Bacterial diversity dynamics in rumen epithelium of wethers fed forage and mixed concentrate forage diets.
Cereal-rich diets used in ruminant intensive production can affect the physiology of the rumen wall but little is known about the changes induced to the epimural bacterial community. Dynamics in epimural bacteria was monitored by PCR-DGGE on four wethers fed forage for 20 weeks (constant diet) and on four wethers fed successively forage, a high concentrate diet (65% wheat) and forage for 4, 8 and 8 weeks, respectively (variable diet group). In the constant diet group, no changes were observed throughout time. In contrast, in the variable diet group, the community tended to differ between high concentrate and forage samples (P=0.06). For both groups, the structure of the community was strongly associated to individual hosts (P=0.001). This difference between individuals could have masked the effect of cereal on the variable diet group. To get more information on the phylotypes present, 120 16S rDNA gene clones were sequenced from the rumen epithelium of a wether fed forage and cereal-rich diets. Firmicutes, Bacteroidetes, and Proteobacteria were the predominant phyla but the bacterial types detected were in general different from those commonly found in rumen contents with a high proportion of Proteobacteria (14%) in the rumen epithelium. In the forage periods, Firmicutes represented nearly 50% of the total epimural community and Bacteroidetes were about 33%, while in the concentrate-rich diet period these percentages were inversed (P<0.05). Clone libraries seemed more sensitive than PCR-DGGE to study the bacteria attached to the rumen epithelium. The role of epimural bacteria on ruminant's health needs further investigation.


Interaction, unscripted.
Quality care for clients with mental illness begins in the classroom. This article describes an effective use of drama with undergraduate nursing students. Because of nursing students' fear and anxiety related to the psychiatric-mental health clinical experience, dramatic presentations are used as a way to expose students to a "real-life" client in a controlled setting. A student volunteer portraying a nurse and the faculty person portraying an individual with a mental illness interact without a written script in front of the class. Following the depiction, the class is encouraged to use the nursing process to plan care for the individual. Students have evaluated the presentations favorably and have indicated that the presentations decrease their anxiety and fear and build their confidence. Practicing nurses could use this kind of simulation as a way to enhance their own communication skills. When faced with a difficult patient, nurses may use drama together in a controlled setting and practice how best to provide quality care to that patient using the nursing process.


Activation of phosphatidylinositol 3-kinase/Akt pathway by androgen through interaction of p85alpha, androgen receptor, and Src.
Recent studies have demonstrated that the cell growth and antiapoptotic actions of androgen could be dissociated from the transcriptional activity of the receptor and were, instead, mediated by activation of a mitogen-activated protein kinase pathway. This finding suggests an important cellular function of androgen receptor (AR) outside the nucleus. In this report, we demonstrate that androgen activates phosphatidylinositol 3-kinase (PI3K) and Akt, including AKT1 and AKT2, in AR-positive cells. Androgen-induced cell growth and survival were inhibited by PI3K inhibitor and dominant-negative Akt. AR interacts with the p85alpha regulatory subunit of PI3K, and its binding affinity is increased after androgen stimulation. The sites of interaction on the two proteins were mapped to the C-terminal Src-homology 2 domain of p85alpha and N terminus of AR. Activation of PI3K/Akt by androgen was inhibited by dominant-negative Src. Neither N-terminal truncated nor proline-rich region-deleted AR mutants, which are unable to bind to p85alpha and Src, respectively, was able to mediate androgen-induced PI3K/Akt activation. AR with deletion of C-terminal region including ligand binding domain, however, retains the ability to activate PI3K/Akt upon androgen stimulation, which supports the notion that nongenomic function of androgen is mediated by its interaction with membrane receptors (1, 3, 4). These findings indicate that a triple complex between AR, p85alpha, and Src is required for androgen-stimulated PI3K/Akt activation, and that the PI3K/Akt pathway, in addition to mitogen-activated protein kinase, mediates androgen-induced cell growth and cell survival.


High-Fidelity Simulation Improves Long-Term Knowledge of Clinical Swallow Evaluation.
Purpose Clinical swallow evaluation (CSE) is a critical skill that speech-language pathologists who manage swallowing impairment must learn. The objective of this mixed-methods study was to determine if using a human patient simulator (HPS) to train speech-language pathology graduate students in CSE improved knowledge, preparedness, and anxiety as compared to traditional instruction alone. Method This was a controlled trial with repeated measures. Participants included graduate students from two cohorts who were enrolled in a swallowing disorders course in consecutive academic years (n = 50). Students in the experimental group participated in a simulation experience in which they performed a CSE on an HPS, generated a treatment plan, and communicated in real time with the HPS, the patient's wife, and a nurse. Quantitative results included quizzes that measured short- and long-term CSE knowledge, and qualitative findings included written feedback from instructors and students. Results Students who participated in simulation training had significantly higher long-term quiz accuracy than the control group, but their short-term quiz scores did not differ. Student ratings of preparedness and anxiety did not differ between the two groups. Many students reported that they appreciated practicing the use of patient-friendly language and preferred clinical simulation over traditional teaching methods. Facilitators reported that simulation increased student engagement and critical thinking skills more than traditional teaching methods. Conclusions CSE simulation provided objective and subjective advantages over traditional teaching methods. Recommendations from students and instructors for improving the CSE simulation training are reported.


NMR imaging in the diagnosis and management of intracranial angiomas.
Fourteen intracranial angiomas were clearly visualized and diagnosed with certainty on fast saturation-recovery images, which highlight blood vessels without the use of contrast media, and on steady-state free-precession images, in which the moving blood leads to removal of signal. Performed as the initial investigation, nuclear magnetic resonance obviates angiography when the site and extent of the angioma would preclude operation, and in other cases provides useful anatomic information complementing the angiogram. When clinical presentation follows hemorrhage the size and position of the associated hematoma can be reliably assessed.


Olea europaea Leaf Extract Improves the Efficacy of Temozolomide Therapy by Inducing MGMT Methylation and Reducing P53 Expression in Glioblastoma.
Unmethylated O6-methylguanine-DNA-methyltransferase (MGMT) promoter leads to Temozolomide (TMZ) resistance in most of the glioblastoma multiforme (GBM) patients. We previously investigated the synergistic effect of Olea europaea leaf extract (OLE) on TMZ cytotoxicity through modulating microRNA expression. To date, knowledge about the effect of OLE on MGMT methylation is insufficient. The aim of the current study was to evaluate the potential modulating effect of OLE on the TMZ response of GBM tumors through MGMT methylation. Exposure to 1 mg/mL OLE caused a significant induction of CpG island methylation in the MGMT gene using Methyl quantitative PCR assay (P < 0.001). In WST-1 analysis, the use of 350 µM TMZ plus 1 mg/mL OLE significantly increased the TMZ response of MGMT unmethylated cells (P = 0.003). Using the comet assay, the impact of 1 mg/mL OLE plus 350 µM TMZ on the formation of DNA strand breaks was significantly higher than that of 450 µM TMZ alone (P < 0.001) and Western blot analysis revealed that, when cells are treated with 1-mg/mL OLE, the total p53 protein levels tended to decrease. The results presented in this study uniquely demonstrated that OLE synergistically increased the TMZ response of GBM tumors by regulating MGMT gene methylation and p53 expression. However, further studies to validate our findings are required.


Urban change analysis and future growth of Istanbul.
This study is aimed at analyzing urban change within Istanbul and assessing the city's future growth potential using appropriate approach modeling for the year 2040. Urban growth is a major driving force of land-use change, and spatial and temporal components of urbanization can be identified through accurate spatial modeling. In this context, widely used urban modeling approaches, such as the Markov chain and logistic regression based on cellular automata (CA), were used to simulate urban growth within Istanbul. The distance from each pixel to the urban and road classes, elevation, and slope, together with municipality and land use maps (as an excluded layer), were identified as factors. Calibration data were obtained from remotely sensed data recorded in 1972, 1986, and 2013. Validation was performed by overlaying the simulated and actual 2013 urban maps, and a kappa index of agreement was derived. The results indicate that urban expansion will influence mainly forest areas during the time period of 2013-2040. The urban expansion was predicted as 429 and 327 km(2) with the Markov chain and logistic regression models, respectively.


Stimulation of poliovirus synthesis in a HeLa cell-free in vitro translation-RNA replication system by viral protein 3CDpro.
The plus-strand RNA genome of poliovirus serves three distinct functions in the life cycle of the virus. The RNA is translated and then replicated, and finally the progeny RNAs are encapsidated. These processes can be faithfully reproduced in a HeLa cell-free in vitro translation-RNA replication system that produces viable poliovirus. We have previously observed a stimulation of virus synthesis when an mRNA, encoding protein 3CD(pro), is added to the translation-RNA replication reactions of poliovirus RNA. Our aim in these experiments was to further define the factors that affect the stimulatory activity of 3CD(pro) in virus synthesis. We observed that purified 3CD(pro) protein also enhances virus synthesis by about 100-fold but has no effect on the translation of the polyprotein. Optimal stimulation is observed only when 3CD(pro) is present early in the incubation period. The stimulation, however, is abolished by a mutation either in the RNA binding domain of 3CD(pro), 3C(pro)R84S/I86A, or by each of two groups of complementary mutations R455A/R456A and D339A/S341A/D349A at interface I in the 3D(pol) domain of 3CD(pro). Surprisingly, virus synthesis is strongly inhibited by the addition of both 3C(pro) and 3CD(pro) at the beginning of incubation. We also examined the effect of other viral or cellular proteins on virus synthesis in the in vitro system. No enhancement of virus synthesis occurred with viral proteins 3BC, 3ABC, 3BCD, 3D(pol), and 3C(pro) or with cellular protein PCBP2. These results suggest that 3CD(pro) has to be present in the reaction at the time the replication complexes are assembled and that both the 3C(pro) and 3D(pol) domains of the protein are required for its activity that stimulates virus production.


FGF8 spliceforms mediate early mesoderm and posterior neural tissue formation in Xenopus.
The relative contributions of different FGF ligands and spliceforms to mesodermal and neural patterning in Xenopus have not been determined, and alternative splicing, though common, is a relatively unexplored area in development. We present evidence that FGF8 performs a dual role in X. laevis and X. tropicalis early development. There are two FGF8 spliceforms, FGF8a and FGF8b, which have very different activities. FGF8b is a potent mesoderm inducer, while FGF8a has little effect on the development of mesoderm. When mammalian FGF8 spliceforms are analyzed in X. laevis, the contrast in activity is conserved. Using a loss-of-function approach, we demonstrate that FGF8 is necessary for proper gastrulation and formation of mesoderm and that FGF8b is the predominant FGF8 spliceform involved in early mesoderm development in Xenopus. Furthermore, FGF8 signaling is necessary for proper posterior neural formation; loss of either FGF8a or a reduction in both FGF8a and FGF8b causes a reduction in the hindbrain and spinal cord domains.


