Stillbirth, hypertensive disorders of pregnancy, and placental pathology.
Stillbirth, preeclampsia, and gestational hypertension (PE/GH) have similar clinical risk factors and redundant placental pathology. We aim to discern if stillbirth with PE/GH has a particular phenotype by comparing stillbirths with and without PE/GH. Secondary analysis of the Stillbirth Collaborative Research Network, a population-based cohort study of all stillbirths and a sample of live births from 2006 to 2008 in five catchment areas. We compared placental pathology between stillbirths and with and without PE/GH, stratified by term or preterm. We also compared placental pathology between stillbirths and live births with PE/GH. 79/518 stillbirths and 140/1200 live births had PE/GH. Amongst preterm stillbirths, there was higher feto-placental ratio in PE/GH pregnancies (OR 1.24 [1.11, 1.37] per unit increase), and there were more parenchymal infarctions (OR 5.77 [3.18, 10.47]). Among PE/GH pregnancies, stillbirths had increased maternal and fetal vascular lesions, including retroplacental hematoma, parenchymal infarction, fibrin deposition, fetal vascular thrombi, and avascular villi. Stillbirth pregnancies are overwhelmingly associated with placental lesions. Parenchymal infarctions are more common in PE/GH preterm stillbirths, but there is significant overlap in lesions found in stillbirths and PE/GH.


Prevalence of latex allergy in spina bifida: genetic and environmental risk factors.
To evaluate the prevalence of latex allergy in a population of children with spina bifida (SB) and to assess the role of early exposure to latex products and others risk factors. SB is related with an higher incidence of latex allergic reactions. These patients received repeated surgical procedures, implant of latex-containing materials and catheterization. Eighty consecutive subjects affected with SB besides answering a questionnaire, underwent a skin-prick test (SPT) to latex and the determination of the specific serum IgE (RAST CAP) to latex. 40% (32/80) of the patients showed a latex sensitization with specific IgE > 0.7 kU/I but only twelve of the 32 sensitized patients (40%) suffered from clinical reactions to latex (urticaria, conjunctivitis, angioedema, rhinitis, bronchial asthma). Number of surgical procedures, but particularly early exposure to latex and familiarity for allergy are correlated with latex allergy (p < 0.01). Latex allergy in SB children is multifactorial situation related with a disease-associated propensity for latex sensitization, early exposure and number of surgical procedures. Prophylactic measures to avoid the exposure, not only in the sanitary environment, through the institution of latex-safe routes and every day, prevent potentially serious allergic reactions.


Implementation of the Quebec mental health reform (2005-2015).
This study evaluates implementation of the Quebec Mental Health (MH) Reform (2005-2015) which aimed to improve accessibility, quality and continuity of care by developing primary care and optimizing integrated service networks. Implementation of MH primary care teams, clinical strategies for consolidating primary care, integration strategies to improve collaboration between primary care and specialized services, and facilitators and barriers related to these measures were examined. Eleven Quebec MH service networks provided the study setting. Networks were identified in consultation with 20 key MH decision makers and selected based on variation in services offered, integration strategies, best practices, and geographic criteria. Data collection included: primary documents, structured questionnaires completed by 25 managers from MH primary care teams and 16 respondent-psychiatrists working in shared-care, and semi-structured interviews with 102 network stakeholders involved in the reform. The study employed a mixed method approach, triangulating the three data sources across networks. While implementation was not fully achieved in most networks, the Quebec reform succeeded in improving primary care services with the creation of adult primary care teams, and one-stop services which increased access to care, mainly for clients with common MH disorders. In terms of clinical strategies implemented, the functions provided by respondent-psychiatrists had a greater impact on the MH primary care teams than on general practitioners (GPs) in medical clinics; whereas the implementation of best practices were indirect outcomes of another reform developed simultaneously by the Quebec substance use disorders program. The main integration strategies used for increasing continuity of care and collaboration between primary care and specialized services were those involving fewer formal procedures such as referrals between teams and organizations. The lack of operational mechanisms and protocols governing new services and structures were important barriers to primary care consolidation and service integration, as was the lack of interest and involvement of most GPs in MH. Successful and sustained healthcare reform requires attention to process and outcomes as well as structural change or service reorganization. Six recommendations for more successful implementation of the Quebec MH Reform are provided, with implications for healthcare reform internationally.


Female exposure to phenols and phthalates and time to pregnancy: the Maternal-Infant Research on Environmental Chemicals (MIREC) Study.
To assess the potential effect of bisphenol A (BPA), triclosan (TCS), and phthalates on women's fecundity, as measured by time to pregnancy (TTP). Pregnancy-based retrospective TTP study. Not applicable. A total of 2,001 women during the first trimester of pregnancy recruited between 2008 and 2011 (the Maternal-Infant Research on Environmental Chemicals (MIREC) Study), with 1,742 women included in the BPA analysis, 1,699 in the TCS analysis, and 1,597 in the phthalates analysis. None. Fecundability odds ratios (FORs) estimated using the Cox model modified for discrete time data. The BPA concentrations were not statistically significantly associated with diminished fecundity either in crude or adjusted models. Women in the highest quartile of TCS (>72 ng/mL) had evidence of decreased fecundity (FOR 0.84; 95% confidence interval, 0.72-0.97) compared with the three lower quartiles as the reference group. Exposure to phthalates was suggestive of a shorter TTP, as indicated by FORs greater than 1, although the 95% confidence interval always included 1. Elevated TCS exposure may be associated with diminished fecundity. BPA and phthalates showed no negative impact; on the contrary, some phthalates might be associated with a shorter time to pregnancy. A major limitation of the study was that only one measurement of exposure was available for each woman after conception. Further research is necessary to test these findings.


Risk factors for sexual victimization among male and female homeless and runaway youth.
Risk factors associated with the likelihood of being sexually victimized by a stranger or friend/acquaintance since being on the street was examined among 372 homeless and runaway youth. Young people were interviewed on the streets and in shelters by outreach workers using a systematic sampling strategy. Youth who engaged in more high-risk behaviors were expected to be at greater risk for sexual victimization by both known and unknown assailants. Results indicated that for females, running from home for the first time at an earlier age was associated with sexual victimization by both a stranger and friend/acquaintance. However, engaging in deviant subsistence strategies, survival sex, and grooming predicted being sexually victimized by a friend/acquaintance. For males, survival sex and grooming predicted stranger sexual victimization, whereas sexual orientation was associated with sexual victimization by a friend/acquaintance. Overall, 35% of the sample had been sexually victimized.


FliN is a major structural protein of the C-ring in the Salmonella typhimurium flagellar basal body.
The Salmonella typhimurium FliN protein has been proposed to form a mutually interacting complex with FliG and FliM, the switch complex, that is required for flagellar morphogenesis and function. We have used affinity chromatography for purification of extended flagellar basal bodies sufficient for quantitative analysis of their protein composition. The belled, extended structure is predominantly comprised of the switch complex proteins; with FliN present in the most copies (111 +/- 13). This explains why single, missense fliN, fliG or fliM mutations, found in many non-motile strains, can alter the belled morphology. Cell lysates from these strains contained the wild-type complement of FliG, FliM and FliN; but the basal bodies lacked the outer, cytoplasmic(C)-ring of the bell and were separated by sedimentation from FliM and FliN. The amount of FliG present in basal bodies from wild-type and one such mutant, FliN100LP, was comparable. These data show that: (1) the mutations define a FliG and FliMFliN multiple contact interface important for motility. (2) FliG is responsible for the increased size of the membrane-embedded MS-ring complex of belled relative to acid-treated basal bodies. (3) FliN, together with FliM, account for most of the C-ring. As a major component of the C-ring, FliN is distinct from the other proteins implicated in axial flagellar protein export. Inner, cytoplasmic rod basal substructure, seen by negative-stain and quick-freeze replica electron microscopy, may gate such export. Lack of connectivity between the cytoplasmic rod and ring substructures places contacts between FliG and FliMFliN at the periphery of the basal body, proximal to the flagellar intramembrane ring particles. This topology is consistent with models where torque results from interaction of circumferential arrays of the switch complex proteins with the ring particles.


Three-Dimensional Electrohydrodynamic Printing and Spinning of Flexible Composite Structures for Oral Multidrug Forms.
A simple method to rapidly customize and to also mass produce oral dosage forms is arguably a current bottleneck in the development of modern personalized medicine. Specifically, delayed-release mechanisms with well-controlled dosage profiles for combinations of traditional Chinese herbal extracts and Western medications are not well established. Herein, we demonstrate a novel multidrug-loaded membrane sandwich with structures infused with ibuprofen (IBU) and Ganoderma lucidum polysaccharide (GLP) using three-dimensional electrohydrodynamic printing and electrospinning techniques. The resulting flexible membrane consists of microscaled, multilayered cellulose acetate (CA) membranes loaded with IBU in the shape of either concentric squares or circles, as the top and bottom layers of a sandwich structure. In between the CA-IBU layers are randomly electrospun polyvinyl pyrrolidone (PVP) layers loaded with GLP. The complete fibrous membrane sandwich can be folded and embedded into a 0-size capsule to achieve oral compliance. Simulated in vitro testing of gastric and intestinal fluids demonstrated a triphasic release profile. There was an immediate release of GLP after gastric juices dissolved the capsule shell and the PVP, followed by the short-term release of 60% of the IBU within an hour afterward, and the remaining IBU was released in a sustained manner following a Fickian diffusion profile. In summary, this multidrug (both hydrophilic and/or hydrophobic) oral system with precision-designed structures should enable personalized therapeutic dosing.


Associations of parental, birth, and early life characteristics with systolic blood pressure at 5 years of age: findings from the Mater-University study of pregnancy and its outcomes.
We examined the associations of a range of parental and early life characteristics with systolic blood pressure at 5 years of age. Information from 3864 children who were followed up prospectively from their mother's first antenatal clinic assessment was used. Maternal age, body mass index, and smoking during pregnancy were all positively associated with offspring systolic blood pressure at 5 years of age. The systolic blood pressure of children whose mothers had smoked throughout pregnancy was on average 0.92 mm Hg (95% CI 0.17 to 1.68) greater than that of children whose mothers had never smoked, after full adjustment. Children who had been breast fed until at least 6 months had lower systolic blood pressure than those who were breast fed for a shorter duration. Paternal body mass index and child's weight, height, and body mass index were all positively associated with blood pressure at age 5. Because childhood blood pressure tracks into adulthood, interventions aimed at early life risk factors, such as quitting smoking during pregnancy, breast feeding, and prevention of obesity in all family members, may be important for reducing the population distribution of blood pressure and thus cardiovascular disease risk.


