The prevalence of antibiotic-resistant bacteria (ARB) in waters of the Lower Ballona Creek Watershed, Los Angeles County, California.
Screening for the prevalence of antibiotic-resistant bacteria (ARB) was done at the Ballona Creek and Wetlands, an urban-impacted wetland system in Los Angeles, California. The goals were (1) to assess the overall prevalence of ARB, and (2) compare differences in ARB abundance and the types of antibiotic resistance (AR) among the following sample types: lagoon water from Del Rey Lagoon, urban runoff from Ballona Creek, and water from the Ballona Wetlands (tidal water flooding in from the adjacent estuary, and ebbing out from the salt marsh). Antibiotic resistance distributions were analyzed using the Kolmogorov-Smirnov test to develop the cumulative frequency of bacteria having resistance of up to eight antibiotics. Distributions from the environmental water samples were compared to unchlorinated secondary effluent from the Hyperion Water Reclamation Plant that was used as comparator samples likely to have an abundance of ARB. As expected, densities of total and ARB were highest in secondary effluent, followed by urban runoff. Samples of water flooding into the wetlands showed similar results to urban runoff; however, a reduction in densities of total and ARB occurred in water ebbing out of the wetlands. During preliminary work to identify ARB species, several bacterial species of relevance to human illness (e.g., Staphylococcus aureus, Enterococcus hirae, Pseudomonas aeruginosa, Aeromonas veronii, Enterobacter cancerogenus, Serratia marcescens, Pseudomonas stutzeri, and Staphylococcus intermedius) were isolated from sampled waters. If wetlands are a sink for ARB, construction and restoration of wetlands can help in the mediation of this human and environmental health concern.


Examining Mechanisms Underlying Fear-Control in the Extended Parallel Process Model.
This investigation sought to advance the extended parallel process model in important ways by testing associations among the strengths of efficacy and threat appeals with fear as well as two outcomes of fear-control processing, psychological reactance and message minimization. Within the context of print ads admonishing against noise-induced hearing loss (NIHL) and the fictitious Trepidosis virus, partial support was found for the additive model with no support for the multiplicative model. High efficacy appeals mitigated freedom threat perceptions across both contexts. Fear was positively associated with both freedom threat perceptions within the NIHL context and favorable attitudes for both NIHL and Trepidosis virus contexts. In line with psychological reactance theory, a freedom threat was positively associated with psychological reactance. Reactance, in turn, was positively associated with message minimization. The models supported reactance preceding message minimization across both message contexts. Both the theoretical and practical implications are discussed with an emphasis on future research opportunities within the fear-appeal literature.


Recognition of auto- and exoantigens by V4-34 gene encoded antibodies.
The antigenic specificities of 24 V4-34-encoded monoclonal antibodies were compared with the amino acid sequence. The specificities were divided into three categories, red blood cells, B lymphocytes and auto/exoantigens. Six anti-I monoclonal antibodies, with multiple substitutions in their VH region, did not bind B lymphocytes or auto/exoantigens. Reactivity to these two antigens segregated with the 16 anti-i monoclonal antibodies, which were derived from the near germline V4-34 gene. All anti-i monoclonal antibodies bound B lymphocytes, albeit with varying intensities. B-cell binding correlated with basic amino acids in the VH-CDR3. Reactivity to auto/exoantigens was demonstrated only by a subset anti-i monoclonal antibodies and did not correlate with B-lymphocyte or i-antigen binding. These anti-ssDNA reactive monoclonal antibodies had basic amino acids in the VH-CDR3, strongly supporting the suggested role of arginine in DNA binding. However, an arginine-rich CDR3 was not enough to ensure DNA reactivity, since six other anti-i monoclonal antibodies that fulfilled this criteria did not bind ssDNA. Thus it is possible that the anti-DNA reactivity of V4-34-encoded monoclonal antibodies is mediated by the classic antigen-binding groove generated by the CDRs of the heavy/light chains. In contrast, anti-B-cell/i-antigen reactivity is mediated, unconventionally, by the V4-34 protein with a dominant influence of the VH-CDR3.


Using Aminocaproic Acid to Reduce Blood Loss After Primary Unilateral Total Knee Arthroplasty.
xtensive blood loss after total knee arthroplasty (TKA) is common, and affected patients often require blood transfusions. Studies suggest that antifibrinolytic agents such as aminocaproic acid (ACA) reduce blood loss and blood transfusion rates in patients undergoing TKA. We conducted a study to evaluate whether a single intravenous 10-g dose of ACA given during primary unilateral TKA would decrease perioperative blood loss, raise postoperative hemoglobin levels, and reduce postoperative blood transfusion rates. We retrospectively reviewed the charts of 50 comparable cemented primary unilateral TKAs. Twenty-five patients had been given a single intraoperative 10-g dose of ACA (antifibrinolytic group), and the other 25 had not been given ACA (control group). Postoperative drain output was decreased significantly (P < .0001) in the antifibrinolytic group (155 mL) compared with the control group (410 mL), as was the number of units of blood transfused after surgery (antifibrinolytic group, 0 units; control group, 10 units; P < .002). There were no adverse events in the antifibrinolytic group. In TKA, perioperative blood loss and blood transfusion rates were reduced significantly in patients given a single intraoperative intravenous 10-g dose of ACA compared with patients not given antifibrinolytics. The positive effects of ACA were obtained without adverse events or complications.


Radiation exposure from computerized tomography and risk of childhood leukemia: Finnish register-based case-control study of childhood leukemia (FRECCLE).
The only well-established risk factors for childhood leukemia are high-dose ionizing radiation and Down syndrome. Computerized tomography is a common source of low-dose radiation. In this study, we examined the magnitude of the risk of childhood leukemia after pediatric computed tomography examinations. We evaluated the association of computed tomography scans with risk of childhood leukemia in a nationwide register-based case-control study. Cases (n=1,093) were identified from the population-based Finnish Cancer Registry and three controls, matched by gender and age, were randomly selected for each case from the Population Registry. Information was also obtained on birth weight, maternal smoking, parental socioeconomic status and background gamma radiation. Data on computed tomography scans were collected from the ten largest hospitals in Finland, covering approximately 87% of all pediatric computed tomography scans. Red bone marrow doses were estimated with NCICT dose calculation software. The data were analyzed using exact conditional logistic regression analysis. A total of 15 cases (1.4%) and ten controls (0.3%) had undergone one or more computed tomography scans, excluding a 2-year latency period. For one or more computed tomography scans, we observed an odds ratio of 2.82 (95% confidence interval: 1.05 - 7.56). Cumulative red bone marrow dose from computed tomography scans showed an excess odds ratio of 0.13 (95% confidence interval: 0.02 - 0.26) per mGy. Our results are consistent with the notion that even low doses of ionizing radiation observably increase the risk of childhood leukemia. However, the observed risk estimates are somewhat higher than those in earlier studies, probably due to random error, although unknown predisposing factors cannot be ruled out.


Self-stigma in psychiatric patients--standardization of the ISMI scale.
Self-stigma in psychiatric patients is an issue deserving both research and therapeutic attention. The objective of the present study was to evaluate the psychometric properties of a Czech version of the Internalized Stigma of Mental Illness (ISMI) scale assessing the levels of self-stigma in individuals with mental disorders. It consists of 29 items classified into 5 subscales, namely alienation, stereotype endorsement, perceived discrimination, social withdrawal and stigma resistance. The study group comprised 369 patients with a mean age of 41.5±13.3 years, of whom 210 (56.6%) were females. The most frequent diagnosis was neurotic disorders (46.1%), followed by affective disorders (18.4%), substance use disorders (13.3%), psychotic disorders (10.8%), personality disorders (9.5%) and organic disorders (1.6%). Reliability of the scale was evaluated by internal consistency analysis (α=0.91), the split-half method (Spearman-Brown coefficient: 0.93) and test-retest at 3 weeks from the first measurement (N=17; r=0.90, p<0.05). Exploratory factor analysis of the scale was performed, its validity was verified and norms were established that were based on T-scores and sten scores for the entire scale and individual subscales. The Czech translation of the ISMI has adequate psychometric properties.


Trends in cause of death after percutaneous coronary intervention.
The impact of changing demographics on causes of long-term death after percutaneous coronary intervention (PCI) remains incompletely defined. We evaluated trends in cause-specific long-term mortality after index PCI performed at a single center from 1991 to 2008. Deaths were ascertained by scheduled prospective surveillance. Cause was determined via telephone interviews, medical records, autopsy reports, and death certificates. Competing-risks analysis of cause-specific mortality was performed using 3 time periods of PCI (1991-1996, 1997-2002, and 2003-2008). Final follow-up was December 31, 2012. A total of 19 077 patients survived index PCI hospitalization, of whom 6988 subsequently died (37%, 4.48 per 100 person-years). Cause was determined in 6857 (98.1%). Across 3 time periods, there was a 33% decline in cardiac deaths at 5 years after PCI (incidence: 9.8%, 7.4%, and 6.6%) but a 57% increase in noncardiac deaths (7.1%, 8.5%, and 11.2%). Only 36.8% of deaths in the recent era were cardiac. Similar trends were observed regardless of age, extent of coronary disease, or PCI indication. After adjustment for baseline variables, there was a 50% temporal decline in cardiac mortality but no change in noncardiac mortality. The decline in cardiac mortality was driven by fewer deaths from myocardial infarction/sudden death (P<0.001) but not heart failure (P=0.85). The increase in noncardiac mortality was primarily attributable to cancer and chronic diseases (P<0.001). This study found a marked temporal switch from predominantly cardiac to predominantly noncardiac causes of death after PCI over 2 decades. The decline in cardiac mortality was independent of changes in baseline clinical characteristics. These findings have implications for patient care and clinical trial design.


Effect of prepaid health plans on a family practice residency.
The rapid growth of prepaid health care plans imposes clinical, financial, and educational changes on residency programs. In the study reported here, the authors examined some perceived and actual effects of such plans on a family medicine center associated with a family medicine residency training program. In the study, 37 residents and 19 faculty members completed a 5-point, 16-item survey covering the effect of the prepaid plans used at the center on the program's practice profile, cost-containment efforts, and education activities over a three-year period. Overall, the respondents agreed with the need for cost containment that accompanied participation in the prepaid plan and agreed that prepaid plans increased the number of patient visits and visits by family members. The residents and faculty members agreed that prepaid patients were more demanding and were seen more often for minor or inappropriate problems. Regarding the educational impact of the prepaid plans, the respondents agreed that they improved their clinical decision-making, and no significant concern regarding limitation of laboratory or consultations was noted. Some of the respondents' perceptions were corroborated by findings in the clinic data base that showed increased numbers of patient visits, more visits by members of the same family, and no significant change in outpatient consultation rates.