Supporting Family Caregivers With Technology for Dementia Home Care: A Randomized Controlled Trial.
The number of persons living with dementia (PLWD) in the United States will reach 16 million by 2050. Behavioral and psychological symptoms of dementia challenge family caregivers and contribute to negative caregiver outcomes such as burden and depression. Available technology can support the delivery of effective interventions to families providing dementia care at home. The Supporting Family Caregivers with Technology for Dementia Home Care (FamTechCare) randomized controlled trial evaluated the effects of a telehealth intervention on caregiver outcomes. The FamTechCare intervention provides tailored dementia-care strategies to in-home caregivers based on video recordings caregivers submit of challenging care situations. An expert team reviews the videos and provides individualized interventions weekly for the experimental group. In the telephone-support attention control group, caregivers receive feedback from an interventionist via the telephone based on caregiver retrospective recall of care challenges. Effects of the intervention on caregiver outcomes, including burden, depression, sleep disturbance, competence, desire to institutionalize the PLWD, and caregiver reaction to behavioral symptoms were evaluated by fitting linear mixed regression models to changes in the outcomes measured at 1 and 3 months. FamTechCare caregivers (n = 42) had greater reductions in depression (p = .012) and gains in competence (p = .033) after 3 months compared to the attention control group (n = 41). Living in rural areas was associated with a reduction in depression for FamTechCare caregivers (p = .002). Higher level of education was associated with greater improvements or lesser declines in burden, competence, and reaction to behavioral symptoms for both the FamTechCare and attention control caregivers. This research demonstrated benefits of using available technology to link families to dementia care experts using video-recording technology. It provides a foundation for future research testing telehealth interventions, tailored based on rich contextual data to support families, including those in rural or remote locations.


Household demand for typhoid fever vaccines in Hue, Vietnam.
The demand function for vaccines against typhoid fever was estimated using stated preference data collected from a random sample of 1065 households in Hue, Vietnam, in 2002. These are the first estimates of private willingness-to-pay (WTP) and demand functions for typhoid vaccines in a developing country. Mean respondent WTP for a single typhoid fever vaccine ranged from USD 2.30 to USD 4.80. Mean household WTP estimates (vaccinating all members of the household) ranged from USD 21 to USD 27. Demand was similar for vaccines with different degrees of effectiveness and intervals of duration. These results suggest a significant potential for private sector provision of typhoid fever vaccines in Hue.


Systemic effects of orally administered p, p'-DDE on immature male Wistar rats during pubertal period.
Systemic effects of p, p'-DDE (1, 1-dichloro-2, 2 bis (p-chlorophenyl) ethylene; DDE) on immature male rats were investigated in pubertal Wistar rats after oral administration of DDE. Special rat chow containing 125 ppm DDE (approximately 10 mg/kg DDE) had been administered daily for 42 d since 6 wk of age and its effects had been observed until 12 wk of age. The administration of DDE did not produce any overt signs of toxicity. Neither physical development nor sexual maturation was affected, and serum biochemistry was not impaired at the dose used in this experiment. Moreover, the male reproductive organs and epididymal sperm count were not affected by the administration of DDE during the pubertal period. Our results showed that even immature male rats were resistant to DDE exposure at the daily dose of ca. 10 mg/kg, but metabolic and immunological changes still remained uncertain. Further investigation should be conducted to reveal all the effects of DDE on immature male rats.


Validity of estimating minute-by-minute energy expenditure of continuous walking bouts by accelerometry.
Objective measurement of physical activity remains an important challenge. For wearable monitors such as accelerometer-based physical activity monitors, more accurate methods are needed to convert activity counts into energy expenditure (EE). The purpose of this study was to examine the accuracy of the refined Crouter 2-Regression Model (C2RM) for estimating EE during the transition from rest to walking and walking to rest. A secondary purpose was to determine the extent of overestimation in minute-by-minute EE between the refined C2RM and the 2006 C2RM. Thirty volunteers (age, 28 ± 7.7 yrs) performed 15 minutes of seated rest, 8 minutes of over-ground walking, and 8 minutes of seated rest. An ActiGraph GT1M accelerometer and Cosmed K4b2 portable metabolic system were worn during all activities. Participants were randomly assigned to start the walking bout at 0, 20, or 40 s into the minute (according to the ActiGraph clock). Acceleration data were analyzed by two methods: 2006 Crouter model and a new refined model. The 2006 Crouter 2-Regression model over-predicted measured kcal kg-1 hr-1 during the first and last transitional minutes of the 20-s and 40-s walking conditions (P < 0.001). It also over-predicted the average EE for a walking bout (4.0 ± 0.5 kcal kg-1 hr-1), compared to both the measured kcal kg-1 hr-1 (3.6 ± 0.7 kcal kg-1 hr-1) and the refined Crouter model (3.5 ± 0.5 kcal kg-1 hr-1) (P < 0.05). The 2006 Crouter 2-regression model over-predicts EE at the beginning and end of walking bouts, due to high variability in accelerometer counts during the transitional minutes. The new refined model eliminates this problem and results in a more accurate prediction of EE during walking.


Accurate placement of a tibial tunnel significantly improves meniscal healing and clinical outcomes at 1 year after medial meniscus posterior root repair.
A medial meniscus posterior root tear results in the loss of meniscal circumferential hoop stress and causes a pathological posteromedial extrusion of the medial meniscus. Although creating a tibial tunnel in the anatomic place improves postoperative medial meniscus posterior extrusion, no studies have evaluated the relationship between tibial tunnel position and clinical outcomes. This study aimed to evaluate how tibial tunnel positioning of medial meniscus posterior root pullout repair affects meniscal healing status and clinical outcomes. Sixty-two patients with 64 medial meniscus posterior root tears (mean age 62.8 ± 7.9 years) who had undergone pullout repairs and second-look arthroscopies were included. All 62 patients were Lachman test negative. Three-dimensional computed tomography images of the tibial surface were evaluated using a rectangular measurement grid to assess the tibial tunnel centre and medial meniscus posterior root attachment centre. Spearman's rank correlation analysis was undertaken to determine displacement distance from the medial meniscus posterior root attachment centre to the tibial tunnel centre and a meniscal healing score, as well as clinical outcomes at 1 year post-repair. Tibial tunnel centres were located more anteriorly and medially than the medial meniscus posterior root attachment centre (mean distance 5.0 ± 2.2 mm). The mean meniscal healing score was 6.7 ± 1.8 of 10 possible points. The 1-year postoperative clinical scores showed significant improvement compared with preoperative scores for all the items. There was a significant negative correlation in the absolute distance between the medial meniscus posterior root attachment centre and the tibial tunnel centre with the meniscal healing score (ρ = - 0.39, p = 0.002). Furthermore, there were significant positive correlations between the distance between the medial meniscus posterior root attachment centre and the tibial tunnel centre in the mediolateral direction and patient-based clinical outcomes (ρ = 0.25-0.43, p < 0.05). Accurate placement of a tibial tunnel, especially in the mediolateral direction, significantly improved meniscal healing and clinical outcomes at 1 year following medial meniscus posterior root repair. Surgeons should create a medial meniscus posterior root tibial tunnel at the anatomic attachment with particular attention to the mediolateral position. Level IV.


Inhospital and Post-discharge Changes in Renal Function After Transcatheter Aortic Valve Replacement.
The aim of this study was to determine the influence of inhospital and post-discharge worsening renal function (WRF) on prognosis after transcatheter aortic valve replacement (TAVR). Severe chronic kidney disease and inhospital WRF are both associated with poor outcomes after TAVR. There are no data available on post-discharge WRF and outcomes. This was a single-center study evaluating all TAVR from June 1, 2008, to June 31, 2014. WRF was defined as an increase in serum creatinine of ≥0.3 mg/dl. Inhospital WRF was measured from day 0 until discharge or day 7 if the hospitalization was >7 days. Post-discharge WRF was measured at 30 days after discharge. Descriptive statistics, Kaplan-Meier time-to-event analysis, and multivariate logistic regression were used. In a series of 208 patients who underwent TAVR, 204 with complete renal function data were used in the inhospital analysis and 168 who returned for the 30-day follow-up were used in the post-discharge analysis. Inhospital WRF was seen in 28%, whereas post-discharge WRF in 12%. Inhospital and post-discharge WRF were associated with lower rates of survival; however, after multivariate analysis, only post-discharge WRF remained a predictor of 1-year mortality (hazard ratio 1.18, p = 0.030 for every 1 mg/dl increase in serum creatinine). In conclusion, the rate of inhospital WRF is higher than the rate of post-discharge WRF after TAVR, and post-discharge WRF is more predictive of mortality than inhospital WRF.


Massive glomus tumor of the stomach: 20-year follow-up and autopsy findings.
A massive glomus tumor of the greater curvature of the stomach was removed in 1961. The patient made an uneventful recovery and died 21 years later of causes unrelated to the tumor. The literature of glomus tumors of the stomach is discussed. The pathology of the lesion is described and the autopsy findings are presented.


Mass turnover and recovery dynamics of a diverse Australian continental radiation.
Trends in global and local climate history have been linked to observed macroevolutionary patterns across a variety of organisms. These climatic pressures may unilaterally or asymmetrically influence the evolutionary trajectory of clades. To test and compare signatures of changing global (Eocene-Oligocene boundary cooling) and continental (Miocene aridification) environments on a continental fauna, we investigated the macroevolutionary dynamics of one of Australia's most diverse endemic radiations, pygopodoid geckos. We generated a time-calibrated phylogeny (>90% taxon coverage) to test whether (i) asymmetrical pygopodoid tree shape may be the result of mass turnover deep in the group's history, and (ii) how Miocene aridification shaped trends in biome assemblages. We find evidence of mass turnover in pygopodoids following the isolation of the Australian continental plate ∼30 million years ago, and in contrast, gradual aridification is linked to elevated speciation rates in the young arid zone. Surprisingly, our results suggest that invasion of arid habitats was not an evolutionary end point. Instead, arid Australia has acted as a source for diversity, with repeated outward dispersals having facilitated diversification of this group. This pattern contrasts trends in richness and distribution of other Australian vertebrates, illustrating the profound effects historical biome changes have on macroevolutionary patterns.


