Effects of Medication Nonadherence and Comorbidity on Health Resource Utilization in Schizophrenia.
Medication nonadherence to antipsychotic drugs, which is commonly seen in patients with schizophrenia who have comorbidities, not only affects the quality of life of individuals suffering from the condition, but can also lead to worsening of disease condition, adverse outcomes, excessive use of health care resources, and higher medical costs. To determine the effect of nonadherence to antipsychotics and related disease comorbidities on medical care utilization with respect to inpatient hospital visits, outpatient visits, office visits, and emergency room (ER) visits. Retrospective, cross-sectional research data was obtained from the Medical Expenditure Panel Surveys (MEPS) for the years 2010-2014. The proportion of days covered (PDC) adherence measure was used to identify and classify individuals as adherent (PDC ≥ 80%) or nonadherent (PDC < 80%). A logistic regression analysis was used to further examine the effect of key study variables and comorbidity on medication nonadherence in patients with schizophrenia. Using the Student's t-test, population characteristics were statistically compared between the adherent and nonadherent populations and between populations with comorbidities and without comorbidities with respect to inpatient, outpatient, office, and ER visits. Of 1.2 million people who reported having schizophrenia in MEPS from 2010 to 2014, as many as 71% were found to be nonadherent to antipsychotic medications (PDC < 80%). Results showed that women (OR = 3.594, 95% CI = 1.33-11.40, P = 0.030) and people with less than 15 years of education (OR = 20.85, 95% CI = 3.91-111.09, P = 0.0005) were more likely to be nonadherent to antipsychotic medications than all other demographics. Compared with the adherent schizophrenia population (n = 353,349), the nonadherent population (n = 868,737) had greater utilization of outpatient visits (0.68 vs. 1.92, P < 0.0001) and office visits (10.95 vs. 18.21, P < 0.0001) but had lower utilization of inpatient visits (0.82 vs. 0.45, P < 0.0001) and ER visits (1.03 vs. 0.79, P = 0.1036). Compared with the schizophrenia population without comorbidities, the population with comorbidities (a classification based on a Charlson Comorbidity Index score of ≥ 1) had greater utilization of inpatient (0.39 vs. 0.76, P < 0.0001); office (13.39 vs. 19.34, P < 0.0001); and ER visits (0.39 vs. 1.41, P < 0.0001) but had lower utilization of outpatient visits (1.86 vs. 1.21, P < 0.0001). Greater medical care resources are used by nonadherent populations with schizophrenia and comorbidities than those without comorbidities. Together, nonadherence and comorbidities pose significant risks to patients with schizophrenia, in clinical and financial terms, and addressing problems stemming from such risks should be an area of priority in schizophrenia management. No outside funding supported this study. The authors have no potential conflicts of interest with respect to the research, authorship, and/or publication of this article.


Spatiotemporal Analysis for COVID-19 Delta Variant Using GIS-Based Air Parameter and Spatial Modeling.
The coronavirus disease of 2019 (COVID-19) pandemic is currently a global challenge, with 210 countries, including Indonesia, seeking to minimize its spread. Therefore, this study aims to determine the spatiotemporal spread pattern of this virus in Surabaya using various data on confirmed cases from 28 April to 26 October 2021. It also aims to determine the relationship between pollutant parameters, such as carbon monoxide (CO), nitrogen dioxide (NO2), sulfur dioxide (SO2), and ozone (O3), as well as the government's high social restrictions policy in Java-Bali. Several methods, such as the weighted mean center, directional distribution, Getis-Ord Gi*, Moran's I, and geographically weighted regression, were used to identify the spatial spread pattern of the virus. The weighted mean center indicated that the epicenter location of the outbreak moved randomly. The directional distribution demonstrated a decrease of 21 km2 at the end of the study phase, which proved that its spread has significantly reduced in Surabaya. Meanwhile, the Getis-Ord Gi* results demonstrated that the eastern and southern parts of the study region were highly infected. Moran's I demonstrate that COVID-19 cases clustered during the spike. The geographically weighted regression model indicated a number of influence zones in the northeast, northwest, and a few in the southwest parts at the peak of R2 0.55. The relationship between COVID-19 cases and air pollution parameters proved that people living at the outbreak's center have low pollution levels due to lockdown. Furthermore, the lockdown policy reduced CO, NO2, SO2, and O3. In addition, increase in air pollutants; namely, NO2, CO, SO2 and O3, was recorded after 7 weeks of lockdown implementation (started from 18 August).


Rebooting the immune system with high-dose cyclophosphamide for treatment of refractory myasthenia gravis.
A small but important proportion of patients with myasthenia gravis (MG) are refractory to conventional immunotherapy. We have treated 12 such patients by "rebooting" the immune system with high-dose cyclophosphamide (Hi Cy, 200 mg/kg), which largely eliminates the mature immune system, while leaving hematopoietic precursors intact. The objective of this report is to describe the clinical and immunologic results of Hi Cy treatment of refractory MG. We have followed 12 patients clinically for 1-9 years, and have analyzed their humoral and cellular immunologic parameters. Hi Cy is safe and effective. All but one of the patients experienced dramatic clinical improvement for variable periods from 5 months to 7.5 years, lasting for more than 1 year in seven of the patients. Two patients are still in treatment-free remission at 5.5 and 7.5 years, and five have achieved responsiveness to immunosuppressive agents that were previously ineffective. Hi Cy typically reduced, but did not completely eliminate, antibodies to the autoantigen AChR or to tetanus or diphtheria toxin; re-immunization with tetanus or diphtheria toxoid increased the antibody levels. Despite prior thymectomy, T cell receptor excision circles, generally considered to reflect thymic emigrant T cells, were produced by all patients. Hi Cy treatment results in effective, but often not permanent, remission in most refractory myasthenic patients, suggesting that the immune system is in fact "rebooted," but not "reformatted." We therefore recommend that treatment of refractory MG with Hi Cy be followed with maintenance immunotherapy.


Emerging trends in ion channel-based assays for predicting the cardiac safety of drugs.
Regulators require prospective new drugs to be tested for potential cardiac effects that are relevant to drug safety. As a result, numerous technologies have been developed to screen drugs for off-target effects that compromise the function of hERG potassium channels, which are associated with acquired long QT syndrome - a disorder with a risk of sudden death as a result of arrhythmia. Automated patch clamp, which is the most important and commonly used technology for the preclinical assessment of the risk of arrhythmia, continues to evolve toward greater sophistication, higher success rates and increased ease of use. Patch clamp advances include the development of population patch clamp and the ability to make more consistent measurements from freshly thawed prevalidated cells. Nevertheless, several discrepancies between hERG testing, QT prolongation and the risk of arrhythmia have been well-publicized. As a result, ion channel screening has been further developed to enable the assaying of currents from a wider variety of cardiac ion channels and the measurement of action potentials from stem cell-derived cardiomyocyte-like cells. Emerging measurement technologies beyond patch clamp include the measurement of field potentials with multi-electrode arrays.


In vivo kinematics of deep lunges and sit-to-stand activities in patients with bicruciate-retaining total knee arthroplasty.
The removal of the cruciate ligaments in total knee arthroplasty (TKA) has been suggested as a potential contributing factor to patient dissatisfaction, due to alteration of the in vivo biomechanics of the knee. Bicruciate retaining (BCR) TKA allows the preservation of the cruciate ligaments, thus offering the potential to reproduce healthy kinematics. The aim of this study was to compare in vivo kinematics between the operated and contralateral knee in patients who have undergone TKA with a contemporary BCR design. A total of 29 patients who underwent unilateral BCR TKA were evaluated during single-leg deep lunges and sit-to-stand tests using a validated computer tomography and fluoroscopic imaging system. In vivo six-degrees of freedom (6DOF) kinematics were compared between the BCR TKA and the contralateral knee. During single-leg deep lunge, BCR TKAs showed significantly less mean posterior femoral translation (13 mm; standard deviation (SD) 4) during terminal flexion, compared with the contralateral knee (16.6 mm, SD 3.7; p = 0.001). Similarly, BCR TKAs showed significantly less mean femoral rollback (11.6 mm (SD 4.5) vs 14.4 mm (SD 4.6); p < 0.043) during sit-to-stand. BCR TKAs showed significantly reduced internal rotation during many parts of the strenuous flexion activities particularly during high-flexion lunge (4° (SD 5.6°) vs 6.5° (SD 6.1°); p = 0.051) and during sit-to-stand (4.5° (SD 6°) vs 6.9° (SD 6.3°); p = 0.048). The contemporary design of BCR TKA showed asymmetrical flexion-extension and internal-external rotation, suggesting that the kinematics are not entirely reproduced during strenuous activities. Future studies are required to establish the importance of patient factors, component orientation and design, in optimizing kinematics in patients who undergo BCR TKA. Cite this article: Bone Joint J 2020;102-B(6 Supple A):59-65.


Elastic properties and breaking strengths of GaS, GaSe and GaTe nanosheets.
Gallium sulphide (GaS), gallium selenide (GaSe), and gallium telluride (GaTe), belonging to the group-III monochalcogenide family, have shown promising optoelectronic performance over graphene and monolayer molybdenum disulphide (MoS2). However, to date, the mechanical properties of these materials have not been investigated, which hinders their utilisation in flexible electronics and optomechanics. Here, we characterize the elastic properties and breaking strengths of suspended two-dimensional (2D) nanosheets of GaS, GaSe, and GaTe, using atomic force microscopy. The 2D Young's modulus values of ∼10 nm thick GaS, GaSe, and GaTe were found to be 1732 ± 154 N m-1, 819 ± 127 N m-1, and 246 ± 160 N m-1, respectively, corresponding to the three-dimensional (3D) Young's modulus values of 173 ± 15 GPa, 81.9 ± 12.7 GPa, and 24.6 ± 16 GPa, respectively. The pre-tension values of these nanosheets were estimated to be 0.34 ± 0.12 N m-1, 0.14 ± 0.04 N m-1, and 0.15 ± 0.03 N m-1 for GaS, GaSe, and GaTe, respectively. GaS nanosheets exhibited the highest Young's modulus (173 GPa) among these nanosheets, which is comparable to that of WS2 and WSe2. A failure characteristic study over these group-III monochalcogenides revealed that these materials can withstand stresses of up to 8 GPa and a maximal strain of 7% before breaking. Altogether, our findings indicate that GaS, GaSe, and GaTe are attractive candidates for use in stretchable electronic applications and in future optomechanical devices.


