Comparing metabolic functionalities, community structures, and dynamics of herbicide-degrading communities cultivated with different substrate concentrations.
Two 4-chloro-2-methylphenoxyacetic acid (MCPA)-degrading enrichment cultures selected from an aquifer on low (0.1 mg liter(-1)) or high (25 mg liter(-1)) MCPA concentrations were compared in terms of metabolic activity, community composition, population growth, and single cell physiology. Different community compositions and major shifts in community structure following exposure to different MCPA concentrations were observed using both 16S rRNA gene denaturing gradient gel electrophoresis fingerprinting and pyrosequencing. The communities also differed in their MCPA-mineralizing activities. The enrichments selected on low concentrations mineralized MCPA with shorter lag phases than those selected on high concentrations. Flow cytometry measurements revealed that mineralization led to cell growth. The presence of low-nucleic acid-content bacteria (LNA bacteria) was correlated with mineralization activity in cultures selected on low herbicide concentrations. This suggests that LNA bacteria may play a role in degradation of low herbicide concentrations in aquifers impacted by agriculture. This study shows that subpopulations of herbicide-degrading bacteria that are adapted to different pesticide concentrations can coexist in the same environment and that using a low herbicide concentration enables enrichment of apparently oligotrophic subpopulations.


Role of Hysterosalpingography in Diagnosing Tubal Blockage - A Prospective Diagnostic Study.
Women with abnormal hysterosalpingography (HSG) are anxious regarding the presence of tubal pathology. It is important to know the predictive value of HSG and the need for subsequent laparoscopy following an abnormal report. In the era of assisted reproductive technology, the role of invasive testing such as diagnostic laparoscopy is being increasingly questioned due to its invasiveness and associated risks. There is a need to explore the positive predictive value (PPV) of HSG in detecting bilateral tubal block in our population as PPV changes with the prevalence of disease. The aim of this study was to evaluate the diagnostic accuracy of HSG in identifying tubal blockage in subfertile women. This was a prospective diagnostic study conducted in the department of reproductive medicine and surgery in a university-level hospital. The study included 199 subfertile women who had undergone HSG earlier and were planned for laparoscopy from April 2017 to January 2021. Findings of HSG and laparoscopy were compared with HSG as index test and laparoscopy as reference test, and the outcomes analysed were PPV of HSG for a bilateral tubal block, bilateral hydrosalpinx, abnormal HSG (unilateral or bilateral tubal block) and agreement between HSG and diagnostic laparoscopy in detecting normal and abnormal findings. Kappa statistics, Stuart-Maxwell tests of marginal homogeneity and prevalence-adjusted bias-adjusted kappa (PABAK) statistics were used. The PPV for a bilateral block with HSG was 20.9% (95% CI: 13.7-29.7). The PPV of HSG for bilateral hydrosalpinx was 50.0% (95% CI: 6.8-93.2). PABAK was estimated to be 0.42 (95% CI: 0.30-0.55), suggestive of moderate agreement between the tests. Findings of laparoscopy in women with at least one patent tube in HSG showed that in 12.3% of cases, the management was likely to change due to the operative findings. The current study showed low PPV for bilateral tubal block diagnosed with HSG which translates into a need for further confirmation by laparoscopy. In one out of every eight women with at least one patent tube on HSG, performing laparoscopy changed the management.


Expression of heart oxytocin receptor and its mRNA in two rat strains with different activity of HPA axis.
Oxytocin (OT) is a neuropeptide acting both as a peripheral hormone and in the brain as neurotransmitter and neuromodulator. In addition to its well-known effects on milk-ejection and uterine contraction, OT was shown to exert neuroendocrine regulation of heart functions. The aim of this study was to investigate the expression of mRNA of OT receptors (OTR) in rat hearts by real-time quantitative PCR (qPCR). The study was performed in Sprague-Dawley (SD) and Lewis (LE) rat strains, the latter having lower activity of HPA axis. We used adult male SD and LE rats. OTR mRNA expression was detected in all heart chambers by comparing their threshold cycle values (CT) to CT of reference gene β-actin. The relative expression ratios were calculated using the 2-ΔΔCT method. The specificity of reaction of primary antibody with OTRs was tested by Western Blot and localization of OTR in the heart compartments was performed by immunofluorescence with commercial OTR specific antibodies. We found expression of OTR mRNA in all heart compartments. The expression of OTR mRNA in both atria (LA, RA) was much higher than in the ventricles (RV, LV). By using two-way ANOVA we found no statistical differences between corresponding compartments of SD and LE rats. Immunohistochemical studies showed that OTR staining is not related to neuronal tissue and findings from left atrium indicate that prevalent localization of OTR is on cell membranes of cardiomyocytes. The finding of expression of OTR mRNA by real-time qPCR and proof of OTR staining by immunohistochemistry in all heart compartments indicate that OT and its receptors may have function as a cardiovascular hormone. The differences in the HPA axis activity, as is exemplified in Sprague-Dawley and Lewis rat strain, do not project in the expression of OTR mRNA under basal condition. The effect of activity of HPA on OTR expression should be studied under stimulated conditions as it was performed in the behavioral studies.