Cost of Treatment in HBeAg-Negative Chronic Hepatitis B Patients with 10-Years Projection Analysis.
Chronic hepatitis B infection has to be treated effectively due to its major complications. The aim of this study is to investigate and compare the therapy alternatives' treatment costs for hepatitis B e antigen (HBeAg) negative patients with 10-year projection analysis according to current reimbursement guideline in Turkey. All testing that should be done before the treatment and during follow-up was determined according to the guideline. Medicinal costs were calculated according to 2014 prices. Cost calculation was performed on 100 hypothetical HBeAg negative, 35 years old patients whose alanine aminotransferase value was >2 X upper limit of the normal laboratory level by taking into consideration the risks of resistance seen during the treatment. Switching to tenofovir scenario was concocted when resistance to lamivudine and telbivudine was developed. In pegylated-interferon non-responders, alternative scenarios have been edited and evaluated. The total cost of patients whose treatment was started with pegylated-interferon and switched to lamivudine and tenofovir due to non-response was found 2,662,504 TL. Treatment cost of patients whose therapy was passed to tenofovir after initiation with lamivudine or telbivudine was 2,444,175 TL and 3,061,869 TL, respectively. From the beginning of treatment in patients taking entecavir or tenofovir, the 10-year cost of treatment was 3,924,960 TL and 3,884,040 TL, respectively. As a result, when assessing the cost of the treatment of chronic hepatitis B not only medicine box costs, but also drug resistance and laboratory testing costs, should be considered. In our country, on the basis of 10 years cost; pegylated-interferon and lamivudine therapy has been found advantageous in patients with low viral load, and tenofovir has been found advantageous in patients with high viral load.


Identifying Direct Coercion in a High Risk Subgroup of Offender Patients With Schizophrenia via Machine Learning Algorithms.
This study aims to explore risk factors for direct coercive measures (seclusion, restraint, involuntary medication) in a high risk subpopulation of offender patients with schizophrenia spectrum disorders. Five hundred sixty nine potential predictor variables were explored in terms of their predictive power for coercion/no coercion in a set of 131 (36.6%) offender patients who experienced coercion and 227 who did not, using machine learning analysis. The dataset was split (70/30%) applying variable filtering, machine learning model building, and selection embedded in nested resampling approach in one subset. The best model was then selected, and the most important variables extracted on the second data subset. In the final model the following variables identified coercion with a balanced accuracy of 73.28% and a predictive power (area under the curve, AUC) of 0.8468: threat of violence, (actual) violence toward others, the application of direct coercive measures during past psychiatric inpatient treatments, the positive and negative syndrome scales (PANSS) poor impulse control, uncooperativeness, and hostility and the total PANSS-score at admission, prescription of haloperidol during inpatient treatment, the daily cumulative olanzapine equivalent antipsychotic dosage at discharge, and the legal prognosis estimated by a team of licensed forensic psychiatrists. Results confirm prior findings, add detail on factors indicative for the use of direct coercion, and provide clarification on inconsistencies. Limitations, clinical relevance, and avenues for future research are discussed.


Thromboelastometry analysis of severe North American pit viper-induced coagulopathy: A case report.
A 51-year-old man presented with rapid onset encephalopathy and respiratory failure after a suspected intravascular envenomation from a North American pit viper. The patient received antivenom and was transferred to a tertiary care facility where he had cardiovascular collapse and persistent coagulopathy requiring 28 vials of Crotalidae polyvalent immune Fab antivenom for initial control and six vials for maintenance. The patient's coagulopathy was monitored using "traditional" measures (platelets, fibrinogen, and prothrombin time/international normalized ratio) and rotational thromboelastometry (ROTEM®). The patient also subsequently developed intestinal necrosis requiring exploratory laparotomy with ileum and colonic resections, and anuric renal failure requiring continuous renal replacement therapy. After coordinated multidisciplinary management, he was discharged to an acute inpatient rehabilitation on hospital day 25 and has since made a full recovery. In the setting of a severe intravascular pit viper envenomation, thromboelastometry correlated well with "traditional" measures. During recovery, ROTEM® demonstrated measurable improvements in the extrinsic coagulation pathway while the INR remained between 1.5 and 1.6. Patient's intestinal necrosis may have resulted from microvascular thrombosis due to Crotalinae venom. The patient's ultimate recovery necessitated a coordinated multidisciplinary effort. ROTEM® abnormalities after North American pit viper envenomation may be more sensitive than "traditional" measures and may have prognostic value to determine the severity of envenomation, but further research to define its utility is required.


Enzyme variation, metabolic flux and fitness: alcohol dehydrogenase in Drosophila melanogaster.
Although there are many in vitro studies of enzyme activity of genetic variants at the Adh locus in D. melanogaster, little is known about the corresponding metabolic activity in living flies. We report here such measurements of the metabolic flux in the conversion of ethanol to the two products, CO2 and lipids, for six different active genotypes, containing the predominant naturally recurring alleles and covering a threefold range of in vitro activity. In adult flies we have found nonsignificant differences between genotypes in metabolic flux when estimates for individual genotypes had standard errors of approximately 10% of the mean value. In vitro activities are, therefore, poor predictors of the physiological consequences of enzyme variation since such determinations ignore the interactions inherent in multienzyme systems. We have no evidence that heterozygote show overdominance either at the enzyme or the flux level. Since fitness differences between genotypes must be generated by physiological differences, investigations of polymorphisms should be based on in vivo studies.


Colchicine for acute gout.
This is an updated Cochrane Review, first published in 2006 and updated in 2014. Gout is one of the most common rheumatic diseases worldwide. Despite the use of colchicine as one of the first-line therapies for the treatment of acute gout, evidence for its benefits and harms is relatively limited. To update the available evidence of the benefits and harms of colchicine for the treatment of acute gout. We updated the search of CENTRAL, MEDLINE, Embase, Clinicaltrials.gov and WHO ICTRP registries to 28 August 2020. We did not impose any date or language restrictions in the search. We considered published randomised controlled trials (RCTs) and quasi-randomised controlled trials (quasi-RCTs) evaluating colchicine therapy compared with another therapy (placebo or active) in acute gout; low-dose colchicine at clinically relevant doses compared with placebo was the primary comparison. The major outcomes were pain, participant global assessment of treatment success (proportion with 50% or greater decrease in pain from baseline up to 32 to 36 hours), reduction of inflammation, function of target joint, serious adverse events, total adverse events and withdrawals due to adverse events. We used standard methodological procedures as expected by Cochrane in this review update. We included four trials (803 randomised participants), including two new trials, in this updated review. One three-arm trial compared high-dose colchicine (52 participants), low-dose colchicine (74 participants) and placebo (59 participants); one trial compared high-dose colchicine with placebo (43 participants); one trial compared low-dose colchicine with non-steroidal anti-inflammatory drugs (NSAIDs) (399 participants); and one trial compared low-dose colchicine with Chuanhu anti-gout mixture (traditional Chinese Medicine compound) (176 participants). We did not identify any trials comparing colchicine to glucocorticoids (by any route). The mean age of participants ranged from 51.2 to 70 years, and trial duration from 48 hours to 12 weeks. Two trials were at low risk of bias, one was possibly susceptible to selection bias (random sequence generation), reporting bias and other bias, and one open-label trial was at high risk of performance and detection bias. For the primary comparison, low-quality evidence from one trial (103 participants, downgraded for imprecision and bias) suggests low-dose colchicine may improve treatment outcome compared to placebo with little or no increased risk of adverse events. The number of people who reported treatment success (50% or greater pain reduction) at 32 to 36 hours was slightly larger with low-dose colchicine (418 per 1000) compared with placebo (172 per 1000; risk ratio (RR) 2.43, 95% confidence interval (CI) 1.05 to 5.64; absolute improvement 25% more reported success (7% more to 42% more, the 95% CIs include both a clinically important and unimportant benefit); relative change of 143% more people reported treatment success (5% more to 464% more). The incidence of total adverse events was 364 per 1000 with low-dose colchicine compared with 276 per 1000 with placebo: RR 1.32, 95% CI 0.68 to 2.56; absolute difference 9% more events with low-dose colchicine (9% fewer to 43% more, the 95% CIs include both a clinically important effect and no effect); relative change of 32% more events (32% fewer to 156% more). No participants withdrew due to adverse events or reported any serious adverse events. Pain, inflammation and function were not reported. Low-quality evidence (downgraded for imprecision and bias) from two trials (124 participants) suggests that high-dose colchicine compared to placebo may improve symptoms, but with increased risk of harms. More participants reported treatment success at 32 to 36 hours with high-dose colchicine (518 per 1000) compared with placebo (240 per 1000): RR 2.16, 95% CI 1.28 to 3.65, absolute improvement 28% (8% more to 46% more); more also had reduced inflammation at this time point with high-dose colchicine (504 per 1000) compared with placebo (48 per 1000): RR 10.50, 95% CI 1.48 to 74.38; absolute improvement 45% greater (22% greater to 68% greater); but more adverse events were reported with high-dose colchicine (829 per 1000 compared with 260 per 1000): RR 3.21, 95% CI 2.01 to 5.11, absolute difference 57% (26% more to 74% more). Pain and function were not reported. Low-quality evidence from a single trial comparing high-dose to low-dose colchicine indicates there may be little or no difference in benefit in terms of treatment success at 32 to 36 hours but more adverse events associated with the higher dose. Similarly, low-quality evidence from a single trial indicates there may also be little or no benefit of low-dose colchicine over NSAIDs in terms of treatment success and pain reduction at seven days, with a similar number of adverse events reported at four weeks follow-up. Reduction of inflammation, function of target joint and withdrawals due to adverse events were not reported in either of these trials, and pain was not reported in the high-dose versus low-dose colchicine trial. We were unable to estimate the risk of serious adverse events for most comparisons as there were few events reported in the trials. One trial (399 participants) reported three serious adverse (one in a participant receiving low-dose colchicine and two in participants receiving NSAIDs), due to reasons unrelated to the trial (low-quality evidence downgraded for bias and imprecision). We found low-quality evidence that low-dose colchicine may be an effective treatment for acute gout when compared to placebo and low-quality evidence that its benefits may be similar to NSAIDs. We downgraded the evidence for bias and imprecision. While both high- and low-dose colchicine improve pain when compared to placebo, low-quality evidence suggests that high-dose (but not low-dose) colchicine may increase the number of adverse events compared to placebo, while low-quality evidence indicates that the number of adverse events may be similar with low-dose colchicine and NSAIDs. Further trials comparing colchicine to placebo or other treatment will likely have an important impact on our confidence in the effect estimates and may change the conclusions of this review. There are no trials reporting the effect of colchicine in populations with comorbidities or in comparison with other commonly used treatments, such as glucocorticoids.