Pro-inflammatory activation of primary microglia and macrophages increases 18 kDa translocator protein expression in rodents but not humans.
The 18kDa Translocator Protein (TSPO) is the most commonly used tissue-specific marker of inflammation in positron emission tomography (PET) studies. It is expressed in myeloid cells such as microglia and macrophages, and in rodent myeloid cells expression increases with cellular activation. We assessed the effect of myeloid cell activation on TSPO gene expression in both primary human and rodent microglia and macrophages in vitro, and also measured TSPO radioligand binding with 3H-PBR28 in primary human macrophages. As observed previously, we found that TSPO expression increases (∼9-fold) in rodent-derived macrophages and microglia upon pro-inflammatory stimulation. However, TSPO expression does not increase with classical pro-inflammatory activation in primary human microglia (fold change 0.85 [95% CI 0.58-1.12], p = 0.47). In contrast, pro-inflammatory activation of human monocyte-derived macrophages is associated with a reduction of both TSPO gene expression (fold change 0.60 [95% CI 0.45-0.74], p = 0.02) and TSPO binding site abundance (fold change 0.61 [95% CI 0.49-0.73], p < 0.0001). These findings have important implications for understanding the biology of TSPO in activated macrophages and microglia in humans. They are also clinically relevant for the interpretation of PET studies using TSPO targeting radioligands, as they suggest changes in TSPO expression may reflect microglial and macrophage density rather than activation phenotype.


International Survey of Trials of Convalescent Plasma to Treat COVID-19 Infection.
The collection and clinical use of COVID-19 convalescent plasma (CCP) as a therapy for COVID-19 infection is under development and early use in many centers worldwide. We conducted an international survey of centers undertaking studies of CCP to provide understanding of the common themes and differences between them. Sixty-four studies in 22 countries were identified from clinical trial registries and personal contacts of the authors. Twenty of the 64 centers (31%) from 12 of 22 countries (55%) responded to the survey. Of the 20 studies, 11 were randomized controlled trials (RCTs), and 9 were case series. Only 4 of the RCTs plan to recruit 400 patients or more, and only 3 RCTs were blinded. The majority of studies will study the effect of CCP on sick patients requiring hospitalization and those requiring critical care, and none is examining the role of CCP in non-infected at-risk individuals. A wide variety of primary and secondary outcomes are being used. The donor eligibility criteria among the studies are very similar, and the use of plasmapheresis for the collection of CCP is almost universal. The planned dose of CCP ranges from as little as 200 mL to well over 1 L, but is 400 to 800 mL or 4 mL/kg or greater in all the RCTs. There is considerable variability in donor antibody testing with no consistency regarding the cut-off for antibody titer for acceptance as CCP or the use of pathogen-inactivation. Our survey provides an understanding of the similarities and differences among the studies of CCP, and that by virtue of their design some studies may be more informative than others.


Acute posterior multifocal placoid pigment epitheliopathy. A follow up study.
Thirteen patients with acute posterior multifocal placoid pigment epitheliopathy (APMPPE) were followed up for a period of from 1 to 20 years (median 5 years 8 months). Five patients developed lesions in the previously unaffected fellow eye during the follow up period. In three patients the condition progressed during a prolonged period without interruption. Widespread severe choroidal atrophy was a late feature in a number of cases. In seven out of thirteen cases, visual acuity remained defective (6/18 or less) in one or both eyes. The prevailing opinion that that APMPPE is self limiting may be incorrect.


Structural dynamics as a contributor to error-prone replication by an RNA-dependent RNA polymerase.
RNA viruses encoding high- or low-fidelity RNA-dependent RNA polymerases (RdRp) are attenuated. The ability to predict residues of the RdRp required for faithful incorporation of nucleotides represents an essential step in any pipeline intended to exploit perturbed fidelity as the basis for rational design of vaccine candidates. We used x-ray crystallography, molecular dynamics simulations, NMR spectroscopy, and pre-steady-state kinetics to compare a mutator (H273R) RdRp from poliovirus to the wild-type (WT) enzyme. We show that the nucleotide-binding site toggles between the nucleotide binding-occluded and nucleotide binding-competent states. The conformational dynamics between these states were enhanced by binding to primed template RNA. For the WT, the occluded conformation was favored; for H273R, the competent conformation was favored. The resonance for Met-187 in our NMR spectra reported on the ability of the enzyme to check the correctness of the bound nucleotide. Kinetic experiments were consistent with the conformational dynamics contributing to the established pre-incorporation conformational change and fidelity checkpoint. For H273R, residues comprising the active site spent more time in the catalytically competent conformation and were more positively correlated than the WT. We propose that by linking the equilibrium between the binding-occluded and binding-competent conformations of the nucleotide-binding pocket and other active-site dynamics to the correctness of the bound nucleotide, faithful nucleotide incorporation is achieved. These studies underscore the need to apply multiple biophysical and biochemical approaches to the elucidation of the physical basis for polymerase fidelity.


Analysis of efficacy of lenvatinib treatment in highly advanced hepatocellular carcinoma with tumor thrombus in the main trunk of the portal vein or tumor with more than 50% liver occupation: A multicenter analysis.
To assess the safety, efficacy, and prognostic impact of clinical factors associated with lenvatinib treatment in highly advanced hepatocellular carcinoma (HCC) with tumor thrombus in the main portal vein trunk (VP4) or tumor with more than 50% liver occupation (tm50%LO). A total of 61 highly advanced HCC patients (41 patients with tm50%LO and 20 patients with VP4) who were treated with lenvatinib at multicenter were enrolled and retrospectively analyzed for treatment outcomes according to their clinical status, including tumor morphology. The most frequent grade ≥3 adverse event in tm50%LO HCC was elevated aspartate aminotransferase (17.1%). Objective response rates were 37.5% and 0% in tm50%LO HCC patients with Child-Pugh grade (CP)-A and CP-B, respectively, and 26.7% and 0% in VP4 HCC patients with CP-A and CP-B, respectively. Estimated median progression-free survival and overall survival were 132 days and 229 days, and 101 days and 201 days in patients with tm50%LO and VP4, respectively. In multivariate analysis, modified albumin-bilirubin grade (hazard ratio 0.372, 95% CI 0.157-0.887; p = 0.0241) and tumor morphology (hazard ratio 0.322, 95% CI 0.116-0.889; p = 0.0287) were independently associated with progression-free survival in patients with tm50%LO HCC. In VP4 HCC, median progression-free survival was worse in CP-B (57 days) than in CP-A patients (137 days, p = 0.0462). Lenvatinib treatment offers a benefit in highly advanced HCC (tm50%LO or VP4) patients with good liver function or nodular-type tumor. The various characteristics identified in this study might be useful as indicators of lenvatinib treatment in highly advanced HCC with tm50%LO or VP4, which are considered very refractory cancers.


Comparison of the self-adjusting file with rotary and hand instrumentation in long-oval-shaped root canals.
The aim of this study was to compare the root canal debridement ability of the self-adjusting file (SAF) with ProFile rotary (PF) and hand filing (HF) instrumentation in long-oval-shaped canals. Extracted human teeth (n = 30) were selected on the basis of a root canal ratio of ≥2.5:1 measured 5 mm from the root apex. Each group (n = 10) was matched with regard to the average root canal ratio. Canals were filled with a radiopaque contrast medium (Vitapex) and instrumented by using SAF, PF, or HF with 20 mL of saline irrigation. Teeth receiving irrigation alone served as controls. Preoperative and postoperative radiographs were taken and submitted to digital subtraction, and the percentage reduction of contrast medium was quantified at 0-5 mm and >5-10 mm from the apex. Values were compared by using 1-way analysis of variance and unpaired t tests. In the 0- to 5-mm segment, the SAF, PF, and HF removed 80.6% ± 14.1%, 84.2% ± 7.7%, and 76.5% ± 10.2% of the contrast medium, respectively (P > .05). In the >5-to 10-mm segment, the SAF, PF, and HF removed 75.5% ± 10.8%, 72.3% ± 12.0%, and 60.9% ± 11.3% of the contrast medium, respectively, with significantly more material removed by SAF compared with HF (P < .05). There was significantly more contrast medium removed by using combined instrumentation and irrigation compared with irrigation alone (P < .01). All 3 techniques removed contrast medium equally well from the 0-to 5-mm segment of long-oval-shaped canals. The SAF performed significantly better than hand filing in the >5-to 10-mm canal segment.


Radiological Mapping of Post-Disaster Nuclear Environments Using Fixed-Wing Unmanned Aerial Systems: A Study From Chornobyl.
In the immediate aftermath following a large-scale release of radioactive material into the environment, it is necessary to determine the spatial distribution of radioactivity quickly. At present, this is conducted by utilizing manned aircraft equipped with large-volume radiation detection systems. Whilst these are capable of mapping large areas quickly, they suffer from a low spatial resolution due to the operating altitude of the aircraft. They are also expensive to deploy and their manned nature means that the operators are still at risk of exposure to potentially harmful ionizing radiation. Previous studies have identified the feasibility of utilizing unmanned aerial systems (UASs) in monitoring radiation in post-disaster environments. However, the majority of these systems suffer from a limited range or are too heavy to be easily integrated into regulatory restrictions that exist on the deployment of UASs worldwide. This study presents a new radiation mapping UAS based on a lightweight (8 kg) fixed-wing unmanned aircraft and tests its suitability to mapping post-disaster radiation in the Chornobyl Exclusion Zone (CEZ). The system is capable of continuous flight for more than 1 h and can resolve small scale changes in dose-rate in high resolution (sub-20 m). It is envisaged that with some minor development, these systems could be utilized to map large areas of hazardous land without exposing a single operator to a harmful dose of ionizing radiation.


Bacteriophage infection of Escherichia coli leads to the formation of membrane vesicles via both explosive cell lysis and membrane blebbing.
Membrane vesicles (MVs) are membrane-bound spherical nanostructures that prevail in all three domains of life. In Gram-negative bacteria, MVs are thought to be produced through blebbing of the outer membrane and are often referred to as outer membrane vesicles (OMVs). We have recently described another mechanism of MV formation in Pseudomonas aeruginosa that involves explosive cell-lysis events, which shatters cellular membranes into fragments that rapidly anneal into MVs. Interestingly, MVs are often observed within preparations of lytic bacteriophage, however the source of these MVs and their association with bacteriophage infection has not been explored. In this study we aimed to determine if MV formation is associated with lytic bacteriophage infection. Live super-resolution microscopy demonstrated that explosive cell lysis of Escherichia coli cells infected with either bacteriophage T4 or T7, resulted in the formation of MVs derived from shattered membrane fragments. Infection by either bacteriophage was also associated with the formation of membrane blebs on intact bacteria. TEM revealed multiple classes of MVs within phage lysates, consistent with multiple mechanisms of MV formation. These findings suggest that bacteriophage infection may be a major contributor to the abundance of bacterial MVs in nature.