Transurethral microwave thermotherapy for treatment of benign prostatic hyperplasia: a preliminary assessment of the Prostalund feedback treatment machine.
The aim of this preliminary study was to evaluate the efficacy of Prostalund feedback treatment (PLFT) using microwave thermotherapy equipment for benign prostate hyperplasia (BPH) in patients with urinary retention and an indwelling urethral catheter. Four unselected patients, 51-68 years old (mean age 60 years), with urinary retention and an indwelling catheter were treated with PLFT under local anaesthesia and sedation. They were considered high-risk patients for treatment by transurethral resection of the prostate (TURP) under spinal or general anaesthesia. Initial assessment included prostate-specific antigen, determination of prostate volume and exclusion of prostate cancer using transrectal ultrasound (TRUS). The patients had the indwelling catheter for 1-2 weeks prior to treatment, the urethral catheters were removed 10-12 days after PLFT. Two patients who failed to void after transurethral microwave thermotherapy underwent urodynamic study to determine the reason for failure. The mean volume of the treated prostate gland was 75.50 cm3 (range 51-150 cm3). Two of the 4 patients were successfully relieved of their indwelling catheter with satisfactory peak flow, residual urine and symptom score, while treatment failed in the remaining 2. The reasons for the failure were identified as a small bladder capacity and neurogenic bladder in a diabetic patient and the presence of a middle lobe with large protruding lateral lobes into the bladder of the other patient. The latter patient had a prostate volume of 150 cm3. 'Salvage' TURP in both patients revealed that PLFT caused extensive necrosis of prostatic tissue and the TURP was then accomplished with minimal blood loss (<200 ml) even in the patient with 150-gram prostate. There were no serious complications such as bleeding, sepsis, rectoprostatic fistula or urine incontinence. The result of this preliminary investigation appears to be satisfactory and indicates that PLFT could be a good, minimally invasive alternative to surgery for BPH patients with retention of urine and an indwelling catheter.


Regulation of dictyostelium discoideum mRNAs specific for prespore or prestalk cells.
Prespore and prestalk cells in Dictyostelium discoideum aggregates can be separated by density gradient centrifugation. Using poly(A+) RNA from the fractionated cells to probe a cDNA library of mRNAs from postaggregation cells, we were able to identify six cDNA clones representing RNAs enriched in prespore or prestalk cells. Remarkably, transcripts of six of seven cDNA clones, previously selected to encode mRNAs present in postaggregating cells but low or absent in growing cells, also are enriched in RNA from either prestalk or prespore cells. By hybridization of cDNA probes to nitrocellulose blots of formaldehyde RNA gels, these 13 mRNA species have been examined with respect to cell type specificity, temporal pattern of accumulation, and affect of disaggregation and cAMP on accumulation. Aggregation-stage mRNAs tend to fit into three different classes. All prespore mRNAs are similar in all aspects of their regulation, while prestalk mRNAs fall into two co-regulated classes. All mRNAs that are present at significant levels during growth and differentiation are found in both cell types at comparable levels. Our results indicate that there is coordinate control of expression of genes specific for the two principal cell types.


Highly linear silicon traveling wave Mach-Zehnder carrier depletion modulator based on differential drive.
We present measurements of the nonlinear distortions of a traveling-wave silicon Mach-Zehnder modulator based on the carrier depletion effect. Spurious free dynamic range for second harmonic distortion of 82 dB·Hz(1/2) is seen, and 97 dB·Hz(2/3) is measured for intermodulation distortion. This measurement represents an improvement of 20 dB over the previous best result in silicon. We also show that the linearity of a silicon traveling wave Mach-Zehnder modulator can be improved by differentially driving it. These results suggest silicon may be a suitable platform for analog optical applications.


In-silico approach to identify novel potent inhibitors against GraR of S. aureus.
With rising antibiotic resistance at alarming rates in S. aureus, a major human pathogen, it is important to identify targets for new antimicrobial therapies. A number of two-component systems (TCS) have been implicated in S. aureus resistance to several antibiotics. The glycopeptide-resistance associated TCS, GraSR, is involved in cationic antimicrobial peptides (CAMPs) resistance through the regulation of mprF, dltABCD, and vraFG operons. GraS is a sensor histidine kinase, while GraR is a response regulator transcription factor, which is potential drug target. In lieu of the significance of GraSR in antibiotic resistance and the lack of structural studies on GraR, we undertook to determine the GraR structure through homology modelling. A series of small molecules were virtually screened and the top-scored molecules were analyzed for different pharmacophore properties and assessed for their binding potency to GraR (IC50). Further, a molecular dynamics simulation study of GraR-ligand complexes revealed that the predicted molecules exhibited good binding affinities at the dimerization interface of GraR. Thus, these molecules could be suitable inhibitors for the GraR-mediated signalling processes, which may be further utilized to develop novel antimicrobial agents against S. aureus.


Identification and characterization of transcript polymorphisms in soybean lines varying in oil composition and content.
Variation in seed oil composition and content among soybean varieties is largely attributed to differences in transcript sequences and/or transcript accumulation of oil production related genes in seeds. Discovery and analysis of sequence and expression variations in these genes will accelerate soybean oil quality improvement. In an effort to identify these variations, we sequenced the transcriptomes of soybean seeds from nine lines varying in oil composition and/or total oil content. Our results showed that 69,338 distinct transcripts from 32,885 annotated genes were expressed in seeds. A total of 8,037 transcript expression polymorphisms and 50,485 transcript sequence polymorphisms (48,792 SNPs and 1,693 small Indels) were identified among the lines. Effects of the transcript polymorphisms on their encoded protein sequences and functions were predicted. The studies also provided independent evidence that the lack of FAD2-1A gene activity and a non-synonymous SNP in the coding sequence of FAB2C caused elevated oleic acid and stearic acid levels in soybean lines M23 and FAM94-41, respectively. As a proof-of-concept, we developed an integrated RNA-seq and bioinformatics approach to identify and functionally annotate transcript polymorphisms, and demonstrated its high effectiveness for discovery of genetic and transcript variations that result in altered oil quality traits. The collection of transcript polymorphisms coupled with their predicted functional effects will be a valuable asset for further discovery of genes, gene variants, and functional markers to improve soybean oil quality.


[Vascularization of the testes of the white rat in the high altitude conditions of the Pamir and Antarctica].
By means of histometric and morphometric methods vascularization of the white rat testes has been studied at adaptation to living conditions in the Alpian Pamir (at the altitude of 4,000 m) and in the Antarctic Continent (at the altitude of 3,488 m). The leading factor in development of the parenchymatous rearrangements of the testes are certain disturbances of the microcirculatory link in the intraorganic bed. Different degree in manifestation of the morphological signs of the circulatory disturbances are revealed: plethora, interstitial tissue edema, degenerative changes in the spermatogenic epithelium cells and hence decreasing activity of spermatogenesis. The disturbances noted are essentially various in different stages of adaptation to the altitude. In contrast to the Pamir series of the experiment, in the group of the animals adapting to the living conditions of "Vostok" station, dilatation of the capillary lumens and signs of moderate plethora accompanying them remain up to the end of the observations.


[Effects of oxygenated free radicals on the electrophysiological activity of the isolated retina of the rat].
Particularly rich in polyunsaturated fatty acids, the retina constitutes an excellent model for testing the functional consequences of membrane lipoperoxidation. The effects could be quantified by measuring the amplitude of the electroretinogram which represents the characteristic response of the retina to its physiological stimulus (light photons). The authors report results obtained on isolated rat retina maintained in survival by perfusion. The membrane lipid peroxidation is induced by the non enzymatic catalytic system (Fe2+ + ascorbate). In the presence of such a system, the amplitude of the b-wave of the electroretinogram rapidly decreases and the survival time is notably shortened. These results are discussed with regards to the mechanism generating the electroretinogram and the specific role played by the disk membrane and plasmic membrane of the rod outer segment. Finally, the authors test the radical scavenger properties of a Ginkgo biloba extract. The results indicate that such an extract develops an antiperoxidative effect probably connected with its flavonoid composition. With this extract the survival time of the isolated retina is significantly increased and this result could be related to the retinal cell membrane disorders observed in diabetes.


Clustering of diet, physical activity and overweight in parents and offspring in South India.
Although there are some studies on familial clustering of metabolic risk factors, there are few on clustering of diet and physical activity. The aim of this study was to identify the extent of familial clustering of weight status, diet and physical activity in urban and rural families in South India. Anthropometric, dietary and physical activity data were obtained for 325 offspring aged 8-21 years and both their parents from urban and rural South India. However, 294 single offspring and their parents were used for analysis. Body mass index (BMI) was computed and individuals categorized as normal, underweight and overweight/obese. Diet was assessed using food frequency questionnaires and physical activity using physical activity questionnaires. Offspring BMI correlated significantly (P<0.001) with the BMIs of both parents (r=0.35, P<0.001). Percent protein, carbohydrate, total fat and saturated fat intakes were significantly correlated (P<0.001) between offspring and parents. Offspring physical activity level (PAL) correlated significantly with the mother (r=0.26, P<0.001), but not the father. A two-step cluster analysis revealed two distinct clusters: obesogenic and nonobesogenic. The obesogenic cluster, which consisted of 83% of families with at least one parent or offspring overweight, had significantly higher intakes of fat, saturated fat and higher sedentary activity levels but lower carbohydrate intakes and sleep duration (all P<0.001) among both offspring and parents. Diet, physical activity and BMI of children, adolescents and young adults in India is associated with parental behavior. Health promotion aimed at reducing obesity in these groups should address familial issues.