Rapid regulatory actions of sex steroids on cell movement through the actin cytoskeleton.
Cell movement is required in relevant physiological processes such as embryonic development, tissue and organ differentiation, inflammation, immune response and wound healing, along with pathological phenomena, such as cancer metastatic spread. Cell motility is tightly controlled by a complex and often redundant array of intracellular signaling pathways largely devoted to the dynamic regulation of the actin cytoskeletal network and of its relationship with the cell membrane and the extracellular matrix. Sex steroids, particularly estrogen and progesterone, are effective regulators of cell migration and tissue organization, and recent evidence indicates that this is in part obtained through the regulation of the cytoskeleton. Intriguingly, many of these regulatory actions related to cell movement are achieved through rapid, non-classical signaling of sex steroid receptors to kinase cascades, independently from nuclear alteration of gene expression or protein synthesis. The identification of the mechanistic basis for these rapid actions on cell cytoskeleton and cell movement has special relevance for the characterization of the effects of sex steroids in physiological conditions, such their role in the control of inflammation, brain or vascular cell remodelling, angiogenesis or wound healing, as well as in the context of pathological conditions such as steroid-sensitive cancer cell invasion and metastasis. This review highlights the physiological and clinical conditions where the regulatory effects on the cytoskeleton and cell movement of sex steroids might have a special importance, as well as the recent advances in the characterization of the mechanisms, providing insights and working hypotheses on possible clinical applications for the modulation of these pathways.


Open-label investigation of rapid initiation of extended-release buprenorphine in patients using fentanyl and fentanyl analogs.
Synthetic opioids, including fentanyl and fentanyl analogs, account for over 70,000 annual overdose deaths in the United States, but there is limited information examining methods of induction and maintenance outcomes for buprenorphine treatment of patients with opioid use disorder (OUD) using these opioids. A secondary analysis of results grouped by fentanyl use status was completed for an open-label study with rapid induction of extended-release buprenorphine in the inpatient research unit. Eligible participants received a single 4 mg dose of transmucosal buprenorphine (BUP-TM) followed by an extended-release buprenorphine 300 mg injection ([BUP-XR]) after approximately 1 h. An extension study continued follow-up up to 6 months (6 monthly injections). Among participants with fentanyl-positive urine samples (FEN+; n = 19), all received BUP-TM, 17 received BUP-XR, 13 elected to receive a second BUP-XR injection, and 10 received all six scheduled injections. Among participants with fentanyl-negative samples (FEN-; n = 7), all received BUP-TM and BUP-XR, four elected to receive a second injection, and two participants received all six scheduled injections. Induction day clinical opioid withdrawal scale (COWS) scores were similar for FEN+ and FEN- groups. In the FEN+ group, mean COWS scores fell to below 5 within 24 h of BUP-XR injection. The treatment of individuals with OUD using fentanyl with a rapid 1-day induction to BUP-XR 300 mg injection is feasible and well-tolerated. A prospective trial of participants grouped by fentanyl use status at induction demonstrates comparable patient retention and clinical response following single-day induction of BUP-XR in participants who are FEN+ and FEN-.


Purification and characterization of DNA polymerase from the archaebacterium Sulfolobus acidocaldarius.
DNA polymerase has been purified about 25,000-fold from the thermoacidophilic archaebacterium Sulfolobus acidocaldarius. On SDS-PAGE the enzyme was observed to have a molecular weight of 100 kDa and to be about 90% pure. The native molecular weight was 108 kDa indicating that the enzyme is composed of a single polypeptide. Activity gel analysis showed an active polypeptide of about 100 kDa. Under conditions promoting proteolysis this polypeptide was degraded to a slightly smaller form of 98 kDa. The enzyme has been characterized in respect to optimal assay conditions, template specificity, sensitivity to inhibitors and associated nuclease activities. The high temperature optimum of 65 degrees C should be emphasized. No substantial similarities have been found with other prokaryotic and eukaryotic DNA polymerases, although the enzyme bears certain resemblances to prokaryotic non-replicative polymerases.


Interleukin 18 binding protein (IL18-BP) inhibits neointimal hyperplasia after balloon injury in an atherosclerotic rabbit model.
Interleukin 18 (IL18) is an interferon (IFN)-gamma-inducing factor and a proinflammatory and proatherogenic cytokine. IL18 binding protein (IL18-BP) functions as an IL18 inhibitor. This study was designed to investigate whether systemic administration of IL18-BP could inhibit neointimal hyperplasia and arterial lipid deposition. New Zealand white, male rabbits were fed with a 21% fat, 0.15% cholesterol diet. The left superficial femoral artery (SFA) was de-endotheliazed with a 2F arterial embolectomy catheter. IL18-BP (5 microg, 10 microg, or 25 microg), or 0.9% saline (control) was administered by i.v. bolus during surgery. Rabbits were followed-up at 2 and 4 weeks. Intima-media (I/M) and lumen-whole artery (L/A) area ratios, and luminal areas were measured. Serum lipid levels, liver enzymes, and kidney function were evaluated. Inflammatory cells were quantified and further verified with immunohistofluorescence staining. The extent of lipid deposition in the artery wall was quantified with Oil Red O (ORO) staining employing Zeiss AxioVision 4.6.3. Image analysis software. Lipid laden cells including macrophages were evaluated by transmission electron microscopy (TEM). Intravenous IL18-BP 5 microg, 10 microg, and 25 microg significantly reduced I/M ratios compared with the control group at both 2 and 4 weeks. There was no significant difference between the 5 microg and 10 microg dose groups. However, at 10 microg, IL18-BP significantly increased L/A ratio more than either the 5 microg IL18-BP or control groups. The high fat diet caused significant elevation of serum lipids at 4 and 6 weeks. IL18-BP had no effect on blood lipid levels. Lipid deposit in the thoracic aorta of the control group at 6 weeks was more than at 4 weeks (P = .025). Administration of IL18-BP inhibited the lipid deposition at 4 weeks (not significant) and 6 weeks (P = .012 to .008) compared with its control group. Lipid laden macrophages (foam cells), as well as endothelial cells and smooth muscle cells were seen in the descending thoracic aorta after 6 weeks of a high fat diet by ORO, immunohistofluorescence staining, and TEM. The lipid laden cells were not seen in either of IL18-BP groups. IL18-BP 10 microg significantly inhibited mono/macro adherence and infiltration in the SFA after balloon-injury at 2 weeks after surgery. A single intravenous dose of IL18-BP significantly decreased arterial neointimal hyperplasia, improved lumen to artery ratio after balloon-injury and also prevented arteriosclerosis progression. A single intravenous dose of IL18BP decreased neointimal hyperplasia and improved arterial L/A ratios in an atherosclerotic balloon-injury animal model. These preliminary results suggest that IL18BP may be a promising molecular approach to inhibit neointimal hyperplasia and arteriosclerosis progression following coronary and peripheral angioplasty.


Systematic review: comparative effectiveness of adjunctive devices in patients with ST-segment elevation myocardial infarction undergoing percutaneous coronary intervention of native vessels.
During percutaneous coronary intervention (PCI), dislodgement of atherothrombotic material from coronary lesions can result in distal embolization, and may lead to increased major adverse cardiovascular events (MACE) and mortality. We sought to systematically review the comparative effectiveness of adjunctive devices to remove thrombi or protect against distal embolization in patients with ST-segment elevation myocardial infarction (STEMI) undergoing PCI of native vessels. We conducted a systematic literature search of Medline, the Cochrane Database, and Web of Science (January 1996-March 2011), http://www.clinicaltrials.gov, abstracts from major cardiology meetings, TCTMD, and CardioSource Plus. Two investigators independently screened citations and extracted data from randomized controlled trials (RCTs) that compared the use of adjunctive devices plus PCI to PCI alone, evaluated patients with STEMI, enrolled a population with 95% of target lesion(s) in native vessels, and reported data on at least one pre-specified outcome. Quality was graded as good, fair or poor and the strength of evidence was rated as high, moderate, low or insufficient. Disagreement was resolved through consensus. 37 trials met inclusion criteria. At the maximal duration of follow-up, catheter aspiration devices plus PCI significantly decreased the risk of MACE by 27% compared to PCI alone. Catheter aspiration devices also significantly increased the achievement of ST-segment resolution by 49%, myocardial blush grade of 3 (MBG-3) by 39%, and thrombolysis in myocardial infarction (TIMI) 3 flow by 8%, while reducing the risk of distal embolization by 44%, no reflow by 48% and coronary dissection by 70% versus standard PCI alone. In a majority of trials, the use of catheter aspiration devices increased procedural time upon qualitative assessment.Distal filter embolic protection devices significantly increased the risk of target revascularization by 39% although the use of mechanical thrombectomy or embolic protection devices did not significantly impact other final health outcomes. Distal balloon or any embolic protection device increased the achievement of MBG-3 by 61% and 20% and TIMI3 flow by 11% and 6% but did not significantly impact other intermediate outcomes versus control. Upon qualitative analysis, all device categories, with exception of catheter aspiration devices, appear to significantly prolong procedure time compared to PCI alone while none appear to significantly impact ejection fraction. Many of the final health outcome and adverse event evaluations were underpowered and the safety of devices overall is unclear due to insufficient amounts of data. In patients with STEMI, for most devices, few RCTs evaluated final health outcomes over a long period of follow-up. Due to insufficient data, the safety of these devices is unclear.


Transfection of BLM into cultured bloom syndrome cells reduces the sister-chromatid exchange rate toward normal.
The gene BLM, mutated in Bloom syndrome (BS), encodes the nuclear protein BLM, which when absent, as it is from most BS cells, results in genomic instability. A manifestation of this instability is an excessive rate of sister-chromatid exchange (SCE). Here we describe the effects on this abnormal cellular phenotype of stable transfection of normal BLM cDNAs into two types of BS cells, SV40-transformed fibroblasts and Epstein-Barr virus (EBV)-transformed lymphoblastoid cells. Clones of BLM-transfected fibroblasts produced normal amounts of BLM by western blot analysis and displayed a normal nuclear localization of the protein by immunofluorescence microscopy. They had a mean of 24 SCEs/46 chromosomes, in contrast to the mean of 69 SCEs in controls transfected only with the vector. BLM-transfected fibroblast clones that expressed highest levels of the BLM protein had lowest levels of SCE. The lymphoblastoid cells transfected with BLM had SCE frequencies of 22 and 42 in two separate experiments in which two different selectable markers were used, in contrast to 57 and 58 in vector-transfected cells; in this type cell, however, the BLM protein was below the level detectable by western blot analysis. These experiments prove that BLM cDNA encodes a functional protein capable of restoring to or toward normal the uniquely characteristic high-SCE phenotype of BS cells.


