simultaneously are involved with long-term and casual "short-term" sex partners. However, the positive effects of having a long-term sex partner generally do not apply over time - in this case, one year after being released from jail. Aside from sexual partners, factors such as employment and housing stability predict whether these young men will experience positive or negative outcomes post-incarceration. This study highlights the importance and potential benefits of health interventions that engage young Black and Latino men who are involved in the criminal justice system in the US, as well as their sex partners, in health promotion programs. The study also confirms the need for programs that address the employment and housing needs of young men after they leave correctional facilities.


First-stage palliation strategy for univentricular heart disease may impact risk for acute kidney injury.
Norwood palliation for patients with single ventricle heart disease is associated with a significant risk for acute kidney injury, which portends a worse prognosis. We sought to investigate the impact of hybrid stage I palliation (Hybrid) on acute kidney injury risk. This study is a single-centre prospective case-control study of seven consecutive neonates with single ventricle undergoing Hybrid palliation. Levels of serum creatinine and four novel urinary biomarkers, namely neutrophil gelatinase-associated lipocalin, interleukin-18, liver fatty acid-binding protein, and kidney injury molecule-1, were obtained before and after palliation. Acute kidney injury was defined as a ⩾50% increase in serum creatinine within 48 hours after the procedure. Data were compared with a contemporary cohort of 12 neonates with single ventricle who underwent Norwood palliation. Patients who underwent Hybrid were more likely to be high-risk candidates (86 versus 25%, p=0.01) compared with those who underwent Norwood. Despite similar preoperative serum creatinine levels, there was a trend towards higher levels of postoperative peak serum creatinine (0.7 [0.63, 0.94] versus 0.56 [0.47, 0.74], p=0.06) and rate of acute kidney injury (67 versus 29%, p=0.17) in the Norwood cohort. Preoperative neutrophil gelatinase-associated lipocalin (58.4 [11, 86.3] versus 6.3 [5, 16.2], p=0.07) and interleukin-18 (30.6 [9.6, 167.2] versus 6.3 [6.3, 16.4], p=0.03) levels were higher in the Hybrid cohort. Nevertheless, longitudinal mixed-effect models demonstrated Hybrid palliation to be a protective factor against increased postoperative levels of neutrophil gelatinase-associated lipocalin (estimate -1.8 [-3.0, -9.0], p<0.001) and liver fatty acid-binding protein (-49.3 [-89.7, -8.8], p=0.018). In this single-centre case-control study, postoperative acute kidney injury risk did not differ significantly by single ventricle stage I treatment strategy; however, postoperative elevation in novel urinary biomarkers, consistent with subclinical kidney injury, was encountered in the Norwood cohort but not in the Hybrid cohort.


Detecting population structure in a high gene-flow species, Atlantic herring (Clupea harengus): direct, simultaneous evaluation of neutral vs putatively selected loci.
In many marine fish species, genetic population structure is typically weak because populations are large, evolutionarily young and have a high potential for gene flow. We tested whether genetic markers influenced by natural selection are more efficient than the presumed neutral genetic markers to detect population structure in Atlantic herring (Clupea harengus), a migratory pelagic species with large effective population sizes. We compared the spatial and temporal patterns of divergence and statistical power of three traditional genetic marker types, microsatellites, allozymes and mitochondrial DNA, with one microsatellite locus, Cpa112, previously shown to be influenced by divergent selection associated with salinity, and one locus located in the major histocompatibility complex class IIA (MHC-IIA) gene, using the same individuals across analyses. Samples were collected in 2002 and 2003 at two locations in the North Sea, one location in the Skagerrak and one location in the low-saline Baltic Sea. Levels of divergence for putatively neutral markers were generally low, with the exception of single outlier locus/sample combinations; microsatellites were the most statistically powerful markers under neutral expectations. We found no evidence of selection acting on the MHC locus. Cpa112, however, was highly divergent in the Baltic samples. Simulations addressing the statistical power for detecting population divergence showed that when using Cpa112 alone, compared with using eight presumed neutral microsatellite loci, sample sizes could be reduced by up to a tenth while still retaining high statistical power. Our results show that the loci influenced by selection can serve as powerful markers for detecting population structure in high gene-flow marine fish species.


Low prevalence of SV40 in Swiss mesothelioma patients after elimination of false-positive PCR results.
The association of simian virus 40 (SV40) with malignant pleural mesothelioma is currently under debate. In some malignancies of viral aetiology, viral DNA can be detected in the patients' serum or plasma. To characterize the prevalence of SV40 in Swiss mesothelioma patients, we optimized a real-time PCR for quantitative detection of SV40 DNA in plasma, and used a monoclonal antibody for immunohistochemical detection of SV40 in mesothelioma tissue microarrays. Real-time PCR was linear over five orders of magnitude, and sensitive to a single gene copy. Repeat PCR determinations showed excellent reproducibility. However, SV40 status varied for independent DNA isolates of single samples. We noted that SV40 detection rates by PCR were drastically reduced by the implementation of strict room compartmentalization and decontamination procedures. Therefore, we systematically addressed common sources of contamination and found no cross-reactivity with DNA of other polyomaviruses. Contamination during PCR was rare and plasmid contamination was infrequent. SV40 DNA was reproducibly detected in only 4 of 78 (5.1%) plasma samples. SV40 DNA levels were low and not consistently observed in paired plasma and tumour samples from the same patient. Immunohistochemical analysis revealed a weak but reproducible SV40 staining in 16 of 341 (4.7%) mesotheliomas. Our data support the occurrence of non-reproducible SV40 PCR amplifications and underscore the importance of proper sample handling and analysis. SV40 DNA and protein were found at low prevalence (5%) in plasma and tumour tissue, respectively. This suggests that SV40 does not appear to play a major role in the development of mesothelioma.


Impact of Combination Chemotherapy in Peritoneal Mesothelioma Hyperthermic Intraperitoneal Chemotherapy (HIPEC): The RENAPE Study.
The introduction of cytoreductive surgery (CRS) in combination with hyperthermic intraperitoneal chemotherapy (HIPEC) improved the prognosis of selected patients with peritoneal mesothelioma (PM). The objective of our study was to evaluate whether different HIPEC agents were associated with different outcomes in patients with PM. From the RENAPE database, we selected all patients with histology-proven PM who underwent CRS + HIPEC from 1989 to 2014. Inclusion criteria were age ≤ 80 years, performance status ≤ 2, and no extraperitoneal metastases. Overall, 249 patients underwent CRS + HIPEC for PM. The HIPEC regimen included five chemotherapeutic agents (CAs), consisting of cisplatin, doxorubicin, mitomycin-C, oxaliplatin, and irinotecan. When considering all CAs (alone or in combination), there was no significant statistical difference in regard to postoperative overall survival (OS). However, OS was better when using two CAs (group 2 drugs) versus one CA (group 1 drug) (p = 0.03). The different CA regimens were equally distributed between the two groups. This association between OS and HIPEC agent, as well as a trend for better progression-free survival, were both observed in the two-drug group versus the one-drug group (p = 0.009) for patients undergoing complete cytoreductive surgery (CC-0) with an epithelioid subtype. This large study seems to show improved OS when combined CAs, especially with platinum-based regimens, are used for HIPEC in patients with PM, but needs to be confirmed by a randomized controlled trial.


Pertussis: a disease affecting all ages.
Bordetella pertussis is a highly contagious bacterium known to cause pertussis (whooping cough) and is transmitted via airborne droplets. Although childhood vaccination has dramatically reduced reported pertussis cases, the incidence of the disease has increased over the past 20 years, most notably in previously immunized adolescents and adults. Pertussis should be suspected in patients of all ages with cough who meet the clinical criteria for the disease. Diagnostic tests currently approved by the U.S. Food and Drug Administration for pertussis infection have low sensitivity. Regardless of test results, physicians should treat clinically suspected pertussis with antimicrobials and report cases to their state health department. A 14-day erythromycin regimen has been the treatment of choice; however, shorter-course macrolide antibiotics (e.g., azithromycin, clarithromycin) may be as effective with fewer adverse effects and better adherence to therapy. The recently recommended tetanus toxoid, reduced diphtheria toxoid, and acellular pertussis (Tdap) vaccine for adolescents and adults may decrease the incidence of pertussis in infants--the group at the greatest risk of pertussis complications.


Hydro-actuation of hybrid carbon nanotube yarn muscles.
Hybrid hydro-responsive actuators are developed by infiltrating carbon nanotube yarns using poly(3,4-ethylenedioxythiophene):poly(styrenesulfonate). These actuators demonstrate impressive rotation and contraction in response to water due to volumetric expansion of the helical arrangement of carbon nanotubes. The total torsional stroke is 3720 revolutions per m and the simultaneously generated contractive strain reaches 24% at a paddle-to-yarn mass ratio of 350. The contraction output can furthermore be significantly enhanced by constraining the rotational motion and it reaches 68% with an applied stress of 1 MPa. Additionally, hybrid yarns exhibit an approximately linear response to humidity changes and show extra capability of electrical actuation, which, combined with the excellent hydro-actuation performance, endow them with great potential for a variety of applications including artificial muscles, hydro-driven generators, moisture switches and microfluidic mixers.