Chromium VI and stomach cancer: a meta-analysis of the current epidemiological evidence.
Chromium VI (hexavalent chromium, Cr(VI)) is an established cause of lung cancer, but its association with gastrointestinal cancer is less clear. The goal of this study was to examine whether the current human epidemiological research on occupationally inhaled Cr(VI) supports the hypothesis that Cr(VI) is associated with human stomach cancer. Following a thorough literature search and review of individual studies, we used meta-analysis to summarise the current epidemiological literature on inhaled Cr(VI) and stomach cancer, explore major sources of heterogeneity, and assess other elements of causal inference. We identified 56 cohort and case-control studies and 74 individual relative risk (RR) estimates on stomach cancer and Cr(VI) exposure or work in an occupation associated with high Cr(VI) exposure including chromium production, chrome plating, leather work and work with Portland cement. The summary RR for all studies combined was 1.27 (95% CI 1.18 to 1.38). In analyses limited to only those studies identifying increased risks of lung cancer, the summary RR for stomach cancer was higher (RR=1.41, 95% CI 1.18 to 1.69). Overall, these results suggest that Cr(VI) is a stomach carcinogen in humans, which is consistent with the tumour results reported in rodent studies.


Prediction of cognitive function by arterial stiffness in the very elderly.
Cognitive function is impaired in elderly subjects, so the aim of the present study was to determine the role of arterial stiffness on cognitive function. Cognitive function and arterial stiffness were assessed by the Mini-Mental State Examination (MMSE) and measurement of the brachial-ankle pulse wave velocity (PWV), respectively. The cross-sectional association of the MMSE score and PWV was studied in 203 subjects (87 men, 116 women), all of whom were 85 years old. Sex distribution, systolic and diastolic blood pressures did not differ between the normal (MMSE score >or=24, n=128) and impaired MMSE groups (MMSE score <24, n=75). In contrast, the PWV was significantly increased in the impaired MMSE group than in the normal MMSE group (25.0+/-0.8 vs 22.9+/-0.5 m/s, p<0.05). In multiple regression analysis, the PWV was also independently and significantly associated with the MMSE score. These results suggest that cognitive function could be predicted by arterial stiffness, as assessed by the PWV, in the very old. Preventing atherosclerosis may play an important role in preserving normal cognitive function until very old age.


Treatment choices and outcomes of patients with manometrically diagnosed achalasia.
This prospective population-based study was designed to evaluate treatment choices in patients with new manometrically diagnosed achalasia and their outcomes. Patients referred to the esophageal function laboratory were enrolled after a new manometric diagnosis of achalasia. Patients completed an initial achalasia symptom score validated questionnaire on their symptom severity, duration, treatment pre-diagnosis and Medical Outcomes Study 36-item Short-Form (SF-36) survey. Treatment decisions were made by the referring physician and the patient. Follow-up questionnaires were completed every 3 months for 1 year. Patients who chose not to undergo treatment at 1-year follow-up completed another questionnaire after 5 years. Between January 2004 and January 2005, 83 of 124 eligible patients were enrolled. Heller myotomy was performed on 31 patients, three patients received botulinum toxin injections, and 25 patients received 29 pneumatic balloon dilatations. Twenty-four patients chose to receive no treatment. Following treatment, patients treated with surgery, dilatation and botulinum toxin had an average improvement in achalasia symptom score of 23 +/- 12.2, 17 +/- 10.9, and 9 +/- 14, respectively. Patients receiving no treatment had worsening symptoms with a symptom score change of -3.5 +/- 11.4. Surgery and dilatation resulted in significant improvement (P < 0.01) relative to no treatment. In univariate logistic regression, symptom severity score (odds ratio [OR] 1.04, 95% confidence interval [CI] 1.00 to 1.08), sphincter tone (OR 1.04, 95% CI 1.00 to 1.09), difficulty swallowing liquids (OR 3.21, 95% 1.15 to 8.99), waking from sleep (OR 2.75, 95% 1.00 to 7.61), and weight loss (OR 5.99, 95% CI 1.93 to 18.58) were all significant in predicting that patients would select treatment. In the multivariate analysis, older age (OR 1.05, 95% CI 1.01 to 1.09) and weight loss (OR 3.91, 95% CI 1.02 to 15.2) were statistically significant for undergoing treatment. At 5 years, five (21%) of those who had initially declined treatment at 1 year ultimately chose a treatment. Patients who finally chose Heller myotomy had lower mental component dimension scores on the SF-36 at 1 year than those who did not. This study shows that almost one third of patients with manometrically diagnosed achalasia choose not to undergo treatment within 1 year of their diagnosis. Patients who are more symptomatic appear to be more likely to undergo treatment by univariate analysis. In multivariate analysis, increasing age and weight loss are predictive of those who will undergo treatment, with weight loss having the greatest influence. Patients who choose not to undergo treatment make lifestyle changes to maintain their quality of life, and only a minority of them ultimately undergo treatment.


Spatial and temporal variability of perfluoroalkyl substances in the Laurentian Great Lakes.
Per- and polyfluoroalkyl substances (PFAS) are a diverse group of fluorinated organic chemicals that have been used in industrial and consumer applications since the 1950s. PFAS are resistant to chemical and biological degradation and are ubiquitous in the environment, including in water, sediment, and biota in the Laurentian Great Lakes. This critical review evaluates the spatial and temporal variability of commonly studied perfluoroalkyl sulfonates (PFSAs) and perfluoroalkyl carboxylates (PFCAs) in the Great Lakes by synthesizing data collected in water, surface sediment, sediment cores, lake trout (Salvelinus namaycush), and herring gull (Larus argentatus) eggs. The lowest PFAS concentrations in all matrices are detected in Lake Superior, which is located in the most pristine region of the Great Lakes Basin. In contrast, higher concentrations are observed in Lakes Erie and Ontario, which are more impacted by industrial activity and wastewater discharge. The distribution of individual PFAS compounds also varies across the lakes in response to changes in PFAS sources, with higher proportions of PFSAs in the eastern lakes. Sediment and biota are enriched in long chain PFSAs and PFCAs relative to concentrations in the water column, as expected based on predicted partitioning behavior. Sediment cores and bioarchives consistently demonstrate that PFAS concentrations increased in the Great Lakes from the initial time points until the early 2000s. The available data indicate that PFOS and PFOA concentrations decline after this period in the upper Great Lakes, but are stable in Lake Ontario. However, these trends depend on the lake, the individual compound, and the organism considered.


Clinical relevance of lymphoblast biological features in children with acute lymphoblastic leukemia.
Improvements in therapy for childhood acute lymphoblastic leukemia (ALL) have led us to reevaluate the prognostic significance of lymphoblast characteristics at diagnosis. From application of univariate and multivariate statistical methods, we determined the relationship of five blast cell features to treatment outcome in 250 patients who were enrolled in two clinical trials at this center from May 1979 through April 1982. Karyotype ploidy, lymphoblast morphology, and immunophenotype were each significantly related to prognosis as measured by time to failure, while periodic acid-Schiff reactivity and glucocorticoid receptor number lacked prognostic implication for this patient population. In addition, clinical features of initial WBC count, age, and race were also significant independent variables in predicting treatment response. By multivariate analysis, both ploidy and morphology contributed prognostic information to a clinical model based on WBC count, age, and race. If the model was adjusted for impact of ploidy, however, French-American-British morphology no longer contributed additional prognostic information. Our findings suggest that many traditional biological features used to estimate prognosis in ALL can be discarded in favor of clinical features (leukocyte count, age, and race) and cytogenetics (ploidy) for planning of future clinical trials.


Measuring patient satisfaction in the pharmacy specialty immunization clinic: a pharmacist-run Immunization Clinic at the Veterans Affairs San Diego Healthcare System.
The aim was to measure patient satisfaction with the Pharmacy Specialty Immunization Clinic (PSIC), a pharmacist-run vaccination clinic. Patient satisfaction was measured using a non-validated instrument containing 10 items with a five-point Likert scale (strongly agree, agree, not sure, disagree and strongly disagree). Patients who were seen at the PSIC and who received at least one vaccination were eligible to take part in the patient satisfaction survey. Priority index, a method used to identify areas where limited resources can be used to maximize patient satisfaction, was calculated for the different items of the instrument to determine areas for quality improvement. This study was conducted at the Veterans Affairs San Diego Healthcare System (VASDHS). A total of 188 (55.1%) out of 341 patients who received at least one vaccine in the PSIC completed the survey. Prior to any encounter with the PSIC, patients perceived that the VASDHS was doing a good job providing vaccinations (92.5% answered agree or strongly agree). This perception continued when asked about overall satisfaction after receiving vaccination through the PSIC (86.9% answered agree or strongly agree). When asked about the time the pharmacist spent with the patient, nearly all answered that the pharmacist spent as much time as necessary (97.8% answered agree or strongly agree). Patient satisfaction with pharmacist counselling was equally well received and reflected good communication between patient and pharmacist (97.8% answered agree or strongly agree). In regard to pharmacist competency, 98.9% (n = 184) of patients agreed that pharmacists in the PSIC administered vaccinations appropriately. Priority index identified access to the vaccine as an area where performance-improvement efforts should be committed to improve patient satisfaction. Patients perceived good overall satisfaction with the pharmacist-run immunization clinic in terms of professionalism and access to vaccination. Priority index identified access to vaccination as a focus for future quality improvement.


The Potential of Streptomyces as Biocontrol Agents against the Rice Blast Fungus, Magnaporthe oryzae (Pyricularia oryzae).
Rice is a staple food source for more than three billion people worldwide. However, rice is vulnerable to diseases, the most destructive among them being rice blast, which is caused by the fungus Magnaporthe oryzae (anamorph Pyricularia oryzae). This fungus attacks rice plants at all stages of development, causing annual losses of approximately 10-30% in various rice producing regions. Synthetic fungicides are often able to effectively control plant diseases, but some fungicides result in serious environmental and health problems. Therefore, there is growing interest in discovering and developing new, improved fungicides based on natural products as well as introducing alternative measures such as biocontrol agents to manage plant diseases. Streptomyces bacteria appear to be promising biocontrol agents against a wide range of phytopathogenic fungi, which is not surprising given their ability to produce various bioactive compounds. This review provides insight into the biocontrol potential of Streptomyces against the rice blast fungus, M. oryzae. The ability of various Streptomyces spp. to act as biocontrol agents of rice blast disease has been studied by researchers under both laboratory and greenhouse/growth chamber conditions. Laboratory studies have shown that Streptomyces exhibit inhibitory activity against M. oryzae. In greenhouse studies, infected rice seedlings treated with Streptomyces resulted in up to 88.3% disease reduction of rice blast. Studies clearly show that Streptomyces spp. have the potential to be used as highly effective biocontrol agents against rice blast disease; however, the efficacy of any biocontrol agent may be affected by several factors including environmental conditions and methods of application. In order to fully exploit their potential, further studies on the isolation, formulation and application methods of Streptomyces along with field experiments are required to establish them as effective biocontrol agents.