Oral Quercetin Supplementation Enhances Adiponectin Receptor Transcript Expression in Polycystic Ovary Syndrome Patients: A Randomized Placebo-Controlled Double-Blind Clinical Trial.
Polycystic ovary syndrome (PCOS), an ovarian-pituitary axis androgen disorder, is a common endocrine disease in women. Obesity-induced androgenesis and imbalance of adipokine secretion may lead to some metabolic features of PCOS. The beneficial effects of polyphenolic compounds such as quercetin have been reported, however, the underlying molecular mechanism is not entirely understood. In the present study, we investigated the effect of quercetin supplementation on the expression of adiponectin receptors at the transcript level in peripheral blood mononuclear cells (PBMC) samples of PCOS patients. In this randomized clinical trial, 84 PCOS subjects were randomly assigned to two groups; the treatment group received 1 g quercetin (two 500 mg capsules) daily for 12 weeks and the control group received placebo. To examine the effect of quercetin supplementation on PCOS patients in addition to biochemical and anthropometric assessments, the expression of ADIPOR1 and ADIPOR2 at the transcript level and AMPK level were determined by quantitative reverse transcription-polymerase chain reaction (RT-qPCR) and ELISA assays respectively. Oral quercetin supplementation significantly increased ADIPOR1 and ADIPOR2 transcript expression by 1.32- and 1.46-fold respecetively (P<0.01). In addition, quercetin supplementation enhanced AMPK level by 12.3% compared with the control group (P<0.05). Oral quercetin supplementation improves the metabolic features of PCOS patients by upregulating the expression of adiponectin receptors and AMPK (Registration Number: IRCT2013112515536N1).


A systematic review on methodological variation in acute:chronic workload research in elite male football players.
Objectives: To investigate and evaluate the methodological variation in research on acute:chronic workload ratio (ACWR) and injury in elite male football players.Methods: Relevant literature was electronically searched on PubMed, Web of Science, ScienceDirect, and Embase. Additional literature was obtained from studies' bibliographies and authors. Cohort studies investigating the effects of ACWR on male elite footballer injuries were included. Information regarding study population, time frame, protocol, injury classification, and statistical analysis were elucidated.Results: Database searches led to 2,689 articles. After full text screening, twelve articles remained. All studies were of poor quality. Five studies had GPS-derived workload measures with consideration of running intensity zones, though little consensus over zone thresholds were found. Nine studies incorporated rated perceived exertion data; heterogeneity in exposure type and data collection timing was observed. All studies applied rolling average ACWRs, exploring 1:2, 1:3, and 1:4-week load ratio windows. Differences in data grouping, inference or regression analysis, and other statistical methods were noted.Conclusion: Existing literature displayed methodological heterogeneity. Future studies should consider consulting guidelines for developing prognostic studies and further examine causal links between workload and injury. From that basis, decisions around ACWR definitions, workload measures, and statistical methods may be more appropriately made.


Role of key amino acids in the transmembrane domain of the Newcastle disease virus fusion protein.
Newcastle disease (ND), caused by the Newcastle disease virus (NDV), is transmitted by poultry with severe infectivity and a high fatality rate. The fusion (F) protein on the NDV envelope facilitates the merger of the viral and host cell membranes with the help of the homologous hemagglutinin-neuraminidase protein (HN). The transmembrane (TM) domains of viral fusion proteins are typically required for fusion, but the key amino acids in NDV F TM domains have not been identified. Site-directed mutagenesis was utilized to change the conserved amino acids at 500, 501, 502, 505, 510, 513, 516, 519, and 520 to alanine. It was found that mutants L519 and V520 had an interrupted protein expression, decreased to below 10%, and mutants A500, I505, V513, and V516 had a hypoactive impact on fusion activity, decreased to 85.38%, 67.05%, 55.38% and 51.13% of wt F, respectively. The results indicated that the TM domain plays a vital part in the fusion activity of the NDV F protein.


The Chloroplast Phylogenomics and Systematics of Zoysia (Poaceae).
The genus Zoysia Willd. (Chloridoideae) is widely distributed from the temperate regions of Northeast Asia-including China, Japan, and Korea-to the tropical regions of Southeast Asia. Among these, four species-Zoysia japonica Steud., Zoysia sinica Hance, Zoysia tenuifolia Thiele, and Zoysia macrostachya Franch. & Sav.-are naturally distributed in the Korean Peninsula. In this study, we report the complete plastome sequences of these Korean Zoysia species (NCBI acc. nos. MF953592, MF967579~MF967581). The length of Zoysia plastomes ranges from 135,854 to 135,904 bp, and the plastomes have a typical quadripartite structure, which consists of a pair of inverted repeat regions (20,962~20,966 bp) separated by a large (81,348~81,392 bp) and a small (12,582~12,586 bp) single-copy region. In terms of gene order and structure, Zoysia plastomes are similar to the typical plastomes of Poaceae. The plastomes encode 110 genes, of which 76 are protein-coding genes, 30 are tRNA genes, and four are rRNA genes. Fourteen genes contain single introns and one gene has two introns. Three evolutionary hotspot spacer regions-atpB~rbcL, rps16~rps3, and rpl32~trnL-UAG-were recognized among six analyzed Zoysia species. The high divergences in the atpB~rbcL spacer and rpl16~rpl3 region are primarily due to the differences in base substitutions and indels. In contrast, the high divergence between rpl32~trnL-UAG spacers is due to a small inversion with a pair of 22 bp stem and an 11 bp loop. Simple sequence repeats (SSRs) were identified in 59 different locations in Z. japonica, 63 in Z. sinica, 62 in Z. macrostachya, and 63 in Z. tenuifolia plastomes. Phylogenetic analysis showed that the Zoysia (Zoysiinae) forms a monophyletic group, which is sister to Sporobolus (Sporobolinae), with 100% bootstrap support. Within the Zoysia clade, the relationship of (Z. sinica, Z japonica), (Z. tenuifolia, Z. matrella), (Z. macrostachya, Z. macrantha) was suggested.


Degradation of mitochondrial DNA in cryoprotectant-treated hard coral (Echinopora spp.) oocytes.
A critical step for successful cryopreservation is to determine the optimal cryoprotectant treatment that can provide protective effects against cryoinjury during freezing and with minimal toxicity. Most cryoprotectants have chemical and osmotic effects when used at high concentrations. Cryoprotectants can damage coral mitochondrial distributions and membrane potentials, which results in reduced ATP production. As mitochondrial DNA (mtDNA) encodes for components of the electron transport chain (ETC) and plays a critical role in ATP synthesis capacity, we determined the effects of cryoprotectants on mtDNA in hard coral (Echinopora spp.) oocytes using quantitative real-time PCR. Our results showed that an insult from a cryoprotectant may be compensated for by the genetic defense mechanisms of these cells. Methanol was found to have the least effect on coral oocytes with regard to their energy status. A single oocyte without cryoprotectant treatment produced an average of 4,220,645 ± 169,990 mtDNA copies, which was greater than that in mammals. However, relatively lower mtDNA copy numbers (<2,000,000) were observed when oocytes were treated with dimethyl sulfoxide (DMSO), propylene glycol (PG), ethylene glycol (EG), or glycerol at a concentration of 3 M for 20 min. These results provide direct evidence that hard coral (Echinopora spp.) oocytes are extremely susceptible to cryoprotectants and support the concerns with regard to the adverse effects of cryoprotectants.


Immuno nanoparticles integrated electrical control of targeted cancer cell development using whole cell bioelectronic device.
Electrical properties of cells determine most of the cellular functions, particularly ones which occur in the cell's membrane. Manipulation of these electrical properties may provide a powerful electrotherapy option for the treatment of cancer as cancerous cells have been shown to be more electronegative than normal proliferating cells. Previously, we used an electrical impedance sensing system (EIS) to explore the responses of cancerous SKOV3 cells and normal HUVEC cells to low intensity (<2 V/cm) AC electric fields, determining that the optimal frequency for SKOV3 proliferation arrest was 200 kHz, without harming the non-cancerous HUVECs. In this study, to determine if these effects are cell type dependant, human breast adenocarcinoma cells (MCF7) were subjected to a range of frequencies (50 kHz-2 MHz) similar to the previously tested SKOV3. For the MCF7, an optimal frequency of 100 kHz was determined using the EIS, indicating a higher sensitivity towards the applied field. Further experiments specifically targeting the two types of cancer cells using HER2 antibody functionalized gold nanoparticles (HER2-AuNPs) were performed to determine if enhanced electric field strength can be induced via the application of nanoparticles, consequently leading to the killing of the cancerous cells without affecting non cancerous HUVECs and MCF10a providing a platform for the development of a non-invasive cancer treatment without any harmful side effects. The EIS was used to monitor the real-time consequences on cellular viability and a noticeable decrease in the growth profile of the MCF7 was observed with the application of the HER2-AuNPs and the electric fields indicating specific inhibitory effects on dividing cells in culture. To further understand the effects of the externally applied field to the cells, an Annexin V/EthD-III assay was performed to determine the cell death mechanism indicating apoptosis. The zeta potential of the SKOV3 and the MCF7 before and after incorporation of the HER2-AuNPs was also obtained indicating a decrease in zeta potential with the incorporation of the nanoparticles. The outcome of this research will improve our fundamental understanding of the behavior of cancer cells and define optimal parameters of electrotherapy for clinical and drug delivery applications.


Monitoring immune modulation by nutrition in the general population: identifying and substantiating effects on human health.
Optimal functioning of the immune system is crucial to human health, and nutrition is one of the major exogenous factors modulating different aspects of immune function. Currently, no single marker is available to predict the effect of a dietary intervention on different aspects of immune function. To provide further guidance on the assessment and interpretation of the modulation of immune functions due to nutrition in the general population, International Life Sciences Institute Europe commissioned a group of experts from academia, government and the food industry to prepare a guidance document. A draft of this paper was refined at a workshop involving additional experts. First, the expert group defined criteria to evaluate the usefulness of immune function markers. Over seventy-five markers were scored within the context of three distinct immune system functions: defence against pathogens; avoidance or mitigation of allergy; control of low-grade (metabolic) inflammation. The most useful markers were subsequently classified depending on whether they by themselves signify clinical relevance and/or involvement of immune function. Next, five theoretical scenarios were drafted describing potential changes in the values of markers compared with a relevant reference range. Finally, all elements were combined, providing a framework to aid the design and interpretation of studies assessing the effects of nutrition on immune function. This stepwise approach offers a clear rationale for selecting markers for future trials and provides a framework for the interpretation of outcomes. A similar stepwise approach may also be useful to rationalise the selection and interpretation of markers for other physiological processes critical to the maintenance of health and well-being.