Antigenic variation in Plasmodium falciparum.
The persistence of the human malaria parasite Plasmodium falciparum during blood stage proliferation in its host depends on the successive expression of variant molecules at the surface of infected erythrocytes. This variation is mediated by the differential control of a family of surface molecules termed PfEMP1 encoded by approximately 60 var genes. Each individual parasite expresses a single var gene at a time, maintaining all other members of the family in a transcriptionally silent state. PfEMP1/var enables parasitized erythrocytes to adhere within the microvasculature, resulting in severe disease. This review highlights key regulatory mechanisms thought to be critical for monoallelic expression of var genes. Antigenic variation is orchestrated by epigenetic factors including monoallelic var transcription at separate spatial domains at the nuclear periphery, differential histone marks on otherwise identical var genes, and var silencing mediated by telomeric heterochromatin. In addition, controversies surrounding var genetic elements in antigenic variation are discussed.


Chemotherapy and radiotherapy for advanced pancreatic cancer.
Pancreatic cancer (PC) is a highly lethal disease with few effective treatment options. Over the past few decades, many anti-cancer therapies have been tested in the locally advanced and metastatic setting, with mixed results. This review attempts to synthesise all the randomised data available to help better inform patient and clinician decision-making when dealing with this difficult disease. To assess the effect of chemotherapy, radiotherapy or both for first-line treatment of advanced pancreatic cancer. Our primary outcome was overall survival, while secondary outcomes include progression-free survival, grade 3/4 adverse events, therapy response and quality of life. We searched for published and unpublished studies in CENTRAL (searched 14 June 2017), Embase (1980 to 14 June 2017), MEDLINE (1946 to 14 June 2017) and CANCERLIT (1999 to 2002) databases. We also handsearched all relevant conference abstracts published up until 14 June 2017. All randomised studies assessing overall survival outcomes in patients with advanced pancreatic ductal adenocarcinoma. Chemotherapy and radiotherapy, alone or in combination, were the eligible treatments. Two review authors independently analysed studies, and a third settled any disputes. We extracted data on overall survival (OS), progression-free survival (PFS), response rates, adverse events (AEs) and quality of life (QoL), and we assessed risk of bias for each study. We included 42 studies addressing chemotherapy in 9463 patients with advanced pancreatic cancer. We did not identify any eligible studies on radiotherapy.We did not find any benefit for chemotherapy over best supportive care. However, two identified studies did not have sufficient data to be included in the analysis, and many of the chemotherapy regimens studied were outdated.Compared to gemcitabine alone, participants receiving 5FU had worse OS (HR 1.69, 95% CI 1.26 to 2.27, moderate-quality evidence), PFS (HR 1.47, 95% CI 1.12 to 1.92) and QoL. On the other hand, two studies showed FOLFIRINOX was better than gemcitabine for OS (HR 0.51 95% CI 0.43 to 0.60, moderate-quality evidence), PFS (HR 0.46, 95% CI 0.38 to 0.57) and response rates (RR 3.38, 95% CI 2.01 to 5.65), but it increased the rate of side effects. The studies evaluating CO-101, ZD9331 and exatecan did not show benefit or harm when compared with gemcitabine alone.Giving gemcitabine at a fixed dose rate improved OS (HR 0.79, 95% CI 0.66 to 0.94, high-quality evidence) but increased the rate of side effects when compared with bolus dosing.When comparing gemcitabine combinations to gemcitabine alone, gemcitabine plus platinum improved PFS (HR 0.80, 95% CI 0.68 to 0.95) and response rates (RR 1.48, 95% CI 1.11 to 1.98) but not OS (HR 0.94, 95% CI 0.81 to 1.08, low-quality evidence). The rate of side effects increased. Gemcitabine plus fluoropyrimidine improved OS (HR 0.88, 95% CI 0.81 to 0.95), PFS (HR 0.79, 95% CI 0.72 to 0.87) and response rates (RR 1.78, 95% CI 1.29 to 2.47, high-quality evidence), but it also increased side effects. Gemcitabine plus topoisomerase inhibitor did not improve survival outcomes but did increase toxicity. One study demonstrated that gemcitabine plus nab-paclitaxel improved OS (HR 0.72, 95% CI 0.62 to 0.84, high-quality evidence), PFS (HR 0.69, 95% CI 0.58 to 0.82) and response rates (RR 3.29, 95% CI 2.24 to 4.84) but increased side effects. Gemcitabine-containing multi-drug combinations (GEMOXEL or cisplatin/epirubicin/5FU/gemcitabine) improved OS (HR 0.55, 95% CI 0.39 to 0.79, low-quality evidence), PFS (HR 0.43, 95% CI 0.30 to 0.62) and QOL.We did not find any survival advantages when comparing 5FU combinations to 5FU alone. Combination chemotherapy has recently overtaken the long-standing gemcitabine as the standard of care. FOLFIRINOX and gemcitabine plus nab-paclitaxel are highly efficacious, but our analysis shows that other combination regimens also offer a benefit. Selection of the most appropriate chemotherapy for individual patients still remains difficult, with clinicopathological stratification remaining elusive. Biomarker development is essential to help rationalise treatment selection for patients.


Language as a multimodal phenomenon: implications for language learning, processing and evolution.
Our understanding of the cognitive and neural underpinnings of language has traditionally been firmly based on spoken Indo-European languages and on language studied as speech or text. However, in face-to-face communication, language is multimodal: speech signals are invariably accompanied by visual information on the face and in manual gestures, and sign languages deploy multiple channels (hands, face and body) in utterance construction. Moreover, the narrow focus on spoken Indo-European languages has entrenched the assumption that language is comprised wholly by an arbitrary system of symbols and rules. However, iconicity (i.e. resemblance between aspects of communicative form and meaning) is also present: speakers use iconic gestures when they speak; many non-Indo-European spoken languages exhibit a substantial amount of iconicity in word forms and, finally, iconicity is the norm, rather than the exception in sign languages. This introduction provides the motivation for taking a multimodal approach to the study of language learning, processing and evolution, and discusses the broad implications of shifting our current dominant approaches and assumptions to encompass multimodal expression in both signed and spoken languages.


Breed differences in plasma calcium, phosphorus and magnesium concentrations of Merino, Dohne Merino and S.A. Mutton Merino sheep with relation to the bent-leg syndrome.
Plasma calcium, phosphorus and magnesium concentrations were determined in ewes and their single and twin lambs from birth for 120 days in Merino, Dohne Merino and South African Mutton Merino sheep. Mineral concentrations as well as the plasma calcium: phosphorus ratio were compared between breeds. Throughout the experimental period, plasma calcium and phosphorus concentrations were higher in both single and twin lambs than in the ewes of all 3 breeds. No breed differences in plasma calcium concentrations were found. Animals of the S.A. Mutton Merino breed had significantly higher (P less than 0.05) plasma phosphorus concentrations than the other 2 breeds. The plasma Ca:P ratio in the Merino and Dohne Merino ram lambs was approximately 1.1:1 and in the S.A. Mutton Merino 0.9:1. This converse plasma Ca:P ratio found in both the S.A. Mutton Merino ewes and lambs is believed to result in an induced plasma ionised calcium deficiency which leads to improper calcification of bone. This is believed to be a contributing factor in the bent-leg syndrome. No difference was recorded in body mass between ram lambs suffering from the bent-leg syndrome and unaffected ram lambs. Plasma magnesium concentrations were not affected by breed or age of the animals.


Integration of data from NIPPON DATA80/90 and National Nutrition Survey in Japan: for cohort studies of representative Japanese on nutrition.
Diet is one of the most important lifestyle factors that a affect healthy life expectancy through onset of various lifestyle-related diseases. Large-scale cohort studies with sufficient baseline nutritional information are scarce. NIPPON DATA80/90 is cohort study of representative Japanese population, and the cohorts also participated in the National Nutrition Survey in Japan (NNSJ) at the baseline. The corresponding datasets could be combined. Individual records of NIPPON DATA and NNSJ were compared and integrated. Intakes of nutrients and food groups for individual participants were calculated by distributing intakes in the each household in NNSJ, considering age and sex of the individuals. The results from an international cooperative epidemiological study (INTERMAP) were utilized to estimate intakes of 75 nutrients for NNSJ80 and 70 nutrients for NNSJ90. Nutrient intakes calculated utilizing INTERMAP data were compared with those in the NNSJ datasets. NIPPON DATA80/90 datasets were enhanced with detailed baseline nutrient intake data (the numbers of participants combined were 10,422 and 8342 for NIPPON DATA80 and 90, respectively). The mean nutrient intakes calculated through utilizing INTEMRAP data and those calculated from the NNSJ datasets were similar, and the calculated values were strongly correlated with those calculated from NNSJ datasets (Pearson's correlation coefficients greater than 0.8 [P < 0.001]). Detailed nutrient intakes (eg, cholesterol, fatty acids, amino acids, and dietary fiber) were complemented. The nutrient intakes calculated from NNSJ datasets for the participants of NIPPON DATA are appropriate as the baseline nutrient intake data. The enhanced cohort datasets are suitable for investigations of baseline dietary habits and the consequent health status.