Sodium intake, angiotensin II receptor blockade, and baroreflex function in conscious rats.
The hypothesis that endogenous angiotensin II (Ang II) chronically supports baroreflex control of lumbar sympathetic nerve activity (LSNA) and heart rate (HR) via AT1 but not AT2 receptors was tested in conscious, normotensive rats. Rats were fed either a sodium deficient diet (LS) to increase circulating Ang II or a high-sodium diet (HS) for 2 to 3 weeks. One to two days after surgery to implant catheters and nerve electrodes, baroreflex curves were produced before and 40 minutes after intravenous administration of the AT1 antagonist losartan (10 mg/kg) or the AT2 antagonist PD123319 (500 micrograms/kg + 50 micrograms.kg-1.min-1). Mean arterial pressure (MAP) after losartan was maintained at basal levels with methoxamine. Forty minutes after losartan in LS rats, LSNA (46 +/- 5 to 22 +/- 1% max) and HR (414 +/- 7 to 387 +/- 8 bpm) were decreased (P < .05). Losartan decreased reflex control of LSNA more in LS than in HS rats (P < .05), as indicated by reductions in maximum LSNA (98 +/- 2 to 78 +/- 3% max) and minimum LSNA (42 +/- 5 to 21 +/- 5% max). Losartan also shifted reflex control of LSNA to a lower pressure in both groups, but the effect was larger in LS rats (-21 +/- 3 [LS] versus -9 +/- 2 [HS] mm Hg at basal LSNA; P < .05). Maximum gain was unaltered in either group. Similarly, losartan reduced maximum HR (534 +/- 6 to 495 +/- 9 bpm) and shifted the HR curve leftward (114 +/- 5 to 105 +/- 4 mm Hg) in LS but not in HS rats. In general, no changes were observed in MAP or baroreflex control of LSNA and HR after PD123319 in LS rats. These results suggest that in conscious, normotensive LS rats, endogenous Ang II supports LSNA and HR over a wide MAP range via AT1 but not AT2 receptors.


Molecular-dynamics approach for studying the nonequilibrium behavior of x-ray-heated solid-density matter.
When matter is exposed to a high-intensity x-ray free-electron-laser pulse, the x rays excite inner-shell electrons leading to the ionization of the electrons through various atomic processes and creating high-energy-density plasma, i.e., warm or hot dense matter. The resulting system consists of atoms in various electronic configurations, thermalizing on subpicosecond to picosecond timescales after photoexcitation. We present a simulation study of x-ray-heated solid-density matter. For this we use XMDYN, a Monte Carlo molecular-dynamics-based code with periodic boundary conditions, which allows one to investigate nonequilibrium dynamics. XMDYN is capable of treating systems containing light and heavy atomic species with full electronic configuration space and three-dimensional spatial inhomogeneity. For the validation of our approach we compare for a model system the electron temperatures and the ion charge-state distribution from XMDYN to results for the thermalized system based on the average-atom model implemented in XATOM, an ab initio x-ray atomic physics toolkit extended to include a plasma environment. Further, we also compare the average charge evolution of diamond with the predictions of a Boltzmann continuum approach. We demonstrate that XMDYN results are in good quantitative agreement with the above-mentioned approaches, suggesting that the current implementation of XMDYN is a viable approach to simulate the dynamics of x-ray-driven nonequilibrium dynamics in solids. To illustrate the potential of XMDYN for treating complex systems, we present calculations on the triiodo benzene derivative 5-amino-2,4,6-triiodoisophthalic acid (I3C), a compound of relevance of biomolecular imaging, consisting of heavy and light atomic species.


Randomized trials in single subjects: the N of 1 study.
Large-scale randomized trials are not available for all disorders, and conventional "trials of therapy" are susceptible to bias. Randomized controlled trials (RCTs) in individual patients (N of 1 RCTs) may provide a solution. In an N of 1 RCT, a patient receives treatments in pairs (one period of the experimental therapy and one period of either an alternative treatment or placebo, in random order), both patient and clinician are kept blind to allocation, and treatment targets are monitored. This type of RCT is useful in chronic, stable conditions in which the proposed treatment has a short half-life. Treatment targets usually include quantitative measurement of symptoms tracked through patient diaries. Pairs of treatment periods are continued until effectiveness is proved or refuted. The N of 1 RCT is potentially of great use in psychopharmacology and in drug development. The practical approach presented here will allow clinicians to conduct their own N of 1 RCTs.


Quantification of infarct size and myocardium at risk: evaluation of different techniques and its implications.
The aim of this study was to evaluate seven methods for quantifying myocardial oedema [2 standard deviation (SD), 3 SD, 5 SD, full width at half maximum (FWHM), Otsu method, manual thresholding, and manual contouring] from T2-weighted short tau inversion recovery (T2w STIR) and also to reassess these same seven methods for quantifying acute infarct size following ST-segment myocardial infarction (STEMI). This study focuses on test-retest repeatability while assessing inter- and intraobserver variability. T2w STIR and late gadolinium enhancement (LGE) are the most widely used cardiovascular magnetic resonance (CMR) techniques to image oedema and infarction, respectively. However, no consensus exists on the best quantification method to be used to analyse these images. This has potential important implications in the research setting where both myocardial oedema and infarct size are increasingly used and measured as surrogate endpoints in clinical trials. Forty patients day 2 following acute reperfused STEMI were scanned for myocardial oedema and infarction (LGE). All patients had a second CMR scan on the same day >6 h apart from the first one. Images were analysed offline by two independent observers using the semi-automated software. Both oedema and LGE were quantified using seven techniques (2 SD, 3 SD, 5 SD, Otsu, FWHM, manual threshold, and manual contouring). Interobserver, intraobserver and test-retest agreement and variability for both infarct size and oedema quantification were assessed. Infarct size and myocardial quantification vary depending on the quantification method used. Overall, manual contouring provided the lowest inter-, intraobserver, and interscan variability for both infarct size and oedema quantification. The FWHM method for infarct size quantification and the Otsu method for myocardial oedema quantification are acceptable alternatives. This study determines that, in acute myocardial infarction (MI), manual contouring has the lowest overall variability for quantification of both myocardial oedema and MI when analysed by experienced observers.


Melaleuca alternifolia anthelmintic activity in gerbils experimentally infected by Haemonchus contortus.
Gastrointestinal parasites are one of the biggest health problems faced in sheep, mainly due to their pathogenicity and resistance to drugs used to control these parasites. Thus, the following study aimed to assess the anthelmintic efficacy of Melaleuca alternifolia against Haemonchus contortus in gerbils (Meriones unguiculatus) experimentally infected. Three treatments were tested: M. alternifolia essential oil, popularly known as tea tree oil (TTO), a solid lipid nanocarrier made with essential oil of Melaleuca (nanoTTO), and terpinen-4-ol (terp-4-ol). In vivo studies were performed by determining the mean worm burden of H. contortus in gerbils TTO (0.75 mL/kg); nanoTTO (0.5 mL/kg) and terp-4-ol (0.5 mL l/kg) were able to reduce 46.36%; 48.64%, and 43.18% worm burden, respectively. H. contortus increased alanine aminotransferase (ALT) and aspartate aminotransferase (AST) levels, as demonstrated by liver injury. It was found that the TTO, nanoTTO, and terp-4-ol were not toxic to liver and kidneys since hepatic and renal functions were not affected. Moreover, terp-4-ol was able to prevent increased levels of seric AST and ALT in infected animals, indicating a hepatoprotective effect. Thus, our results indicate that TTO, nanoTTO, and terp-4-ol are safe and efficient against H. contortus infection in gerbils, and possibly the terp-4-ol may be considered the compound present in the Melaleuca alternifolia responsible for parasitic action against H. contortus.


HIV-1 Reverse Transcriptase Still Remains a New Drug Target: Structure, Function, Classical Inhibitors, and New Inhibitors with Innovative Mechanisms of Actions.
During the retrotranscription process, characteristic of all retroviruses, the viral ssRNA genome is converted into integration-competent dsDNA. This process is accomplished by the virus-coded reverse transcriptase (RT) protein, which is a primary target in the current treatments for HIV-1 infection. In particular, in the approved therapeutic regimens two classes of drugs target RT, namely, nucleoside RT inhibitors (NRTIs) and nonnucleoside RT inhibitors (NNRTIs). Both classes inhibit the RT-associated polymerase activity: the NRTIs compete with the natural dNTP substrate and act as chain terminators, while the NNRTIs bind to an allosteric pocket and inhibit polymerization noncompetitively. In addition to these two classes, other RT inhibitors (RTIs) that target RT by distinct mechanisms have been identified and are currently under development. These include translocation-defective RTIs, delayed chain terminators RTIs, lethal mutagenesis RTIs, dinucleotide tetraphosphates, nucleotide-competing RTIs, pyrophosphate analogs, RT-associated RNase H function inhibitors, and dual activities inhibitors. This paper describes the HIV-1 RT function and molecular structure, illustrates the currently approved RTIs, and focuses on the mechanisms of action of the newer classes of RTIs.


Cell damage caused by vaginal Candida albicans isolates from women with different symptomatologies.
The present study aimed to characterize cell damage caused by vaginal Candida albicans isolates from women with different symptomatologies. It was evaluated 12 clinical isolates of C. albicans from vaginal samples: 4 from asymptomatic women (AS), 4 from women with a single episode of vulvovaginal candidiasis (VVC) and 4 from women with recurrent vulvovaginal candidiasis (RVVC). We evaluated the ability of C. albicans to adhere to human cervical cancer cells (SiHa), the yeast-SiHa cell interactions and cell damage. All of the clinical isolates presented a high adhesion capacity on SiHa cells. However, clinical isolates from symptomatic women (VVC and RVVC) had higher filamentation after contact (24 h) with SiHa cells and a greater capacity to cause cell damage (>80 %). Clinical isolates from symptomatic women had greater potential to invade SiHa cells, suggesting that they are more pathogenic than AS isolates.


Molecular analysis of natural populations of Populus nigra L intermingled with cultivated hybrids.
In this study six simple sequence repeats (SSR or microsatellites) were selected for their ability to fingerprint a total of 60 commercial clones of Populus deltoides Marsh. and Populus x canadensis Moench (typically derived from crosses between Populus nigra L and P. deltoides) and to characterize a natural population of P. nigra growing along the Ticino river in the North of Italy. Out of six SSRs used, four microsatellite loci were found to have alleles which were species-specific to P. deltoides and could therefore be used as markers for introgression of P. deltoides into P. nigra. In the studied region hybrid poplars and P. deltoides commercial clones are cultivated as monoclonal stands close to the area where black poplar has its natural habitat. SSR analysis was performed to investigate whether there was evidence of introgression between the natural population and the monoclonal plantations of hybrids and P. deltoides clones cultivated in the surrounding area. Three stages of the natural population were analysed: a group of old trees about a hundred years old, a younger population (aged 2-30 years) and the seedlings of three females of this population. Alleles specific to P. deltoides were detected only in the old cohort of the natural population, while no introgression was observed in the younger individuals and their progenies. These results were also confirmed by isozyme analysis of loci PGI-B, PGM and LAP-A, which were previously identified as diagnostic for P. nigra, P. deltoides and P.xcanadensis.