Regulation of freezing tolerance and flowering in temperate cereals: the VRN-1 connection.
In winter wheat (Triticum spp.) and barley (Hordeum vulgare) varieties, long exposures to nonfreezing cold temperatures accelerate flowering time (vernalization) and improve freezing tolerance (cold acclimation). However, when plants initiate their reproductive development, freezing tolerance decreases, suggesting a connection between the two processes. To better understand this connection, we used two diploid wheat (Triticum monococcum) mutants, maintained vegetative phase (mvp), that carry deletions encompassing VRN-1, the major vernalization gene in temperate cereals. Homozygous mvp/mvp plants never flower, whereas plants carrying at least one functional VRN-1 copy (Mvp/-) exhibit normal flowering and high transcript levels of VRN-1 under long days. The Mvp/- plants showed reduced freezing tolerance and reduced transcript levels of several cold-induced C-REPEAT BINDING FACTOR transcription factors and COLD REGULATED genes (COR) relative to the mvp/mvp plants. Diploid wheat accessions with mutations in the VRN-1 promoter, resulting in high transcript levels under both long and short days, showed a significant down-regulation of COR14b under long days but not under short days. Taken together, these studies suggest that VRN-1 is required for the initiation of the regulatory cascade that down-regulates the cold acclimation pathway but that additional genes regulated by long days are required for the down-regulation of the COR genes. In addition, our results show that allelic variation in VRN-1 is sufficient to determine differences in freezing tolerance, suggesting that quantitative trait loci for freezing tolerance previously mapped on this chromosome region are likely a pleiotropic effect of VRN-1 rather than the effect of a separate closely linked locus (FROST RESISTANCE-1), as proposed in early freezing tolerance studies.


Innovative Approaches for Estimating the Levels of Tobacco-Specific Nitrosamines in Cured Tobacco Samples.
Tobacco-specific nitrosamines (TSNAs), mainly the 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone (NNK), are known carcinogens. Part of the NNK found in smoke is provided from matrix-bound NNK, and its determination is extremely relevant. However, the reference extraction procedure of matrix-bound NNK is time-consuming and labor-intensive and has a limited analytical capacity. Three different methodologies were proposed to predict matrix-bound NNK: simple linear regression (LR) with soluble NNK; multiple linear regression (MLR) considering soluble NNK and characteristic parameters of the samples; and orthogonal partial least-squares (O-PLS) regression using high-throughput screening by flow injection analysis coupled to high-resolution mass spectrometry (HTS-FIA-HRMS) data. Simple linear regression showed a high influence of matrix and leaf origin. Although an existing linearity trend has been observed ( R2 = 0.62) for the global model, higher correlation values were achieved for matrix and country segregation models. Multiple linear regression predicted matrix-bound NNK with more satisfactory efficiency than simple linear regression models. The coefficients of determination were 0.87 and 0.94 for flue-cured Virginia and air-cured Burley, respectively. However, this method has a limited application, since previous information about the sample is required. The proposed method based on HTS-FIA-HRMS and O-PLS has shown the most suitable performance in the prediction of matrix-bound NNK, with errors comparable to the reference method, and a higher throughput. In addition, this approach allows to determine other soluble nitrosamines, namely N'-nitrosoanatabine, N'-nitrosoanabasine, and N-nitrosonornicotine, with relative percentage errors between 5.25 and 11.98%. Therefore, the third approach is the best method for a large number of cured tobacco for accuracy in determination of TSNAs.


Ecotoxicity of single-wall carbon nanotubes to freshwater snail Lymnaea luteola L.: Impacts on oxidative stress and genotoxicity.
Mammalian studies have raised concerns about the toxicity of carbon nanotubes, but there is very limited data on ecogenotoxicity to aquatic organisms. The aim of this study was to determine eco-geno toxic effects of single walled carbon nanotubes (SWCNTs) in fresh water snail, Lymnea luteola (L. luteola). A static test system was used to expose L. luteola to a freshwater control, 0.05, 0.15, 0.30, 0.46 mg/L SWCNTs for up to 4 days. SWCNTs changed a significant reduction in glutathione, glutathione-S-transferase, and glutathione peroxidase with in hepatopancreas of L. luteola. Lipid peroxidation (LPO) and catalase showed dose- and time-dependent and statistically significant increase in hepatopancreas during SWCNTs exposure compared with control. However, a significant (p < 0.01) induction in DNA damage was observed by the comet assay in hepatopancreas cells treated with SWCNTs. These results demonstrate that SWCNTs are ecogenotoxic to freshwater snail L. luteola. The oxidative stress and comet assay can successfully be used as sensitive tools of aquatic pollution biomonitoring.


Gene expression analysis in radiotherapy patients and C57BL/6 mice as a measure of exposure to ionizing radiation.
Dose assessment after radiological disasters is imperative to decrease mortality through rationally directed medical intervention. Our goal was to identify biomarkers capable of qualitative (nonirradiated/irradiated) and/or quantitative (dose) assessment of radiation exposure. Using real-time quantitative PCR, biodosimetry genes were identified in blood samples from cancer patients undergoing total-body irradiation. Time- (5, 12, 23, 48 h) and dose- (0-8 Gy) dependent changes in gene expression were examined in C57BL/6 mice. A training set was used to derive weighted voting classification algorithms (nonirradiated/irradiated) and continuous regression (dose assessment) models that were tested in a separate validation set of mice. Of eight biodosimetry genes identified in cancer patients ( ACTA2 , BBC3 , CCNG1 , CDKN1A , GADD45A , MDK , SERPINE1 , Tnfrsf10b ), expression of BBC3 , CCNG1 , CDKN1A , SERPINE1 and Tnfrsf10b was significantly (P < 0.05) increased in irradiated mice. CCNG1 and CDKN1A expression segregated irradiated mice from controls with an accuracy, specificity and sensitivity of 96.3, 100.0 and 94.4%, respectively, at 48 h. Multiple linear regression analysis predicted doses for the 0-, 1-, 2-, 4-, 6- and 8-Gy treatment groups as 0.0 ± 0.2, 1.6 ± 1.0, 2.9 ± 1.4, 5.1 ± 2.0, 5.3 ± 0.7 and 10.5 ± 5.6 Gy, respectively. These results suggest that gene expression analysis could be incorporated into biodosimetry protocols for qualitative and quantitative assessment of radiation exposure.


Late gene therapy limits the restoration of retinal function in a mouse model of retinitis pigmentosa.
Retinitis pigmentosa is an inherited photoreceptor degeneration that begins with rod loss followed by cone loss. This cell loss greatly diminishes vision, with most patients becoming legally blind. Gene therapies are being developed, but it is unknown how retinal function depends on the time of intervention. To uncover this dependence, we utilize a mouse model of retinitis pigmentosa capable of artificial genetic rescue. This model enables a benchmark of best-case gene therapy by removing variables that complicate answering this question. Complete genetic rescue was performed at 25%, 50%, and 70% rod loss (early, mid and late, respectively). Early and mid treatment restore retinal output to near wild-type levels. Late treatment retinas exhibit continued, albeit slowed, loss of sensitivity and signal fidelity among retinal ganglion cells, as well as persistent gliosis. We conclude that gene replacement therapies delivered after 50% rod loss are unlikely to restore visual function to normal. This is critical information for administering gene therapies to rescue vision.


INGUINAL REPAIR VIA ROBOTIC ASSISTED TECHNIQUE: LITERATURE REVIEW.
Inguinal hernia is one of the most frequent surgical diseases. Currently, with the advantages of minimally invasive surgery, new questions arise: what will be the best approach for correction of inguinal hernia? Is there real benefit to the robotic approach? To compile results of the published studies that used the robot-assisted technique in the repair of inguinal hernia, analyzing its limitations, complications and comparing it with those of the pre-existing techniques. The review was performed from the Medline database with the following descriptors: (inguinal hernia repair OR hernioplasty OR hernia) AND (robot OR robotic OR robotic assisted) being retrieved 391 articles. After verification of the titles and abstracts, we identified eight series of cases congruent with the objectives of this review. Three reviewers participated in the extraction and selection of results. Comparative studies showed an increase in surgical time in relation to the open and videolaparoscopic approach. The complications present similar rates with the other repair routes. This technique has been shown to be effective for the correction of inguinal hernia, but the benefits of using robotic surgery are unclear. So, there is a need for randomized studies comparing laparoscopic to robotic repair.


Genetic analysis of ColN plasmid determinants for colicin production, release, and immunity.
Colicin N was identified as the 39,000-molecular-weight protein encoded by the 4,900-base-pair, multiple copy number, amplifiable plasmid ColN -284. Its production was controlled by the SOS regulatory circuit and by catabolite repression. Colicin accumulated intracellularly to ca. 10(6) molecules per cell after growth for 2 to 3 h in medium containing 0.5 microgram of mitomycin C per ml and was then released as the cells underwent partial lysis. Strains carrying pColN -284 and its derivatives exhibited low-level immunity to colicin N and were fully sensitive to all other colicins tested. Regions of the plasmid responsible for colicin N activity (cna), for mitomycin-induced lysis ( cnl ), and for colicin N immunity ( cni ) were localized and characterized by cloning, transposon Tn5 and hydroxylamine mutagenesis, and restriction endonuclease deletion and mapping analysis. The results are discussed in terms of both the organization of the cna, cnl , and cni genes and the respective role of cnl expression and colicin N production in mitomycin sensitivity, colicin export, and induced partial lysis of ColN + cells.