Mixed infections with Chlamydia and porcine epidemic diarrhea virus - a new in vitro model of chlamydial persistence.
Chlamydiae induce persistent infections, which have been associated with a wide range of chronic diseases in humans and animals. Mixed infections with Chlamydia and porcine epidemic diarrhea virus (PEDV) may result in generation of persistent chlamydial infections. To test this hypothesis, an in vitro model of dual infection with cell culture-adapted PEDV and Chlamydia abortus or Chlamydia pecorum in Vero cells was established. Infected cultures were investigated by immunofluorescence (IF), transmission electron microscopy (TEM) and re-infection experiments. By IF, Chlamydia-infected cells showed normal inclusions after 39 hpi. Dual infections with Chlamydia abortus revealed a heterogenous mix of inclusion types including small inclusions consisting of aberrant bodies (ABs), medium-sized inclusions consisting of ABs and reticulate bodies and normal inclusions. Only aberrant inclusions were observable in dual infection experiments with Chlamydia pecorum and PEDV. TEM examinations of mixed infections with Chlamydia abortus and Chlamydia pecorum revealed aberrant chlamydial inclusions containing reticulate-like, pleomorphic ABs, which were up to 2 microm in diameter. No re-differentiation into elementary bodies (EBs) was detected. In re-infection experiments, co-infected cells produced fewer EBs than monoinfected cells. In the present study we confirm that PEDV co-infection alters the developmental cycle of member species of the family Chlamydiaceae, in a similar manner to other well-described persistence induction methods. Interestingly, this effect appears to be partially species-specific as Chlamydia pecorum appears more sensitive to PEDV co-infection than Chlamydia abortus, as evidenced by TEM and IF observations of a homogenous population of aberrant inclusions in PEDV - Chlamydia pecorum co-infections.


Two fractions of rough endoplasmic reticulum from rat liver. I. Recovery of rapidly sedimenting endoplasmic reticulum in association with mitochondria.
Low-speed centrifugation (640 g) of rat liver homogenates, prepared with a standard ionic medium, yielded a pellet from which a rapidly sedimenting fraction of rough endoplasmic reticulum (RSER) was recovered free of nuclei. This fraction contained 20-25% of cellular RNA and approximately 30% of total glucose-6-phosphatase (ER marker) activity. A major portion of total cytochrome c oxidase (mitochondrial marker) activity was also recovered in this fraction, with the remainder sedimenting between 640 and 6,000 g. Evidence is provided which indicates that RSER may be intimately associated with mitochondria. Complete dissociation of ER from mitochondria in the RSER fraction required very harsh conditions. Sucrose density gradient centrifugation analysis revealed that 95% dissociation could be achieved when the RSER fraction was first resuspended in buffer containing 500 mM KCl and 20 mM EDTA, and subjected to shearing. Excluding KCl, EDTA, or shearing from the procedure resulted in incomplete separation. Both electron microscopy and marker enzyme analysis of mitochondria purified by this procedure indicated that some structural damage and leakage of proteins from matrix and intermembrane compartments had occurred. Nevertheless, when mitochondria from RSER and postnuclear 6,000-g pellet fractions were purified in this way fromanimals injected with [35S]methionine +/- cycloheximide, mitochondria from the postnuclear 6,000-g pellet were found to incorporate approximately two times more cytoplasmically synthesized radioactive protein per milligram mitochondrial protein (or per unit cytochrome c oxidase activity) than did mitochondria from the RSER fraction. Mitochondria-RSER associations, therefore, do not appear to facilitate enhanced incorporation of mitochondrial proteins which are newly synthesized in the cytoplasm.


Comparison of Two Highly Discriminatory Typing Methods to Analyze Aspergillus fumigatus Azole Resistance.
Aspergillus fumigatus molecular typing has become increasingly more important for detecting outbreaks as well as for local and global epidemiological investigations and surveillance. Over the years, many different molecular methods have been described for genotyping this species. Some outstanding approaches are based on microsatellite markers (STRAf assay, which is the current gold standard), or based on sequencing data (TRESP typing improved in this work with a new marker and was renamed TRESPERG). Both methodologies were used to type a collection of 212 A. fumigatus isolates that included 70 azole resistant strains with diverse resistance mechanisms from different geographic locations. Our results showed that both methods are totally reliable for epidemiological investigations showing similar stratification of the A. fumigatus population. STRAf assay offered higher discriminatory power (D = 0.9993) than the TRESPERG typing method (D = 0.9972), but the latter does not require specific equipment or skilled personnel, allowing for a prompt integration into any clinical microbiology laboratory. Among azole resistant isolates, two groups were differentiated considering their resistance mechanisms: cyp51A single point mutations (G54, M220, or G448), and promoter tandem repeat integrations with or without cyp51A modifications (TR34/L98H, TR46/Y121F/A289T, or TR53). The genotypic differences were assessed to explore the population structure as well as the genetic relationship between strains and their azole resistance profile. Genetic cluster analyses suggested that our A. fumigatus population was formed by 6-7 clusters, depending on the methodology. Also, the azole susceptible and resistance population showed different structure and organization. The combination of both methodologies resolved the population structure in a similar way to what has been described in whole-genome sequencing works.


Psychometric evaluation and comparison of three retrospective, multi-item measures of childhood sexual abuse.
To evaluate and compare the psychometric qualities of three retrospective, multi-item measures of childhood sexual abuse (CSA): the CSA Frequency Scale, CSA Count Index, and CSA Multiple Characteristics Index. Two samples of women 20-50 years old who experienced CSA were recruited from a family practice clinic (N=132) and the community (N=19). The measures were designed to represent the three most commonly used approaches for multi-item measurement of past CSA, as determined by a review of literature. Items were derived from a self-administered instrument from a larger study. The CSA Frequency Scale was evaluated for test-retest reliability, internal consistency, and criterion-related validity. The CSA Count and CSA Multiple Characteristics Indexes were evaluated for content specification, indicator specification, indicator collinearity, external validity, criterion-related validity, and test-retest reliability. Criterion-related validity variables were (a) physical and psychosocial symptomatology and (b) depression. Upon evaluation, all three measures met criteria for satisfactory quality, with one exception: external validity for both indexes. When compared across test-retest reliability and criterion-related validity results, none of the measures proved superior. Intraclass correlation coefficients for test-retest reliability ranged from .92 to .94; correlations with the criterion-related validity variables ranged from r=.36 to .41. The results support the continued use of established scales similar to the CSA Frequency scale. In addition, the results suggest further development and standardization of indexes similar to the CSA Count and Multiple Characteristics Indexes is warranted.


'We are still obsessed by this idea of abstinence': A critical analysis of UK news media representations of proposals to introduce drug consumption rooms in Glasgow, UK.
Drug consumptions rooms (DCRs) are a well-established and evidence-based harm reduction response to drug use. Recently, a consortium led by health services in Glasgow, United Kingdom (UK), proposed piloting a DCR. In this article, we examine how the proposals were represented in news media reporting, and the possible effects of such reporting. A quantitative content and qualitative thematic analysis of UK news media (n = 174 articles) representations of the proposals to introduce DCRs in the city of Glasgow, UK, was conducted. Analysis was informed by Bacchi's (2009, 2012, 2017) approach to policy analysis, 'What's the problem represented to be?' Competing representations of the 'problem' of injecting drug use (IDU) were contested by a range of actors with different political visions. The applicability of the 'evidence base', potential benefits of DCRs to both users and the public, and the associated economic costs, were presented in differing ways depending on the underlying assumptions and presumptions of the arguments constructed (e.g. harm reduction vs recovery). As a result, a number of conflicting subject positions were presented that may have implications for the way that people who inject drugs (PWID) see themselves, and how they are viewed and treated by society. Whilst proponents positioned DCRs within a discourse of public health, an underlying rhetoric of abstinence and recovery underpinned the arguments against DCRs. It was this latter discourse that underpinned the UK Government's rejection of the proposals, which the Scottish Government were prevented from overruling within the political constraints of their devolved powers, with the lived effect of people who use drugs (PWUD) being denied access to public health services that mitigate harm. We conclude that attempts to introduce and gain public and political support for harm reduction responses such as DCRs through the news media face challenges within the historical and political context of prohibitionist UK drugs policy.