Synthesis and biological evaluation of bupropion analogues as potential pharmacotherapies for smoking cessation.
Bupropion (2a) analogues were synthesized and tested for their ability to inhibit monoamine uptake and to antagonize the effects of human alpha3beta4*, alpha4beta2, alpha4beta4, and alpha1* nAChRs. The analogues were evaluated for their ability to block nicotine-induced effects in four tests in mice. Nine analogues showed increased monoamine uptake inhibition. Similar to 2a, all but one analogue show inhibition of nAChR function selective for human alpha3beta4*-nAChR. Nine analogues have higher affinity at alpha3beta4*-nAChRs than 2a. Four analogues also had higher affinity for alpha4beta2 nAChR. Analogues 2r, 2m, and 2n with AD(50) values of 0.014, 0.015, and 0.028 mg/kg were 87, 81, and 43 times more potent than 2a in blocking nicotine-induced antinociception in the tail-flick test. Analogue 2x with IC(50) values of 31 and 180 nM for DA and NE, respectively, and with IC(50) of 0.62 and 9.8 microm for antagonism of alpha3beta4 and alpha4beta2 nAChRs had the best overall in vitro profile relative to 2a.


The multifaceted role of the microenvironment in liver metastasis: biology and clinical implications.
The liver is host to many metastatic cancers, particularly colorectal cancer, for which the last 2 decades have seen major advances in diagnosis and treatment. The liver is a vital organ, and the extent of its involvement with metastatic disease is a major determinant of survival. Metastatic cells arriving in the liver via the bloodstream encounter the microenvironment of the hepatic sinusoid. The interactions of the tumor cells with hepatic sinusoidal and extrasinusoidal cells (endothelial, Kupffer, stellate, and inflammatory cells) determine their fate. The sinusoidal cells can have a dual role, sometimes fatal to the tumor cells but also facilitatory to their survival and growth. Adhesion molecules participate in these interactions and may affect their outcome. Bone marrow-derived cells and chemokines also play a part in the early battle for survival of the metastases. Once the tumor cells have arrested and survived the initial onslaught, tumors can grow within the liver in 3 distinct patterns, reflecting differing host responses, mechanisms of vascularization, and proteolytic activity. This review aims to present current knowledge of the interactions between the host liver cells and the invading metastases that has implications for the clinical course of the disease and the response to treatment.


Lipo-chitooligosaccharide signaling in endosymbiotic plant-microbe interactions.
The arbuscular mycorrhizal (AM) and the rhizobia-legume (RL) root endosymbioses are established as a result of signal exchange in which there is mutual recognition of diffusible signals produced by plant and microbial partners. It was discovered 20 years ago that the key symbiotic signals produced by rhizobial bacteria are lipo-chitooligosaccharides (LCO), called Nod factors. These LCO are perceived via lysin-motif (LysM) receptors and activate a signaling pathway called the common symbiotic pathway (CSP), which controls both the RL and the AM symbioses. Recent work has established that an AM fungus, Glomus intraradices, also produces LCO that activate the CSP, leading to induction of gene expression and root branching in Medicago truncatula. These Myc-LCO also stimulate mycorrhization in diverse plants. In addition, work on the nonlegume Parasponia andersonii has shown that a LysM receptor is required for both successful mycorrhization and nodulation. Together these studies show that structurally related signals and the LysM receptor family are key components of both nodulation and mycorrhization. LysM receptors are also involved in the perception of chitooligosaccharides (CO), which are derived from fungal cell walls and elicit defense responses and resistance to pathogens in diverse plants. The discovery of Myc-LCO and a LysM receptor required for the AM symbiosis, therefore, not only raises questions of how legume plants discriminate fungal and bacterial endosymbionts but also, more generally, of how plants discriminate endosymbionts from pathogenic microorganisms using structurally related LCO and CO signals and of how these perception mechanisms have evolved.


Biochemical and structural studies on the high affinity of Hsp70 for ADP.
The molecular chaperone 70-kDa heat shock protein (Hsp70) is driven by ATP hydrolysis and ADP-ATP exchange. ADP dissociation from Hsp70 is reportedly slow in the presence of inorganic phosphate (P(i) ). In this study, we investigated the interaction of Hsp70 and its nucleotide-binding domain (NBD) with ADP in detail, by isothermal titration calorimetry measurements and found that Mg(2+) ion dramatically elevates the affinity of Hsp70 for ADP. On the other hand, P(i) increased the affinity in the presence of Mg(2+) ion, but not in its absence. Thus, P(i) enhances the effect of the Mg(2+) ion on the ADP binding. Next, we determined the crystal structures of the ADP-bound NBD with and without Mg(2+) ion. As compared with the Mg(2+) ion-free structure, the ADP- and Mg(2+) ion-bound NBD contains one Mg(2+) ion, which is coordinated with the β-phosphate group of ADP and associates with Asp10, Glu175, and Asp199, through four water molecules. The Mg(2+) ion is also coordinated with one P(i) molecule, which interacts with Lys71, Glu175, and Thr204. In fact, the mutations of Asp10 and Asp199 reduced the affinity of the NBD for ADP, in both the presence and the absence of P(i) . Therefore, the Mg(2+) ion-mediated network, including the P(i) and water molecules, increases the affinity of Hsp70 for ADP, and thus the dissociation of ADP is slow. In ADP-ATP exchange, the slow ADP dissociation might be rate-limiting. However, the nucleotide-exchange factors actually enhance ADP release by disrupting the Mg(2+) ion-mediated network.


Characterisation of physiological and immunological responses in beef cows to abrupt weaning and subsequent housing.
Weaning involves the permanent separation of the calf from the dam and has been shown to be stressful for both. The objectives of this study were to characterise the effect of i) abrupt weaning and ii) subsequent housing on the extended physiological and immunological responses of beef cows. At weaning (day (d) 0, mean age of calf (s.d.) 212 (24.5) d), cows were abruptly separated from their calves and returned to the grazing area. After 35 d at pasture, cows were housed in a slatted floor shed and offered grass silage ad libitum plus a mineral-vitamin supplement daily. Rectal body temperature was recorded and blood samples were obtained on i) d 0 (weaning), 2, 7, 14, 21, 28, 35 and subsequently on ii) d 0 (housing), 2, 7, 14 and 21 for physiological, haematological and immunological measurements. Post-weaning, concentration of cortisol and dehydroepiandrosterone were unchanged (P > 0.05). Rectal body temperature, neutrophil number and neutrophil: lymphocyte ratio increased (P < 0.01) on d 2 compared with pre-weaning baseline. Lymphocyte and neutrophil number decreased (P < 0.05) on d 2 to 7 and d 7 to 21, respectively, compared with pre-weaning baseline. Interferon-gamma production decreased (P < 0.05) on d 2 compared with pre-weaning baseline. An increase (P < 0.05) in acute phase proteins, fibrinogen and haptoglobin was evident on d 2 to 35 compared with pre-weaning baseline. Concentration of glucose increased on d 2 to 28, whereas non-esterified fatty acid decreased on d 2 to 35 compared with pre-weaning baseline. Post-housing, concentrations of cortisol, rectal body temperature, total leukocyte number, and glucose were unchanged (P > 0.05). On d 2 post-housing, neutrophil number and neutrophil: lymphocyte ratio increased (P < 0.05), whereas lymphocyte number and concentrations of dehydroepiandrosterone, fibrinogen and non-esterified fatty acid decreased (P < 0.05) compared with pre-housing baseline. Concentration of haptoglobin increased (P < 0.05) on d 14 to 21 post-housing. A transitory increase in neutrophil number and decrease in lymphocyte number, increased neutrophil:lymphocyte ratio coupled with decreased interferon-gamma production, and increased concentration of acute phase proteins indicate a stress response in cows post-weaning, whereas post-housing, changes were less marked.


Bradykinin relaxation in small porcine retinal arterioles.
To study changes in the spontaneous diameter of small retinal arterioles and bradykinin (BK)-induced vasodilation during inhibition of the synthesis of nitric oxide (NO), prostaglandins (PGs), and cytochrome P450 2C8/9-dependent endothelial-derived hyperpolarizing factor (EDHF). Forty-eight isolated porcine arterioles with a diameter of approximately 70 microm were mounted in a double-barreled pipette system placed in an organ bath, and diameter changes were studied under isobaric conditions. After an equilibration period, the arterioles were incubated with inhibitors of the synthesis of NO, PGs, or cytochrome P450 2C8/9-dependent EDHF, and spontaneous diameter changes were studied. Subsequently, the arterioles were precontracted, and the diameter was assessed after addition of BK in cumulative concentrations. Inhibition of NOS elicited a significant decrease in the spontaneous diameter of the vessels (P = 0.028), whereas no change in the spontaneous diameter was induced by inhibition of PG or cytochrome P450 2C8/9 dependent EDHF synthesis (P = 0.35 and P = 0.75, respectively). The vasodilating effect of BK was decreased by inhibition of NO (P = 0.002) but not by inhibition of prostaglandin or cytochrome P450 2C8/9-dependent EDHF synthesis (P = 0.82 and P = 0.94, respectively). The results suggest the presence of a spontaneous release of NO, which keeps the retinal microcirculation dilated under normal conditions. The finding of BK-induced relaxation being dependent on the NO synthase (NOS), but not on PGs or cytochrome P450 2C8/9-dependent EDHF may be of importance for understanding the microcirculatory effects of pharmacologic compounds affecting the BK metabolism, such as angiotensin-converting enzyme (ACE) inhibitors.


Head-to-Head Comparison of the Penetration Efficiency of Lipid-Based Nanoparticles into Tumor Spheroids.
Most tumor-targeted drug delivery systems must overcome a large variety of physiological barriers before reaching the tumor site and diffuse through the tight network of tumor cells. Many studies focus on optimizing the first part, the accumulation of drug carriers at the tumor site, ignoring the penetration efficiency, i.e., a measure of the ability of a drug delivery system to overcome tumor surface adherence and uptake. We used three-dimensional (3D) tumor spheroids in combination with light-sheet fluorescence microscopy in a head-to-head comparison of a variety of commonly used lipid-based nanoparticles, including liposomes, PEGylated liposomes, lipoplexes, and reconstituted high-density lipoproteins (rHDL). Whilst PEGylation of liposomes only had minor effects on the penetration efficiency, we show that lipoplexes are mainly associated with the periphery of tumor spheroids, possibly due to their positive surface charge, leading to fusion with the cells at the spheroid surface or aggregation. Surprisingly, the rHDL showed significantly higher penetration efficiency and high accumulation inside the spheroid. While these findings indeed could be relevant when designing novel drug delivery systems based on lipid-based nanoparticles, we stress that the used platform and the detailed image analysis are a versatile tool for in vitro studies of the penetration efficiency of nanoparticles in tumors.