Childhood obesity intervention studies: A narrative review and guide for investigators, authors, editors, reviewers, journalists, and readers to guard against exaggerated effectiveness claims.
Being able to draw accurate conclusions from childhood obesity trials is important to make advances in reversing the obesity epidemic. However, obesity research sometimes is not conducted or reported to appropriate scientific standards. To constructively draw attention to this issue, we present 10 errors that are commonly committed, illustrate each error with examples from the childhood obesity literature, and follow with suggestions on how to avoid these errors. These errors are as follows: using self-reported outcomes and teaching to the test; foregoing control groups and risking regression to the mean creating differences over time; changing the goal posts; ignoring clustering in studies that randomize groups of children; following the forking paths, subsetting, p-hacking, and data dredging; basing conclusions on tests for significant differences from baseline; equating "no statistically significant difference" with "equally effective"; ignoring intervention study results in favor of observational analyses; using one-sided testing for statistical significance; and stating that effects are clinically significant even though they are not statistically significant. We hope that compiling these errors in one article will serve as the beginning of a checklist to support fidelity in conducting, analyzing, and reporting childhood obesity research.


Mobile Phone Use and Acceptability for the Delivery of Mental Health Information Among Perinatal Adolescents in Nigeria: Survey Study.
There are several barriers that may hamper adolescent mothers' utilization of available health interventions for perinatal depression. Innovative treatment approaches are needed to increase adolescent mothers' access to mental health care for improved maternal and child health outcomes. Mobile phones have the potential to serve as important conduits to mental health care in Africa. However, mobile phone use patterns and needs among young mothers in Nigeria are not well documented. This study sought to determine the prevalence of mobile phone use among perinatal adolescents and report patterns of use, as well as to assess the openness of young mothers to mobile health (mHealth) mental health interventions. We surveyed 260 adolescent mothers (ages 16-19 years) in their perinatal or postnatal periods of pregnancies in 33 primary health care clinics in Ibadan, Oyo State, Nigeria in 2020. Respondents were included if they were pregnant with a gestation age of greater than or equal to 4 weeks, or had babies (which they had birthed) that were younger than 12 months. The total study sample consisted of 260 adolescent mothers with a mean age of 18.4 (SD 0.88) years. The majority of the respondents (233/260, 89.6%) owned mobile phones (eg, keypad, keypad and internet, smartphones); 22 (8.5%) of the 260 mothers had access to phones that belonged to relatives who lived in the same household, while 5 (1.9%) had access only to public paid phones. Only 23% (54/233) of phone owners (which is 20.5% of the total study population) had smartphones. On average, respondents reported first using mobile phones at 15.5 (SD 2.06) years old. The majority of respondents (222/260, 85.4%) reported using their phones for an average of 45 minutes daily for calls to family members. Facebook was the social media platform that was most often used among respondents who had phones with internet access (122/146 minutes per day, 83.4%). The majority of the sample responded as being "interested" and "very interested" in the use of mobile phones for preventive interventions (250/260, 96.2%) and treatment (243/260, 93.5%) information on mental illness such as depression and "hearing voices." Half of the respondents (126/233, 50.4%) preferred to receive such information in the form of text messages. Findings from this study suggest that the vast majority of perinatal adolescents in Nigeria own and use mobile phones and that they are interested in leveraging these devices for prevention, treatment, and informational campaigns focused on mental health. The use of smartphones in this population is relatively low, and health intervention through text messages were favored by the women.


Influence of different approaches to the cerebellopontine angle on the function of the intermediate nerve.
Surgery in the cerebellopontine angle (CPA) is a standard procedure at many centers. Outcome is focused mainly on preservation of hearing and facial function postoperatively. The nonmotor part of the facial nerve, the intermediate nerve, is nearly neglected nowadays. A retrospective study was designed, including a questionnaire that was sent to 178 patients who had undergone surgery between 2000 and 2004. Data were obtained in 156 cases. The questionnaire was divided into five parts assessing the presence of increased tearing, reduced tearing, salivation disturbances, increased nasal secretions, and abnormalities in taste. Finally, a self-assessment of patient symptoms was analyzed. Postoperatively, 70 patients (45%) experienced crocodile tears, whereas 62 (40%) had dry eyes. Disturbances in taste were noted in six patients (4%) preoperatively and in 52 patients (33%) after surgery. Increased nasal secretion was noted in 68 patients (44%) postoperatively. An analysis of the correlation between the surgical approach used and disturbances in the intermediate nerve revealed a strong tendency to a higher rate of symptoms following the middle fossa approach (p = 0.071, chi-square test). Data in this study demonstrate the clinical importance of nonmotor defects associated with CPA tumor removal. More than 50% of the patients in this study reported postoperative crocodile tears, dry eyes, nasal secretions, and/or taste disturbances. These sequelae may affect both short- and long-term postoperative quality of life to the same extent as deafness or transient facial paresis. The risk of injury seems to be higher after the middle fossa approach.


Cytoarchitecture of mouse and rat cingulate cortex with human homologies.
A gulf exists between cingulate area designations in human neurocytology and those used in rodent brain atlases with a major underpinning of the former being midcingulate cortex (MCC). The present study used images extracted from the Franklin and Paxinos mouse atlas and Paxinos and Watson rat atlas to demonstrate areas comprising MCC and modifications of anterior cingulate (ACC) and retrosplenial cortices. The laminar architecture not available in the atlases is also provided for each cingulate area. Both mouse and rat have a MCC with neurons in all layers that are larger than in ACC and layer Va has particularly prominent neurons and reduced neuron densities. An undifferentiated ACC area 33 lies along the rostral callosal sulcus in rat but not in mouse and area 32 has dorsal and ventral subdivisions with the former having particularly large pyramidal neurons in layer Vb. Both mouse and rat have anterior and posterior divisions of retrosplenial areas 29c and 30, although their cytology is different in rat and mouse. Maps of the rodent cingulate cortices provide for direct comparisons with each region in the human including MCC and it is significant that rodents do not have a posterior cingulate region composed of areas 23 and 31 like the human. It is concluded that rodents and primates, including humans, possess a MCC and this homology along with those in ACC and retrosplenial cortices permit scientists inspired by human considerations to test hypotheses on rodent models of human diseases.


[Nugent and Spiegel criteria for the diagnosis of bacterial vaginosis. Analysis of discordant specimens by the Ison and Hay method].
Vaginal infection is the commonest cause of genital symptoms and has obstetric and gynecological implications. To compare the Nugent and Spiegel methods for the diagnosis of bacterial vaginosis (BV) and to analyze discordant specimens using Ison and Hay (Ison/Hay) criteria. After discarding cases with Candidiasis, deficient specimens or those lacking bacteria, a total of 348 Gram-stained smears vaginal specimens received for the diagnosis of BV, were analyzed. Vaginal microbiota was classified as normal in 203 and 237 samples (58 and 68% of samples), according to Nugent and Spiegel criteria, respectively One hundred and five (30%) and 111 samples (32%), were classified as VB according to Nugent and Spiegel criteria, respectively. Both criteria were concordant in 308 samples (88.5%). The 40 (11.5%) discordant specimens were classified as intermediate microflora by the Nugent system and as normal or BV by Spiegel. Among these, the Ison/Hay procedure identified four categories of microbiota. Ten (25%) specimens were classified as grade II microbiota, confirming their categorization by Nugent as intermediate microbiota, six (15%) were classified in the III category, confirming the diagnosis of BV by Spiegel, 13 (32.5%) corresponded to the category III, that does not exist in the Nugent and Spiegel categorization systems. Finally, 11 specimens could not be assigned to one category due to microscopic limitations to distinguish bacterial morphotypes. The systems proposed by Spiegel, Nugent and Ison/Hay are comparable for the diagnosis of BV. However, we recommend the use of Ison/Hay procedure to evaluate vaginal microbiota, due to its wider range of categories, allowing a better discrimination of the vaginal microbiota.


Research note: effects of age and immunization interval on the anamnestic response to T-cell-dependent and T-cell-independent antigens in chickens.
Measurement of a humoral anamnestic response is a common procedure used to evaluate immunological competence. The purposes of the present study were to determine the optimum time period between primary and secondary immunizations needed to obtain the maximum antibody titer and to determine whether this interval differs with the age of the chicken or with the type of inducing antigen. The anamnestic responses of birds of two ages to a T-cell-dependent antigen (SRBC) and to a T-cell-independent antigen (Brucella abortus, BA) were evaluated. Birds of 4 wk or 6 mo of age were injected i.v. with either BA or SRBC, and a secondary injection was given 2, 4, 6, or 8 wk later. Agglutination titers for total and mercaptoethanol (ME)-resistant antibodies were determined from serum samples collected at 3, 6, and 9 days postsecondary immunization. Generally, titers were highest at 6 days postsecondary immunization. For both antigens, the chick total antibody levels at Day 6 postsecondary immunization were higher at 4-, 6-, and 8-wk immunization intervals than at 2-wk intervals; adult titers were independent of immunization interval. There was an interaction between age of birds and type of antigen.


Dexmedetomidine Promotes Hippocampal Neurogenesis and Improves Spatial Learning and Memory in Neonatal Rats.
Dexmedetomidine (Dex) is a highly selective α2-adrenoceptor agonist used as an off-label medication for pediatric sedation and analgesia. Recently, Dex was reported to exhibit neuroprotective efficacy in several brain injury models. Here we investigate whether neonatal Dex administration promotes hippocampal neurogenesis and enhances hippocampus-dependent spatial learning and memory under physiological conditions. Postnatal day 7 (P7) pups were administered saline (vehicle control) or Dex (10, 20, or 40 µg/kg) by intraperitoneal injection. Neurogenesis and astrogenesis were examined in brain slices by BrdU immunostaining on P8 and changes in the expression levels of GDNF, NCAM, CREB, PSD95, and GAP43 were assessed by Western blotting on P35, respectively. Open field and Morris water maze (MWM) tests were conducted from P28 to P36 in order to assess effects on general motor activity and spatial learning, respectively. Dexmedetomidine at 20 µg/kg significantly enhanced neurogenesis and astrogenesis in hippocampus and upregulated GDNF, NCAM, CREB, PSD95, and GAP43 compared to vehicle and other Dex doses. Moreover, 20 µg/kg Dex-injected rats showed no changes in motor or anxiety-like behavior but performed better in the MWM test compared to all other groups. Neonatal injection of Dex (20 µg/kg) enhances spatial learning and memory in rat pups, potentially by promoting hippocampal neurogenesis and synaptic plasticity via activation of GDNF/NCAM/CREB signaling.