Inactivation of Btk by insertion of lacZ reveals defects in B cell development only past the pre-B cell stage.
Bruton's tyrosine kinase (Btk) is a cytoplasmic protein kinase that is defective in X-linked agammaglobulinaemia in man and in X-linked immunodeficiency in the mouse. There is controversy regarding the stages of B cell development that are dependent on Btk function. To determine the point in B cell differentiation at which defects in Btk become apparent, we generated a mouse model by inactivating the Btk gene through an in-frame insertion of a lacZ reporter by homologous recombination in embryonic stem cells. The phenomenon of X-chromosome inactivation in Btk+/- heterozygous female mice enabled us to evaluate the competition between B cell progenitors expressing wild-type Btk and those expressing the Btk-/lacZ allele in each successive step of development. Although Btk was already expressed in pro-B cells, the first selective disadvantage only became apparent at the transition from small pre-B cells to immature B cells in the bone marrow. A second differentiation arrest was found during the maturation from IgD(low)IgM(high) to IgD(high)IgM(low) stages in the periphery. Our results show that Btk expression is essential at two distinct differentiation steps, both past the pre-B cell stage.


Iron oxide nanoparticles induce human microvascular endothelial cell permeability through reactive oxygen species production and microtubule remodeling.
Engineered iron nanoparticles are being explored for the development of biomedical applications and many other industry purposes. However, to date little is known concerning the precise mechanisms of translocation of iron nanoparticles into targeted tissues and organs from blood circulation, as well as the underlying implications of potential harmful health effects in human. The confocal microscopy imaging analysis demonstrates that exposure to engineered iron nanoparticles induces an increase in cell permeability in human microvascular endothelial cells. Our studies further reveal iron nanoparticles enhance the permeability through the production of reactive oxygen species (ROS) and the stabilization of microtubules. We also showed Akt/GSK-3beta signaling pathways are involved in iron nanoparticle-induced cell permeability. The inhibition of ROS demonstrate ROS play a major role in regulating Akt/GSK-3beta - mediated cell permeability upon iron nanoparticle exposure. These results provide new insights into the bioreactivity of engineered iron nanoparticles which can inform potential applications in medical imaging or drug delivery. Our results indicate that exposure to iron nanoparticles induces an increase in endothelial cell permeability through ROS oxidative stress-modulated microtubule remodeling. The findings from this study provide new understandings on the effects of nanoparticles on vascular transport of macromolecules and drugs.


Neoadjuvant chemotherapy without radiotherapy for patients with locally advanced rectal cancer. Oncologic outcomes.
the standard treatment for locally advanced rectal cancer is neoadjuvant chemo-radiotherapy, surgery and adjuvant chemotherapy. Only 50% of patients receive the adjuvant treatment due to the surgical complications and toxicity of radiotherapy. Recently, neoadjuvant chemotherapy has been investigated in the locally advanced rectal cancer setting, with the aim of guaranteeing an uninterrupted systemic treatment. The objective of the present study was to assess the safety and efficacy of neoadjuvant chemotherapy in locally advanced rectal cancer. patients treated with neoadjuvant chemotherapy and surgery were identified from a prospective database of patients with rectal cancer (cII-III). The primary outcomes were the assessment of the number of R0 resections, the degree of pathologic response, patterns of recurrence and overall and disease-free survival. Treatment schedule: patients received 6-8 cycles of oxaliplatin and fluoropyrimides based chemotherapy. twenty-seven patients who received neoadjuvant chemotherapy were identified. Twenty-six anterior resections and one Hartmann intervention were performed. An R0 resection was performed in 27 (100%) patients and no involvement of the circumferential margin was observed. Complete pathologic response (ypT0N0) was confirmed in four (14.8%) patients. The median follow-up was 35 months (range: 10-81) and four distant recurrences were recorded. Overall and disease-free survival at five years was 85% and 84.7%, respectively. Twenty-seven (100%) patients received all the cycles of chemotherapy, with a mean of six cycles (range 5-8) per patient. neoadjuvant chemotherapy is a promising alternative in the locally advanced rectal cancer setting and further phase III clinical trials are clearly warranted.


Hundreds of putatively functional small open reading frames in Drosophila.
The relationship between DNA sequence and encoded information is still an unsolved puzzle. The number of protein-coding genes in higher eukaryotes identified by genome projects is lower than was expected, while a considerable amount of putatively non-coding transcription has been detected. Functional small open reading frames (smORFs) are known to exist in several organisms. However, coding sequence detection methods are biased against detecting such very short open reading frames. Thus, a substantial number of non-canonical coding regions encoding short peptides might await characterization. Using bio-informatics methods, we have searched for smORFs of less than 100 amino acids in the putatively non-coding euchromatic DNA of Drosophila melanogaster, and initially identified nearly 600,000 of them. We have studied the pattern of conservation of these smORFs as coding entities between D. melanogaster and Drosophila pseudoobscura, their presence in syntenic and in transcribed regions of the genome, and their ratio of conservative versus non-conservative nucleotide changes. For negative controls, we compared the results with those obtained using random short sequences, while a positive control was provided by smORFs validated by proteomics data. The combination of these analyses led us to postulate the existence of at least 401 functional smORFs in Drosophila, with the possibility that as many as 4,561 such functional smORFs may exist.


Chasing the dragon: the cultural metamorphosis of opium in the United States, 1825-1935.
Many things to many people, opium has played a role in the emergence of several power bases in the United States. In turn, these bases of power have shaped what opium is for the rest of us. Allopathic medicine brought opium and its derivatives under its control around the turn of the century, promulgating "addiction theory" and addiction clinics as part of its rise to preeminence among rival forms of medicine. Opium also played a role in the U.S.'s international economic and imperialistic ascendance. When politicians began to deploy a new discourse on opium early in this century, they were able to appropriate medical rhetoric. As the politics of opium heated up, some doctors were able to exploit the emerging politically inspired discourse to generate a subtly different medical knowledge of opiates and addiction while establishing a new subdiscipline with the political support of lawmakers and state institutions.


Influence of bicycle seat pressure on compression of the perineum: a MRI analysis.
It is a common belief that bicycle seat pressure compresses neurovascular tissues in the perineum and may lead to perineal and penile pathologies in male cyclists. The purpose of this study was to examine the effect bicycle seat pressure has on compression of the perineal cavernous spaces, which house the penile neurovascular tissues. A second purpose was to identify where peak cavernous compression occurs in relation to a bicycle seat. Five males were assessed for compression of the corpus spongiosum and corpora cavernosa with and without bicycle seat pressure using MRI. Seat pressure was applied using a custom loading device designed to replicate seat pressure recorded during stationary bicycling. The distance between a horizontal midline of the seat and the point of peak cavernous space compression was made on sagittal plane images. Diameter measurements of the cavernous spaces at the point of peak compression were made on coronal plane images. Results revealed that peak cavernous space compression occurred below the pubic symphysis, 40.7(+/-11.4) mm anterior to the midline of the seat. Corpus spongiosum values in the unloaded condition were 148% greater than the loaded condition (p=0.008). Similarly, the left and right corpora cavernosa values for the unloaded condition were 252% and 232% greater, respectively, than the loaded condition (p=0.02-0.03). Cavernous spaces that house penile arteries and nerves were compressed maximally below the pubic symphysis. Because this location of peak compression was not different between subjects, it may be a universal impingement zone that limits blood flow and neural activity to and from the penis. This information can be used to optimize seat design and thus reduce perineal injuries.


Guidelines for differential diagnosis of suspected multiple sclerosis.
This article comments on guidelines published by the International Advisory Committee on Clinical Trials in MS in relation to exclusion of alternative diagnoses when multiple sclerosis (MS) is suspected. The guidelines cover recommendations from three working groups, each with a different remit: identification of 'red flags' for alternative diagnoses to MS; design of diagnostic algorithms for common clinically isolated syndromes; and differentiation of MS from other idiopathic inflammatory demyelinating diseases. The major red flags identified in the study provide a useful list of diseases that need to be considered in the differential diagnosis of MS. The definition of clinically isolated syndromes is broadened, and the panel suggests that a stricter definition of encephalopathy should be set for acute disseminated encephalomyelitis. The guidelines demonstrate the potential complexity of MS diagnosis--especially during the early stages of disease--and give useful guidance, especially for the non-MS neurologist, as to how to approach the diagnosis in a systematic way.