Survivin blockade sensitizes rhabdomyosarcoma cells for lysis by fetal acetylcholine receptor-redirected T cells.
Cellular immunotherapy may provide a strategy to overcome the poor prognosis of metastatic and recurrent rhabdomyosarcoma (RMS) under the current regimen of polychemotherapy. Because little is known about resistance mechanisms of RMS to cytotoxic T cells, we investigated RMS cell lines and biopsy specimens for expression and function of immune costimulatory receptors and anti-apoptotic molecules by RT-PCR, Western blot analysis, IHC, and cytotoxicity assays using siRNA or transfection-modified RMS cell lines, together with engineered RMS-directed cytotoxic T cells specific for the fetal acetylcholine receptor. We found that costimulatory CD80 and CD86 were consistently absent from all RMSs tested, whereas inducible T-cell co-stimulator ligand (ICOS-L; alias B7H2) was expressed by a subset of RMSs and was inducible by tumor necrosis factor α in two of five RMS cell lines. Anti-apoptotic survivin, along with other inhibitor of apoptosis (IAP) family members (cIAP1, cIAP2, and X-linked inhibitor of apoptosis protein), was overexpressed by RMS cell lines and biopsy specimens. Down-regulation of survivin by siRNA or pharmacologically in RMS cells increased their susceptibility toward a T-cell attack, whereas induction of ICOS-L did not. Treatment of RMS-bearing Rag(-/-) mice with fetal acetylcholine receptor-specific chimeric T cells delayed xenograft growth; however, this happened without definitive tumor eradication. Combined blockade of survivin and application of chimeric T cells in vivo suppressed tumor proliferation during survivin inhibition. In conclusion, survivin blockade provides a strategy to sensitize RMS cells for T-cell-based therapy.


Long-term Outcome of Epilepsy and Cortical Malformations Due to Abnormal Migration and Postmigrational Development: A Cohort Study.
To evaluate the long-term outcomes of patients with epilepsy and malformations of cortical development (MCD). We conducted a historical cohort study of patients with epilepsy and MCD due to impaired neuronal migration and postmigration organization with a follow-up period of ≥5 years. For each patient, MCD was classified after accurate neuroimaging reappraisal by an expert neuroradiologist. The primary outcome was remission, defined as a period of seizure freedom ≥5 years at any time from epilepsy onset. We used Kaplan-Meier estimates for survival analysis and univariate and multivariate Cox regression analyses to evaluate baseline variables as possible factors associated with remission. The cohort included 71 patients (M/F 31/40) with a 17-year median follow-up (1,506 person-years). About half (49.3%) had heterotopia, 35.2% polymicrogyria, 7% lissencephaly, and 8.5% the combination of 2 MCD. The mean age at seizure onset was 12.4 ± 7.2 years. Intellectual disability and neurologic deficits were observed in 30.4% and 40.9%, respectively. More than 60% of patients had refractory epilepsy. In 3 patients who underwent epilepsy surgery, MCD diagnosis was confirmed by histology. At the last visit, 44% of patients had been seizure-free during the previous year, but none of them had stopped antiseizure medication. Thirty patients achieved remission (42.2%) at some point in their disease history, whereas 41 individuals (57.8%) had never been in remission for ≥5 years. The cumulative remission rate was 38% by 20 years from inclusion. In the Cox model, unilateral distribution of MCD (hazard ratio [HR] 2.68, 95% CI 1.04-6.92) and a low seizure frequency at onset (HR 5.01, 95% CI 1.12-22.5) were significantly associated with remission. Patients with epilepsy and MCD showed a remission rate of 38% by 20 years from onset. Unilateral distribution of the MCD is associated with a 3-fold probability of achieving remission. About 40% of patients showed a drug-sensitive condition with risk of relapse during their epilepsy course. This study provides Class II evidence that in patients with epilepsy and MCD, unilateral MCD and low seizure frequency at onset are associated with achieving epilepsy remission.


Young women smokers' response to using plain cigarette packaging: qualitative findings from a naturalistic study.
The aim of this study was to explore in-depth the response of young women smokers (18-35 years) to using dark brown 'plain' cigarette packs in naturalistic settings. Participants were recruited in six towns and cities in Scotland to take part in a naturalistic study, where they used plain cigarette packs for a week. Participants completed a number of questionnaires during the study period (reported elsewhere), and a sub-sample participated in post-study telephone interviews to explore their experiences of using the plain packs. Of the 187 participants who completed the study, 23 were randomly selected to participate in the post-study interviews. Within the interviews a semi-structured topic guide was used to assess perceptions of the plain pack, feelings created by the pack, feelings about smoking, and avoidant and smoking behaviour. The brown (plain) packs were perceived negatively due to the colour, the undesirable image the pack conveyed, and the reaction from others. The plain packs were also associated with negative feelings, such as embarrassment, discomfort and guilt. Some participants also commented that they felt differently about the product, considered to be less enjoyable or more harmful, when using the plain packs, and were less interested in, or felt more negatively about, smoking. A number of participants said that they had engaged in avoidant behavior with the plain packs, such as hiding it, due to their negative thoughts about the packs and the reaction of others. Some participants also mentioned cessation-related behaviours when using the plain packs, such as forgoing cigarettes, stubbing cigarettes out early and thinking about quitting, largely due to the decreased enjoyment of smoking. The experience of using cigarettes in plain packs prompted a range of negative responses from young women smokers, who are a crucial target group for tobacco control interventions.


How to Measure Behavioral Spillovers: A Methodological Review and Checklist.
A growing stream of literature at the interface between economics and psychology is currently investigating 'behavioral spillovers' in (and across) different domains, including health, environmental, and pro-social behaviors. A variety of empirical methods have been used to measure behavioral spillovers to date, from qualitative self-reports to statistical/econometric analyses, from online and lab experiments to field experiments. The aim of this paper is to critically review the main experimental and non-experimental methods to measure behavioral spillovers to date, and to discuss their methodological strengths and weaknesses. A consensus mixed-method approach is then discussed which uses between-subjects randomization and behavioral observations together with qualitative self-reports in a longitudinal design in order to follow up subjects over time. In particular, participants to an experiment are randomly assigned to a treatment group where a behavioral intervention takes place to target behavior 1, or to a control group where behavior 1 takes place absent any behavioral intervention. A behavioral spillover is empirically identified as the effect of the behavioral intervention in the treatment group on a subsequent, not targeted, behavior 2, compared to the corresponding change in behavior 2 in the control group. Unexpected spillovers and additional insights (e.g., drivers, barriers, mechanisms) are elicited through analysis of qualitative data. In the spirit of the pre-analysis plan, a systematic checklist is finally proposed to guide researchers and policy-makers through the main stages and features of the study design in order to rigorously test and identify behavioral spillovers, and to favor transparency, replicability, and meta-analysis of studies.


Meniscal repair in anterior cruciate ligament reconstruction: a long-term outcome study.
To study the long-term outcome of patients who have undergone inside-out, vertical stacked mattress suture repair of meniscal tears combined with anterior cruciate ligament (ACL) reconstruction. From a database of ACL reconstructed patients, consecutive patients undergoing concomitant meniscal repair and ACL reconstruction between 1991 and 1999 were identified. Patients with previous ACL or meniscal pathology were excluded. Two age- and sex-matched cohorts who had undergone combined ACL reconstruction and menisectomy and who had undergone ACL reconstruction with normal menisci were identified for comparison. Outcome scoring included IKDC and Lysholm scores for the meniscal repair group. Two sample t-tests and chi-square tests were used to compare the IKDC subjective scores, with a minimum level of significance set at 5% (P = 0.05). Fourty-four patients were identified for analysis with a median follow-up of 10 years (7.7-12.6). Patients undergoing ACL reconstruction combined with meniscal repair had a mean IKDC of 84.2 compared with a mean score of 70.5 (P = 0.008) in patients who had undergone menisectomy and 88.2 (P = 0.005) in patients with intact menisci. 86.2% of patients with ACL reconstruction and meniscal repair had Lysholm score of between 80 and 100%. Sixty-nine percent follow-up was achieved. Survival analysis by life table method shows a best case of 89% 10-years survival. This study demonstrates that good long-term outcomes can be obtained in patients up to over 12 years after combined ACL reconstruction and meniscal repair. Improved functional scores can be achieved when compared with ACL reconstruction and menisectomy. The authors advocate repair of meniscal tears during ACL reconstruction unless there is complex tearing, radial tearing or plastic deformation of the remaining meniscus. III.


Phytoplankton-bacterial interactions mediate micronutrient colimitation at the coastal Antarctic sea ice edge.
Southern Ocean primary productivity plays a key role in global ocean biogeochemistry and climate. At the Southern Ocean sea ice edge in coastal McMurdo Sound, we observed simultaneous cobalamin and iron limitation of surface water phytoplankton communities in late Austral summer. Cobalamin is produced only by bacteria and archaea, suggesting phytoplankton-bacterial interactions must play a role in this limitation. To characterize these interactions and investigate the molecular basis of multiple nutrient limitation, we examined transitions in global gene expression over short time scales, induced by shifts in micronutrient availability. Diatoms, the dominant primary producers, exhibited transcriptional patterns indicative of co-occurring iron and cobalamin deprivation. The major contributor to cobalamin biosynthesis gene expression was a gammaproteobacterial population, Oceanospirillaceae ASP10-02a. This group also contributed significantly to metagenomic cobalamin biosynthesis gene abundance throughout Southern Ocean surface waters. Oceanospirillaceae ASP10-02a displayed elevated expression of organic matter acquisition and cell surface attachment-related genes, consistent with a mutualistic relationship in which they are dependent on phytoplankton growth to fuel cobalamin production. Separate bacterial groups, including Methylophaga, appeared to rely on phytoplankton for carbon and energy sources, but displayed gene expression patterns consistent with iron and cobalamin deprivation. This suggests they also compete with phytoplankton and are important cobalamin consumers. Expression patterns of siderophore- related genes offer evidence for bacterial influences on iron availability as well. The nature and degree of this episodic colimitation appear to be mediated by a series of phytoplankton-bacterial interactions in both positive and negative feedback loops.