Psychophysical assessment of timing in individuals with autism.
Perception of time, in the seconds to minutes range, is not well characterized in autism. The required interval timing system (ITS) develops at the same stages during infancy as communication, social reciprocity, and other cognitive and behavioral functions. The authors used two versions of a temporal bisection procedure to study the perception of duration in individuals with autism and observed quantifiable differences and characteristic patterns in participants' timing functions. Measures of timing performance correlated with certain autism diagnostic and intelligence scores, and parents described individuals with autism as having a poor sense of time. The authors modeled the data to provide a relative assessment of ITS function in these individuals. The implications of these results for the understanding of autism are discussed.


Long-term prognostic assessment of 185 newly diagnosed gliomas: Grade III glioma showed prognosis comparable to that of Grade II glioma.
We evaluated the prognoses of newly diagnosed gliomas through WHO Grades II, III and IV to assess the overall tendency of treatment results for glioma in our institute. Furthermore, statistical analysis was performed to determine factors influencing the prognosis. A total of 185 newly diagnosed glioma patients were operated on from 2000 to 2006. The primary endpoint was the overall survival from the date of surgery. The factors assessed as to whether they influenced the prognosis were the WHO grades of sex, age, location of the lesion, pre-operative Karnofsky Performance Status (KPS), extent of resection and whether or not radiation therapy was performed. The WHO grades influenced the survival significantly (P < 0.0001). The Grades II and III showed no statistically significant difference in survival (P = 0.174), whereas Grades III and IV showed a significant difference (P < 0.0001). The factor influencing survival as well as the grades was the KPS (P < 0.0001). The comparison of survival over WHO grades in the same KPS group was performed for 2 KPS groups (KPS = 100, KPS 80-90), and these also showed significant differences (P = 0.0009 and 0.0143, respectively). Despite the different distributions of the KPS, the Grade III glioma patients showed survival comparable to that of the Grade II. On the other hand, the Grade IV glioma patients showed significantly poorer survival compared with Grade II or III.


Proton-pump inhibitors in patients requiring antiplatelet therapy: new FDA labeling.
Proton-pump inhibitors (PPIs) are recommended for patients who require antiplatelet therapy and have a history of upper gastrointestinal bleeding. Proton-pump inhibitors should also be considered for patients receiving antiplatelet therapy who have other risk factors for gastrointestinal bleeding, including use of aspirin. Thus, evidence of pharmacokinetic and pharmacodynamic interactions between PPIs and consequent impaired effectiveness of the antiplatelet agent clopidogrel has caused concern. Here, we discuss comparative studies suggesting that the extent to which a PPI reduces exposure to the active metabolite of clopidogrel and attenuates its antithrombotic effect differs among PPIs. Although a clinically meaningful effect of the interaction between PPIs and clopidogrel on cardiovascular outcomes has not been established, these studies provided the basis for recent changes in US Food and Drug Administration (FDA) labeling for several PPIs and clopidogrel. New labeling suggests that PPI use among patients taking clopidogrel be limited to pantoprazole, rabeprazole, lansoprazole, or dexlansoprazole. Because comparative studies indicate that omeprazole and esomeprazole have a greater effect on the CYP2C19-mediated conversion of clopidogrel to its active metabolite and, consequently, clopidogrel's effect on platelet reactivity, FDA labeling recommends avoiding omeprazole and esomeprazole in patients taking clopidogrel. Even a 12-hour separation of dosing does not appear to prevent drug interactions between omeprazole and clopidogrel.


Pleiotropy informed adaptive association test of multiple traits using genome-wide association study summary data.
Genetic variants associated with disease outcomes can be used to develop personalized treatment. To reach this precision medicine goal, hundreds of large-scale genome-wide association studies (GWAS) have been conducted in the past decade to search for promising genetic variants associated with various traits. They have successfully identified tens of thousands of disease-related variants. However, in total these identified variants explain only part of the variation for most complex traits. There remain many genetic variants with small effect sizes to be discovered, which calls for the development of (a) GWAS with more samples and more comprehensively genotyped variants, for example, the NHLBI Trans-Omics for Precision Medicine (TOPMed) Program is planning to conduct whole genome sequencing on over 100 000 individuals; and (b) novel and more powerful statistical analysis methods. The current dominating GWAS analysis approach is the "single trait" association test, despite the fact that many GWAS are conducted in deeply phenotyped cohorts including many correlated and well-characterized outcomes, which can help improve the power to detect novel variants if properly analyzed, as suggested by increasing evidence that pleiotropy, where a genetic variant affects multiple traits, is the norm in genome-phenome associations. We aim to develop pleiotropy informed powerful association test methods across multiple traits for GWAS. Since it is generally very hard to access individual-level GWAS phenotype and genotype data for those existing GWAS, due to privacy concerns and various logistical considerations, we develop rigorous statistical methods for pleiotropy informed adaptive multitrait association test methods that need only summary association statistics publicly available from most GWAS. We first develop a pleiotropy test, which has powerful performance for truly pleiotropic variants but is sensitive to the pleiotropy assumption. We then develop a pleiotropy informed adaptive test that has robust and powerful performance under various genetic models. We develop accurate and efficient numerical algorithms to compute the analytical P-value for the proposed adaptive test without the need of resampling or permutation. We illustrate the performance of proposed methods through application to joint association test of GWAS meta-analysis summary data for several glycemic traits. Our proposed adaptive test identified several novel loci missed by individual trait based GWAS meta-analysis. All the proposed methods are implemented in a publicly available R package.


Control of a tyrosyl radical mediated protein cross-linking reaction by electrostatic interaction.
Herein, we demonstrate the control of protein heteroconjugation via a tyrosyl coupling reaction by using electrostatic interaction. Aspartic acid and arginine were introduced to a tyrosine containing peptide tag (Y-tag) to provide electrostatic charge. Designed negatively or positively charged Y-tags were tethered to the C-terminus of Escherichia coli alkaline phosphatase (BAP) and streptavidin (SA), and these model proteins were subjected to horseradish peroxidase (HRP) treatment. The negatively charged Y-tags showed low reactivity due to repulsive interactions between the Y-tags with the negatively charged BAP and SA. In contrast, the positively charged Y-tags showed high reactivity, indicating that the electrostatic interaction between Y-tags and proteins significantly affects the tyrosyl radical mediated protein cross-linking. From the heteroconjugation reaction of BAP and SA, the SA with the positively charged Y-tags exhibited favorable cross-linking toward negatively charged BAP, and the BAP-SA conjugates prepared from BAP with GY-tag (GGGGY) and SA with RYR-tag (RRYRR) had the best performance on a biotin-coated microplate. Encompassing the reactive tyrosine residue with arginine residues reduced the reactivity against HRP, enabling the modulation of cross-linking reaction rates with BAP-GY. Thus, by introducing a proper electrostatic interaction to Y-tags, it is possible to kinetically control the heteroconjugation behavior of proteins, thereby maximizing the functions of protein heteroconjugates.


Integrating Refined Kano Model and QFD for Service Quality Improvement in Healthy Fast-Food Chain Restaurants.
People are paying greater attention to health. To maintain a good health status and obtain food fast, customers may go to healthy fast-food chain restaurants such as Subway more often than before in China and Taiwan. Healthy fast-food chain restaurants come with a healthy spin, seeking to differentiate themselves from other fast-food restaurants. This paper combined the refined Kano model and the quality function deployment (QFD) method. The refined Kano model was used to understand how customers perceive service attributes developed based on DINESERV measurements. QFD was employed to describe the relationships among the critical service attributes and corresponding improvements as well as to identify the priority for these improvements. The analysis results revealed that providing limited offers (due to periods, seasons, and regions) should be at the top of their improvement list, followed by staff suggestions for ingredients, and a temperature display to enhance the image of fresh ingredients. Other improvement actions include providing regular launches of new flavors/products, designing new and attractive slogans, and providing restaurant apps.


Empathy as a neuropsychological heuristic in social decision-making.
Decision-making in social dilemmas is suggested to rely on three factors: the valuation of a choice option, the relative judgment of two or more choice alternatives, and individual factors affecting the ease at which judgments and decisions are made. Here, we test whether empathy-an individual's relative ability to understand others' thoughts, emotions, and intentions-acts as an individual factor that alleviates conflict resolution in social decision-making. We test this by using a framed, iterated prisoners' dilemma (PD) game in two settings. In a behavioral experiment, we find that individual differences in empathic ability (the Empathy Quotient, EQ) were related to lower response times in the PD game, suggesting that empathy is related to faster social choices, independent of whether they choose to cooperate or defect. In a subsequent neuroimaging experiment, using functional magnetic resonance imaging, we find that EQ is positively related to individual differences in the engagement of brain structures implemented in mentalizing, including the precuneus, superior temporal sulcus, and dorsolateral prefrontal cortex. These results suggest that empathy is related to the individual difference in the engagement of mentalizing in social dilemmas and that this is related to the efficiency of decision-making in social dilemmas.


The physiology of learning and memory: role of peptides and stress.
The neuropeptides, as well as their respective receptors, are widely distributed throughout the mammalian central nervous system. During learning and memory processes, besides structural synaptic remodeling, changes are observed at molecular and metabolic levels with the alterations in neurotransmitter and neuropeptide synthesis and release. While there is consensus that brain cholinergic neurotransmission plays a critical role in the processes related to learning and memory, it is also well known that these functions are influenced by a tremendous number of neuropeptides and non-peptide molecules. Arginine vasopressin (AVP), oxytocin, angiotensin II, insulin, growth factors, serotonin (5-HT), melanin concentrating hormone, histamine, bombesin and gastrin-releasing peptide (GRP), glucagon-like peptide-1 (GLP-1), cholecystokinin (CCK), dopamine, corticotropin releasing factor (CRF) have modulatory effects on learning and memory. Among these peptides CCK, 5-HT and CRF play strategic roles in the modulation of memory processes under stressful conditions. CRF is accepted as the main neuropeptide involved in both physical and emotional stress, with a protective role during stress, possibly through the activation of the hypothalamo-pitiuitary (HPA) axis. The peptide CCK has been proposed to facilitate memory processing and CCK-like immunoreactivity in the hypothalamus was observed upon stress exposure, suggesting that CCK may participate in the central control of stress response and stress-induced memory dysfunction. On the other hand, 5-HT appears to play a role in behaviors that involve a high cognitive demand and stress exposure activates serotonergic systems in a variety of brain regions. The physiological role and therapeutic efficacy of various neuropeptides and the impact of stress exposure in the acquisition and consolidation of memory will be reviewed thoroughly.