United States Guild Certified Feldenkrais Teachers®: a survey of characteristics and practice patterns.
Feldenkrais Method® teachers help students improve function and quality of life through verbally and manually guided lessons. The reasons people seek Feldenkrais® lessons are poorly understood. Similarly, little is known about practice characteristics and patterns. To address these knowledge gaps, we conducted an extensive survey of United States Guild Certified Feldenkrais Teachers®. We invited all Feldenkrais Teachers to participate in this survey delivered in web-based or print formats. We obtained overall and question-specific response rates, descriptive statistics, chi-square tests of response bias, and performed qualitative thematic review of comments. Overall response rate was 30.5% (392/1287). Ninety percent of responders had college degrees in diverse fields; 12.5% had credentials outside health care, 36.9% held conventional health care licenses, and 23.1% had complementary and alternative medicine credentials. Mean age was 55.7 years; most teachers were women (83%). California (n = 100) and New York (n = 34) had the most teachers. Forty-five percent of teachers earned ≤ 20% of their gross income from their practices, while 26% earned > 80%. Most saw < 10 students/week for individual lessons and < 10 students/week for group lessons. Students were mostly women (71.1%) and 45-64 years old. The primary reason students sought Feldenkrais lessons was pain. A quarter of students self-referred, a fifth were referred by conventional health care providers, and two-thirds paid for services directly. Themes from comments included: beliefs that Feldenkrais training had important personal and professional benefits for teachers; recognition of the challenges of operating small businesses and succinctly describing the Feldenkrais Method; the variety of practice approaches; and a deep commitment to the Feldenkrais Method. Most Feldenkrais Teachers were well educated, often held additional credentials, were located in the West, were women, were older than 50 years, and had part-time practices. Most students were women, were adults, came from various referral sources, and paid directly for services. Teachers and students utilized the Feldenkrais Method in diverse settings and applications. These findings may foster practice development by Feldenkrais Teachers, improve communication between health care consumers and providers and assist decision-making, and stimulate more research concerning the Feldenkrais Method.


Comparison of unfractioned and low molecular weight heparin for prophylaxis of coagulopathies in 52 horses with colic: a randomised double-blind clinical trial.
Unfractioned heparin (UFH) is widely used for prophylaxis of coagulation disorders, especially in colic-affected horses. However, it is accompanied by certain side effects. To compare the efficacy and side effects of unfractioned and low molecular weight heparin (LMWH) in horses with colic. The study was carried out as a randomised, double-blind, controlled clinical trial. Fifty-two horses with colic were treated subcutaneously with either UFH (heparin calcium, 150 iu/kg bwt initially, followed by 125 iu/kg bwt q. 12 h for 3 days and then 100 iu/kg bwt q. 12 h) or LMWH (dalteparin, 50 iu/kg bwt q. 24 h). All horses underwent daily physical examination including assessment of jugular veins, local reaction to heparin injections, haematological evaluation and coagulation profiles over up to 9 days. The type of heparin used did not affect the general behaviour and condition. There were significantly more jugular vein changes in horses treated with UFH. Packed cell volume decreased significantly within the first few days of UFH treatment, but did not change significantly in horses treated with LMWH. Activated partial thromboplastin time (aPTT) and thrombin time (TT) were prolonged in horses treated with UFH but not in those treated with LMWH. It was concluded that, in comparison to UFH, LMWH has markedly fewer side effects in horses. Therefore, LMWH is recommended for prophylaxis of coagulation disorders in colic patients.


In vivo electroporation of DNA into the wing epidermis of the butterfly, Bicyclus anynana.
The direct transfer of genes into differentiated insect tissues is a useful method of determining gene function because it circumvents the need to perform germ line transformations and of having information on tissue-specific gene promoters. Here in vivo gene delivery is achieved through electroporation of a reporter gene into the pupal forewing of the butterfly Bicyclus anynana (Butler) (Lepidoptera: Nymphalidae). Plasmids containing the coding sequence for enhanced green fluorescent protein (EGFP), driven by the Drosophila heat-shock promoter hsp70, were successfully expressed in epidermal cells after plasmid injection followed by electroporation and heat shock. EGFP expression was restricted to the vicinity of the injection and electroporation site, but the number of transformed cells varied from a few to over 5000 cells. Electroporation parameters were optimized in order to maximize the number of transformed cells while minimizing the extent of damage to the adult wing. While certain electrical parameters were well tolerated by the wing tissue, the physical damage caused by the insertion of the tungsten electrodes led to frequent disruptions of the adult wing pattern around the puncture sites. While this technique can be useful to test the correct expression of marker genes (such as EGFP) in newly build plasmids immediately following their injection, its potential use in testing the function of candidate genes in wing pattern formation is limited.