Maturation of cyclopean visual evoked potential phase in preterm and full-term infants.
P1 is the major positive component of pattern-reversal visual evoked potentials (PR-VEPs). The rapid decrease of its latency correlates with the progressive myelination in the developing infant brain, which affects signal transmission in the visual system. An age-dependent phase shift, analogous to P1 peak latency, can be observed in dynamic random dot correlogram (DRDC)-evoked VEPs (DRDC-VEPs), a method used to assess binocular function. Our goal was to study the relationship between cyclopean DRDC-VEP phases and PR-VEP P1 latencies in full-term and preterm infants so as to further explore the experience dependence of early binocular developmental processes. DRDC-VEPs and PR-VEPs were recorded in 128 full-term and 47 preterm healthy infants and toddlers. DRDC stimuli were presented on the red and green channels of a CRT monitor while infants wore red-green goggles for dichoptic viewing. Reliability of VEP responses was assessed by the statistic. Logistic function was fit to the phase and latency data as a function of age, and goodness of fit was assessed by analysis of residuals. The phase shift of DRDC-VEPs and the rapid decrease of P1 latencies occur at identical postconceptual ages. A correlation also was found between P1 latencies and DRDC-VEP phases. Although development of binocularity is an extremely experience-dependent process, our data suggest that DRDC-VEP phase and P1 latency mature independently from visual experience. We propose that both the phase shift and decreasing P1 latency are indicators of myelination and increasingly faster signal transmission in the developing visual system.


The anti-ghrelin Spiegelmer NOX-B11-3 blocks ghrelin- but not fasting-induced neuronal activation in the hypothalamic arcuate nucleus.
The hypothalamic arcuate nucleus (Arc) is the presumed target site for the orexigenic hormone ghrelin, which is secreted from the stomach during fasting. Ghrelin directly activates Arc neurones. Similar to exogenous ghrelin, overnight food deprivation also induces c-Fos expression in the Arc of mice. In the present study, we tested the role of endogenous ghrelin in the fasting-induced c-Fos expression in the Arc of mice. We used NOX-B11-3, the latest generation of the recently developed ghrelin-binding compounds, so-called RNA Spiegelmers (SPM) to block endogenous ghrelin action during food deprivation. The specificity and potency of this compound was also tested in electrophysiological and immunohistological experiments. In electrophysiological in vitro single cell recordings, NOX-B11-3 effectively blocked the excitatory effect of ghrelin in the medial Arc (ArcM) of rats whereas the biologically inactive control SPM had no effect. Furthermore, NOX-B11-3 (15 mg/kg i.p.) potently suppressed ghrelin-induced (25 microg/kg s.c., 12 h after SPM injection) c-Fos expression in the Arc. However, when injected at the beginning of a 14-h fasting period, the same dose of NOX-B11-3 had no effect on the c-Fos expression in the Arc of mice. These results demonstrate that NOX-B11-3 is a long-acting compound, which effectively blocks the effect of exogenous ghrelin on neuronal activity in the Arc under in vitro and in vivo conditions. Furthermore, increased ghrelin signalling does not appear to be a necessary factor for the activation of Arc neurones during food deprivation or other fasting-related signals might have masked or compensated for the loss of the ghrelin effect.


A phase 2 study of lenalidomide, rituximab, cyclophosphamide, and dexamethasone (LR-CD) for untreated low-grade non-Hodgkin lymphoma requiring therapy.
Patients with indolent non-Hodgkin lymphoma (NHL) have multiple treatment options yet there is no consensus as to the best initial therapy. Lenalidomide, an immunomodulatory agent, has single agent activity in relapsed lymphoma. This trial was conducted to assess feasibility, efficacy, and safety of adding lenalidomide to rituximab, cyclophosphamide, and dexamethasone (LR-CD) in untreated indolent NHL patients requiring therapy. This was a single institution phase II trial. Treatment consisted of IV rituximab 375 mg/m2 day 1; oral lenalidomide 20 mg days 1-21; cyclophosphamide 250 mg/m2 days 1, 8, and 15; and dexamethasone 40 mg days 1, 8, 15, and 22 of a 28-day cycle. Treatment continued 2 cycles beyond best response for a maximum of 12 cycles without rituximab maintenance. Thirty-three patients were treated. Median age was 68 (43-83 years). 39% had stage IV disease. Histologic subtypes included 8 follicular lymphoma (FL), 7 marginal zone lymphoma (MZL) (1 splenic, 2 extranodal, and 4 nodal), 15 Waldenström's macroglobulinemia (WM), 1 lymphoplasmacytic lymphoma, 1 small lymphocytic lymphoma, and 1 low-grade B-cell lymphoma with plasmacytic differentiation (unable to be classified better as MZL or LPL). Hematologic toxicity was the most common adverse event. Median time of follow-up was 23.4 months (range 1.8-50.9). The overall response rate was 87.9%, with 30.3% complete response. The median duration of response was 38.7 months. The median progression free survival was 39.7 months, while median overall survival (OS) has not yet been reached. Lenalidomide can be safely added to a simple regimen of rituximab, oral cyclophosphamide, and dexamethasone and is an effective combination as initial therapy for low-grade B-cell NHL.


Treatment of Malignant Pleural Mesothelioma: American Society of Clinical Oncology Clinical Practice Guideline.
Purpose To provide evidence-based recommendations to practicing physicians and others on the management of malignant pleural mesothelioma. Methods ASCO convened an Expert Panel of medical oncology, thoracic surgery, radiation oncology, pulmonary, pathology, imaging, and advocacy experts to conduct a literature search, which included systematic reviews, meta-analyses, randomized controlled trials, and prospective and retrospective comparative observational studies published from 1990 through 2017. Outcomes of interest included survival, disease-free or recurrence-free survival, and quality of life. Expert Panel members used available evidence and informal consensus to develop evidence-based guideline recommendations. Results The literature search identified 222 relevant studies to inform the evidence base for this guideline. Recommendations Evidence-based recommendations were developed for diagnosis, staging, chemotherapy, surgical cytoreduction, radiation therapy, and multimodality therapy in patients with malignant pleural mesothelioma. Additional information is available at www.asco.org/thoracic-cancer-guidelines and www.asco.org/guidelineswiki .


Common and distinct neural substrates of the money illusion in win and loss domains.
People often evaluate money based on its face value and overlook its real purchasing power, a phenomenon known as the money illusion. In the present study, using functional magnetic resonance imaging (fMRI) combined with a gambling task, we examined the neural signatures of the money illusion in both win and loss domains. Behavioral results showed that self-reported satisfaction with outcomes was modulated by the face value but not the true value of money in both win and loss domains. At the neural level, activity in the posterior insula was associated with the true value of money in the win domain, but not in the loss domain. Importantly, we found that the ventral striatum, ventromedial prefrontal cortex (vmPFC) and amygdala encoded the money illusion in both domains, indicating a domain-general rather than domain-specific neural signature. Moreover, participants with a larger degree of money illusion at the behavioral level showed stronger functional connectivity between the ventral striatum and ventral anterior cingulate cortex (vACC) in the win domain, but stronger functional connectivity between the ventral striatum and amygdala in the loss domain. Our findings highlight the overlapping and distinct neural substrates underlying the money illusion in the context of wins and losses.


Ultrasonographic evaluation of reticular motility in 144 cattle with vagal indigestion.
The reticular motility in 144 cattle with vagal indigestion was evaluated by ultrasonography; 97 had proximal functional stenosis (reticulo-omasal stenosis) and 47 had distal functional stenosis (pyloric stenosis). A 3.5 MHz linear transducer was used to measure the frequency, amplitude, duration and speed of the reticular contractions per three minutes while the animals were standing and unsedated. The mean (sd) number of contractions per three minutes was 4.3 (2.09) with a range from 0 to 12; nine of the cattle had one or two contractions, 17 had three contractions, 59 had four or five contractions, 37 had more than five contractions and 12 had reticular atony. The cattle with proximal functional stenosis had 4.6 (2.01) contractions per three minutes, significantly more (P<0.05) than those with distal functional stenosis (3.6 [2.16]). The position, contour and size of the reticulum, the amplitude and speed of the contractions and the area surrounding the reticulum did not differ significantly between the two groups.


Genome-wide association studies of serum magnesium, potassium, and sodium concentrations identify six Loci influencing serum magnesium levels.
Magnesium, potassium, and sodium, cations commonly measured in serum, are involved in many physiological processes including energy metabolism, nerve and muscle function, signal transduction, and fluid and blood pressure regulation. To evaluate the contribution of common genetic variation to normal physiologic variation in serum concentrations of these cations, we conducted genome-wide association studies of serum magnesium, potassium, and sodium concentrations using approximately 2.5 million genotyped and imputed common single nucleotide polymorphisms (SNPs) in 15,366 participants of European descent from the international CHARGE Consortium. Study-specific results were combined using fixed-effects inverse-variance weighted meta-analysis. SNPs demonstrating genome-wide significant (p<5 x 10(-8)) or suggestive associations (p<4 x 10(-7)) were evaluated for replication in an additional 8,463 subjects of European descent. The association of common variants at six genomic regions (in or near MUC1, ATP2B1, DCDC5, TRPM6, SHROOM3, and MDS1) with serum magnesium levels was genome-wide significant when meta-analyzed with the replication dataset. All initially significant SNPs from the CHARGE Consortium showed nominal association with clinically defined hypomagnesemia, two showed association with kidney function, two with bone mineral density, and one of these also associated with fasting glucose levels. Common variants in CNNM2, a magnesium transporter studied only in model systems to date, as well as in CNNM3 and CNNM4, were also associated with magnesium concentrations in this study. We observed no associations with serum sodium or potassium levels exceeding p<4 x 10(-7). Follow-up studies of newly implicated genomic loci may provide additional insights into the regulation and homeostasis of human serum magnesium levels.


Inter-relationship between visual symptoms, activity limitation and psychological functioning in patients with diabetic retinopathy.
We explored the direct and indirect impact of restrictions in daily living activities on the relationship between perceived visual symptoms associated with diabetic retinopathy (DR) and psychological functioning. In this prospective, cross-sectional study, 514 tertiary patients with DR (mean age±SD, 60.4±12.6 years; 64% male) answered questions related to nine domains of DR-specific quality of life. These were classified into the following predictor, mediating and outcome variables: visual symptoms (predictor variable); activity limitation, driving, lighting, social restriction, inconvenience, mobility (mediating variables); and emotional distress and concerns (outcome variables). Direct and indirect relationships between study variables were assessed using path analysis, using interval-level person measures derived from Rasch analyses of the study questionnaires. We found no direct effect of DR-related visual symptoms on emotional distress or concerns. Rather, the association between visual symptoms and emotional distress was mediated (all p<0.05) by mobility (indirect effect=0.07), inconvenience (indirect effect=0.28), activity limitation (indirect effect=0.13) and social restriction (indirect effect=0.11). Similarly, the relationship between DR-related visual symptoms and concerns was mediated by inconvenience (indirect effect=0.36) and social restriction (indirect effect=0.11). Lighting and driving did not have a mediating role. The link between vision and psychological functioning outcomes such as emotional distress and concern is complex in patients with DR and mediated by limitations in daily living activities and social factors. Interventions to enhance daily functioning and social interaction may be effective in reducing emotional distress associated with DR-related vision impairment.