Factors associated with nutritional risk in patients receiving haemodialysis assessed by Nutritional Risk Screening 2002 (NRS2002).
Nutritional Risk Screening 2002 is recommended as a screening tool to identify patients at risk of undernutrition for all patients in hospitals by the European Society of Clinical Nutrition and Metabolism. Nutritional risk is associated with increased morbidity and mortality in patients, and it is common among patients on haemodialysis. Factors associated with nutritional risk that could facilitate the screening/diagnostic procedures are warranted. Identification of factors that are associated with nutritional risk in patients with end-stage renal disease treated with haemodialysis. Single-centre, cross-sectional study in patients receiving haemodialysis (n = 53) were screened for nutritional risk using Nutritional Risk Screening 2002. Associations were made with data on dietary intake by 24-h dietary recall, and measurement of body composition, anthropometric measurements and biochemical variables. Nutritional risk was common among patients on haemodialysis (26%), and was associated with low energy and protein intake, and low pre-albumin concentrations also after adjustments for age and sex. Nutritional risk was neither associated with diabetes nor duration of dialysis treatment. Measurement of pre-albumin and dietary assessment using a 24-h dietary recall can support the identification of patients receiving haemodialysis at nutritional risk.


The current functional state of local neuronal circuits controls the magnitude of a BOLD response to incoming stimuli.
The purpose of this study was to determine how the history-dependent activation state of neuronal networks controls fMRI signals to incoming stimuli. Simultaneous electrophysiological and blood oxygen level-dependent (BOLD) responses were monitored during stimulation of the perforant pathway with low, high, and again low intensity but, otherwise identical pulse trains. Under three different anesthetics (alpha-chloralose, medetomidine, isoflurane) consecutive low intensity stimulation trains, set just below the threshold for population spike generation to single pulses, yielded a stable BOLD response, although at different magnitudes. The first high intensity train increased the BOLD response under all anesthetics and generated population spikes, with varying amplitudes and latencies (alpha-chloralose, metedomidine) or in a regular pattern (isoflurane). Concurrent to the second high intensity train, the BOLD response became minimal, then slowly increasing with subsequent trains (alpha-chloralose, metedomidine), or immediately rising to a stable level (isoflurane). Second train population spikes became regularized, but at low amplitudes and long latencies that were slowly reversed across trains (alpha-chloralose, medetomidine); while under isoflurane, amplitude and latencies became stabilized with the second train. In comparison to initial stimulation, the final low intensity stimulation trains failed to produce BOLD responses (alpha-chloralose, medetomidine), or left the response unchanged (isoflurane), only reaching stable potentiation of population spikes when under isoflurane. Therefore, the fate of BOLD responses depends on whether a new stable functional state of the intrinsic network can be reached after high intensity stimulation.


Phosphate deprivation induces transfer of DGDG galactolipid from chloroplast to mitochondria.
In many soils plants have to grow in a shortage of phosphate, leading to development of phosphate-saving mechanisms. At the cellular level, these mechanisms include conversion of phospholipids into glycolipids, mainly digalactosyldiacylglycerol (DGDG). The lipid changes are not restricted to plastid membranes where DGDG is synthesized and resides under normal conditions. In plant cells deprived of phosphate, mitochondria contain a high concentration of DGDG, whereas mitochondria have no glycolipids in control cells. Mitochondria do not synthesize this pool of DGDG, which structure is shown to be characteristic of a DGD type enzyme present in plastid envelope. The transfer of DGDG between plastid and mitochondria is investigated and detected between mitochondria-closely associated envelope vesicles and mitochondria. This transfer does not apparently involve the endomembrane system and would rather be dependent upon contacts between plastids and mitochondria. Contacts sites are favored at early stages of phosphate deprivation when DGDG cell content is just starting to respond to phosphate deprivation.


Phylogenetic analysis of the triterpene cyclase protein family in prokaryotes and eukaryotes suggests bidirectional lateral gene transfer.
Functional constraints to modifications in triterpene cyclase amino acid sequences make them good candidates for evolutionary studies on the phylogenetic relatedness of these enzymes in prokaryotes as well as in eukaryotes. In this study, we used a set of identified triterpene cyclases, a group of mainly bacterial squalene cyclases and a group of predominantly eukaryotic oxidosqualene cyclases, as seed sequences to identify 5288 putative triterpene cyclase homologues in publicly available databases. The Cluster Analysis of Sequences software was used to detect groups of sequences with increased pairwise sequence similarity. The sequences fall into two main clusters, a bacterial and a eukaryotic. The conserved, informative regions of a multiple sequence alignment of the family were used to construct a neighbour-joining phylogenetic tree using the AsaturA and maximum likelihood phylogenetic tree using the PhyML software. Both analyses showed that most of the triterpene cyclase sequences were similarly grouped to the accepted taxonomic relationships of the organism the sequences originated from, supporting the idea of vertical transfer of cyclase genes from parent to offspring as the main evolutionary driving force in this protein family. However, a small group of sequences from three bacterial species (Stigmatella, Gemmata and Methylococcus) grouped with an otherwise purely eukaryotic cluster of oxidosqualene cyclases, while a small group of sequences from seven fungal species and a sequence from the fern Adiantum grouped consistently with a cluster of otherwise purely bacterial squalene cyclases. This suggests that lateral gene transfer may have taken place, entailing a transfer of oxidosqualene cyclases from eukaryotes to bacteria and a transfer of squalene cyclase from bacteria to an ancestor of the group of Pezizomycotina fungi.


Immunoglobulin deposition in the cornea after application of autologous serum.
A 47-year-old man with a history of multiple corneal allografts for recurrent herpes simplex keratitis developed a subtotal nonhealing corneal epithelial defect. The patient was treated with hourly drops of autologous serum. A ringlike infiltrate was subsequently observed, followed by reepithelialization of the graft. The patient later suffered allograft rejection of the cornea and recurrence of the epithelial defect, and a repeated penetrating keratoplasty was performed. Examination of the excised button demonstrated a total epithelial defect, changes compatible with allograft rejection, and, in addition, eosinophilic granular deposits within the superficial corneal stroma that corresponded to the "immune ring" observed clinically. Immunoperoxidase staining was positive for IgG, IgM, IgA, and kappa and lambda light chains. These pathologic changes lend credence to the hypothesis that the precorneal tear film may be a source of immunoglobulin that becomes deposited within the stroma.


Early development of mechanisms of change as a predictor of subsequent change and treatment outcome: The case of working alliance.
Advanced statistical tools have created the opportunity to systematically examine the effect of early trajectories in predictors of therapeutic change, such as early alliance development patterns, on outcome. To date, however, these methods have been used almost exclusively to examine the effect of the development of early symptoms on later ones. Development patterns of alliance early in treatment, and their association with treatment outcome, have received much theoretical attention, but few systematic examinations have been conducted so far. We integrated exploratory cluster analysis with the accumulated theoretical and empirical knowledge on patterns of alliance development to identify distinct patterns of early alliance development across the 1st 4 sessions of treatment in a sample of 166 patients receiving psychotherapy. Three patterns of early alliance development were identified: early gradual strengthening, early repaired rupture, and early unrepaired rupture. The gradual strengthening and the repaired rupture patterns early in treatment predicted alliance strengthening later in treatment, whereas the unrepaired rupture pattern early in treatment predicted alliance stability later in treatment. The effect of early alliance development patterns on treatment outcome was moderated by pretreatment interpersonal problems: For patients with better interpersonal functioning at intake, the gradual strengthening and the repaired rupture patterns showed better outcomes, whereas for those with poorer interpersonal functioning at intake, the early unrepaired rupture pattern showed better outcomes. Findings suggest that early alliance development patterns affect treatment process and outcome. (PsycINFO Database Record


Setting the trajectory: racial disparities in newborn telomere length.
To explore racial differences in newborn telomere length (TL) and the effect moderation of the sex of the infant while establishing the methodology for the use of newborn blood spots for TL analyses. Pregnant mothers were recruited from the Greater New Orleans area. TL was determined via monochrome multiplex quantitative real-time polymerase chain reaction on DNA extracted from infant blood spots. Demographic data and other covariates were obtained via maternal report before the infant's birth. Birth outcome data were obtained from medical records and maternal report. Black infants weighed significantly less than white infants at birth and had significantly longer TL than white infants (P=.0134), with the strongest effect observed in black female infants. No significant differences in gestational age were present. Significant racial differences in TL were present at birth in this sample, even after we controlled for a range of birth outcomes and demographic factors. Because longer initial TL is predictive of more rapid TL attrition across the life course, these findings provide evidence that, even at birth, biological vulnerability to early life stress may differ by race and sex.


Redesigning the procaspase-8 dimer interface for improved dimerization.
Caspase-8 is a cysteine directed aspartate-specific protease that is activated at the cytosolic face of the cell membrane upon receptor ligation. A key step in the activation of caspase-8 depends on adaptor-induced dimerization of procaspase-8 monomers. Dimerization is followed by limited autoproteolysis within the intersubunit linker (IL), which separates the large and small subunits of the catalytic domain. Although cleavage of the IL stabilizes the dimer, the uncleaved procaspase-8 dimer is sufficiently active to initiate apoptosis, so dimerization of the zymogen is an important mechanism to control apoptosis. In contrast, the effector caspase-3 is a stable dimer under physiological conditions but exhibits little enzymatic activity. The catalytic domains of caspases are structurally similar, but it is not known why procaspase-8 is a monomer while procaspase-3 is a dimer. To define the role of the dimer interface in assembly and activation of procaspase-8, we generated mutants that mimic the dimer interface of effector caspases. We show that procaspase-8 with a mutated dimer interface more readily forms dimers. Time course studies of refolding also show that the mutations accelerate dimerization. Transfection of HEK293A cells with the procaspase-8 variants, however, did not result in a significant increase in apoptosis, indicating that other factors are required in vivo. Overall, we show that redesigning the interface of procaspase-8 to remove negative design elements results in increased dimerization and activity in vitro, but increased dimerization, by itself, is not sufficient for robust activation of apoptosis.


[Application of calixarenes for DNA transfection in cells].
First results of non-ionic and poly-cationic calixarenes utilization for gene transfection are presented and analyzed in this survey. State of the art in the field of scientific searching for new non-viral vectors are shown in the general form. The prospects of supramolecular concept in design agents for transfection are demonstrated. Some relationships between calixarene architecture and calixarene ability to promote gene transfection are revealed, namely: formation of supramolecular self-assembled aggregates at water media facilitates hierarchical formation of complexes with DNA molecules. Latter particles will effectively transfect genes if they are nano-sized and positively-charged.