Tobacco mutants with a decreased number of functional nia genes compensate by modifying the diurnal regulation of transcription, post-translational modification and turnover of nitrate reductase.
Although nitrate reductase (NR. EC 184.108.40.206) is thought to control the rate of nitrate assimilation, mutants with 40-45% of wildtype (WT) NR activity (NRA) grow as fast as the WT. We have investigated how tobacco (Nicotiana tabacum L. cv. Gatersleben) mutants with one or two instead of four functional nia genes compensate. (i) The nia transcript was higher in the leaves of the mutants. However, the diurnal rhythm was retained in the mutants, with a maximum at the end of the night and a strong decline during the photoperiod. (ii) Nitrate reductase protein and NRA rose to a maximum after 3-4 h light in WT leaves, and then decreased by 50-60% during the second part of the photoperiod and the first part of the night. Leaves of mutants contained 40-60% less NR protein and NRA after 3-4 h illumination, but NR did not decrease during the photoperiod. At the end of the photoperiod the WT and the mutants contained similar levels of NR protein and NRA. (iii) Darkening led to a rapid inactivation of NR in the WT and the mutants. However, in the mutants, this inactivation was reversed after 1-3 h darkness. Calyculin A prevented this reversal. When magnesium was included in the assay to distinguish between the active and inactive forms of NR, mutants contained 50% more activity than the WT during the night. Conversion of [15N]-nitrate to organic compounds in leaves in the first 6 h of the night was 60% faster in the mutants than in the WT. (iv) Growth of WT plants in enhanced carbon dioxide prevented the decline of NRA during the second part of the photoperiod, and led to reactivation of NR in the dark. (v) Increased stability of NR in the light and reversal of dark-inactivation correlated with decreased levels of glutamine in the leaves. When glutamine was supplied to detached leaves it accelerated the breakdown of NR, and led to inactivation of NR, even in the light. (vi) Diurnal changes were also investigated in roots. In the WT, the amount of nia transcript rose to a maximum after 4 h illumination and then gradually decreased. The amplitude of the changes in transcript amount was smaller in roots than in leaves, and there were no diurnal changes in NRA. In mutants, nia transcript levels were high through the photoperiod and the first part of the night. The NRA was 50% lower during the day but rose during the night to an activity almost as high as in the WT. The rate of [15N]-nitrate assimilation in the roots of the mutants resembled that in the WT during the first 6 h of the night. (vii) Diurnal changes were also compared in Nia30(145) transformants with very low NRA, and in nitrate-deficient WT plants. Both sets of plants had similar low growth rates. Nitrate reductase did not show a diurnal rhythm in leaves or roots of Nia30(145), the leaves contained very low glutamine, and NR did not inactivate in the dark. Nitrate-deficient WT plants were watered each day with 0.2 mM nitrate. After watering, there was a small peak of nia transcript NR protein and NRA and, slightly later, a transient increase of glutamine and other amino acids in the leaves. During the night glutamine was low, and NR did not inactivate. In the roots, there was a very marked increase of nitrate, nia transcript and NRA 2-3 h after the daily watering with 0.2 mM nitrate. (viii) It is concluded that WT plants have excess capacity for nitrate assimilation. They only utilise this potential capacity for a short time each day, and then down-regulate nitrate assimilation in response, depending on the conditions, to accumulation of the products of nitrate assimilation or exhaustion of external nitrate. Genotypes with a lower capacity for nitrate assimilation compensate by increasing expression of NR and weakening the feedback regulation, to allow assimilation to continue for a longer period each day.


IRF5 is increased in labouring myometrium and regulates pro-labour mediators.
Preterm birth continues to be the leading cause of neonatal mortality and morbidities that can extend into adult life. Few treatment options stem from our incomplete understanding of the mechanisms of human labour and delivery. Activation of the inflammatory response in gestational tissues by inflammation and/or infection leads to the production of pro-inflammatory and pro-labour mediators, thus preterm birth. Interferon regulatory factor 5 (IRF5) has recently emerged as an important pro-inflammatory transcription factor involved in acute and chronic inflammation. The aims of this study were to determine the expression of IRF5 in human myometrium from labouring and non-labouring women, and whether IRF5 is involved in the genesis of pro-inflammatory and pro-labour mediators induced by pro-inflammatory cytokines or toll-like receptor (TLR) ligands. IRF5 mRNA and protein expression was significantly higher in human myometrium after spontaneous term labour, compared to non-labouring tissues. IRF5 mRNA expression was also significantly higher in primary myometrial cells treated with the pro-inflammatory cytokines IL1B or TNF. In primary myometrial cells, IRF5 knockdown by siRNA (siIRF5) was associated with significantly decreased expression and or secretion of pro-inflammatory cytokines (IL1A, IL6), chemokines (CXCL8, CCL2), adhesion molecules (ICAM1, VCAM1) and contraction-associated proteins PTGS2, PGF2α and PTGFR when in the presence of IL1B, TNF, fsl-1 (TLR2/6 ligand) or flagellin (TLR5 ligand). siIRF5-transfected cells also displayed decreased NF-κB RELA transcriptional activity in the presence of these preterm birth mediators. Our study suggests a novel role for IRF5 in the regulation of the inflammatory response in human myometrium.


Evaluation of pharmacological evidence for forensic purposes.
A thorough analysis of a case that involves a medication that may have caused or contributed to an adverse outcome, or a comparison of two compounds in a patent dispute, requires consideration of many processes that affect the clinical effects of a medication. These include its chemical structure, its pharmacological actions (pharmacodynamics), the pharmaceutical formulation, and its absorption, distribution, metabolism, and excretion (pharmacokinetics). They also include analysis of clinical details, including the diagnosis, the quality of the prescribing decisions, the accuracy of the prescription, dispensing, and administration of medications, and how appropriately the case was managed, including monitoring. A causality assessment should be attempted for both the general case and the particular case. Knowledge of the systems that describe a medication's mechanisms of action (EIDOS) and the dose-relationships and time-courses of adverse outcomes and individual susceptibilities to them (DoTS) can inform several aspects of the analysis. Reports should be written in clear English and should not contain statements that rely on expertise that the expert does not possess.


Comparison of therapeutic and subtherapeutic nasal continuous positive airway pressure for obstructive sleep apnoea: a randomised prospective parallel trial.
Nasal continuous positive airway pressure (NCPAP) is widely used as a treatment for obstructive sleep apnoea. However, to date there are no randomised controlled trials of this therapy against a well-matched control. We undertook a randomised prospective parallel trial of therapeutic NCPAP for obstructive sleep apnoea compared with a control group on subtherapeutic NCPAP. Men with obstructive sleep apnoea, defined as an Epworth sleepiness score of 10 or more and ten or more dips per h of more than 4% SaO2 caused by obstructive sleep apnoea on overnight sleep study, were randomly assigned therapeutic NCPAP or subtherapeutic NCPAP (about 1 cm H2O) for 1 month. Primary outcomes were subjective sleepiness (Epworth sleepiness score), objective sleepiness (maintenance of wakefulness test), and SF-36 questionnaire measurements of self-reported functioning and well-being. 107 men entered the study: 53 received subtherapeutic NCPAP and 54 therapeutic NCPAP. Use of NCPAP by the two treatment groups was similar: 5.4 h (therapeutic) and 4.6 h (subtherapeutic) per night. Subtherapeutic NCPAP did not alter the overnight number of SaO2 dips per h compared with baseline, and thus acted as a control. Therapeutic NCPAP was superior to subtherapeutic NCPAP in all primary outcome measures. The Epworth score was decreased from a median of 15.5 to 7.0 on therapeutic NCPAP, and from 15.0 to 13.0 on subtherapeutic NCPAP (between treatments, p<0.0001). Mean maintenance-of-wakefulness time increased from 22.5 to 32.9 min on therapeutic NCPAP and, not significantly, from 20.0 to 23.5 min on subtherapeutic NCPAP (between treatments p<0.005). Effect sizes for SF-36 measures of energy and vitality were 1.68 (therapeutic) and 0.97 (subtherapeutic) NCPAP (between treatments p<0.0001). For mental summary score, the corresponding values were 1.02 and 0.4 (between treatments p=0.002). Therapeutic NCPAP reduces excessive daytime sleepiness and improves self-reported health status compared with a subtherapeutic control. Compared with controls, the effects of therapeutic NCPAP are large and confirm previous uncontrolled clinical observations and the results of controlled trials that used an oral placebo.