Hsp90 inhibitors reduce influenza virus replication in cell culture.
The viral RNA polymerase complex of influenza A virus consists of three subunits PB1, PB2 and PA. Recently, the cellular chaperone Hsp90 was shown to play a role in nuclear import and assembly of the trimeric polymerase complex by binding to PB1 and PB2. Here we show that Hsp90 inhibitors, geldanamycin or its derivative 17-AAG, delay the growth of influenza virus in cell culture resulting in a 1-2 log reduction in viral titre early in infection. We suggest that this is caused by the reduced half-life of PB1 and PB2 and inhibition of nuclear import of PB1 and PA which lead to reduction in viral RNP assembly. Hsp90 inhibitors may represent a new class of antiviral compounds against influenza viruses.


NTP Toxicology and Carcinogenesis Studies of C.I. Acid Orange 3 (CAS No. 6373-74-6) in F344/N Rats and B6C3F1 Mice (Gavage Studies).
C.I. Acid Orange 3 is a dinitrodiphenylamine derivative used exclusively as a dye (up to 0.2%) in semipermanent hair coloring products. This study was one of a series on semipermanent hair dyes, which included HC Blue No. 1 (NTP TR 271), HC Blue No. 2 (NTP TR 293), HC Red No. 3 (NTP TR 281), and C.I. Disperse Blue 1 (NTP TR 299). Toxicology and carcinogenesis studies of C.I. Acid Orange 3 (90% pure, containing 10% water for short-term studies and containing 6%-8% water and 2%-4% acetone for 2-year studies) were conducted by administering the dye in corn oil by gavage to F344/N rats and B6C3F1 mice of each sex for 14 days, 13 weeks, or 2 years. Fourteen-Day and Thirteen-Week Studies: In the 14-day studies (at 94-1,500 mg/kg in rats and 62-1,000 mg/kg in mice), no compound-related deaths or body weight changes were observed and no adverse effects were observed at necropsy. In the 13-week studies (at 94-1,500 mg/kg in rats and 31-2,000 mg/kg in mice), compound-related kidney lesions were observed in rats and mice of each sex. These lesions included variable degrees of degeneration and necrosis of epithelial cells in the proximal convoluted tubules, regeneration of tubular epithelium, and granular casts in the tubules. In a few female rats of the highest dose group, necrosis of the renal papillae and suppurative inflammation were also observed. Mean body weights were generally comparable among groups of rats and mice. Mice receiving 2,000 mg/kg had body weights 11%-12% lower than those of vehicle controls. Five of 10 female rats that received the highest dose of 1,500 mg/kg died before the end of the study, but no compound-related deaths occurred in male rats or mice of either sex. Based on these results, 2-year studies of C.I. Acid Orange 3 were conducted by administering the dye by gavage in corn oil at 0, 375, or 750 mg/kg to groups of 50 F344/N rats of each sex, 5 days per week for 103 weeks. Groups of 50 male B6C3F1 mice were administered 0, 125, or 250 mg/kg C.I. Acid Orange 3 on the same schedule, and groups of 50 female B6C3F1 mice were administered 0, 250, or 500 mg/kg. These doses were selected on the basis of the nature and severity of the renal lesions in both species. Body Weights and Survival in the Two-Year Studies: Mean body weights of high dose rats were generally more than 10% lower than those of vehicle controls after week 52 for males and week 70 for females. Mean body weights forlow dose groups were comparable to those of vehicle controls. The survival of high dose male (after week 33) and female (after week 14) rats was lower (P<0.05) than that of vehicle controls and was attributed to nephrotoxicity (final survival-- male: vehicle control, 36/50; low dose, 30/50; high dose, 0/50; female: 43/50; 34/50; 7/50). Mean body weights of dosed male and female mice were lower than those of vehicle controls (high dose, 5%-11% after week 74; low dose, 7%-17% after week 48). Survival of both the low dose (after week 102) and high dose (after week 100) groups of male mice was lower than that of the vehicle controls (final survival: 38/50; 25/50; 26/50). Although survival was lower than usual, no notable differences in survival were observed between groups of female mice (final survival: 23/50; 23/50; 24/50). Nonneoplastic and Neoplastic Lesions in the Two-Year Studies: For both species, the kidney was the major target organ for C.I. Acid Orange 3. These findings are summarized in the accompanying table. The incidences of renal pelvic epithelial hyperplasia were increased in dosed rats of each sex. No renal neoplasms were observed in dosed male rats, but a tubular cell adenocarcinoma was observed in a vehicle control male rat. Six transitional cell carcinomas of the kidney were observed in high dose female rats; kidney transitional cell neoplasms have not been observed in 1,697 corn oil vehicle control female F344/N rats. Nonneoplastic lesions characteristic of secondary renal hyperparathyroidism or secondary to uremia also occurred in dosed rats. The lesions included parathyroid hyperplasia, fibrous dysplasia of boia also occurred in dosed rats. The lesions included parathyroid hyperplasia, fibrous dysplasia of bone, erosion and ulcers of the glandular stomach, and mineralization of the aorta and glandular stomach. Epithelial hyperplasia of the urinary bladder was observed in one low dose and three high dose female mice. A squamous cell carcinoma was seen in the urinary bladder of one low dose female mouse. Even though no squamous cell urinary bladder neoplasms have been observed in 1,665 corn oil vehicle control female B6C3F1 mice, this single neoplasm in a low dose animal was not considered to be related to the administration of C.I. Acid Orange 3. Genetic Toxicology: C.I. Acid Orange 3 was mutagenic with and without exogenous metabolic activation in Salmonella typhimurium strains TA97; TA98; and TA100; no mutagenicity was observed for strain TA1535. Audit: The data, documents, and pathology materials from the 2-year studies of C.I. Acid Orange 3 have been audited. The audit findings show that the conduct of the studies is documented adequately and support the data and results given in this Technical Report. Conclusions: Under the conditions of these 2-year gavage studies, there was no evidence of carcinogenic activity of C.I. Acid Orange 3 for male F344/N rats administered 375 mg/kg; because of a marked reduction in survival and no indication of carcinogenicity, the 750 mg/kg group was considered to be inadequate for assessment of carcinogenic activity. There was clear evidence of carcinogenic activity of C.I. Acid Orange 3 for female F344/N rats as shown by the occurrence of transitional cell carcinomas of the kidney in the 750 mg/kg group; this group had reduced survival and chemically related nonneoplastic lesions of the kidney. There was no evidence of carcinogenic activity of C.I. Acid Orange 3 for male B6C3F1 mice administered 125 or 250 mg/kg or for female B6C3F1 mice administered 250 or 500 mg/kg. Nonneoplastic lesions of the kidney were observed in both dose groups of both sexes of rats and mice. Synonyms: 2-anilino-5-(2,4-dinitroanilino)-benzenesulfonic acid, monosodium salt; 5[(2,4-dinitrophenol)amine]-2-(phenylamine)-benzenesulfonic acid, monosodium salt; C.I. 10385; Tetracid Light Yellow 2R


Sanctioning of Illegal and Dangerous Ruck Cleanouts During the 2018 Super Rugby Competition.
Rugby is a high contact sport that results in many injuries. The majority of injuries at senior elite levels result from contact phases during match-play. It is estimated that 10% of all match injuries are associated with the ruck in professional and community rugby. Surveillance of legal and illegal ruck cleanouts and the sanctions imposed by the on-field referees will help identify whether referees are actually enforcing the law according to the laws of the game, which will consequently contribute to the creation and implementation of further injury prevention strategies. Players should play the game in accordance with the laws of the game and be mindful of their own safety and that of others. Coaches and trainers of the game have the responsibility to ensure that players are prepared in a manner that comply with the World Rugby (WR) laws of the game and safe practices. Laws and law amendments are fundamental to the development of sport and introduced for a variety of reasons. The aim of this study was to investigate the rate of sanctioning of illegal and dangerous ruck cleanouts during the 2018 Super Rugby competition by using Nacsport Basic+ video software; 120 round robin matches from the 2018 Super Rugby competition were coded and analyzed. The analysis of the intra reliability showed an almost perfect (>0.95) agreement between all the performance indicators. In total, 22,281 ruck cleanouts were coded of which 9% (n = 2111) were illegal ruck cleanouts and 93% were not sanctioned by the referees; 57% (1087 out of 1953) of the illegal ruck cleanouts not sanctioned by the referees were deemed dangerous. The majority of dangerous illegal ruck cleanouts not sanctioned by the referees were "shoulder charge" (88%, n = 280), "neck roll" (86%, n = 100), and "contact above the shoulder" (81%, n = 201). To aid injury prevention efforts in rugby, future research studies should investigate why on-field referees are not sanctioning all illegal and dangerous ruck cleanouts according to WR Laws of the Game.


Smoking reduction, smoking cessation, and incidence of fatal and non-fatal myocardial infarction in Denmark 1976-1998: a pooled cohort study.
To analyse the effects of smoking reduction and smoking cessation on incidence of myocardial infarction after adjustment for established cardiovascular risk factors. Prospective cohort study with record linkage to mortality and hospital registers. The association of individual change in smoking with myocardial infarction was examined in Cox proportional hazard analyses with continuous heavy smokers (> or =5 cigarettes/day) as reference. Pooled data from three population studies conducted in Copenhagen, Denmark. 10 956 men and 8467 women with complete information on smoking habits at two examinations five to ten years apart were followed up from the second examination for a first hospital admission or death from myocardial infarction. Mean duration of follow up was 13.8 years. A total of 643 participants who were heavy smokers at baseline reduced their daily tobacco consumption by at least 50% without quitting between first and second examination, and 1379 participants stopped smoking. During follow up 1658 men and 521 women experienced a fatal or non-fatal myocardial infarction. After adjustment for cardiovascular risk factors, people who stopped smoking had a decreased risk of myocardial infarction, hazard ratio 0.71 (95% confidence intervals 0.59 to 0.85). Smoking reduction was not associated with reduced risk of myocardial infarction, hazard ratio 1.15 (95% confidence intervals 0.94 to 1.40). These associations remained unchanged after controlling for baseline illness in different ways. Smoking cessation in healthy people reduces the risk of a subsequent myocardial infarction, whereas this study provides no evidence of benefit from reduction in the amount smoked.