Enhanced Uptake and Endosomal Release of LbL Microcarriers Functionalized with Reversible Fusion Proteins.
The efficient application of smart drug-delivery systems requires further improvement of their cellular uptake and in particular their release from endolysosomal compartments into the cytoplasm of target cells. The usage of virus proteins allows for such developments, as viruses have evolved efficient entry mechanisms into the cell, mediated by their fusion proteins. In our investigations, the transferability of the glycoprotein G which is a fusion protein of the vesicular stomatitis virus (VSV-G) onto the surface of a layer-by-layer (LbL) designed microcarrier was investigated. The assembly of VSV-G as a reversible viral fusion protein onto LbL microcarriers indeed induced an enhanced uptake rate on Vero cells as well as a fast and efficient release of the intact carriers from endolysosomes into the cytoplasm. Additionally, neither virus-associated effects on cellular viability nor activation of an interferon response were detected. Our study emphasizes the suitability of VSV-G as an efficient surface functionalization of drug-delivery systems.


Phosphorylation of serine 303 is a prerequisite for the stress-inducible SUMO modification of heat shock factor 1.
The heat shock response, which is accompanied by a rapid and robust upregulation of heat shock proteins (Hsps), is a highly conserved protection mechanism against protein-damaging stress. Hsp induction is mainly regulated at transcriptional level by stress-inducible heat shock factor 1 (HSF1). Upon activation, HSF1 trimerizes, binds to DNA, concentrates in the nuclear stress granules, and undergoes a marked multisite phosphorylation, which correlates with its transcriptional activity. In this study, we show that HSF1 is modified by SUMO-1 and SUMO-2 in a stress-inducible manner. Sumoylation is rapidly and transiently enhanced on lysine 298, located in the regulatory domain of HSF1, adjacent to several critical phosphorylation sites. Sumoylation analyses of HSF1 phosphorylation site mutants reveal that specifically the phosphorylation-deficient S303 mutant remains devoid of SUMO modification in vivo and the mutant mimicking phosphorylation of S303 promotes HSF1 sumoylation in vitro, indicating that S303 phosphorylation is required for K298 sumoylation. This finding is further supported by phosphopeptide mapping and analysis with S303/7 phosphospecific antibodies, which demonstrate that serine 303 is a target for strong heat-inducible phosphorylation, corresponding to the inducible HSF1 sumoylation. A transient phosphorylation-dependent colocalization of HSF1 and SUMO-1 in nuclear stress granules provides evidence for a strictly regulated subnuclear interplay between HSF1 and SUMO.


Wave propagation in a graded-index taper.
We present a model of a graded-index taper for which the field solutions can be obtained directly by separation of variables. Those fields tightly concentrated about the axis of the taper are given very accurately by remarkably simple expressions which clearly illustrate the influence of the tapering. Frequent comparisons with a geometrical optics analysis demonstrate the link between the ray and field approaches and also assist in the physical interpretation of several results. Examples of the application of these field solutions are also described.


Semi-commercial-scale production of Japanese B encephalitis virus vaccines from tissue culture.
This study was undertaken to modify and develop procedures for tissue culture-inactivated Japanese B encephalitis (JBE) virus vaccine production in large quantities. Various types of glass bottles were tried and, considering many advantages, long cylindrical roller (CR) bottles were selected. Several variables were investigated including number and volume of trypsinized cells to be seeded, volume of growth medium required for optimum cell growth, amount of calf serum, and volume of harvest medium for a high-titer virus yield. A good confluent cell sheet in CR bottles was obtained within a week by increasing the calf serum from 4 to 10% and when such tissue in a CR bottle was inoculated with 45,000 viral mean tissue culture infective doses directly into the medium, the cytopathological effects (CPE) appeared on day 5. High-titer virus yields were obtained when the harvests were made at 4(+) CPE using medium 199 with 2% human albumin at pH 8.3 to 8.5. No appreciable gain in titer was found from such harvests by blending to release intracellular virions. The production methods finally adopted gave consistently good results, and several inactivated JBE virus vaccine lots with minimum immunizing doses, ranging from 0.005 to 0.017 ml, were prepared using a large number of CR bottles in a simulated commercial-scale production system.


Glucose Improves the Efficacy of Photobiomodulation in Changing ATP and ROS Levels in Mouse Fibroblast Cell Cultures.
In this study, we tested the idea that photobiomodulation-the application of red to near infrared light (~λ = 600-1300 nm) to body tissues-is more effective in influencing cell metabolism when glucose is readily available. To this end, we used a mouse fibroblast (L-929) cell culture model and had two sets of conditions: non-stressed (10% FBS (foetal bovine serum)) and stressed (1% FBS), both either with or without glucose. We treated (or not) cells with photobiomodulation using an 810 nm laser at 15 mW/cm2 (~7.2 J/cm2). Our results showed that photobiomodulation was neither cytotoxic nor effective in enhancing measures of cell viability and proliferation, together with protein levels in any of the cell cultures. Photobiomodulation was, however, effective in increasing adenosine triphosphate (ATP) and decreasing reactive oxygen species (ROS) levels and this was-most importantly-only in conditions where glucose was present; corresponding cultures that did not contain glucose did not show these changes. In summary, we found that the benefits of photobiomodulation, in particular in changing ATP and ROS levels, were induced only when there was glucose available. Our findings lay a template for further explorations into the mechanisms of photobiomodulation, together with having considerable experimental and clinical implications.


Inhibition and excitation of primate spinothalamic tract neurons by stimulation in region of nucleus reticularis gigantocellularis.
1. Electrical stimulation in the region of the nucleus reticularis gigantocellularis (NGc) in anesthetized monkeys inhibited or excited spinothalamic tract neurons in the lumbosacral and cervical enlargements. 2. The descending effects were generally more pronounced for activity evoked by cutaneous A-delta-fibers than for activity produced by large myelinated cutaneous afferents. Nevertheless, the responses to all types of natural stimuli used could be inhibited or facilitated. 3. The excitation from repeated brief stimulus trains to the NGc sometimes increased progressively, suggesting the existence of a positive feedback system. Occasionally, repeated stimulation of the NGc produced a progressively greater inhibition. 4. The threshold stimulus strength to elicit the inhibitory and excitatory actions was usually less than 50 microA, and in some cases less than 25 microA. The inhibition and excitation increased as the stimulus intensity was raised above the threshold value, or as the number and/or frequency of pulses in the stimulus train was increased. 5. The strongest inhibition and excitation was produced by stimulation within the NGc on either side of the brain stem. There was no obvious topographic organization of inhibitory and excitatory zones. 6. Dorsolateral tractotomies in the high cervical spinal cord did not prevent the effects of NGc stimulation, indicating that the inhibitory and excitatory pathways descend in the ventral parts of the white matter. 7. It is suggested that the inhibition and excitation are mediated by the medullary reticulospinal system.


Large and small herbivores have strong effects on tundra vegetation in Scandinavia and Alaska.
Large and small mammalian herbivores are present in most vegetated areas in the Arctic and often have large impacts on plant community composition and ecosystem functioning. The relative importance of different herbivores and especially how their specific impact on the vegetation varies across the Arctic is however poorly understood.Here, we investigate how large and small herbivores influence vegetation density and plant community composition in four arctic vegetation types in Scandinavia and Alaska. We used a unique set of exclosures, excluding only large (reindeer and muskoxen) or all mammalian herbivores (also voles and lemmings) for at least 20 years.We found that mammalian herbivores in general decreased leaf area index, NDVI, and abundance of vascular plants in all four locations, even though the strength of the effect and which herbivore type caused these effects differed across locations. In three locations, herbivore presence caused contrasting plant communities, but not in the location with lowest productivity. Large herbivores had a negative effect on plant height, whereas small mammalian herbivores increased species diversity by decreasing dominance of the initially dominating plant species. Above- or belowground disturbances caused by herbivores were found to play an important role in shaping the vegetation in all locations.Synthesis: Based on these results, we conclude that both small and large mammalian herbivores influence vegetation in Scandinavia and Alaska in a similar way, some of which can mitigate effects of climate change. We also see important differences across locations, but these depend rather on local herbivore and plant community composition than large biogeographical differences among continents.


Drug-Induced Liver Injury during Antidepressant Treatment: Results of AMSP, a Drug Surveillance Program.
Drug-induced liver injury is a common cause of liver damage and the most frequent reason for withdrawal of a drug in the United States. The symptoms of drug-induced liver damage are extremely diverse, with some patients remaining asymptomatic. This observational study is based on data of Arzneimittelsicherheit in der Psychiatrie, a multicenter drug surveillance program in German-speaking countries (Austria, Germany, and Switzerland) recording severe drug reactions in psychiatric inpatients. Of 184234 psychiatric inpatients treated with antidepressants between 1993 and 2011 in 80 psychiatric hospitals, 149 cases of drug-induced liver injury (0.08%) were reported. The study revealed that incidence rates of drug-induced liver injury were highest during treatment with mianserine (0.36%), agomelatine (0.33%), and clomipramine (0.23%). The lowest probability of drug-induced liver injury occurred during treatment with selective serotonin reuptake inhibitors ([0.03%), especially escitalopram [0.01%], citalopram [0.02%], and fluoxetine [0.02%]). The most common clinical symptoms were nausea, fatigue, loss of appetite, and abdominal pain. In contrast to previous findings, the dosage at the timepoint when DILI occurred was higher in 7 of 9 substances than the median overall dosage. Regarding liver enzymes, duloxetine and clomipramine were associated with increased glutamat-pyruvat-transaminase and glutamat-oxalat-transaminase values, while mirtazapine hardly increased enzyme values. By contrast, duloxetine performed best in terms of gamma-glutamyl-transferase values, and trimipramine, clomipramine, and venlafaxine performed worst. Our findings suggest that selective serotonin reuptake inhibitors are less likely than the other antidepressants, examined in this study, to precipitate drug-induced liver injury, especially in patients with preknown liver dysfunction.