Mechanical factors relate to pain in knee osteoarthritis.
Pain experienced by people with knee osteoarthritis is related to psychosocial factors and damage to articular tissues and/or the pain pathway itself. Mechanical factors have been speculated to trigger this pain experience; yet mechanics have not been identified as a source of pain in this population. The purpose of this study was to identify whether mechanics could explain variance in pain intensity in people with knee osteoarthritis. Data from 53 participants with physician-diagnosed knee osteoarthritis (mean age=68.5 years; standard deviation=8.6 years) were analyzed. Pain intensity was reported on the Western Ontario and McMaster Universities Osteoarthritis Index. Mechanical measures included weight-bearing varus-valgus alignment, body mass index and isokinetic quadriceps torque. Gait analysis captured the range of adduction-abduction angle, range of flexion-extension angle and external knee adduction moment during level walking. Pain intensity was significantly related to the dynamic range of flexion-extension during gait and body mass index. A total of 29% of the variance in pain intensity was explained by mechanical variables. The range of flexion-extension explained 18% of variance in pain intensity. Body mass index added 11% to the model. The knee adduction moment was unrelated to pain intensity. The findings support that mechanical factors are related to knee osteoarthritis pain. Because limitations in flexion-extension range of motion and body size are modifiable factors, future research could examine whether interventions targeting these mechanics would facilitate pain management.


Lead isotope ratios for bullets, forensic evaluation in a Bayesian paradigm.
Forensic science is a discipline concerned with collection, examination and evaluation of physical evidence related to criminal cases. The results from the activities of the forensic scientist may ultimately be presented to the court in such a way that the triers of fact understand the implications of the data. Forensic science has been, and still is, driven by development of new technology, and in the last two decades evaluation of evidence based on logical reasoning and Bayesian statistic has reached some level of general acceptance within the forensic community. Tracing of lead fragments of unknown origin to a given source of ammunition is a task that might be of interest for the Court. Use of data from lead isotope ratios analysis interpreted within a Bayesian framework has shown to be suitable method to guide the Court to draw their conclusion for such task. In this work we have used isotopic composition of lead from small arms projectiles (cal. .22) and developed an approach based on Bayesian statistics and likelihood ratio calculation. The likelihood ratio is a single quantity that provides a measure of the value of evidence that can be used in the deliberation of the court.


Aldosterone and renin in relation to surrogate measures of sympathetic activity: the SABPA study.
Hypertension, particularly in black populations, is often accompanied by augmented sympathetic nervous system activity and suppressed renin activity, indicative of possible blood pressure (BP) dysregulation. The potential role of the interrelationship between the renin-angiotensin-aldosterone system (RAAS) and the sympathetic nervous system in the context of low-renin conditions is unclear. We therefore explored whether surrogate measures of sympathetic activity [noradrenaline, 24-hour heart rate (HR) and percentage (%) dipping of night-time HR] relate to renin, aldosterone and aldosterone-to-renin ratio (ARR) in black and white South Africans. We included black (n = 127) and white (n = 179) males and females aged 20-63 years. We measured 24-hour BP and HR, and calculated night-time dipping. We determined renin and aldosterone levels in plasma and calculated ARR. Noradrenaline and creatinine levels were determined in urine and the noradrenaline:creatinine ratio was calculated. More blacks had low renin levels (80.3%) compared to whites (58.7%) (p < 0.001). In univariate and after multivariate analyses the following significant associations were evident in only the black group: HR dipping was associated negatively with aldosterone level (β = -0.18, p = 0.024) and ARR (β = -0.20, p = 0.011), while 24-hour HR was associated positively with renin level (β = 0.20, p = 0.024). Additionally, there was a borderline significant positive association between noradrenaline:creatinine ratio and aldosterone level (β = 0.19, p = 0.051). The observed associations between surrogate measures of sympathetic nervous system activity and components of the RAAS in the black group suggest that the adverse effects of aldosterone and its ratio to renin on the cardiovascular system may be coupled to the effects of the sympathetic nervous system.


Cryoethics: seeking life after death.
Cryonic suspension is a relatively new technology that offers those who can afford it the chance to be 'frozen' for future revival when they reach the ends of their lives. This paper will examine the ethical status of this technology and whether its use can be justified. Among the arguments against using this technology are: it is 'against nature', and would change the very concept of death; no friends or family of the 'freezee' will be left alive when he is revived; the considerable expense involved for the freezee and the future society that will revive him; the environmental cost of maintaining suspension; those who wish to use cryonics might not live life to the full because they would economize in order to afford suspension; and cryonics could lead to premature euthanasia in order to maximize chances of success. Furthermore, science might not advance enough to ever permit revival, and reanimation might not take place due to socio-political or catastrophic reasons. Arguments advanced by proponents of cryonics include: the potential benefit to society; the ability to cheat death for at least a few more years; the prospect of immortality if revival is successful; and all the associated benefits that delaying or avoiding dying would bring. It emerges that it might be imprudent not to use the technology, given the relatively minor expense involved and the potential payoff. An adapted and more persuasive version of Pascal's Wager is presented and offered as a conclusive argument in favour of utilizing cryonic suspension.


A molecular shift register based on electron transfer.
An electronic shift-register memory at the molecular level is described. The memory elements are based on a chain of electron-transfer molecules and the information is shifted by photoinduced electron-transfer reactions. This device integrates designed electronic molecules onto a very large scale integrated (silicon microelectronic) substrate, providing an example of a "molecular electronic device" that could actually be made. The design requirements for such a device and possible synthetic strategies are discussed. Devices along these lines should have lower energy usage and enhanced storage density.


Optical properties of in situ eye lenses measured with X-ray Talbot interferometry: a novel measure of growth processes.
The lens, a major optical component of the eye, has a gradient refractive index, which is required to provide sufficient refractive power and image quality. The refractive index variations across the lens are dependent on the distributions and concentrations of the varying protein classes. In this study, we present the first measurements of the refractive index in the in situ eye lens from five species using a specially constructed X-ray Talbot grating interferometer. The measurements have been conducted in two planes: the one containing the optic axis (the sagittal plane) and the plane orthogonal to this (the equatorial plane). The results show previously undetected discontinuities and fluctuations in the refractive index profile that vary in different species. These may be linked to growth processes and may be the first optical evidence of discrete developmental stages.


Enhancement of G-CSF-induced stem cell mobilization by antibodies against the beta 2 integrins LFA-1 and Mac-1.
The beta 2 integrins leukocyte function antigen-1 (LFA-1, CD11a) and macrophage antigen-1 (Mac-1, CD11b) have been reported to play a role in the attachment of CD34(+) cells to stromal cells in the bone marrow. When administered prior to interleukin-8 (IL-8), anti-LFA-1 antibodies completely prevent the IL-8-induced mobilization of hematopoietic stem cells in mice. Here, we studied the role of anti-beta 2 integrin antibodies in granulocyte colony-stimulating factor (G-CSF)-induced mobilization of hematopoietic progenitor cells. Administration of antibodies against the alpha chain of LFA-1 or against the alpha chain of Mac-1 followed by daily injections of G-CSF for more than 1 day resulted in a significant enhancement of mobilization of hematopoietic progenitor cells when compared with mobilization induced by G-CSF alone. Also, the number of late (day 28) cobblestone area-forming cells in vitro was significantly higher after mobilization with anti-LFA-1 antibodies followed by 5 microg G-CSF for 5 days than with G-CSF alone (119 +/- 34 days vs 17 +/- 14 days), indicating mobilization of repopulating stem cells. Pretreatment with blocking antibodies to intercellular adhesion molecule-1 (ICAM-1; CD54), a ligand of LFA-1 and Mac-1, did not result in an effect on G-CSF-induced mobilization, suggesting that the enhancing effect required an interaction of the beta 2 integrins and one of their other ligands. Enhancement of mobilization was not observed in LFA-1-deficient (CD11a) mice, indicating that activated cells expressing LFA-1 mediate the synergistic effect, rather than LFA-1-mediated adhesion.