Effect of thiols, zinc, and redox conditions on Hg uptake in Shewanella oneidensis.
Mercury uptake in bacteria represents a key first step in the production and accumulation of methylmercury in biota. Previous experiments with mercury methylating bacteria have shown that Hg uptake is enhanced by some thiols, in particular cysteine, and that it is an energy-dependent process through heavy metal transporters [Schaefer et al. Environ. Sci. Technol. 2014, 48, 3007]. In this study, we examine Hg uptake in the nonmethylating facultative aerobe, Shewanella oneidensis, under both anaerobic and aerobic conditions. Our results demonstrate similar characteristics of the Hg uptake system to those of the Hg methylating strains: (1) uptake is enhanced in the presence of some thiols but not others; (2) uptake is energy dependent as evidenced by inhibition by a protonophore, and (3) uptake is inhibited by high Zn(II) concentrations. Initial cellular uptake rates in S. oneidensis were remarkably similar under aerobic and fumarate-reducing conditions. These data support a similar Hg(II) uptake mechanism within the proteobacteria of accidental Hg(II) transport through heavy metal transporters with similar rates of uptake but differences in the ability to take up Hg bound to different thiols.


Changes in the Oscillatory Electric Potential on the Olfactory Epithelium and in Reproductive Hormone Levels during the Breeding Season in the Toad (Bufo japonicus).
A slow transient electric potential change (electro-olfactogram, EOG) can be recorded through an Ag-AgCl electrode placed on the olfactory epithelium in response to stimulation with an air stream to the tissue in toads (Bufo japonicus). During the breeding season, oscillatory potential changes (OSC) superimpose on the EOG. In the present study the OSC amplitude was found to be highly correlated with the migratory behavior. Since toads track the route to and from the breeding pond using olfactory cues along the migration route, the enhanced OSC should be responsible for the breeding migration.A significant positive correlation was found between plasma gonadotropin levels and the OSC amplitude in males captured during the breeding migration. There was no significant relationship between plasma gonadotropin levels and the OSC amplitude in female toads during the breeding season, but there was a significant correlation between plasma progesterone levels and the OSC amplitude. In males, hypophysectomy just before the breeding season decreased the OSC amplitude. And testis weight was also positively correlated with the OSC amplitude in January. These results suggest that the appearance of the OSC is related to the timing of the activation of the reproductive system. However, treatment of toads with hCG (human chorionic gonadotropin), testosterone, estradiol or progesterone in the non-breeding season did not induce a significant change in the OSC amplitude. Other factor or factors may be required in activation of the olfactory system of the toad in a non-reproductive stage together with the hormones of the gonadal axis.


Facilitators and barriers to the delivery of palliative care to children with life-limiting and life-threatening conditions: a qualitative study of the experiences and perceptions of healthcare professionals.
To understand healthcare system facilitators and barriers to the delivery of palliative care for children with life-limiting and life-threatening conditions and their family members. Focus groups with children's palliative care professionals. Data were analysed using thematic analysis. Four regions of England (West Midlands, South West, Yorkshire and Humber, and London) from December 2017 to June 2018. Healthcare professionals (doctors, nurses and allied healthcare professionals) working in children's palliative care services. A total of 71 healthcare professionals participated in the focus groups. Three overarching themes were identified which influenced whether and when children were referred to and started to receive palliative care: (1) the unspoken background of clinical uncertainty which often delayed palliative care; (2) the cultural 'collusion of immortality', where conversations about the possibility of dying can be avoided or deferred; and (3) the role of paediatric palliative care teams in 'illuminating the blind spot' of palliative care as well as providing hands-on care. Palliative care is a holistic approach to care that focuses on quality of life for people living with life-limiting and life-threatening conditions that can be delivered alongside active treatment. There is a need to prioritise and integrate this into healthcare services for children more effectively if improvements in care are to be realised. While more specialist paediatric palliative care services are needed, the unspoken background of clinical uncertainty needs to be addressed together with the collusion of immortality within healthcare culture and organisations.


Protein misfolding and temperature up-shift cause G1 arrest via a common mechanism dependent on heat shock factor in Saccharomycescerevisiae.
Accumulation of misfolded proteins in the cell at high temperature may cause entry into a nonproliferating, heat-shocked state. The imino acid analog azetidine 2-carboxylic acid (AZC) is incorporated into cellular protein competitively with proline and can misfold proteins into which it is incorporated. AZC addition to budding yeast cells at concentrations sufficient to inhibit proliferation selectively activates heat shock factor (HSF). We find that AZC treatment fails to cause accumulation of glycogen and trehalose (Msn2/4-dependent processes) or to induce thermotolerance (a protein kinase C-dependent process). However, AZC-arrested cells can accumulate glycogen and trehalose and can acquire thermotolerance in response to a subsequent heat shock. We find that AZC treatment arrests cells in a viable state and that this arrest is reversible. We find that cells at high temperature or cells deficient in the ubiquitin-conjugating enzymes Ubc4 and Ubc5 are hypersensitive to AZC-induced proliferation arrest. We find that AZC treatment mimics temperature up-shift in arresting cells in G1 and represses expression of CLN1 and CLN2. Mutants with reduced G1 cyclin-Cdc28 activity are hypersensitive to AZC-induced proliferation arrest. Expression of the hyperstable Cln3-2 protein prevents G1 arrest upon AZC treatment and temperature up-shift. Finally, we find that the EXA3-1 mutation, encoding a defective HSF, prevents efficient G1 arrest in response to both temperature up-shift and AZC treatment. We conclude that nontoxic levels of misfolded proteins (induced by AZC treatment or by high temperature) selectively activate HSF, which is required for subsequent G1 arrest.


Collagen-binding activity of Prevotella intermedia measured by a microtitre plate adherence assay.
The ability of Prevotella intermedia to bind type I collagen was investigated. A simple method in which bacterial cells were allowed to attach to collagen-coated microtitre plate wells was used to characterize the activity. All strains of P. intermedia tested, as well as those of the closely related species Prevotella nigrescens, showed a capacity to attach to the collagen film. Exponential-phase cultures of P. intermedia demonstrated a greater binding capacity than older cells. Attachment to the collagen film was inhibited by the presence of EDTA, type I and IV collagen, denatured collagen (gelatin), fibrinogen or fibronectin. Pretreatment of bacterial cells with heat (60 degrees C, 30 min) or proteinase K also inhibited the binding. The collagen-binding activity could be solubilized from the bacterial cell surface by incubation with Zwittergent 3-14, a zwitterionic detergent. The collagen-binding capacity of P. intermedia demonstrated in the present study represents a mechanism of colonization allowing these bacteria to attach to a tissue matrix.


Estimation of the spatial profile of neuromodulation and the temporal latency in motor responses induced by focused ultrasound brain stimulation.
This study investigates the spatial profile and the temporal latency of the brain stimulation induced by the transcranial application of pulsed focused ultrasound (FUS). The site of neuromodulation was detected using 2-deoxy-2-[¹⁸F]fluoro-D-glucose PET immediately after FUS sonication on the unilateral thalamic area of Sprague-Dawley rats. The latency of the stimulation was estimated by measuring the time taken from the onset of the stimulation of the appropriate brain motor area to the corresponding tail motor response. The brain area showing elevated glucose uptake from the PET image was much smaller (56±10% in diameter, 24±6% in length) than the size of the acoustic focus, which is conventionally defined by the full-width at half-maximum of the acoustic intensity field. The spatial dimension of the FUS-mediated neuromodulatory area was more localized, approximated to be full-width at 90%-maximum of the acoustic intensity field. In addition, the time delay of motor responses elicited by the FUS sonication was 171±63 (SD) ms from the onset of sonication. When compared with latencies of other nonultrasonic neurostimulation techniques, the longer time delay associated with FUS-mediated motor responses is suggestive of the nonelectrical modes of neuromodulation, making it a distinctive brain stimulation method.


Adropin: An endocrine link between the biological clock and cholesterol homeostasis.
Identify determinants of plasma adropin concentrations, a secreted peptide translated from the Energy Homeostasis Associated (ENHO) gene linked to metabolic control and vascular function. Associations between plasma adropin concentrations, demographics (sex, age, BMI) and circulating biomarkers of lipid and glucose metabolism were assessed in plasma obtained after an overnight fast in humans. The regulation of adropin expression was then assessed in silico, in cultured human cells, and in animal models. In humans, plasma adropin concentrations are inversely related to atherogenic LDL-cholesterol (LDL-C) levels in men (n = 349), but not in women (n = 401). Analysis of hepatic Enho expression in male mice suggests control by the biological clock. Expression is rhythmic, peaking during maximal food consumption in the dark correlating with transcriptional activation by RORα/γ. The nadir in the light phase coincides with the rest phase and repression by Rev-erb. Plasma adropin concentrations in nonhuman primates (rhesus monkeys) also exhibit peaks coinciding with feeding times (07:00 h, 15:00 h). The ROR inverse agonists SR1001 and the 7-oxygenated sterols 7-β-hydroxysterol and 7-ketocholesterol, or the Rev-erb agonist SR9009, suppress ENHO expression in cultured human HepG2 cells. Consumption of high-cholesterol diets suppress expression of the adropin transcript in mouse liver. However, adropin over expression does not prevent hypercholesterolemia resulting from a high cholesterol diet and/or LDL receptor mutations. In humans, associations between plasma adropin concentrations and LDL-C suggest a link with hepatic lipid metabolism. Mouse studies suggest that the relationship between adropin and cholesterol metabolism is unidirectional, and predominantly involves suppression of adropin expression by cholesterol and 7-oxygenated sterols. Sensing of fatty acids, cholesterol and oxysterols by the RORα/γ ligand-binding domain suggests a plausible functional link between adropin expression and cellular lipid metabolism. Furthermore, the nuclear receptors RORα/γ and Rev-erb may couple adropin synthesis with circadian rhythms in carbohydrate and lipid metabolism.