Regulatory Review Duration and Differences in Submission Times of Drugs in the United States and Europe, 2011 to 2020.
The speed of drug regulatory agencies in the United States and Europe is often a source of discussion. The objective of this research was to assess regulatory review duration of first and supplementary indications approved between 2011 and 2020 in the United States and Europe (European Union [EU] and Switzerland) and differences in submission times between the United States and Europe. Descriptive statistics were applied to review times between the jurisdictions and across the therapeutic areas. A regression analysis was done to estimate the association between approval agency and review times. The primary analysis cohort included 241 drugs approved in the United States, the EU, and Switzerland. Of these, 128 drugs had supplemental indications (331 in total) in the United States and 87 had supplemental indications (206 in total) in the EU. Overall median review duration from submission to approval subtracting the clock stop period was 39 weeks in the United States, 44 weeks in the EU, and 44 weeks in Switzerland. When review times within each drug were compared, the European Medicines Agency took a median of 3.7 weeks (IQR, -6.7 to 14.9 weeks) longer than the U.S. Food and Drug Administration and Swissmedic a median of 0.3 weeks (IQR, -10.6 to 15.3 weeks) longer. Median total review duration for supplemental indications was 26 weeks in the United States and 40 weeks in the EU. Applications were submitted a median of 1.3 and 17.9 weeks later in the EU and Switzerland, respectively, than in the United States. The regression analysis showed small differences in submission times between the United States and the EU (-2.1 weeks [95% CI, -11.7 to 7.6 weeks]) and larger differences between the United States and Switzerland (33.0 weeks [CI, 23.1 to 42.8 weeks]). It would be beneficial for patients if differences in submission times between the United States and Europe continue to be minimized.


Effect of different ascent profiles on performance at 4,200 m elevation.
Two groups of sea level residents were studied at the summit of Mauna Kea (4,200 m elevation) following ascent by vehicle. "Commuters" spent 6 h at the summit, while "shiftworkers" lived on the mountain for 5 d. Although PaO2 levels were lower in commuters, they experienced fewer altitude sickness symptoms than shiftworkers on the first day at 4,200 m. After 5 d, shiftworkers reported fewer symptoms and performed better at tests of numerate memory and psychomotor ability than commuters. At high altitude, pulse rates were increased in both groups, but only shiftworkers exhibited an elevation in systemic blood pressure. Arterial-alveolar oxygen tension gradients were not increased at 4,200 m. Despite frequent and rapid ascents and descents, with minimal provision for acclimatization, high altitude pulmonary and cerebral oedema were uncommon.


Role of A-Kinase Anchoring Protein 1 in Retinal Ganglion Cells: Neurodegeneration and Neuroprotection.
A-Kinase anchoring protein 1 (AKAP1) is a multifunctional mitochondrial scaffold protein that regulates mitochondrial dynamics, bioenergetics, and calcium homeostasis by anchoring several proteins, including protein kinase A, to the outer mitochondrial membrane. Glaucoma is a complex, multifactorial disease characterized by a slow and progressive degeneration of the optic nerve and retinal ganglion cells (RGCs), ultimately resulting in vision loss. Impairment of the mitochondrial network and function is linked to glaucomatous neurodegeneration. Loss of AKAP1 induces dynamin-related protein 1 dephosphorylation-mediated mitochondrial fragmentation and loss of RGCs. Elevated intraocular pressure triggers a significant reduction in AKAP1 protein expression in the glaucomatous retina. Amplification of AKAP1 expression protects RGCs from oxidative stress. Hence, modulation of AKAP1 could be considered a potential therapeutic target for neuroprotective intervention in glaucoma and other mitochondria-associated optic neuropathies. This review covers the current research on the role of AKAP1 in the maintenance of mitochondrial dynamics, bioenergetics, and mitophagy in RGCs and provides a scientific basis to identify and develop new therapeutic strategies that could protect RGCs and their axons in glaucoma.


Relapsing polychondritis: a chameleon among orphan diseases.
Relapsing polychondritis (RPC) is a rare disease with recurrent episodes of inflammation of cartilage tissue leading to fibrosis and organ damage. Despite unknown etiology, there is some evidence of a genetic predisposition. The clinical presentation is heterogeneous and an association with other autoimmune disorders such as rheumatoid arthritis or different forms of vasculitis has been described. All organ systems containing cartilage can be affected, such as ear, nose, joints, trachea, aorta, and coronary arteries. Given the broad spectrum of potential manifestations, a variety of medical specialists may be involved in the management of RPC patients. As establishing the diagnosis of RPC may be difficult, an interdisciplinary approach may be preferable. Treatment options include glucocorticoids, dapsone, disease-modifying antirheumatic drugs, and biologics. Prognosis is as heterogeneous as the clinical picture, depending on the severity of organ damage. In this paper we give an overview of the current knowledge with regard to pathogenesis, clinical picture, diagnosis, and therapy of RPC.


Adenomyomatous hyperplasia of the human gallbladder.
Adenomyomatous hyperplasia of the human gallbladder, an infrequently observed alteration, has a controversial pathogenesis and clinical significance. In the nine gallbladders and patient profiles discussed, the morphologic alterations and associated clinical phenomena are compared to other mucosal-stromal reactions of the gastrointestinal tract. It is suggested that adenomyomatous hyperplasia represents a reaction to injury that occurs after an initial structural degenerative process and is followed by a proliferative repair.


The link between chronic thromboembolic pulmonary hypertension and sarcoidosis: association or visual masquerade?
Chronic thromboembolic pulmonary hypertension (CTEPH) and sarcoidosis are recognized causes of pulmonary hypertension according to the World Health Organization classification scheme. This case series describes seven patients with sarcoidosis with a mean age of 61 who developed pulmonary hypertension. They were found to have CTEPH, diagnosed by either CT pulmonary angiography or a lung ventilation perfusion scan. They all underwent confirmatory right heart catheterization showing elevated mean pulmonary artery pressures (mean of 42 mmHg - normal less than 25 mmHg). Sarcoidosis has been previously shown to be associated with increased rates of venous thromboembolic disease. In these cases, patients with sarcoidosis later developed CTEPH and this may be another mechanism in which sarcoidosis can lead to pulmonary hypertension. (Sarcoidosis Vasc Diffuse Lung Dis 2017; 34: 352-355).


Composition, physicochemical properties of pea protein and its application in functional foods.
Field pea is one of the most important leguminous crops over the world. Pea protein is a relatively new type of plant proteins and has been used as a functional ingredient in global food industry. Pea protein includes four major classes (globulin, albumin, prolamin, and glutelin), in which globulin and albumin are major storage proteins in pea seeds. Globulin is soluble in salt solutions and can be further classified into legumin and vicilin. Albumin is soluble in water and regarded as metabolic and enzymatic proteins with cytosolic functions. Pea protein has a well-balanced amino acid profile with high level of lysine. The composition and structure of pea protein, as well as the processing conditions, significantly affect its physical and chemical properties, such as hydration, rheological characteristics, and surface characteristics. With its availability, low cost, nutritional values and health benefits, pea protein can be used as a novel and effective alternative to substitute for soybean or animal proteins in functional food applications.


Using Approximate Bayesian Computation to infer sex ratios from acoustic data.
Population sex ratios are of high ecological relevance, but are challenging to determine in species lacking conspicuous external cues indicating their sex. Acoustic sexing is an option if vocalizations differ between sexes, but is precluded by overlapping distributions of the values of male and female vocalizations in many species. A method allowing the inference of sex ratios despite such an overlap will therefore greatly increase the information extractable from acoustic data. To meet this demand, we developed a novel approach using Approximate Bayesian Computation (ABC) to infer the sex ratio of populations from acoustic data. Additionally, parameters characterizing the male and female distribution of acoustic values (mean and standard deviation) are inferred. This information is then used to probabilistically assign a sex to a single acoustic signal. We furthermore develop a simpler means of sex ratio estimation based on the exclusion of calls from the overlap zone. Applying our methods to simulated data demonstrates that sex ratio and acoustic parameter characteristics of males and females are reliably inferred by the ABC approach. Applying both the ABC and the exclusion method to empirical datasets (echolocation calls recorded in colonies of lesser horseshoe bats, Rhinolophus hipposideros) provides similar sex ratios as molecular sexing. Our methods aim to facilitate evidence-based conservation, and to benefit scientists investigating ecological or conservation questions related to sex- or group specific behaviour across a wide range of organisms emitting acoustic signals. The developed methodology is non-invasive, low-cost and time-efficient, thus allowing the study of many sites and individuals. We provide an R-script for the easy application of the method and discuss potential future extensions and fields of applications. The script can be easily adapted to account for numerous biological systems by adjusting the type and number of groups to be distinguished (e.g. age, social rank, cryptic species) and the acoustic parameters investigated.


Improving cellular adhesion on scaffolds for transplantation: synthesising a poly(MMA-co-PEGM) network.
Electrospun fibrous matrices prepared from methacrylate-based copolymers are investigated as a tool for retinal pigment epithelium (RPE) transplantation in the treatment of degenerative retinal diseases. Human RPE cells were used to probe the cell-surface interactions on these copolymer matrices. For the first time, simple changes in chemical functionality have been found to induce gel formation of these methacrylate backbone copolymers in vitro. This effect is shown to significantly improve RPE cell adhesion and survival.


Mapping of QTL for chicken body weight, carcass composition, and meat quality traits in a slow-growing line.
Slow-growing chicken lines are valuable genetic resources for the development of well-perceived alternative free-range production. While there is no constraint on increasing growth rate, breeding programs have to evolve in order to include new traits improving the positioning of such lines in the growing market for parts and processed products. In this study, we used dense genotyping to fine map QTL for chicken growth, body composition, and meat quality traits in view of developing new tools for selection of a slow-growing line. The dataset included a total of 836 birds (10 sires, 87 dams, 739 descendants) and 40,203 SNP. QTL for the 15 traits analyzed were detected by 3 different methods, i.e., linkage and linkage disequilibrium haplotype-based analysis (LDLA), family-based single marker association (FASTA), and Bayesian multi-marker regression (Bayes Cπ). After filtering for QTL redundancy, we found 16, 16, and 9 QTL when using the FASTA, LDLA, and Bayes Cπ methods, respectively, with a threshold of 2.49 × 10-5 for FASTA and LDLA, and a Bayes factor of 150 for the Bayes Cπ analysis. They comprised 17 QTL for body weight, 9 QTL for body composition, and 15 QTL for breast meat quality or behavior at slaughter. The 3 methods agreed in the detection of highly significant QTL such as that detected on GGA24 for body weight at 3, 6, and 9 wk, and the 2 QTL detected on GGA17 and GGA18 for breast meat yield. Several significant QTL were also detected for the different components of breast meat quality. This study provided new locations for investigation in order to improve our understanding of the genetic architecture of growth, carcass composition, and meat quality in the chicken and to develop molecular tools for the selection of these traits in a slow-growing line.