Caring for caregivers: experiences and evaluation of interventions for a palliative care team.
Palliative care is a fairly recent development in The Netherlands. The first palliative unit in a hospital setting was the Palliative Care Unit (PCU) in the Dr Daniel den Hoed Clinic, a specialized oncology hospital, now part of the Academic Hospital Rotterdam (AZR-Daniel). This PCU was designed to function as an academic laboratory, where palliative care concepts are being developed, put into practice, and evaluated. It's mission was to put the patient first in mind, heart and soul. Right from the start, special attention was paid to the implementation a system of 'care for caregivers' in order to support the staff of the PCU in caring for the terminally ill and to prevent burnout. A process evaluation has been carried out with a special focus on the effects of the implementation of the new unit on the wellbeing of the interdisciplinary team of caregivers during the first year of its existence and on the experiences with a residential training preceding the start of patient care activities and the weekly staff support meetings. In the second half of the first year, the PCU staff experienced increased levels of job-related stress. Most stress was attributed to organizational factors, related to management issues but also to lack of institutional support. Teamwork also contributed to stress within the interdisciplinary team. Direct patient care activities also had some impact on experienced stress levels, through a heavy work-load of complex care, a shortage of staff and an experienced lack of competence. While the staff support activities were designed to give emotional support and deal with death and bereavement, these issues did not arise as major problems. As a consequence, the staff support in the PCU was perhaps less successful than it could have been. However, the high stress levels of this innovative palliative team warrant attention. We discuss how support meetings could be a more effective way of caring for staff. They should be accompanied by adequate resources, a supportive management structure, and an extensive educational program.


Trapped radiation belts of saturn: first look.
Pioneer 11 has made the first exploration of the magnetosphere and trapped radiation belts of Saturn. Saturn's magnetosphere is intermediate in size between Earth's and Jupiter's, with trapped particle intensities comparable to Earth's. The outer region of Saturn's magnetosphere contains lower energy radiation and is variable with time; the inner region contains higher energy particles. The pitch angle distributions show a remarkable variety of field-aligned and locally mirroring configurations. The moons and especially the rings of Saturn are effective absorbers of trapped particles; underneath the rings, the trapped radiation is completely absorbed. We confirm the discovery of a new ring, called the F ring, a new division, the Pioneer division, and a moon, called 1979 S 2. The latter has probably been seen from Earth. There may be evidence for more bodies like 1979 S 2, but at this stage the interpretation of the data is ambiguous. Using particle diffusion rates, we estimate that the cross-sectional area of the F ring is > 7 x 10(13) square centimeters and that the opacity is > 10(-5). Cosmic-ray albedo neutron decay should be looked into as a source of energetic particles in the inner magnetosphere of Saturn.


Elastic wave propagation and scattering in solids with uniaxially aligned cracks.
In this article, elastic wave propagation and scattering in a solid medium permeated by uniaxially aligned penny-shaped microcracks are studied. The crack alignment refers to the case in which the unit normals of all cracks are randomly oriented within a plane of isotropy. The analysis is restricted to the limit of the noninteraction approximation among individual cracks. Explicit expressions for attenuations and wave speeds of the shear horizontal, quasilongitudinal, and quasishear vertical waves are obtained using stochastic wave theory in a generalized dyadic approach. The ensemble average elastic wave response is governed by the Dyson equation, which is solved in terms of the anisotropic elastic Green's dyadic. The analysis of expressions is limited to frequencies below the geometric optics limit. The resulting attenuations are investigated in terms of the directional, frequency, and damage dependence. In particular, the attenuations are simplified considerably within the low frequency Rayleigh regime. Finally, numerical results are presented and discussed in terms of the relevant dependent parameters.


Expression of ICAM-1 in airway epithelium after acute ozone exposure in the mouse.
We investigated the time course and regional distribution of the expression of intercellular adhesion molecule-1 (ICAM-1) on airway epithelial cells and the polymorphonuclear leukocyte (PMN) inflammatory response in the lung after acute exposure to ozone (O3). C57BL/6J mice were exposed to air or 2 ppm O3 for 3 h and killed immediately or 3, 6, 9, or 21 h after exposure. Expression of ICAM-1 was examined by immunohistochemical staining of frozen sections. PMN influx was evaluated by lavage and by histochemical staining of myeloperoxidase (MPO) and measurement of tissue MPO activity. ICAM-1 expression exhibited regional selectivity and temporal patterns that were unique to each region. Upregulation of ICAM-1 expression on the epithelial cells in the trachea, and to a lesser extent in the lobar and segmental bronchi, was observed 3-9 h after exposure and remained present at 21 h. Enhanced ICAM-1 expression in bronchioles and terminal bronchiole/alveolar duct regions was evident earlier (immediately to 3 h after exposure) but returned to baseline levels by 21 and 9 h, respectively. Maximal ICAM-1 expression and PMN influx in the lung parenchyma were concurrently observed at 3 h, followed by transepithelial migration of PMNs to the airway lumen. These results demonstrate regional variations in airway inflammatory activity and are supportive of the notion that upregulation of ICAM-1 on the airway epithelium may play a role in local regulation of PMN influx to the airways after acute O3 exposure.


[Lymphoedema and elephantiasis due to filariasis. Pathogenesis and clinical aspects (author's transl)].
Adult lymphatic filariae (Wuchereria bancrofti, Brugia malayi) can cause blocking of lymphatics producing obliterating endolymphitis lesions. The subsequent extravasation of lymph (or chyle when the obstruction is canal) is at the origin of the formation of lymphedema or elephantiasis, in which the main histological finding is great hypertrophy of collagen elements. This theory involving filaria only is not the full picture, and bacterial infection, mainly by streptococci, is an important factor. The association of filaria with microbes is particularly dangerous because the presence of the latter, or its toxins, causes death of local microfilariae and even adult worms, which are known to be more harmful dead than alive. The progression of the disease, especially in cases with lymphedema, which mainly affects the limbs and the genital organs, depends on three factors: the species of filaria, the degree of transmission, and the receptivity of the patient to the parasite. Large differences are found according to the region involved, and in the same endemic zone, according to the individuals affected. However, they almost always occur progressively in areas where there have been recurrent attacks of acute lymphangitis.


Dissecting cancer through mathematics: from the cell to the animal model.
This Timeline article charts progress in mathematical modelling of cancer over the past 50 years, highlighting the different theoretical approaches that have been used to dissect the disease and the insights that have arisen. Although most of this research was conducted with little involvement from experimentalists or clinicians, there are signs that the tide is turning and that increasing numbers of those involved in cancer research and mathematical modellers are recognizing that by working together they might more rapidly advance our understanding of cancer and improve its treatment.


BldD is a direct regulator of key developmental genes in Streptomyces coelicolor A3(2).
BldD is a transcription factor required for aerial hyphae formation in the filamentous bacterium Streptomyces coelicolor. Three targets of BldD regulation were discovered by a number of means, including examination of bld gene interdependence, selective enrichment of chromosomal DNA fragments bound by BldD and searching the promoter regions of known developmental genes for matches to a previously characterized BldD binding site. The three BldD targets identified were the developmental sigma factor genes, whiG and bldN, and a previously uncharacterized gene, designated bdtA, encoding a putative transcription factor. In each target gene, the sequences bound by BldD were characterized by electrophoretic mobility shift and DNase I footprinting assays, and their alignment suggested AGTgA (n)m TCACc as a consensus BldD operator. The in vivo effect of mutation in bldD on the expression of these three target genes was assessed using S1 nuclease protection assays. In each case, target gene expression was upregulated during early colony development in the bldD background, suggesting that, in the wild type, BldD acts to repress premature expression of whiG, bldN and bdtA during vegetative growth.


Preparation of Bioscaffolds Delivering Stem Cells for Neural Regeneration.
Bioscaffolds have been proven for their feasibility in neural repair. Neural conduits have been investigated in the repair of wounded peripheral nerve and spinal cord. These conduits support axonal growth by providing structural guidance. Induced pluripotent stem cells (iPSCs) that are induced from a patient's own somatic cells have demonstrated significant neural cell differentiation capability and can circumvent immune system rejection. The combinatorial implantation of neural conduits and iPSCs may significantly enhance neural regeneration. The repair of nerves and spinal cords using biodegradable multichannel collagen conduits has been reported in our previous studies. In this review, we describe a method to fabricate a collagen neural conduit containing iPSC-derived neural cells.


Dissociation between short-term unloading and resistance training effects on skeletal muscle Na+,K+-ATPase, muscle function, and fatigue in humans.
Physical training increases skeletal muscle Na+,K+-ATPase content (NKA) and improves exercise performance, but the effects of inactivity per se on NKA content and isoform abundance in human muscle are unknown. We investigated the effects of 23-day unilateral lower limb suspension (ULLS) and subsequent 4-wk resistance training (RT) on muscle function and NKA in 6 healthy adults, measuring quadriceps muscle peak torque; fatigue and venous [K+] during intense one-legged cycling exercise; and skeletal muscle NKA content ([3H]ouabain binding) and NKA isoform abundances (immunoblotting) in muscle homogenates (α1-3, β1-2) and in single fibers (α1-3, β1). In the unloaded leg after ULLS, quadriceps peak torque and cycling time to fatigue declined by 22 and 23%, respectively, which were restored with RT. Whole muscle NKA content and homogenate NKA α1-3 and β1-2 isoform abundances were unchanged with ULLS or RT. However, in single muscle fibers, NKA α3 in type I (-66%, P = 0.006) and β1 in type II fibers (-40%, P = 0.016) decreased after ULLS, with other NKA isoforms unchanged. After RT, NKA α1 (79%, P = 0.004) and β1 (35%, P = 0.01) increased in type II fibers, while α2 (76%, P = 0.028) and α3 (142%, P = 0.004) increased in type I fibers compared with post-ULLS. Despite considerably impaired muscle function and earlier fatigue onset, muscle NKA content and homogenate α1 and α2 abundances were unchanged, thus being resilient to inactivity induced by ULLS. Nonetheless, fiber type-specific downregulation with inactivity and upregulation with RT of several NKA isoforms indicate complex regulation of muscle NKA expression in humans.


[Comparative characteristic of surgical approaches for aneurysms of thoracoabdominal aorta].
The surgical approaches for aneurysms of thoracoabdominal aorta are analyzed. Both thoracoretroperitoneal and thoracolaparotomic approaches are appropriate; they are up to quality of visualization and manipulation on low-thoracic part, visceral branches and bifurcation of aorta that is verified by experimental studies (wound depth 11-15 cm, angle of surgical action 100-110 degrees, angle of surgical action axis 80-90 degrees). The level of thoracotomy and type of approach depends on variant of lesion and planned reconstruction. Modified thoracophrenopararectal approach (wound depth 11.4+/-0.2 cm, angle of surgical action 97.0+/-3.7 degrees, angle of surgical action axis 88.0+/-2.0 degrees) is optimal for manipulations on aortic isthmus.