AAA proteases of mitochondria: quality control of membrane proteins and regulatory functions during mitochondrial biogenesis.
An ubiquitous and conserved proteolytic system regulates the stability of mitochondrial inner membrane proteins. Two AAA proteases with catalytic sites at opposite membrane surfaces form a membrane-integrated quality control system and exert crucial functions during the biogenesis of mitochondria. Their activity is modulated by another membrane-protein complex that is composed of prohibitins. Peptides generated upon proteolysis in the matrix space are transported across the inner membrane by an ATP-binding cassette transporter. The function of these conserved components is discussed in the present review.


Phototropin influence on eyespot development and regulation of phototactic behavior in Chlamydomonas reinhardtii.
The eyespot of Chlamydomonas reinhardtii is a light-sensitive organelle important for phototactic orientation of the alga. Here, we found that eyespot size is strain specific and downregulated in light. In a strain in which the blue light photoreceptor phototropin was deleted by homologous recombination, the light regulation of the eyespot size was affected. We restored this dysfunction in different phototropin complementation experiments. Complementation with the phototropin kinase fragment reduced the eyespot size, independent of light. Interestingly, overexpression of the N-terminal light, oxygen or voltage sensing domains (LOV1+LOV2) alone also affected eyespot size and phototaxis, suggesting that aside from activation of the kinase domain, they fulfill an independent signaling function in the cell. Moreover, phototropin is involved in adjusting the level of channelrhodopsin-1, the dominant primary receptor for phototaxis within the eyespot. Both the level of channelrhodopsin-1 at the onset of illumination and its steady state level during the light period are downregulated by phototropin, whereas the level of channelrhodopsin-2 is not significantly altered. Furthermore, a light intensity-dependent formation of a C-terminal truncated phototropin form was observed. We propose that phototropin is a light regulator of phototaxis that desensitizes the eyespot when blue light intensities increase.


Treatment of renal stones with flexible ureteroscopy in preschool age children.
The aim of the study is to evaluate the efficacy and safety of retrograde intrarenal surgery (RIRS) to treat renal stones in preschool age (<7 years) children. From September 2005 to May 2013, a total of 65 children (31 boys and 34 girls) with 72 renal stones were treated using RIRS. Patients were considered stone-free when the absence of residual fragments was observed on imaging studies. In the presence of residual calculi >4 mm, a second-stage RIRS was performed. The pre-operative, operative and post-operative data of the patients were retrospectively analyzed. A total of 65 patients with a mean age of 4.31 ± 1.99 years (6 months-7 years) were included in the present study. The mean stone size was 14.66 ± 6.12 mm (7-30 mm). The mean operative time was 46.47 ± 18.27 min. In 5 (7.69%) patients, the initial procedure failed to reach the renal collecting system and ended with the insertion of a pigtail stent. The stone-free rates were 83.07 and 92.3% after the first and second procedures, respectively. Complications were observed in 18 (27.7%) patients and classified according to the Clavien system. Post-operative hematuria (Clavien I) occurred in 6 (9.2%) patients, post-operative urinary tract infection with fever (Clavien II) was observed in 10 (15.4%) patients, and ureteral wall injury (Clavien III) was noted in 2 (3%) patients. RIRS is an effective and safe procedure that can be used to manage renal stones in preschool age children.


Chronic ethanol (EtOH) consumption differentially alters gray and white matter EtOH methyl ¹H magnetic resonance intensity in the primate brain.
In vivo magnetic resonance spectroscopy (MRS) has previously been used to directly monitor brain ethanol (EtOH). It has been proposed that the EtOH methyl ¹H resonance intensity is larger in EtOH-tolerant individuals than in sensitive individuals. To characterize the relationship between long-term EtOH exposure and the brain EtOH MRS intensity, we present data from a longitudinal experiment conducted using nonhuman primate subjects. In vivo MRS was used to measure the gray matter (GM) and white matter (WM) EtOH methyl ¹H MRS intensity in 18 adult male rhesus macaques at 4 time points throughout the course of a chronic drinking experiment. Time points were prior to EtOH drinking, following a 3-month EtOH induction procedure, and following 6, and 12 subsequent months of 22 h/d of "open access" to EtOH (4% w/v) and water. The EtOH methyl ¹H MRS intensity, which we observed to be independent of age over the range examined, increased with chronic EtOH exposure in GM and WM. In GM, MRS intensity increased from naïve level following the EtOH induction period (90 g/kg cumulative EtOH intake). In WM, MRS intensity was not significantly different from the EtOH-naïve state until after 6 months of 22-hour free access (110 to 850 g/kg cumulative intake range). The WM MRS intensity in the EtOH-naïve state was positively correlated with future drinking, and the increase in WM MRS intensity was negatively correlated with the amount of EtOH consumed throughout the experiment. Chronic exposure to EtOH is associated with brain changes that result in differential increases in EtOH MRS intensity in GM and WM. The EtOH-naïve WM MRS intensity pattern is consistent with its previously proposed relationship to innate tolerance to the intoxicating effects of EtOH. EtOH-dependent MRS intensity changes in GM required less EtOH exposure than was necessary to produce changes in WM. Within WM, an unexpected, potentially age dependent, enhanced sensitivity to EtOH in light drinkers relative to heavy drinkers was observed.


Gold Nanorods for Light-Based Lung Cancer Theranostics.
Gold nanorods (AuNRs) have the potential to be used in photoacoustic (PA) imaging and plasmonic photothermal therapy (PPTT) due to their unique optical properties, biocompatibility, controlled synthesis, and tuneable surface plasmon resonances (SPRs). Conventionally, continuous-wave (CW) lasers are used in PPTT partly due to their small size and low cost. However, if pulsed-wave (PW) lasers could be used to destroy tissue then combined theranostic applications, such as PA-guided PPTT, would be possible using the same laser system and AuNRs. In this study, we present the effects of AuNR size on PA response, PW-PPTT efficacy, and PA imaging in a tissue-mimicking phantom, as a necessary step in the development of AuNRs towards clinical use. At equivalent NP/mL, the PA signal intensity scaled with AuNR size, indicating that overall mass has an effect on PA response, and reinforcing the importance of efficient tumour targeting. Under PW illumination, all AuNRs showed toxicity at a laser fluence below the maximum permissible exposure to skin, with a maximum of 80% cell-death exhibited by the smallest AuNRs, strengthening the feasibility of PW-PPTT. The theranostic potential of PW lasers combined with AuNRs has been demonstrated for application in the lung.


20 GHz spacing multi-wavelength generation of Brillouin-Raman fiber laser in a hybrid linear cavity.
We demonstrate a tunable multi-wavelength Brillouin-Raman fiber laser with 20 GHz wavelength spacing. The setup is arranged in a linear cavity by employing 7.2 and 11 km dispersion compensating fibers (DCF) in addition to a 30 cm Bismuth-oxide erbium doped fiber. In this experiment, for the purpose of increasing the Stokes lines, it is necessary to optimize Raman pump power and Brillouin pump power together with its corresponding wavelengths. At the specific Brillouin pump wavelength, it is found that the longer length of 11 km DCF with optimized parameters results in larger number of Stokes combs and optical signal to noise ratios (OSNRs). In this case, a total of 195 Brillouin Stokes combs are produced across 28 nm bandwidth at Brillouin pump power of -2 dBm and Raman pump power of 1000 mW. In addition, all Brillouin Stokes signals exhibit an average OSNR of 26 dB.


Acute mechanical injury of the human intervertebral disc: link to degeneration and pain.
Excessive mechanical loading or acute trauma to intervertebral discs (IVDs) is thought to contribute to degeneration and pain. However, the exact mechanisms by which mechanical injury initiates and promotes degeneration remain unclear. This study investigates biochemical changes and extracellular matrix disruption in whole-organ human IVD cultures following acute mechanical injury. Isolated healthy human IVDs were rapidly compressed by 5% (non-injured) or 30% (injured) of disc height. 30% strain consistently cracked cartilage endplates, confirming disc trauma. Three days post-loading, conditioned media were assessed for proteoglycan content and released cytokines. Tissue extracts were assessed for proteoglycan content and for aggrecan integrity. Conditioned media were applied to PC12 cells to evaluate if factors inducing neurite growth were released. Compared to controls, IVD injury caused significant cell death. Injury also caused significantly reduced tissue proteoglycan content with a reciprocal increase of proteoglycan content in culture media. Increased aggrecan fragmentation was observed in injured tissue due to increased matrix metalloproteinase and aggrecanase activity. Injured-IVD conditioned media contained significantly elevated interleukin (IL)-5, IL-6, IL-7, IL-8, MCP-2, GROα, and MIG, and ELISA analysis showed significantly increased nerve growth factor levels compared to non-injured media. Injured-disc media caused significant neurite sprouting in PC12 cells compared to non-injured media. Acute mechanical injury of human IVDs ex vivo initiates release of factors and enzyme activity associated with degeneration and back pain. This work provides direct evidence linking acute trauma, inflammatory factors, neo-innervation and potential degeneration and discogenic pain in vivo.


RNA enzymes as tools for gene ablation.
Ribozymes have the potential to ablate the expression of any gene in a sequence-specific manner and, therefore, may be useful as therapeutic molecules or as tools for the analysis of gene function. Although a number of reports have described ribozymes that are effective in inhibiting gene expression, few studies have attempted, systematically, to analyze the features of ribozymes that affect their potency within cells. Experimental observations suggest that emerging rules governing ribozyme potency in cells can be understood in terms of the competitive interactions between RNA-binding proteins, complementary RNAs and their internal secondary structure.


Lateral Transorbital Endoscopic Access to the Hippocampus, Amygdala, and Entorhinal Cortex: Initial Clinical Experience.
Transorbital approaches traditionally have focused on skull base and cavernous sinus lesions medial to the globe. Lateral orbital approaches to the temporal lobe have not been widely explored despite several theoretical advantages compared to open craniotomy. Recently, we demonstrated the feasibility of the lateral transorbital technique in cadaveric specimens with endoscopic visualization. We describe our initial clinical experience with the endoscope-assisted lateral transorbital approach to lesions in the temporal lobe. Two patients with mesial temporal lobe pathology presenting with seizures underwent surgery. The use of a transpalpebral or Stallard-Wright eyebrow incision enabled access to the intraorbital compartment, and a lateral orbital wall 'keyhole' opening permitted visualization of the anterior temporal pole. This approach afforded adequate access to the surgical target and surrounding structures and was well tolerated by the patients. To the best of our knowledge, this report constitutes the first case series describing the endoscope-assisted lateral transorbital approach to the temporal lobe. We discuss the limits of exposure, the nuances of opening and closing, and comparisons to open craniotomy. Further prospective investigation of this approach is warranted for comparison to traditional approaches to the mesial temporal lobe.