Optimization of lycopene extraction from tomato cell suspension culture by response surface methodology.
Radioisotope-labeled lycopene is an important tool for biomedical research but currently is not commercially available. A tomato cell suspension culture system for the production of radioisotope-labeled lycopene was previously developed in our laboratory. In the current study, the goal was to optimize the lycopene extraction efficiency from tomato cell cultures for preparatory high-performance liquid chromatography (HPLC) separation. We employed response surface methodology (RSM), which combines fractional factorial design and a second-degree polynomial model. Tomato cells were homogenized with ethanol, saponified by KOH, and extracted with hexane, and the lycopene content was analyzed by HPLC-PDA. We varied five factors at five levels: ethanol volume (1.33-4 mL/g); homogenization period (0-40 s/g); saturated KOH solution volume (0-0.67 mL/g); hexane volume (1.67-3 mL/g); and vortex period (5-25 s/g). Ridge analysis by SAS suggested that the optimal extraction procedure to extract 1 g of tomato cells was at 1.56 mL of ethanol, 28 s homogenization, 0.29 mL of KOH, 2.49 mL of hexane, and 17.5 s vortex. These optimal conditions predicted by RSM were confirmed to enhance lycopene yield from standardized tomato cell cultures by more than 3-fold.


How Can eHealth Technology Address Challenges Related to Multimorbidity? Perspectives from Patients with Multiple Chronic Conditions.
Patient eHealth technology offers potential support for disease self-management, but the value of existing applications for patients with multiple chronic conditions (MCCs) is unclear. To understand self-management and health care navigation challenges that patients face due to MCCs and to identify opportunities to support these patients through new and enhanced eHealth technology. After administering a screening survey, we conducted 10 focus groups of 3-8 patients grouped by age, sex, and common chronic conditions. Patients discussed challenges associated with having MCCs and their use of (and desires from) technology to support self-management. Three investigators used standard content analysis methods to code the focus group transcripts. Emergent themes were reviewed with all collaborators, and final themes and representative quotes were validated with a sample of participants. Fifty-three individuals with ≥3 chronic conditions and experience using technology for health-related purposes. Focus group participants had an average of five chronic conditions. Participants reported using technology most frequently to search for health information (96%), communicate with health care providers (92%), track medical information (83%), track medications (77%), and support decision-making about treatment (55%). Three themes emerged to guide eHealth technology development: (1) Patients with MCCs manage a high volume of information, visits, and self-care tasks; (2) they need to coordinate, synthesize, and reconcile health information from multiple providers and about different conditions; (3) their unique position at the hub of multiple health issues requires self-advocacy and expertise. Focus groups identified desirable eHealth resources and tools that reflect these themes. Although patients with multiple health issues use eHealth technology to support self-care for specific conditions, they also desire tools that transcend disease boundaries. By addressing the holistic needs of patients with MCCs, eHealth technology can advance health care from a disease-centered to a patient-centered model.


Molecular systematics and historical biogeography of the Nocomis biguttatus species group (Teleostei: Cyprinidae): nuclear and mitochondrial introgression and a cryptic Ozark species.
The Nocomis biguttatus species group ranges widely across North America from the Red River in Oklahoma and Arkansas north to Minnesota and east-west from Wyoming to Ontario. The group includes three traditionally recognized allopatric species: the wide-ranging N. biguttatus and two geographically more restricted species, N. asper from the western Ozarks (Arkansas River system) and two disjunct locations in the Red River system, and N. effusus from the Green, Cumberland, and lower Tennessee rivers. Separate analyses of the mitochondrial cytb gene and two nuclear genes (S7 intron 1 and a portion of the gene for growth hormone, GH), each resolved a cryptic species previously treated as N. biguttatus from the southern Ozarks (White River). Relationships among the four species were unresolved because of conflicts between cytb and S7 and a lack of resolution for GH. A previously indicated N. biguttatus-N. effusus sister-relationship appears to reflect past hybridization and mtDNA capture by N. effusus. Nocomis biguttatus includes four primary cytb clades with unresolved inter-relationships. A Northern Ozarks-Great Plains-Upper Midwest Clade and an Ohio River-Eastern Great Lakes Clade presumably represent late Quaternary dispersal from glacial refugia in, respectively, the northern Ozarks and an unglaciated portion of the Ohio River system. Other clades include one from the Meramec River and a Black River-St. Francis River Clade. There was evidence in N. effusus for a phylogeographic break between the lower Tennessee River and the Green-Cumberland basins. Geographic structure is weak in N. asper, indicating relatively recent contact between now disjunct populations in the Arkansas and Red river basins. The Blue River population of N. asper appears to reflect late Pleistocene or Holocene hybridization and genetic swamping of a resident native population of N. biguttatus by an invading population of N. asper. This postulates past occurrence of N. biguttatus far south of its present range.


Conformation and ion channel activity of lymphotoxin at neutral and low pH.
Lymphotoxin (LT or TNF-beta), a T cell-derived lymphokine with partial homology to TNF-alpha, was found to bind to dimyristoylphosphatidylcholine vesicles in a pH-dependent manner: binding increased with decreasing pH. Binding was not limited to surface association with phospholipid head groups because studies with a photoreactive membrane-restricted probe revealed protein penetration of the hydrocarbon core of the bilayer. Intramembranous photolabeling of the trimeric form of LT demonstrated maintenance of quaternary structure upon bilayer insertion. The efficiency of insertion was greatly enhanced with gel-phase bilayers compared with fluid phase bilayers even though the binding efficiency was much lower. Hence, binding and insertion represent two distinct physical processes. Intrinsic fluorescence and dye binding assays showed that the acid-facilitated membrane interactions stemmed from acid-induced changes in protein conformation. The acquisition of hydrophobic characteristics through these conformational changes supplies a physical explanation for LT conversion from a water-soluble form to a membrane-embedded structure. Moreover, the use of vesicle-embedded LT to prepare planar bilayers vs the addition of soluble LT subsequent to bilayer formation demonstrated that LT exhibits channel activity and that low pH-induced membrane insertion precedes and is distinct from expression of voltage-dependent ion gating. LT's ability to associate intimately with lipid vesicles and form ion channels mirrors the behavior of TNF-alpha. Thus, although LT and TNF-alpha are secreted by different cell types, the conservation of membrane binding, insertion, and channel-forming activities suggests a functional role in response induction.


Assessing the associations of sodium intake with long-term all-cause and cardiovascular mortality in a hypertensive cohort.
Although higher sodium intake is known to increase blood pressure, its association with cardiovascular mortality is less established. We examined the association of baseline sodium intake in a hypertensive cohort with all-cause and cardiovascular mortality over a mean follow-up of 18.6 years. Three thousand five hundred five subjects were participants in a worksite hypertension program. Sodium intake was estimated by 24-hour urine excretion. Mortality data were obtained from the U.S. National Death Index. Unadjusted and multivariable-adjusted associations between sodium quartiles (quartile I (QI) to quartile IV (QIV)) and mortality were assessed using Cox models. Estimated mean ± SD sodium intake was 130±69 mmol overall (55±20 mmol in QI; 220±56 mmol in QIV). Baseline systolic blood pressure did not vary significantly between groups. Last available mean systolic blood pressure was highest in QI and lowest in QIV (137±16 vs. 134±14 mm Hg; P = 0.009). Overall there were 1,013 deaths (399 cardiovascular). Unadjusted models exhibited significant inverse relationships between sodium and mortality outcomes. In adjusted models, sodium intake was not significantly associated with cardiovascular mortality (QI vs. QIV: hazard ratio (HR) = 1.00; 95% confidence interval (CI) = 0.71-1.42; P = 0.99). A borderline significant direct association with all-cause mortality was observed (QI vs. QIV: HR = 0.81; 95% CI = 0.66-1.00; P = 0.05) driven partly by noncardiovascular deaths. Our study found no significant association between sodium intake and cardiovascular outcomes, although a significant association with all-cause mortality was observed. Although these findings suggest that sodium may not have a strong relationship with cardiovascular mortality, the inconsistent results cast doubt on whether a single measurement can reliably predict mortality over a prolonged follow-up period.


Current results of the surgical management of aneurysms of the basilar apex.
To provide current information regarding the expected clinical outcomes and sources of morbidity and mortality in the modern surgical management of basilar apex aneurysms. A retrospective review was conducted of 303 cases of such aneurysms that were treated surgically during 18 years at one institution. Postoperative angiography was performed in 81% of the cases. Clinical grading using the Glasgow Outcome Scale was conducted at the time of hospital discharge and for 91% of the surviving patients at 6 months after surgery. The preoperative parameters that were linked statistically to poor clinical outcome were identified through the use of single and multivariate analyses. More than 80% of the patients were operated on using some modification of the trans-sylvian exposure, and temporary arterial occlusion was used routinely. Good outcomes (Glasgow Outcome Scale scores of 4 or 5) were achieved in 76% of the patients at the time of discharge and in 81% of the patients at 6 months after surgery. There was no incidence of postoperative subarachnoid hemorrhage. Residual aneurysm was revealed by postoperative angiography in 6% of the cases. Factors found to be statistically linked to poor outcome included poor admission grade (Hunt and Hess Grades IV and V), patient age older than 65 years, computed tomographic demonstration of thick basal cistern clot, aneurysm size greater than 20 mm, and symptoms attributable to brain stem compression. Direct microsurgical repair of basilar apex aneurysms should result in good clinical outcomes in 80 to 85% of cases, with reliable prevention of subarachnoid bleeding and routine elimination/reduction of symptoms secondary to mass effect. Those patients who are at high risk for poor outcomes can be identified by the presence of certain clinical, radiographic, and demographic features before undergoing surgery and can be considered for alternative or adjunctive modes of therapy if long-term efficacy of such treatment is demonstrated.


Palmar arch revascularization for arterial occlusion of the distal upper extremity.
During a period of nine years, 10 patients, ages 42 to 66 years (55 +/- 7, mean +/- SD) underwent upper extremity bypass for ischemic changes to the hand not responsive to conservative management. Patients were referred from the vascular department at the authors' institution. Preoperative angiograms were performed and attempts at angioplasty or intravenous attempts to dissolve clots were carried out (with tissue plasminogen activator-tPA) when appropriate. Patients with persistent upper extremity ischemia and an obvious occlusion with reconstitution in the hand were candidates for upper extremity bypass to the palmar arch. All patients had upper extremity bypasses performed with reverse saphenous vein grafts. The proximal anastomoses (end-to-side) were performed by either the vascular or plastic surgery team, while all distal anastomoses (end-to-side) were performed by plastic surgery team microscopic magnification to the deep or superficial palmar arch. Postoperative follow-up ranged from 3 months to 3 years. The bypass graft to the hand resulted in improved pain and resolution of tissue ischemia in all cases. Patients with preoperative ulcers were completely healed by 3 months. The results are in accordance with previous studies demonstrating that improved blood flow afforded by the procedure can improve the healing of recalcitrant ulcers and mitigate the symptoms of ischemic changes. In addition, end-to-side anastomosis to the palmar arch offers significant advantages, in that the continuity of the arch is maintained with all possible outflow vessels, and the problems associated with size discrepancy in the anastomosed vessels are eliminated.