Contemporary Outcomes After Partial Resection of Infected Aortic Grafts.
Aortic graft infection remains a considerable clinical challenge, and it is unclear which variables are associated with adverse outcomes among patients undergoing partial resection. A retrospective, multi-institutional study of patients who underwent partial resection of infected aortic grafts from 2002 to 2014 was performed using a standard database. Baseline demographics, comorbidities, operative, and postoperative variables were recorded. The primary outcome was mortality. Descriptive statistics, Kaplan-Meier (KM) survival analysis, and Cox regression analysis were performed. One hundred fourteen patients at 22 medical centers in 6 countries underwent partial resection of an infected aortic graft. Seventy percent were men with median age 70 years. Ninety-seven percent had a history of open aortic bypass graft: 88 (77%) patients had infected aortobifemoral bypass, 18 (16%) had infected aortobiiliac bypass, and 1 (0.8%) had an infected thoracic graft. Infection was diagnosed at a median 4.3 years post-implant. All patients underwent partial resection followed by either extra-anatomic (47%) or in situ (53%) vascular reconstruction. Median follow-up period was 17 months (IQR 1, 50 months). Thirty-day mortality was 17.5%. The KM-estimated median survival from time of partial resection was 3.6 years. There was no significant survival difference between those undergoing in situ reconstruction or extra-anatomic bypass (P = 0.6). During follow up, 72% of repairs remained patent and 11% of patients underwent major amputation. On univariate Cox regression analysis, Candida infection was associated with increased risk of mortality (HR 2.4; P = 0.01) as well as aortoenteric fistula (HR 1.9, P = 0.03). Resection of a single graft limb only to resection of abdominal (graft main body) infection was associated with decreased risk of mortality (HR 0.57, P = 0.04), as well as those with American Society of Anesthesiologists classification less than 3 (HR 0.35, P = 0.04). Multivariate analysis did not reveal any factors significantly associated with mortality. Persistent early infection was noted in 26% of patients within 30 days postoperatively, and 39% of patients were found to have any post-repair infection during the follow-up period. Two patients (1.8%) were found to have a late reinfection without early persistent postoperative infection. Patients with any post-repair infection were older (67 vs. 60 years, P = 0.01) and less likely to have patent repairs during follow up (59% vs. 32%, P = 0.01). Patients with aortoenteric fistula had a higher rate of any post-repair infection (63% vs. 29%, P < 0.01) CONCLUSION: This large multi-center study suggests that patients who have undergone partial resection of infected aortic grafts may be at high risk of death or post-repair infection, especially older patients with abdominal infection not isolated to a single graft limb, or with Candida infection or aortoenteric fistula. Late reinfection correlated strongly with early persistent postoperative infection, raising concern for occult retained infected graft material.


REM sleep reduction effects on depression syndromes.
Thirty-four endogenous and 18 reactive, depressed patients (hospitalized and nonschizophrenic) were treated in a double-blind, crossover study of the hypothesis that rapid eye movement (REM) sleep reduction (by awakenings) relieves depression. In the endogenous group-but not in the reactive group-subjects deprived of REM sleep for three weeks improved significantly more than control subjects awakened from non-REM sleep. Therapeutic efficacy of REM sleep reduction appeared similar to reported efficacy of imipramine hydrochloride treatment of depression. Eight of nine endogenous patients, unimproved by REM sleep deprivation, did not improve with imipramine. Results suggested (1) that substantial REM sleep reduction has antidepressant activity, and (2) since imipramine and other drug antidepressants reduce REM sleep much more so than nonantidepressant drugs, that an antidepressant "mechanism" of drugs resides in their capacity to substantially reduce REM sleep.


The Diagnostic Value of Capillary Refill Time for Detecting Serious Illness in Children: A Systematic Review and Meta-Analysis.
Capillary refill time (CRT) is widely recommended as part of the routine assessment of unwell children. To determine the diagnostic value of capillary refill time for a range of serious outcomes in children. We searched Medline, Embase and CINAHL from inception to June 2014. We included studies that measured both capillary refill time and a relevant clinical outcome such as mortality, dehydration, meningitis, or other serious illnesses in children aged up to 18 years of age. We screened 1,265 references, of which 24 papers were included in this review. Where sufficient studies were available, we conducted meta-analysis and constructed hierarchical summary ROC curves. Meta-analysis on the relationship between capillary refill time and mortality resulted in sensitivity of 34.6% (95% CI 23.9 to 47.1%), specificity 92.3% (88.6 to 94.8%), positive likelihood ratio 4.49 (3.06 to 6.57), and negative likelihood ratio 0.71 (0.60 to 0.84). Studies of children attending Emergency Departments with vomiting and diarrhea showed that capillary refill time had specificity of 89 to 94% for identifying 5% dehydration, but sensitivity ranged from 0 to 94%. This level of heterogeneity precluded formal meta-analysis of this outcome. Meta-analysis was not possible for other outcomes due to insufficient data, but we found consistently high specificity for a range of outcomes including meningitis, sepsis, admission to hospital, hypoxia, severity of illness and dengue. Our results show that capillary refill time is a specific sign, indicating that it can be used as a "red-flag": children with prolonged capillary refill time have a four-fold risk of dying compared to children with normal capillary refill time. The low sensitivity means that a normal capillary refill time should not reassure clinicians.


The Complete Mitochondrial Genome of Delia antiqua and Its Implications in Dipteran Phylogenetics.
Delia antiqua is a major underground agricultural pest widely distributed in Asia, Europe and North America. In this study, we sequenced and annotated the complete mitochondrial genome of this species, which is the first report of complete mitochondrial genome in the family Anthomyiidae. This genome is a double-stranded circular molecule with a length of 16,141 bp and an A+T content of 78.5%. It contains 37 genes (13 protein-coding genes, 22 tRNAs and 2 rRNAs) and a non-coding A+T rich region or control region. The mitochondrial genome of Delia antiqua presents a clear bias in nucleotide composition with a positive AT-skew and a negative GC-skew. All of the 13 protein-coding genes use ATN as an initiation codon except for the COI gene that starts with ATCA. Most protein-coding genes have complete termination codons but COII and ND5 that have the incomplete termination codon T. This bias is reflected in both codon usage and amino acid composition. The protein-coding genes in the D. antiqua mitochondrial genome prefer to use the codon UUA (Leu). All of the tRNAs have the typical clover-leaf structure, except for tRNASer(AGN) that does not contain the dihydrouridine (DHU) arm like in many other insects. There are 7 mismatches with U-U in the tRNAs. The location and structure of the two rRNAs are conservative and stable when compared with other insects. The control region between 12S rRNA and tRNAIle has the highest A+T content of 93.7% in the D. antiqua mitochondrial genome. The control region includes three kinds of special regions, two highly conserved poly-T stretches, a (TA)n stretch and several G(A)nT structures considered important elements related to replication and transcription. The nucleotide sequences of 13 protein-coding genes are used to construct the phylogenetics of 26 representative Dipteran species. Both maximum likelihood and Bayesian inference analyses suggest a closer relationship of D. antiqua in Anthomyiidae with Calliphoridae, Calliphoridae is a paraphyly, and both Oestroidea and Muscoidea are polyphyletic.


Curcuma oil attenuates accelerated atherosclerosis and macrophage foam-cell formation by modulating genes involved in plaque stability, lipid homeostasis and inflammation.
In the present study, the anti-atherosclerotic effect and the underlying mechanism of curcuma oil (C. oil), a lipophilic fraction from turmeric (Curcuma longa L.), was evaluated in a hamster model of accelerated atherosclerosis and in THP-1 macrophages. Male golden Syrian hamsters were subjected to partial carotid ligation (PCL) or FeCl3-induced arterial oxidative injury (Ox-injury) after 1 week of treatment with a high-cholesterol (HC) diet or HC diet plus C. oil (100 and 300 mg/kg, orally). Hamsters fed with the HC diet were analysed at 1, 3 and 5 weeks following carotid injury. The HC diet plus C. oil-fed group was analysed at 5 weeks. In hyperlipidaemic hamsters with PCL or Ox-injury, C. oil (300 mg/kg) reduced elevated plasma and aortic lipid levels, arterial macrophage accumulation, and stenosis when compared with those subjected to arterial injury alone. Similarly, elevated mRNA transcripts of matrix metalloproteinase-2 (MMP-2), MMP-9, cluster of differentiation 45 (CD45), TNF-α, interferon-γ (IFN-γ), IL-1β and IL-6 were reduced in atherosclerotic arteries, while those of transforming growth factor-β (TGF-β) and IL-10 were increased after the C. oil treatment (300 mg/kg). The treatment with C. oil prevented HC diet- and oxidised LDL (OxLDL)-induced lipid accumulation, decreased the mRNA expression of CD68 and CD36, and increased the mRNA expression of PPARα, LXRα, ABCA1 and ABCG1 in both hyperlipidaemic hamster-derived peritoneal and THP-1 macrophages. The administration of C. oil suppressed the mRNA expression of TNF-α, IL-1β, IL-6 and IFN-γ and increased the expression of TGF-β in peritoneal macrophages. In THP-1 macrophages, C. oil supplementation prevented OxLDL-induced production of TNF-α and IL-1β and increased the levels of TGF-β. The present study shows that C. oil attenuates arterial injury-induced accelerated atherosclerosis, inflammation and macrophage foam-cell formation.