Method for evaluating drip-rate accuracy of intravenous flow-regulating devices.
The effects of patient movement and position on the drip-rate accuracy of several i.v. flow-regulating devices were investigated. Intravenous infusion sites were established in 20 healthy adult volunteers. All the subjects received 5% dextrose injection through the same type of i.v. tubing from 500-mL bags hung from standard i.v. fluid poles. The flow-regulating devices tested were the IVAC 280, which served as the control device; a roller clamp; the Dial-A-Flo; the Exacdrop; and the 3M IV Flow Regulator. Drip rates were present at 40 drops/min and were measured before and after the subjects moved among the supine, sitting, and standing positions and walking. The drip rate was reset to 40 drops/min after each position change. Changing position from supine to sitting did not affect mean drip rates for the IVAC 280 and 3M IV Flow Regulator devices but significantly decreased the rates for the roller clamp, Dial-A-Flo, and Exacdrop. The change from sitting to standing did not affect the IVAC 280 and 3M IV Flow Regulator drip rates but significantly decreased the rates for the other devices. None of the rates was dramatically affected when the subjects went from standing to walking, although the effect achieved significance for the roller clamp and Exacdrop devices. The change from walking to the supine position did not affect the drip rates for the IVAC 280 and 3M IV Flow Regulator but significantly increased the rates for the other devices. The drip-rate accuracy of the roller clamp, Dial-A-Flo, and Exacdrop devices was significantly affected when subjects changed positions.(ABSTRACT TRUNCATED AT 250 WORDS)


[50 Years ago: First Successful Treatment of Acute Allograft Rejection after a Human Heart Transplantation].
The first clinical use of the "Munich antilymphocyte globulin" (ALG) at the occasion of the first successful human heart transplantation is briefly described. The cardiac transplantation was carried out by Christiaan Barnard and his team in Cape Town, South Africa, in 1968. The patient developed an acute allograft rejection which could be successfully reversed within three weeks using the intravenous administration of ALG. This event can be regarded as the beginning of a success story of ALG in its use as a powerful immunosuppressive agent in all categories of clinical organ transplantation.


Autocrine regulation of mda-7/IL-24 mediates cancer-specific apoptosis.
A noteworthy aspect of melanoma differentiation-associated gene-7/interleukin-24 (mda-7/IL-24) as a cancer therapeutic is its ability to selectively kill cancer cells without harming normal cells. Intracellular MDA-7/IL-24 protein, generated from an adenovirus expressing mda-7/IL-24 (Ad.mda-7), induces cancer-specific apoptosis by inducing an endoplasmic reticulum (ER) stress response. Secreted MDA-7/IL-24 protein, generated from cells infected with Ad.mda-7, induces growth inhibition and apoptosis in surrounding noninfected cancer cells but not in normal cells, thus exerting an anti-tumor "bystander" effect. The present studies reveal a provocative finding that recombinant MDA-7/IL-24 protein can robustly induce expression of endogenous mda-7/IL-24, which generates the signaling events necessary for bystander killing. To evaluate the mechanism underlying this positive autocrine feedback loop, we show that MDA-7/IL-24 protein induces stabilization of its own mRNA without activating its promoter. Furthermore, this posttranscriptional effect depends on de novo protein synthesis. As a consequence of this autocrine feedback loop MDA-7/IL-24 protein induces sustained ER stress as evidenced by expression of ER stress markers (BiP/GRP78, GRP94, GADD153, and phospho-eIF2alpha) and reactive oxygen species production, indicating that both intracellular and secreted proteins activate similar signaling pathways to induce apoptosis. Thus, our results clarify the molecular mechanism by which secreted MDA-7/IL-24 protein (generated from Ad.mda-7-infected cells) exerts cancer-specific killing.


Wnt signaling pathway in mammary gland development and carcinogenesis.
The signaling pathway mediated by Wingless-type (Wnt) proteins is highly conserved in evolution. This pivotal pathway is known to regulate cell fate decisions, cell proliferation, morphology, migration, apoptosis, differentiation and stem cell self-renewal. It currently includes the canonical or Wnt/beta-catenin pathway in which Wnt proteins bind to 'frizzled' receptors, which leads to downstream activation of gene transcription by beta-catenin. Second, the noncanonical or beta-catenin-independent pathways are now known to be mediated by three possible mechanisms: (1) the Wnt/Ca(2+) pathway, (2) the Wnt/G protein signaling pathway, and (3) the Wnt/PCP or planar cell polarity pathway. Wnt signaling is implicated at several stages of mammary gland growth and differentiation, and possibly in the involution of mammary gland following lactation. Recent evidence suggests the role of Wnt signaling in human breast cancer involves elevated levels of nuclear and/or cytoplasmic beta-catenin using immunohistochemistry, overexpression or downregulation of specific Wnt proteins, overexpression of CKII and sFRP4, downregulation of WIF-1 and sFRP1, as well as amplification of DVL-1. Further research is required to determine how Wnt signaling is involved in the development of different histological types of breast cancer and whether it promotes the viability of cancer stem cells or not.


Molecular cloning and characterization of a novel sequence, vof-16, with enhanced expression in permanent ischemic rat brain.
We reported previously that chronic hypoperfusion induced by permanent occlusion of the bilateral common carotid arteries (2VO) in rats caused progressive cognitive deficits and neuronal damage in the hippocampus and the white matter. These changes are similar to those observed in human dementia. Reverse transcription-polymerase chain reaction (RT-PCR) differential display was carried out to identify mRNAs encoding the intrinsic factors involved in permanent ischemia from the 2VO rat brain. Over 20 clones which showed different expression levels in 2VO and sham-operated rats were isolated. One of these, named vof-16, was markedly enhanced the expression by 2VO. The whole sequence of vof-16 mRNA was 2098 nt. The distribution of vof-16 transcripts was examined by RT-PCR and in situ hybridization. The results revealed that vof-16 was abundant in the hippocampus, the tenia tecta, the piriform cortex and the area around the aorta. The expression levels of vof-16 in 2VO and sham-operated rat hippocampus were determined by a quantitative PCR method. The expression was abundant in the hippocampus of rats with cognitive impairment induced by 2VO. In contrast, the expression levels of vof-16 were lower in the 2VO rats with no impairment and in sham-operated rats. These results suggest that the expression levels of vof-16 may be related to the cognitive impairment induced by chronic ischemia after 2VO.


Longitudinal child-oriented dietary intervention: Association with parental diet and cardio-metabolic risk factors. The Special Turku Coronary Risk Factor Intervention Project.
Background The child-oriented dietary intervention given in the prospective Special Turku Coronary Risk Factor Intervention Project (STRIP) has decreased the intake of saturated fat and lowered serum cholesterol concentration in children from infancy until early adulthood. In this study, we investigated whether the uniquely long-term child-oriented intervention has affected also secondarily parental diet and cardio-metabolic risk factors. Methods The STRIP study is a longitudinal, randomized infancy-onset atherosclerosis prevention trial continued from the child's age of 8 months to 20 years. The main aim was to modify the child's diet towards reduced intake of saturated fat. Parental dietary intake assessed by a one-day food record and cardio-metabolic risk factors were analysed between the child's ages of 9-19 years. Results Saturated fat intake of parents in the intervention group was lower [mothers: 12.0 versus 13.9 daily energy (E%), p < 0.0001; fathers: 12.5 versus 13.9 E%, p < 0.0001] and polyunsaturated fat intake was higher (mothers: 6.1 versus 5.4 E%, p < 0.0001; fathers: 6.3 versus 5.9 E%, p = 0.0003) compared with the control parents. Maternal total and low-density lipoprotein cholesterol concentrations were lower in the intervention compared with the control group (mean ± SE 5.02 ± 0.04 versus 5.14 ± 0.04 mmol/l, p = 0.04 and 3.19 ± 0.04 versus 3.30 ± 0.03 mmol/l, p = 0.03, respectively). Paternal cholesterol values did not differ between the intervention and control groups. Other cardio-metabolic risk factors were similar in the study groups. Conclusions Child-oriented dietary intervention shifted the dietary fat intakes of parents closer to the recommendations and tended to decrease total and low-density lipoprotein cholesterol in the intervention mothers. Dietary intervention directed to children benefits also parents.


Nurse practitioners' and certified nurse midwives' experiences providing comprehensive early abortion care in New England, USA.
Access to abortion is linked to better maternal health and is essential to advancing women's economic and social equality. Globally, nurses and midwives have been identified as key for increasing access to this service. In this qualitative study we describe the experiences of nurse practitioners and nurse midwives who provide comprehensive early abortion care in the New England region of the United States. In contrast to much of the existing literature from around the world on nurses who deliver abortion care, we found participants' experiences providing comprehensive early abortion was generally positive. The study has implications for theory development, research, and policy.


Human embryonic stem cells express elevated levels of multiple pro-apoptotic BCL-2 family members.
Two of the greatest challenges in regenerative medicine today remain (1) the ability to culture human embryonic stem cells (hESCs) at a scale sufficient to satisfy clinical demand and (2) the ability to eliminate teratoma-forming cells from preparations of cells with clinically desirable phenotypes. Understanding the pathways governing apoptosis in hESCs may provide a means to address these issues. Limiting apoptosis could aid scaling efforts, whereas triggering selective apoptosis in hESCs could eliminate unwanted teratoma-forming cells. We focus here on the BCL-2 family of proteins, which regulate mitochondrial-dependent apoptosis. We used quantitative PCR to compare the steady-state expression profile of all human BCL-2 family members in hESCs with that of human primary cells from various origins and two cancer lines. Our findings indicate that hESCs express elevated levels of the pro-apoptotic BH3-only BCL-2 family members NOXA, BIK, BIM, BMF and PUMA when compared with differentiated cells and cancer cells. However, compensatory expression of pro-survival BCL-2 family members in hESCs was not observed, suggesting a possible explanation for the elevated rates of apoptosis observed in proliferating hESC cultures, as well as a mechanism that could be exploited to limit hESC-derived neoplasms.


Expression vectors and gene transfer.
The development of DNA cloning techniques, together with the possibility of reintroducing cloned DNA fragments into the genome of a living organism, has led to an extraordinary growth of our knowledge in molecular biology over the past twenty years. In the present paper a brief overview of the vectors and techniques used in transforming different groups of organisms is given. The importance and applications of genetic engineering for each group (yeasts, plants, Drosophila and mammals) will be discussed.