Should we exclude acute stroke patients with previous intracerebral hemorrhage from receiving intravenous thrombolysis?
Current guidelines have contraindicated history of intracerebral hemorrhage for intravenous recombinant tissue plasminogen activator. This study aimed to investigate the safety and effectiveness of intravenous recombinant tissue plasminogen activator for patients who had previous intracerebral hemorrhage on history or initial brain magnetic resonance imaging. Using a prospective multicenter stroke registry database, we identified acute ischemic stroke patients treated with intravenous recombinant tissue plasminogen activator within 4.5 h of onset. Previous intracerebral hemorrhage was defined as having a clinical history or evidence of old intracerebral hemorrhage on initial brain magnetic resonance imaging. Associations of previous intracerebral hemorrhage with symptomatic hemorrhagic transformation during hospitalization and functional outcome and mortality at discharge and three months were analyzed. Among 1495 patients who were treated with intravenous recombinant tissue plasminogen activator, 73 (4.9%) had previous intracerebral hemorrhage; 9 on history only, 61 on magnetic resonance imaging only and 3 on both. Of those 1495 patients, 71 (4.7%) experienced symptomatic hemorrhagic transformation; 6.8% in patients with previous intracerebral hemorrhage and 4.6% in those without previous intracerebral hemorrhage. Multivariable logistic regression analysis showed that previous intracerebral hemorrhage did not significantly increase the risk of symptomatic hemorrhagic transformation (odds ratio 1.08, 95% confidence interval 0.39-2.96) mortality, and most of functional outcome measures Previous intracerebral hemorrhage may neither increase the risk of symptomatic hemorrhagic transformation nor alter major clinical outcomes in acute ischemic stroke patients receiving intravenous recombinant tissue plasminogen activator. This study suggests reconsideration of prior history of intracerebral hemorrhage as an exclusion criterion for intravenous recombinant tissue plasminogen activator administration in acute ischemic stroke.


Plasma membrane calcium ATPase isoform 4 inhibits vascular endothelial growth factor-mediated angiogenesis through interaction with calcineurin.
Vascular endothelial growth factor (VEGF) has been identified as a crucial regulator of physiological and pathological angiogenesis. Among the intracellular signaling pathways triggered by VEGF, activation of the calcineurin/nuclear factor of activated T cells (NFAT) signaling axis has emerged as a critical mediator of angiogenic processes. We and others previously reported a novel role for the plasma membrane calcium ATPase (PMCA) as an endogenous inhibitor of the calcineurin/NFAT pathway, via interaction with calcineurin, in cardiomyocytes and breast cancer cells. However, the functional significance of the PMCA/calcineurin interaction in endothelial pathophysiology has not been addressed thus far. Using in vitro and in vivo assays, we here demonstrate that the interaction between PMCA4 and calcineurin in VEGF-stimulated endothelial cells leads to downregulation of the calcineurin/NFAT pathway and to a significant reduction in the subsequent expression of the NFAT-dependent, VEGF-activated, proangiogenic genes RCAN1.4 and Cox-2. PMCA4-dependent inhibition of calcineurin signaling translates into a reduction in endothelial cell motility and blood vessel formation that ultimately impairs in vivo angiogenesis by VEGF. Given the importance of the calcineurin/NFAT pathway in the regulation of pathological angiogenesis, targeted modulation of PMCA4 functionality might open novel therapeutic avenues to promote or attenuate new vessel formation in diseases that occur with angiogenesis.


The Best Age for Pregnancy and Undue Pressures.
In western countries we assist at the paradox that fertility is socially discouraged by a mindset that depicts fertility as a resource to exploit as late as possible. So, couples have high expectative about the advantages of delayed parenthood, but they are scarcely informed about its risks. Scientific data suggests to anticipate the first pregnancy, but social pressures impose to wait, though delayed childbearing can provoke sterility and a greater gap between generations. The best age to become parents should be autonomously decided by a couple, under the condition of being a free informed choice and not a social imposition, but currently this is not guaranteed to western women and men.


Tai Chi for rheumatoid arthritis.
Rheumatoid arthritis (RA) is a chronic, systemic, inflammatory, autoimmune disease that results in joint deformity and immobility of the musculoskeletal system. The major goals of treatment are to relieve pain, reduce inflammation, slow down or stop joint damage, prevent disability, and preserve or improve the person's sense of well-being and ability to function. Tai Chi, interchangeably known as Tai Chi Chuan, is an ancient Chinese health-promoting martial art form that has been recognized in China as an effective arthritis therapy for centuries. This is an update of a review published in 2004. To assess the benefits and harms of Tai Chi as a treatment for people with rheumatoid arthritis (RA). We updated the search of CENTRAL, MEDLINE, Embase, and clinical trial registries from 2002 to September 2018. We selected randomized controlled trials and controlled clinical trials examining the benefits (ACR improvement criteria or pain, disease progression, function, and radiographic progression), and harms (adverse events and withdrawals) of exercise programs with Tai Chi instruction or incorporating principles of Tai Chi philosophy. We included studies of any duration that included control groups who received either no therapy or alternate therapy. We used standard methodological procedures expected by Cochrane. Adding three studies (156 additional participants) to the original review, this update contains a total of seven trials with 345 participants. Participants were mostly women with RA, ranging in age from 16 to 80 years, who were treated in outpatient settings in China, South Korea, and the USA. The majority of the trials were at high risk of bias for performance and detection bias, due to the lack of blinding of participants or assessors. Almost 75% of the studies did not report random sequence generation, and we judged the risk of bias as unclear for allocation concealment in the majority of studies. The duration of the Tai Chi programs ranged from 8 to 12 weeks.It is uncertain whether Tai Chi-based exercise programs provide a clinically important improvement in pain among Tai Chi participants compared to no therapy or alternate therapy. The change in mean pain in control groups, measured on visual analog scale (VAS 0 to 10 score, reduced score means less pain) ranged from a decrease of 0.51 to an increase of 1.6 at 12 weeks; in the Tai Chi groups, pain was reduced by a mean difference (MD) of -2.15 (95% confidence interval (CI) -3.19 to -1.11); 22% absolute improvement (95% CI, 11% to 32% improvement); 2 studies, 81 participants; very low-quality evidence, downgraded for imprecision, blinding and attrition bias.There was very low-quality evidence, downgraded for, blinding, and attrition, that was inconclusive for an important difference in disease activity, measured using Disease Activity Scale (DAS-28-ESR) scores (0 to 10 scale, lower score means less disease activity), with no change in the control group and 0.40 reduction (95% CI -1.10 to 0.30) with Tai Chi; 4% absolute improvement (95% CI 11% improvement to 3% worsening); 1 study, 43 participants.For the assessment of function, the change in mean Health Assessment Questionnaire (HAQ; 0 to 3 scale, lower score means better function) ranged from 0 to 0.1 in the control group, and reduced by MD 0.33 in the Tai Chi group (95% CI -0.79 to 0.12); 11% absolute improvement (95% CI 26% improvement to 4% worsening); 2 studies, 63 participants; very low-quality evidence, downgraded for imprecision, blinding, and attrition. We are unsure of an important improvement, as the results were inconclusive.Participants in Tai Chi programs were less likely than those in a control group to withdraw from studies at 8 to 12 weeks (19/180 in intervention groups versus 49/165 in control groups; risk ratio (RR) 0.40 (95% CI 0.19 to 0.86); absolute difference 17% fewer (95% CI 30% fewer to 3% fewer); 7 studies, 289 participants; low-quality evidence, downgraded for imprecision and blinding.There were no data available for radiographic progression. Short-term adverse events were not reported by group, but in two studies there was some narrative description of joint and muscle soreness and cramps; long-term adverse events were not reported. It is uncertain whether Tai Chi has any effect on clinical outcomes (joint pain, activity limitation, function) in RA, and important effects cannot be confirmed or excluded, since all outcomes had very low-quality evidence. Withdrawals from study were greater in the control groups than the Tai Chi groups, based on low-quality evidence. Although the incidence of adverse events is likely to be low with Tai Chi, we are uncertain, as studies failed to explicitly report such events. Few minor adverse events (joint and muscle soreness and cramps) were described qualitatively in the narrative of two of the studies. This updated review provides minimal change in the conclusions from the previous review, i.e. a pain outcome.


Regulation of yeast replicative life span by TOR and Sch9 in response to nutrients.
Calorie restriction increases life span in many organisms, including the budding yeast Saccharomyces cerevisiae. From a large-scale analysis of 564 single-gene-deletion strains of yeast, we identified 10 gene deletions that increase replicative life span. Six of these correspond to genes encoding components of the nutrient-responsive TOR and Sch9 pathways. Calorie restriction of tor1D or sch9D cells failed to further increase life span and, like calorie restriction, deletion of either SCH9 or TOR1 increased life span independent of the Sir2 histone deacetylase. We propose that the TOR and Sch9 kinases define a primary conduit through which excess nutrient intake limits longevity in yeast.


Colorectal cancer liver metastases organoids retain characteristics of original tumor and acquire chemotherapy resistance.
Colorectal cancer (CRC) liver metastasis is highly unfavorable for patient outcome and is a leading cause of cancer-related death. Pre-clinical research of CRC liver metastasis predominately utilizes CRC cell lines grown in tissue culture. Here, we demonstrate that CRC liver metastases organoids derived from human specimens recapitulate some aspects of human disease. Human CRC liver metastases pathological specimens were obtained following patient consent. Tumor disaggregates were plated and organoids were allowed to expand. CRC markers were identified by immunofluorescence. Stem cell genes were analysed by QPCR and flow cytometry. Response to drug therapy was quantified using time-lapse imaging and MATLAB analysis. Organoids showed global expression of the epithelial marker, EpCAM and the adenocarcinoma marker, CEA CAM1. Flow cytometry analysis demonstrated that organoids express the stem cell surface markers CD24 and CD44. Finally, we demonstrated that CRC liver metastases organoids acquire chemotherapy resistance and can be utilized as surrogates for drug testing. These data demonstrate that CRC liver metastases organoids recapitulate some aspects of human disease and may provide an invaluable resource for investigating novel drug therapies, chemotherapy resistance and mechanism of metastasis.