Nondamaging Retinal Laser Therapy: Rationale and Applications to the Macula.
Retinal photocoagulation and nondamaging laser therapy are used for treatment of macular disorders, without understanding of the response mechanism and with no rationale for dosimetry. To establish a proper titration algorithm, we measured the range of tissue response and damage threshold. We then evaluated safety and efficacy of nondamaging retinal therapy (NRT) based on this algorithm for chronic central serous chorioretinopathy (CSCR) and macular telangiectasia (MacTel). Retinal response to laser treatment below damage threshold was assessed in pigmented rabbits by expression of the heat shock protein HSP70 and glial fibrillary acidic protein (GFAP). Energy was adjusted relative to visible titration using the Endpoint Management (EpM) algorithm. In clinical studies, 21 eyes with CSCR and 10 eyes with MacTel were treated at 30% EpM energy with high spot density (0.25-diameter spacing). Visual acuity, retinal and choroidal thickness, and subretinal fluid were monitored for 1 year. At 25% EpM energy and higher, HSP70 was expressed acutely in RPE, and GFAP upregulation in Müller cells was observed at 1 month. Damage appeared starting at 40% setting. Subretinal fluid resolved completely in 81% and partially in 19% of the CSCR patients, and visual acuity improved by 12 ± 3 letters. Lacunae in the majority of MacTel patients decreased while preserving the retinal thickness, and vision improved by 10 letters. Heat shock protein expression in response to hyperthermia helps define the therapeutic window for NRT. Lack of tissue damage enables high-density treatment to boost clinical efficacy, therapy in the fovea, and retreatments to manage chronic diseases.


Role of polymorphic and monomorphic human arylamine N-acetyltransferases in determining sulfamethoxazole metabolism.
Sulfonamides are associated with a variety of adverse reactions, some of which have been linked with the classical acetylator phenotypes. Although the slow acetylator phenotype has been identified as a risk factor for hypersensitivity reactions to sulfamethoxazole (SMX), the disposition of this compound appears not to be affected by the acetylation polymorphism in vivo in humans. We therefore investigated the acetylation of SMX by monomorphic (NAT1) and polymorphic (NAT2) arylamine N-acetyltransferases in humans with the objective of determining their role in the metabolism of SMX. SMX was acetylated by both NAT1 and NAT2. Km values determined in hepatic cytosol for NAT1- and NAT2-mediated acetylation of SMX were 1.2 mM and approximately 5 mM, respectively, at an acetyl coenzyme A concentration of 100 microM. Mononuclear leukocytes, which contain only NAT1, had a Km value of 1.2 mM. Km values determined with recombinant NAT1 and NAT2 proteins expressed in Escherichia coli were 1.5 mM and approximately 15 mM, respectively. The higher affinity of NAT1 for SMX indicates that acetylation by this enzyme will predominate at therapeutic plasma concentrations, in agreement with the observed in vivo monomorphic acetylation of SMX. NAT1 may be the primary determinant of SMX systemic metabolic clearance. However, in the hepatocyte NAT2 variation may be an important competitive pathway which influences the extent of oxidative metabolism of SMX to its reactive hydroxylamine metabolite. Therefore, variation in both monomorphic and polymorphic N-acetyltransferases may play a role in determining susceptibility to sulfamethoxazole toxicity.


Implicating genes, pleiotropy, and sexual dimorphism at blood lipid loci through multi-ancestry meta-analysis.
Genetic variants within nearly 1000 loci are known to contribute to modulation of blood lipid levels. However, the biological pathways underlying these associations are frequently unknown, limiting understanding of these findings and hindering downstream translational efforts such as drug target discovery. To expand our understanding of the underlying biological pathways and mechanisms controlling blood lipid levels, we leverage a large multi-ancestry meta-analysis (N = 1,654,960) of blood lipids to prioritize putative causal genes for 2286 lipid associations using six gene prediction approaches. Using phenome-wide association (PheWAS) scans, we identify relationships of genetically predicted lipid levels to other diseases and conditions. We confirm known pleiotropic associations with cardiovascular phenotypes and determine novel associations, notably with cholelithiasis risk. We perform sex-stratified GWAS meta-analysis of lipid levels and show that 3-5% of autosomal lipid-associated loci demonstrate sex-biased effects. Finally, we report 21 novel lipid loci identified on the X chromosome. Many of the sex-biased autosomal and X chromosome lipid loci show pleiotropic associations with sex hormones, emphasizing the role of hormone regulation in lipid metabolism. Taken together, our findings provide insights into the biological mechanisms through which associated variants lead to altered lipid levels and potentially cardiovascular disease risk.


Quantification of chemotherapeutic target gene mRNA expression in human breast cancer biopsies: comparison of real-time reverse transcription-PCR vs. relative quantification reverse transcription-PCR utilizing DNA sequencer analysis of PCR products.
The solid tumor mRNA expression of genes related to the mechanism of action of certain antineoplastic agents is often predictive of clinical efficacy. We report here on the development of a rapid and practical real-time RT-PCR method to quantify genetic expression in solid tumors. The genes examined are related to the intracellular pharmacology of gemcitabine and cisplatin, two drugs that are used in the treatment of several types of advanced cancer. We evaluated target gene mRNA levels from breast tumor samples using two quantitative RT-PCR methods: 1) an improved relative RT-PCR method using fluorescence-labeled primers, automated PCR set up, and GeneScan analysis software; and 2) real-time RT-PCR with redesigned primers using an ABI 7900HT instrument, with additional postprocessing of the data to adjust for efficiency differences across the target genes. Using these methods, we quantified mRNA expression levels of deoxycytidine kinase (dCK), deoxycytidylate deaminase (dCDA), the M1 and M2 subunits of ribonucleotide reductase (RRM1, RRM2), and excision cross complementation group 1 (ERCC1) in 35 human "fresh" frozen breast cancer biopsies. While both assay methods were substantially more rapid than traditional RT-PCR, real-time RT-PCR appeared to be superior to the amplification end-point measurement in terms of precision and high throughput, even when a DNA sequencer was used to assess fluorescence-labeled PCR products. This reproducible, highly sensitive real-time RT-PCR method for the detection and quantification of the mRNAs for dCK, dCDA, RRM1, RRM2, and ERCC1 in human breast cancer biopsies appears to be more informative and less time-consuming than either classical radioisotope-dependent RT-PCR or the technique utilizing GeneScan analysis described herein. By allowing the measurement of intratumoral target gene expression, these new methods may prove useful in predicting the clinical utility of gemcitabine- and platinum-containing chemotherapy programs in patients with solid tumors.


Comparison of outpatient versus inpatient transurethral prostate resection for benign prostatic hyperplasia: Comparative, prospective bi-centre study.
We compare the symptomatic relief with urodynamic parameter change and operative safety of the outpatient transurethral resection in saline (TURIS-V) technique with inpatient transurethral resection of the prostate (TURP) for the management of benign prostatic hyperplasia (BPH). This prospective cohort comparison study enrolled patients who needed BPH surgery. Between January 2010 and June 2011, outpatient TURIS-V was performed at 1 centre and the results of the treatment were compared with inpatient TURP performed at a separate hospital. Preoperative characteristics, including prostate volume, were similar in both groups. Perioperative data and any treatment complications were recorded. The analysis compared postoperative outcomes, including a 6-month postoperative International Prostate Symptom Score (IPSS), a quality of life (QoL) evaluation and a record of any changes in uroflowmetry findings, between the 2 groups. In the TURIS-V patient group, 75 patients agreed to be in the study. Of these, 69 ultimately complete the study. In the TURP group, 76 patients agreed and 71 of these completed the study. Both study groups were well-matched for age, IPSS, QoL and uroflowmetry findings. The TURIS-V group experienced both shorter operation times (54.6 vs. 74.8 minutes) and shorter catheterization times (2.2 vs. 4.2 days) when compared to the TURP group. There were comparable improvements in the 6-month postoperative IPSS, QoL, and uroflowmetry findings between the 2 groups. There were also equally low incidence rates of procedural complications. Both TURIS-V and TURP relieve lower urinary tract symptoms in a similar way, with great efficacy and safety. Overall, TURIS-V had shorter operative and catheterization times compared to TURP. Notwithstanding the paper's limitations (non- randomized cohort comparison with possible selection or surgeon bias and small heterogeneous sample size), TURIS-V can be performed safely even in an outpatient setting.


Does stable sitting influence upper limb function in children with cerebral palsy?
Seyhan K, Kerem-Günel M. Does stable sitting influence upper limb function in children with cerebral palsy? Turk J Pediatr 2019; 61: 79-84. Bilateral spastic cerebral palsy (BSCP) patients frequently need to use various sitting devices for body control and function. The aim of this study was to investigate whether the use of a belt to position the pelvis in an adjustable chair would affect upper limb function in preschool children with BSCP. Fortyone children with BSCP [mean age 44 ±11, range 18-60 months] classified according to the Gross Motor Function Classification System (GMFCS), as level III (n=21) and level IV (n=20) were fitted with a hip-positioning belt. Upper limb functions were assessed by Quality of Upper Extremity Skills Test (QUEST). The median scores of the following upper extremity functions increased significantly by wearing the hip positioning belt: dissociated movements, grasping, weight bearing and protective extension. The total QUEST score increased from 56.7 (±46.3) to 66.1 (±39.2) (p < 0.001). The portable and adaptable hip-positioning belt may be used in daily life to improve upper limb activity in preschool children with moderate to severe BSCP.