The natural history of tree nut allergy.
Although 20% of children outgrow peanut allergy, the natural history of tree nut (TN) allergy has not been well studied. The goals of the study were to estimate the proportion of children who outgrow TN allergy and examine predictors of outgrowing it. Patients with TN allergy, defined as a history of reaction on ingestion and evidence of TN-specific IgE (TN-IgE) or positive TN-specific IgE level but no history of ingestion, were evaluated. If all current TN-IgE levels were less than 10 kilounits of antibody (kU(A))/L, double-blind, placebo-controlled food challenges were offered. Patients who had undergone open TN challenges as part of routine clinical care were also included. Two hundred seventy-eight patients with TN allergy were identified. One hundred one (36%) had a history of acute reactions, 12 (12%) of whom had reactions to multiple TNs and 73 (63%) of whom had a history of moderate-to-severe reactions. Nine of 20 patients who had previously reacted to a TN passed challenges, so that 9 (8.9%; 95% CI, 4% to 16%) of 101 patients with a history of prior TN reactions outgrew TN allergy. Fourteen of 19 who had never ingested TNs but had detectable TN-specific IgE levels passed challenges. One hundred sixty-one did not meet the challenge criteria, and 78 met the criteria but declined challenges. Looking at specific TN-IgE cutoffs, 58% with TN-IgE levels of 5 kU(A)/L or less and 63% with TN-IgE levels of 2 kU(A)/L or less passed challenges. Approximately 9% of patients outgrow TN allergy, including some who had prior severe reactions. Although ideal cutoffs for challenge cannot be firmly recommended on the basis of these data, patients aged 4 years or older with all TN-IgE levels of 5 kU(A)/L or less should be considered for challenge.


Reactivity of T cells from women with antibodies to the human platelet antigen (HPA)-1a to peptides encompassing the HPA-1 polymorphism.
The human platelet antigen-1a (HPA-1a) is the most common alloantigenic target in fetal and neonatal alloimmune thrombocytopenia (NAIT). Treatment currently depends on the outcome in previous pregnancies. HPA-1 specific T cell responses were determined in 14 HPA-1a alloimmunized women during or after pregnancies affected by NAIT. Peripheral blood mononuclear cells were incubated with peptides encompassing the Leu33Pro polymorphism (residues 20-39 and 24-45 in both Leu33 (HPA-1a) and Pro33 (HPA-1b) forms) or control recall antigens in the presence of autologous sera and T cell proliferation was measured by (3)H-thymidine incorporation. Control antenatal and postpartum sera suppressed T cell proliferation and use of such sera was avoided. Most patients (86%) responded to the HPA-1a peptides with 64% also having weaker T cell proliferation to the HPA-1b peptides; 14% had no activity towards any peptide despite responding to control antigens. Administration of IVIG during pregnancy appeared to reduce T cell reactivity to HPA-1 peptides. Postnatal anti-HPA-1a T cell responses from women who had a severe history of NAIT (an intracranial haemorrhage in a previous fetus) were greater than those from women with a mild history. This assay may have the potential to predict disease severity if performed prior to or early in pregnancy.


Enrolling vulnerable, uninsured but eligible children in public health insurance: association with health status and primary care access.
Given that more than two thirds of uninsured children in California are eligible for public health insurance coverage, this study examined differences in primary care access and health status between uninsured but eligible (UBE) children and those who were insured. Using data on 19485 children from the 2001 California Health Interview Survey, this study examined differences in primary care access and health status for UBE children versus those who were enrolled in public coverage. Results are stratified by profiles of other risk factors (RF) for poor access: nonwhite, low income, low parent education, and non-English speaking. UBE children were less likely than publicly enrolled children to have a physician visit in the past year, dental visit in the past year, and a regular source of care. On the basis of differences between the UBE children and enrollees in the prevalence of each dependent measure, UBE children with multiple RFs experienced greater disparities than UBE children with fewer RFs. For example, enrollees were more likely than UBE children to have a regular source of care among children with 2, 3, or 4 RFs (differences of 26, 26, and 25 percentage points, respectively) compared with 1 RF (19 percentage points) and 0 RFs (12 percentage points). A similar pattern was found for dental visits but not physician visits. Although there was no difference in health status between UBE children and enrollees overall, enrollees were more likely than UBE children to have excellent/very good health status among children with 2 RFs (difference of 9 percentage points), 3 RFs (12 percentage points), and 4 RFs (11 percentage points). This study demonstrates that UBE children in California have poorer access to care compared with enrollees, and those with the highest levels of risk have poorer health status. This suggests that providing insurance to these children (and particularly those with multiple RFs) may lead to improved access and health for these vulnerable children.


Direct α-C-H amination using various amino agents by selective oxidative copper catalysis: a divergent access to functional quinolines.
Herein, we present, for the first time, direct dehydrogenative α-C-H amination of tetrahydroquinolines (THQs) using various amino agents by selective aerobic copper catalysis, which enables divergent access to 2-aminoquinolines, the core structures of numerous functional products. In which, the catalyst system preferentially oxidizes the tetrahydroquinolines between two amino reactants, and the presence of TEMPO significantly enhances the capability of the first oxidation of THQs and makes it a kinetically controlled process, thus favoring the C-N bond-forming step. The developed chemistry features broad substrates, excellent functional tolerance, high chemo-selectivity, and no need for pre-preparation of specific aminating agents, which offers a practical way for diverse and atom-economic synthesis of 2-aminoquinolines that are difficult to prepare or inaccessible with the existing C-H amination approaches.


High glycemic load diet, milk and ice cream consumption are related to acne vulgaris in Malaysian young adults: a case control study.
The role of dietary factors in the pathophysiology of acne vulgaris is highly controversial. Hence, the aim of this study was to determine the association between dietary factors and acne vulgaris among Malaysian young adults. A case-control study was conducted among 44 acne vulgaris patients and 44 controls aged 18 to 30 years from October 2010 to January 2011. Comprehensive acne severity scale (CASS) was used to determine acne severity. A questionnaire comprising items enquiring into the respondent's family history and dietary patterns was distributed. Subjects were asked to record their food intake on two weekdays and one day on a weekend in a three day food diary. Anthropometric measurements including body weight, height and body fat percentage were taken. Acne severity was assessed by a dermatologist. Cases had a significantly higher dietary glycemic load (175 ± 35) compared to controls (122 ± 28) (p < 0.001). The frequency of milk (p < 0.01) and ice-cream (p < 0.01) consumptions was significantly higher in cases compared to controls. Females in the case group had a higher daily energy intake compared to their counterparts in the control group, 1812 ± 331 and 1590 ± 148 kcal respectively (p < 0.05). No significant difference was found in other nutrient intakes, Body Mass Index, and body fat percentage between case and control groups (p > 0.05). Glycemic load diet and frequencies of milk and ice cream intake were positively associated with acne vulgaris.


Discriminant subspace analysis: a Fukunaga-Koontz approach.
The Fisher Linear Discriminant (FLD) is commonly used in pattern recognition. It finds a linear subspace that maximally separates class patterns according to the Fisher Criterion. Several methods of computing the FLD have been proposed in the literature, most of which require the calculation of the so-called scatter matrices. In this paper, we bring a fresh perspective to FLD via the Fukunaga-Koontz Transform (FKT). We do this by decomposing the whole data space into four subspaces with different discriminability, as measured by eigenvalue ratios. By connecting the eigenvalue ratio with the generalized eigenvalue, we show where the Fisher Criterion is maximally satisfied. We prove the relationship between FLD and FKT analytically, and propose a unified framework to understanding some existing work. Furthermore, we extend our our theory to Multiple Discriminant Analysis (MDA). This is done by transforming the data into intra- and extra-class spaces, followed by maximizing the Bhattacharyya distance. Based on our FKT analysis, we identify the discriminant subspaces of MDA/FKT, and propose an efficient algorithm, which works even when the scatter matrices are singular, or too large to be formed. Our method is general and may be applied to different pattern recognition problems. We validate our method by experimenting on synthetic and real data.


De novo transcriptome assembly, functional annotation and differential gene expression analysis of juvenile and adult E. fetida, a model oligochaete used in ecotoxicological studies.
Earthworms are sensitive to toxic chemicals present in the soil and so are useful indicator organisms for soil health. Eisenia fetida are commonly used in ecotoxicological studies; therefore the assembly of a baseline transcriptome is important for subsequent analyses exploring the impact of toxin exposure on genome wide gene expression. This paper reports on the de novo transcriptome assembly of E. fetida using Trinity, a freely available software tool. Trinotate was used to carry out functional annotation of the Trinity generated transcriptome file and the transdecoder generated peptide sequence file along with BLASTX, BLASTP and HMMER searches and were loaded into a Sqlite3 database. To identify differentially expressed transcripts; each of the original sequence files were aligned to the de novo assembled transcriptome using Bowtie and then RSEM was used to estimate expression values based on the alignment. EdgeR was used to calculate differential expression between the two conditions, with an FDR corrected P value cut off of 0.001, this returned six significantly differentially expressed genes. Initial BLASTX hits of these putative genes included hits with annelid ferritin and lysozyme proteins, as well as fungal NADH cytochrome b5 reductase and senescence associated proteins. At a cut off of P = 0.01 there were a further 26 differentially expressed genes. These data have been made publicly available, and to our knowledge represent the most comprehensive available transcriptome for E. fetida assembled from RNA sequencing data. This provides important groundwork for subsequent ecotoxicogenomic studies exploring the impact of the environment on global gene expression in E. fetida and other earthworm species.


Trends in asthma prevalence, health care use, and mortality in the United States, 2001-2010.
Asthma prevalence increased from 2001 to 2010: An estimated 25.7 million persons had asthma in 2010. Certain demographic groups had higher asthma prevalence: children aged 0–17 years, females, black persons, persons of multiple race, Puerto Rican persons, and persons with a family income below the poverty level. This report examines rates for asthma outcomes (health care encounters and death) for persons with asthma rather than for the general population. Rates for the general population represent the burden of asthma in the United States. Rates for the population with asthma take into account changes in asthma prevalence over time and differences in asthma prevalence among demographic groups. From 2001 to 2009, rates for ED visits and hospitalizations per 100 persons with asthma remained stable, while rates for asthma visits in primary care settings (physician offices or hospital outpatient departments) and asthma deaths declined. For the period 2007–2009, asthma visit rates (per 100 persons with asthma) in primary care settings for black persons were similar to those for white persons, but rates for asthma ED visits, hospitalizations, and death (per 1,000) were higher. Compared with adults, children aged 0–17 years had a higher rate for asthma visits in primary care settings and EDs, but had a similar hospitalization rate and a lower asthma death rate.


Peptidylarginine Deiminase Isozyme-Specific PAD2, PAD3 and PAD4 Inhibitors Differentially Modulate Extracellular Vesicle Signatures and Cell Invasion in Two Glioblastoma Multiforme Cell Lines.
Glioblastoma multiforme (GBM) is an aggressive adult brain tumour with poor prognosis. Roles for peptidylarginine deiminases (PADs) in GBM have recently been highlighted. Here, two GBM cell lines were treated with PAD2, PAD3 and PAD4 isozyme-specific inhibitors. Effects were assessed on extracellular vesicle (EV) signatures, including EV-microRNA cargo (miR21, miR126 and miR210), and on changes in cellular protein expression relevant for mitochondrial housekeeping (prohibitin (PHB)) and cancer progression (stromal interaction molecule 1 (STIM-1) and moesin), as well as assessing cell invasion. Overall, GBM cell-line specific differences for the three PAD isozyme-specific inhibitors were observed on modulation of EV-signatures, PHB, STIM-1 and moesin protein levels, as well as on cell invasion. The PAD3 inhibitor was most effective in modulating EVs to anti-oncogenic signatures (reduced miR21 and miR210, and elevated miR126), to reduce cell invasion and to modulate protein expression of pro-GBM proteins in LN229 cells, while the PAD2 and PAD4 inhibitors were more effective in LN18 cells. Furthermore, Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways for deiminated proteins relating to cancer, metabolism and inflammation differed between the two GBM cell lines. Our findings highlight roles for the different PAD isozymes in the heterogeneity of GBM tumours and the potential for tailored PAD-isozyme specific treatment.


Feed your gut with caution!
The intestinal epithelium, a single layer of cells organized into crypts and villi, is an important component of host defense against microbial community and its noxious metabolites. To preserve the epithelium integrity under constant exposure of intense physiological and environmental stressors, this surface is continuously and rapidly renewed by pluripotent intestinal epithelial stem cells that reside in the base of crypts. A recent study by Kaiko et al. shows that structural design of intestinal crypts protects the colonic epithelial stem/progenitor cells from proliferation-suppressing metabolites derived from the gut microbiota. Among these metabolites, butyrate is identified as the most potent inhibitor of colonic epithelial stem/progenitor cells proliferation. By rapidly metabolizing the butyrate, the colonocytes at the crypt further institute a metabolic barrier to protect the crypt-resident stem cells that fuel its renewal and repair.


Association of viral infection and appendicitis.
What causes appendicitis is not known; however, studies have suggested a relationship between viral diseases and appendicitis. Building on evidence of cyclic patterns of appendicitis with apparent outbreaks consistent with an infectious etiology, we hypothesized that there is a relationship between population rates of appendicitis and several infectious diseases. Epidemiologic study. The National Hospital Discharge Survey Estimated US hospitalized population. International Classification of Diseases, Ninth Revision, Clinical Modification discharge diagnosis codes of the National Hospital Discharge Survey were queried from 1970 to 2006 to identify admissions for appendicitis, influenza, rotavirus, and enteric infections. Cointegration analysis of time series data was used to determine if the disease incidence trends for these various disease entities varied over time together. Rates of influenza and nonperforating appendicitis declined progressively from the late 1970s to 1995 and rose thereafter, but influenza rates exhibited more distinct seasonal variation than appendicitis rates. Rotavirus infection showed no association with the incidence of nonperforating appendicitis. Perforating appendicitis showed a dissimilar trend to both nonperforating appendicitis and viral infection. Hospital admissions for enteric infections substantially increased over the years but were not related to appendicitis cases. Neither influenza nor rotavirus are likely proximate causes of appendicitis given the lack of a seasonal relationship between these disease entities. However, because of significant cointegration between the annual incidence rates of influenza and nonperforated appendicitis, it is possible that these diseases share common etiologic determinates, pathogenetic mechanisms, or environmental factors that similarly affect their incidence.


Measurement properties of multidimensional patient-reported outcome measures in neurodisability: a systematic review of evaluation studies.
To identify and appraise the quality of studies that primarily assessed the measurement properties of English language versions of multidimensional patient-reported outcome measures (PROMs) when evaluated with children with neurodisability, and to summarize this evidence. MEDLINE, Embase, PsycINFO, CINAHL, AMED, and the National Health Service Economic Evaluation Database were searched. The methodological quality of the papers was assessed using the COnsensus-based Standards for selection of health Measurement INstruments checklist. Evidence of content validity, construct validity, internal consistency, test-retest reliability, proxy reliability, responsiveness, and precision was extracted and judged against standardized reference criteria. We identified 48 studies of mostly fair to good methodological quality: 37 papers for seven generic PROMs (CHIP, CHQ, CQoL, KIDSCREEN, PedsQL, SLSS, and YQOL), seven papers for two chronic-generic PROMs (DISABKIDS and Neuro-QOL), and four papers for three preference-based measures (HUI, EQ-5D-Y, and CHSCS-PS). On the basis of this appraisal, the DISABKIDS appears to have more supportive evidence in samples of children with neurodisability. The overall lack of evidence for responsiveness and measurement error is a concern when using these instruments to measure change, or to interpret the findings of studies in which these PROMs have been used to assess change.


Functional MRI Preprocessing in Lesioned Brains: Manual Versus Automated Region of Interest Analysis.
Functional magnetic resonance imaging (fMRI) has significant potential in the study and treatment of neurological disorders and stroke. Region of interest (ROI) analysis in such studies allows for testing of strong a priori clinical hypotheses with improved statistical power. A commonly used automated approach to ROI analysis is to spatially normalize each participant's structural brain image to a template brain image and define ROIs using an atlas. However, in studies of individuals with structural brain lesions, such as stroke, the gold standard approach may be to manually hand-draw ROIs on each participant's non-normalized structural brain image. Automated approaches to ROI analysis are faster and more standardized, yet are susceptible to preprocessing error (e.g., normalization error) that can be greater in lesioned brains. The manual approach to ROI analysis has high demand for time and expertise, but may provide a more accurate estimate of brain response. In this study, commonly used automated and manual approaches to ROI analysis were directly compared by reanalyzing data from a previously published hypothesis-driven cognitive fMRI study, involving individuals with stroke. The ROI evaluated is the pars opercularis of the inferior frontal gyrus. Significant differences were identified in task-related effect size and percent-activated voxels in this ROI between the automated and manual approaches to ROI analysis. Task interactions, however, were consistent across ROI analysis approaches. These findings support the use of automated approaches to ROI analysis in studies of lesioned brains, provided they employ a task interaction design.


Outcomes of tethered cord repair with a layered soft tissue closure.
Previous studies have demonstrated the benefit of releasing symptomatic tethered cords; however, complications such as seroma, cerebrospinal fluid leak, and infection continue to plague these patients. We propose that composite tissue closure of tethered cord repairs yields superior outcomes and that a collaborative effort between neurosurgery and plastic surgery may result in enhanced structural and functional results. This is a retrospective study comprised of consecutive patients with tethered cord syndrome by 2 neurosurgeons and 2 plastic surgeons between 1994 and 2008 at a single institution. All consecutive patients who underwent tethered cord release by neurosurgery and subsequent composite tissue closure with fascial and musculofascial flaps by plastic surgery were included. Data were collected by retrospective chart review and analyzed using parametric methods. A total of 86 consecutive patients were included in this study, with follow-up ranged from 12 to 144 months (average follow-up, 29 months). There were no statistical differences in follow-up time, comorbidities, or surgeon when comparing hospital readmission or reoperation. There was no statistical difference in complications when comparing the different flap closures. We had a 1.2% infection rate, a 4.7% readmission rate, and a 3.5% reoperation rate. We believe that local soft tissue rearrangement improves the closure by providing an additional layer of vascularized tissue between the skin and the spinal cord. We believe our series represents a significant sample size compared with those previously reported for an experience that achieves multilayered soft tissue closure after tethered cord repair. Our results support the idea that neurosurgeons should consider consultation of plastic surgeons when treating patients with tethered cord syndrome surgically.


Identification of bile acid precursors as endogenous ligands for the nuclear xenobiotic pregnane X receptor.
Sterol 27-hydroxylase (CYP27A1) is required for bile acid synthesis by both the classical and alternate pathways. Cyp27a1(-/-) mice exhibit a dramatic increase in the activity of cytochrome P450 3A (CYP3A), which catalyzes side-chain hydroxylations of bile acid intermediates, thereby facilitating their excretion in the bile and urine. We examine the role of the nuclear xenobiotic receptor PXR (pregnane X receptor) in this process. We demonstrate that expression of Cyp3a11 and other established PXR target genes is increased in the Cyp27a1(-/-) mice. WhenCyp27a1(-/-) mice are fed a diet containing either cholic acid or chenodeoxycholic acid, expression of CYP7A1, which catalyzes the rate-limiting step in bile acid biosynthesis, is strongly suppressed. In parallel, the induction of Cyp3a11 observed in these mice is reversed, suggesting that bile acid intermediates serve as PXR activators. In support of this hypothesis, three potentially toxic sterols (7alpha-hydroxy-4-cholesten-3-one, 5beta-cholestan-3alpha,7alpha,12alpha-triol, and 4-cholesten-3-one), including two that are known to accumulate in Cyp27a1(-/-) mice, are efficacious activators of mouse PXR. All three compounds are more potent activators of mouse PXR than of human PXR, which may explain in part why humans who lack functional CYP27A1 do not display a corresponding increase in CYP3A activity and are stricken with the disease cerebrotendinous xanthomatosis. Taken together, these results reveal the existence of a feedforward regulatory loop by which potentially toxic bile acid intermediates activate PXR and induce their own metabolism. In addition, this study demonstrates that animal models with alterations in gene expression can be used to identify endogenous ligands for orphan nuclear receptors.


The NeuroIMAGE study: a prospective phenotypic, cognitive, genetic and MRI study in children with attention-deficit/hyperactivity disorder. Design and descriptives.
Attention-deficit/hyperactivity disorder (ADHD) is a persistent neuropsychiatric disorder which is associated with impairments on a variety of cognitive measures and abnormalities in structural and functional brain measures. Genetic factors are thought to play an important role in the etiology of ADHD. The NeuroIMAGE study is a follow-up of the Dutch part of the International Multicenter ADHD Genetics (IMAGE) project. It is a multi-site prospective cohort study designed to investigate the course of ADHD, its genetic and environmental determinants, its cognitive and neurobiological underpinnings, and its consequences in adolescence and adulthood. From the original 365 ADHD families and 148 control (CON) IMAGE families, consisting of 506 participants with an ADHD diagnosis, 350 unaffected siblings, and 283 healthy controls, 79 % participated in the NeuroIMAGE follow-up study. Combined with newly recruited participants the NeuroIMAGE study comprehends an assessment of 1,069 children (751 from ADHD families; 318 from CON families) and 848 parents (582 from ADHD families; 266 from CON families). For most families, data for more than one child (82 %) and both parents (82 %) were available. Collected data include a diagnostic interview, behavioural questionnaires, cognitive measures, structural and functional neuroimaging, and genome-wide genetic information. The NeuroIMAGE dataset allows examining the course of ADHD over adolescence into young adulthood, identifying phenotypic, cognitive, and neural mechanisms associated with the persistence versus remission of ADHD, and studying their genetic and environmental underpinnings. The inclusion of siblings of ADHD probands and controls allows modelling of shared familial influences on the ADHD phenotype.


Immune suppression of the echinoderm Asterias rubens (L.) following long-term ocean acidification.
We compared effects of exposure to predict near-future (2100) ocean acidification (OA; pH 7.7) and normal seawater (Control; pH 8.1) on immune and stress responses in the adult sea star Asterias rubens. Analyses were made after one week and after six months of continuous exposure. Following one week exposure to acidified water, the pH of coelomic fluid was significantly reduced. Levels of the chaperon Hsp70 were elevated while key cellular players in immunity, coelomocytes, were reduced by approximately 50%. Following long-term exposure (six months) levels of Hsp70 returned to control values, whereas immunity was further impaired, evidenced by the reduced phagocytic capacity of coelomocytes and inhibited activation of p38 MAP-kinase. Such impacts of reduced seawater pH may have serious consequences for resistance to pathogens in a future acidified ocean.


14-3-3 cooperates with LKB1 to regulate the activity and localization of QSK and SIK.
The LKB1 tumour suppressor kinase phosphorylates and activates a number of protein kinases belonging to the AMP-activated protein kinase (AMPK) subfamily. We have used a modified tandem affinity purification strategy to identify proteins that interact with AMPKalpha, as well as the twelve AMPK-related kinases that are activated by LKB1. The AMPKbeta and AMPKgamma regulatory subunits were associated with AMPKalpha, but not with any of the AMPK-related kinases, explaining why AMP does not influence the activity of these enzymes. In addition, we identified novel binding partners that interacted with one or more of the AMPK subfamily enzymes, including fat facets/ubiquitin specific protease-9 (USP9), AAA-ATPase-p97, adenine nucleotide translocase, protein phosphatase 2A holoenzyme and isoforms of the phospho-protein binding adaptor 14-3-3. Interestingly, the 14-3-3 isoforms bound directly to the T-loop Thr residue of QSK and SIK, after these were phosphorylated by LKB1. Consistent with this, the 14-3-3 isoforms failed to interact with non-phosphorylated QSK and SIK, in LKB1 knockout muscle or in HeLa cells in which LKB1 is not expressed. Moreover, mutation of the T-loop Thr phosphorylated by LKB1, prevented QSK and SIK from interacting with 14-3-3 in vitro. Binding of 14-3-3 to QSK and SIK, enhanced catalytic activity towards the TORC2 protein and the AMARA peptide, and was required for the cytoplasmic localization of SIK and for localization of QSK to punctate structures within the cytoplasm. To our knowledge, this study provides the first example of 14-3-3 binding directly to the T-loop of a protein kinase and influencing its catalytic activity and cellular localization.


Dipole radiation within one-dimensional anisotropic microcavities: a simulation method.
We present a simulation method for light emitted in uniaxially anisotropic light-emitting thin film devices. The simulation is based on the radiation of dipole antennas inside a one-dimensional microcavity. Any layer in the microcaviy can be uniaxially anisotropic with an arbitrary orientation of the optical axis. A plane wave expansion for the field of an elementary dipole inside an anisotropic medium is derived from Maxwell's equations. We employ the scattering matrix method to calculate the emission by dipoles inside an anisotropic microcavity. The simulation method is applied to calculate the emission of dipole antennas in a number of cases: a dipole antenna in an infinite medium, emission into anisotropic slab waveguides and waveguides in liquid crystals. The dependency of the intensity and the polarization on the direction of emission is illustrated for a number of anisotropic microcavities.


Computations on the primary photoreaction of Br2 with CO2: stepwise vs concerted addition of Br atoms.
It was proposed previously that Br2-sensitized photolysis of liquid CO2 proceeds through a metastable primary photoproduct, CO2Br2. Possible mechanisms for such a photoreaction are explored here computationally. First, it is shown that the CO2Br radical is not stable in any geometry. This rules out a free-radical mechanism, for example, photochemical splitting of Br2 followed by stepwise addition of Br atoms to CO2-which in turn accounts for the lack of previously observed Br2+CO2 photochemistry in gas phases. A possible alternative mechanism in liquid phase is formation of a weakly bound CO2:Br2 complex, followed by concerted photoaddition of Br2. This hypothesis is suggested by the previously published spectroscopic detection of a binary CO2:Br2 complex in the supersonically cooled gas phase. We compute a global binding-energy minimum of -6.2 kJ mol(-1) for such complexes, in a linear geometry. Two additional local minima were computed for perpendicular (C2v) and nearly parallel asymmetric planar geometries, both with binding energies near -5.4 kJ mol(-1). In these two latter geometries, C-Br and O-Br bond distances are simultaneously in the range of 3.5-3.8 Å, that is, perhaps suitable for a concerted photoaddition under the temperature and pressure conditions where Br2 + CO2 photochemistry has been observed.


Stock assessment of fishery target species in Lake Koka, Ethiopia.
Effective management is essential for small-scale fisheries to continue providing food and livelihoods for households, particularly in developing countries where other options are often limited. Studies on the population dynamics and stock assessment on fishery target species are thus imperative to sustain their fisheries and the benefits for the society. In Lake Koka (Ethiopia), very little is known about the vital population parameters and exploitation status of the fishery target species: tilapia Oreochromis niloticus, common carp Cyprinus carpio and catfish Clarias gariepinus. Our study, therefore, aimed at determining the vital population parameters and assessing the status of these target species in Lake Koka using length frequency data collected quarterly from commercial catches from 2007-2012. A total of 20,097 fish specimens (distributed as 7,933 tilapia, 6,025 catfish and 6,139 common carp) were measured for the analysis. Von Bertalarffy growth parameters and their confidence intervals were determined from modal progression analysis using ELEFAN I and applying the jackknife technique. Mortality parameters were determined from length-converted catch curves and empirical models. The exploitation status of these target species were then assessed by computing exploitation rates (E) from mortality parameters as well as from size indicators i.e., assessing the size distribution of fish catches relative to the size at maturity (Lm), the size that provides maximum cohort biomass (Lopt) and the abundance of mega-spawners. The mean value of growth parameters L∞, K and the growth performance index ø' were 44.5 cm, 0.41/year and 2.90 for O. niloticus, 74.1 cm, 0.28/year and 3.19 for C. carpio and 121.9 cm, 0.16/year and 3.36 for C. gariepinus, respectively. The 95 % confidence intervals of the estimates were also computed. Total mortality (Z) estimates were 1.47, 0.83 and 0.72/year for O. niloticus, C. carpio and C. gariepinus, respectively. Our study suggest that O. niloticus is in a healthy state, while C. gariepinus show signs of growth overfishing (when both exploitation rate (E) and size indicators were considered). In case of C. carpio, the low exploitation rate encountered would point to underfishing, while the size indicators of the catches would suggest that too small fish are harvested leading to growth overfishing. We concluded that fisheries production in Lake Koka could be enhanced by increasing E toward optimum level of exploitation (Eopt) for the underexploited C. carpio and by increasing the size at first capture (Lc) toward the Lopt, range for all target species.


A requirement for phosphatidylinositol 3-kinase in pseudopod extension.
Phagocytosis requires actin assembly and pseudopod extension, two cellular events that coincide spatially and temporally. The signal transduction events underlying both processes may be distinct. We tested whether phagocytic signaling resembles that of growth factor receptors, which induce actin polymerization via activation of phosphatidylinositol 3-kinase (PI 3-kinase). Fcgamma receptor-mediated phagocytosis was accompanied by a rapid increase in the accumulation of phosphatidylinositol 3,4,5-trisphosphate in vivo, and addition of wortmannin (WM) or LY294002, two inhibitors of PI 3-kinase(s), inhibited phagocytosis but not Fcgamma receptor-directed actin polymerization. However, both compounds prevented maximal pseudopod extension, suggesting that PI 3-kinase inhibition produced a limitation in membrane required for pseudopod extension. Availability of plasma membrane was not limiting for phagocytosis, because blockade of ingestion in the presence of WM was not overcome by reducing the number of particles adhering to macrophages. However, decreasing bead size, and hence the magnitude of pseudopod extension required for particle engulfment, relieved the inhibition of phagocytosis in the presence of WM or LY294002 by up to 80%. The block in phagocytosis of large particles occurred before phagosomal closure, because both compounds inhibited spreading of macrophages on substrate-bound IgG. Macrophage spreading on IgG was accompanied by exocytic insertion of membrane from an intracellular source, as measured by the dye FM1-43. These results indicate that one or more isoforms of PI 3 kinase are required for maximal pseudopod extension but not phagocytosis per se. We suggest that PI 3-kinase is required for coordinating exocytic membrane insertion and pseudopod extension.


Transactivation of elements in the human endogenous retrovirus W family by viral infection.
Aberrant expression of human endogenous retrovirus (HERV) elements in the W family has previously been associated with schizophrenia, multiple sclerosis and preeclampsia. Little is know regarding the basal expression, transcriptional regulation and functional significance of individual HERV-elements. Since viral infections have previously been reported to transactivate retroviral long terminal repeat regions we examined the basal expression of HERV-W elements and following infections by influenza A/WSN/33 and Herpes simplex 1 viruses in human cell-lines. Relative levels of transcripts encoding HERV-W elements and cellular genes were analyzed by qPCR methods. An analysis of amplicon melting temperatures was used to detect variations in the frequencies of amplicons in discrete ranges of such melting temperatures. These frequency-distributions were taken as proxy markers for the repertoires of transcribed HERV-W elements in the cells. We report cell-specific expression patterns of HERV-W elements during base-line conditions. Expressed elements include those with intact regulatory long terminal repeat regions (LTRs) as well as elements flanked by truncated LTRs. Subsets of HERV-W elements were transactivated by viral infection in the different cell-lines. Transcriptional activation of these elements, including that encoding syncytin, was dependent on viral replication and was not induced by antiviral responses. Serum deprivation of cells induced similar changes in the expression of HERV-W elements suggesting that the observed phenomena are, in part, an effect of cellular stress. We found that HERV-W elements, including elements lacking regulatory LTRs, are expressed in cell-specific patterns which can be modulated by environmental influences. This brings into light that mechanisms behind the regulation of expression of HERV-W elements are more complex than previously assumed and suggests biological functions of these transcripts.


The prevalence of mental disorders in adults in different level general medical facilities in Kenya: a cross-sectional study.
The possibility that a significant proportion of the patients attending a general health facility may have a mental disorder means that psychiatric conditions must be recognised and managed appropriately. This study sought to determine the prevalence of common psychiatric disorders in adult (aged 18 years and over) inpatients and outpatients seen in public, private and faith-based general hospitals, health centres and specialised clinics and units of general hospitals. This was a descriptive cross-sectional study conducted in 10 health facilities. All the patients in psychiatric wards and clinics were excluded. Stratified and systematic sampling methods were used. Informed consent was obtained from all study participants. Data were collected over a 4-week period in November 2005 using various psychiatric instruments for adults. Descriptive statistics were generated using SPSS V. 11.5. A total of 2,770 male and female inpatients and outpatients participated in the study. In all, 42% of the subjects had symptoms of mild and severe depression. Only 114 (4.1%) subjects had a file or working diagnosis of a psychiatric condition, which included bipolar mood disorder, schizophrenia, psychosis and depression. The 4.1% clinician detection rate for mental disorders means that most psychiatric disorders in general medical facilities remain undiagnosed and thus, unmanaged. This calls for improved diagnostic practices in general medical facilities in Kenya and in other similar countries.


Treatment of obstruction of the superior vena cava by combination chemotherapy with and without irradiation in small-cell carcinoma of the bronchus.
In a randomised prospective trial of chemotherapy with and without radiotherapy in small-cell carcinoma of the bronchus, 37 of 366 patients presented with obstruction of the superior vena cava at the time of diagnosis. In the study all patients received four cycles of combination chemotherapy over a period of 12 weeks and, provided there was not progressive disease, then received either radiotherapy to the mediastinum and primary tumour followed by eight further courses of chemotherapy or eight cycles of chemotherapy alone. Of the 37 patients presenting with superior vena caval obstruction, nine had relapsed and treatment was stopped during or after the initial four cycles of chemotherapy. Of the remainder, 15 patients received radiotherapy plus chemotherapy and 13 chemotherapy alone. After four cycles of chemotherapy (12 weeks) 21 of the 37 patients had initial complete relief of symptoms secondary to superior vena caval obstruction, 10 had substantial but partial relief, and six had no relief. Twelve patients developed recurrence of superior vena caval obstruction, of whom six had received radiotherapy and four chemotherapy alone; two relapsed in the initial 12 weeks of the study. The median survival of the patients with superior vena caval obstruction allocated at diagnosis to either treatment arm was identical. The survival of patients with obstruction was similar to that of other patients in the main study. Chemotherapy is an effective treatment for superior vena caval obstruction and there appears to be no additional advantage in giving radiotherapy after 12 weeks of cyclical chemotherapy.


Psychotherapy for Military-Related PTSD: A Review of Randomized Clinical Trials.
Posttraumatic stress disorder (PTSD) is a disabling psychiatric disorder common among military personnel and veterans. First-line psychotherapies most often recommended for PTSD consist mainly of "trauma-focused" psychotherapies that involve focusing on details of the trauma or associated cognitive and emotional effects. To examine the effectiveness of psychotherapies for PTSD in military and veteran populations. PubMed, PsycINFO, and PILOTS were searched for randomized clinical trials (RCTs) of individual and group psychotherapies for PTSD in military personnel and veterans, published from January 1980 to March 1, 2015. We also searched reference lists of articles, selected reviews, and meta-analyses. Of 891 publications initially identified, 36 were included. Two trauma-focused therapies, cognitive processing therapy (CPT) and prolonged exposure, have been the most frequently studied psychotherapies for military-related PTSD. Five RCTs of CPT (that included 481 patients) and 4 RCTs of prolonged exposure (that included 402 patients) met inclusion criteria. Focusing on intent-to-treat outcomes, within-group posttreatment effect sizes for CPT and prolonged exposure were large (Cohen d range, 0.78-1.10). CPT and prolonged exposure also outperformed waitlist and treatment-as-usual control conditions. Forty-nine percent to 70% of participants receiving CPT and prolonged exposure attained clinically meaningful symptom improvement (defined as a 10- to 12-point decrease in interviewer-assessed or self-reported symptoms). However, mean posttreatment scores for CPT and prolonged exposure remained at or above clinical criteria for PTSD, and approximately two-thirds of patients receiving CPT or prolonged exposure retained their PTSD diagnosis after treatment (range, 60%-72%). CPT and prolonged exposure were marginally superior compared with non-trauma-focused psychotherapy comparison conditions. In military and veteran populations, trials of the first-line trauma-focused interventions CPT and prolonged exposure have shown clinically meaningful improvements for many patients with PTSD. However, nonresponse rates have been high, many patients continue to have symptoms, and trauma-focused interventions show marginally superior results compared with active control conditions. There is a need for improvement in existing PTSD treatments and for development and testing of novel evidence-based treatments, both trauma-focused and non-trauma-focused.


Readability of pediatric biomedical research informed consent forms.
Informed consent forms are used in biomedical research as a mechanism to convey study information to potential subjects so that they may arrive at a decision concerning their willingness to participate. Although the Department of Health and Human Services Regulations for the Protection of Human Subjects require the presentation of specific study information at a level that is easily understood, according to research concerning adult biomedical consent forms, the typical form is not readily comprehensible. Unfortunately, no data exist concerning the readability of informed consent forms that are used in the context of pediatric biomedical research. In the present study, readability analyses were conducted on a large sample (N = 238) of pediatric biomedical informed consent forms obtained during a 10-year period from a large midwestern children's hospital. For the entire sample, results derived from two readability estimates (Fry grade equivalent and Flesch Reading Ease methods) indicated that the consent forms were written at the college graduate level. Although there was a linear increase in the length of the consent document during the 10-year period evaluated, expanded length was not associated with improved readability. According to analyses, a differential pattern of reading difficulty was associated with specific sections of the informed consent document. Findings are highly consistent with those from studies of adult biomedical consent form and document that the purpose of the informed consent form is being compromised, in part, by a readability factor. Suggestions for solving this critical problem are advanced.


Bitter melon protects against ER stress in LS174T colonic epithelial cells.
Bitter Melon (BM) has been used as a functional food in traditional Chinese and Indian medicine for many generations and has gained a great deal of attention due to its apparent benefits in moderating some of the pathogenic processes in a variety of inflammatory conditions. BM extract (BME) has been shown to possess strong anti-oxidant properties. In addition, it can ameliorate oxidative stress and potentially ER stress. There is increasing evidence that oxidative and ER stress are major contributors for intestinal secretory cell dysfunction which leads to local inflammation and disease pathogenesis that are hallmarks of inflammatory bowel diseases (IBD). Hence, the search for potential therapeutics against ER stress and oxidative stress in intestinal epithelial secretory cells may provide valuable resources for the management of IBD. The aim of the present study was to investigate the effects of BME in ameliorating ER stress in colonic epithelial cells. Human colonic adenocarcinoma LS174T cells were used for the assessment of BME effects on colonic epithelial cells in vitro. Cell viability was assessed using trypan blue exclusion and the effect of BME in ameliorating tunicamycin (TM)-induced ER stress was determined by analysing the mRNA expression of the common ER stress markers; ATF6, XBP1, GRP78, CHOP and PERK by quantitative RT-PCR and GRP78 and CHOP by western blot. In the absence of ER stress, BME exhibited no cell toxicity up to 2.0% w/v and no significant effect on the basal mRNA expression of ER stress markers in LS174T cells. In contrast, pre-treatment of LS174T cells with BME followed by induction of ER stress resulted in a significant decrease in mRNA expression of ATF6, XBP1, GRP78, CHOP and PERK and protein expression of GRP78 and CHOP. Co-treatment during induction of ER stress and post- treatment following induction of ER Stress in LS174T cells resulted in a lower but still significant reduction in mRNA expression levels of most ER stress markers. This is one of the first studies demonstrating the efficacy of BME in reducing expression of ER stress markers in colonic epithelial cells suggesting the potential of BME as a dietary intervention in ameliorating ER stress and oxidation in IBD. Interestingly, while the most significant effect was seen with pre-treatment of cells with BME there was a reduced but still significant effect when co-treated or even post-treated. This suggests that BME may even be effective in modulating ER stress in the face of an existing cell stress environment.


Airway epithelium expresses interleukin-18.
Interleukin (IL)-18 is an interferon (IFN)-gamma-inducing cytokine suggested to be important in regulating inflammatory responses. This study investigated the pulmonary expression of IL-18 under conditions characterized by T-helper (Th)1 (lipopolysaccharide (LPS) treatment/sarcoidosis) and Th2 (ovalbumin (OVA) challenge/asthma) cytokine production. In situ hybridization and immunocytochemistry were used to determine the number of cells expressing IL-18, IFN-gamma, IL-5 and major basic protein (MBP) within lung tissue from Balb/c mice stimulated with LPS, OVA and in normal control mice. Bronchial biopsies from patients with sarcoidosis, asthma and control individuals were also examined. IL-18 was localized primarily to airway epithelium and mononuclear cells. Constitutive expression was observed within the lungs of control mice. Animals challenged with LPS exhibited more IL-18 messenger ribonucleic acid (mRNA)-positive and IFN-gamma immunoreactive cells, compared to control mice (p<0.01). OVA-challenged mice had fewer IL-18 mRNA positive and more IL-5 and MBP immunoreactive cells, compared to control mice (p<0.01). Similarly, constitutive expression of IL-18 protein was observed within the airway epithelium of control individuals, with more positive cells found within sarcoidosis tissue (p<0.01) and fewer within asthmatic tissue (p<0.01), compared to controls. These results demonstrate the expression of interleukin-18 within airway epithelium and the regulation of this cytokine under conditions of both T-helper1 and T-helper2 cytokine production.