The impact of lay health advisors on cardiovascular health promotion: using a community-based participatory approach.
Overweight and obesity, lack of exercise, and exposure to tobacco smoke are clearly identified behavioral risk factors for cardiovascular disease. These problems disproportionately affect some communities. To address these problems within one community of underserved Latinos, participatory research methods were used to design an outreach program through a collaborative partnership between UCLA School of Nursing, Los Angeles County Department of Health Services, and members of the target community. Latina lay health advisors (LHAs) (n = 12) from the community were recruited and trained to teach 3 classes on healthy nutrition, physical activity, and maintaining smoke-free environments. Classes were offered in Spanish to adult Latinos recruited through the LHAs' social networks. A questionnaire on lifestyle behaviors was completed at baseline and 1 month following the last class. Exploratory open-ended questions on successes and challenges of applying new knowledge were also asked at follow-up. Results of paired t-tests on 256 participants showed significant increases in scores from baseline to follow-up in overall lifestyle behaviors and all 3 behavioral subsets (nutrition, physical activity, smoke-free behavior). Qualitative findings are also presented. Findings support utilizing LHAs as a feasible and effective healthcare delivery strategy for cardiovascular community health promotion, especially among immigrant populations.


Preconception use of cART by HIV-positive pregnant women increases the risk of infants being born small for gestational age.
The benefits of combination anti-retroviral therapy (cART) in HIV-positive pregnant women (improved maternal health and prevention of mother to child transmission [pMTCT]) currently outweigh the adverse effects due to cART. As the variety of cART increases, however, the question arises as to which type of cART is safest for pregnant women and women of childbearing age. We studied the effect of timing and exposure to different classes of cART on adverse birth outcomes in a large HIV cohort in the Netherlands. We included singleton HEU infants registered in the ATHENA cohort from 1997 to 2015. Multivariate logistic regression analysis for single and multiple pregnancies was used to evaluate predictors of small for gestational age (SGA, birth weight <10th percentile for gestational age), low birth weight and preterm delivery. A total of 1392 children born to 1022 mothers were included. Of these, 331 (23.8%) children were SGA. Women starting cART before conception had an increased risk of having a SGA infant compared to women starting cART after conception (OR 1.35, 95% CI 1.03-1.77, p = 0.03). The risk for SGA was highest in women who started a protease inhibitor-(PI) based regimen prior to pregnancy, compared with women who initiated PI-based cART during pregnancy. While the association of preterm delivery and preconception cART was significant in univariate analysis, on multivariate analysis only a non-significant trend was observed (OR 1.39, 95% CI 0.94-1.92, p = 0.06) in women who had started cART before compared to after conception. In multivariate analysis, the risk of low birth weight (OR 1.34, 95% CI 0.94-1.92, p = 0.11) was not significantly increased in women who had started cART prior to conception compared to after conception. In our cohort of pregnant HIV-positive women, the use of cART prior to conception, most notably a PI-based regimen, was associated with intrauterine growth restriction resulting in SGA. Data showed a non-significant trend in the risk of PTD associated with preconception use of cART compared to its use after conception. More studies are needed with regard to the mechanisms taking place in the placenta during fetal growth in pregnant HIV-positive women using cART. It will only be with this knowledge that we can begin to understand the potential impact of HIV and cART on the fetus, in order to be able to determine the optimal individualised drug regimen for HIV-infected women of childbearing age.


Lectin affinity capillary electrophoresis in glycoform analysis applying the partial filling technique.
The study of protein glycosylation and its significance in biological interactions is a field of growing interest. This work demonstrates a lectin-based separation of protein glycoforms of alpha1-acid glycoprotein (AGP or orosomucoid) with capillary electrophoresis. Glycoform analysis was performed with a "partial filling technique" with the lectin Concanavalin A (Con A) as affinity ligand. Con A separated human AGP into two peaks; the first peak included AGP glycoforms without biantennary glycans, and the second peak represented the fraction that had one or more biantennary glycans. The applicability of the method was demonstrated with the analysis of AGP from clinical samples and AGP treated with N-glycosidase F. The AGP separation was also used as a reporter system to estimate the dissociation constant (KD) between Con A and a competing sugar.


Selective depletion of macrophages reveals distinct, opposing roles during liver injury and repair.
Macrophages perform both injury-inducing and repair-promoting tasks in different models of inflammation, leading to a model of macrophage function in which distinct patterns of activation have been proposed. We investigated macrophage function mechanistically in a reversible model of liver injury in which the injury and recovery phases are distinct. Carbon tetrachloride---induced liver fibrosis revealed scar-associated macrophages that persisted throughout recovery. A transgenic mouse (CD11b-DTR) was generated in which macrophages could be selectively depleted. Macrophage depletion when liver fibrosis was advanced resulted in reduced scarring and fewer myofibroblasts. Macrophage depletion during recovery, by contrast, led to a failure of matrix degradation. These data provide the first clear evidence that functionally distinct subpopulations of macrophages exist in the same tissue and that these macrophages play critical roles in both the injury and recovery phases of inflammatory scarring.


Kidney stones.
The age of peak incidence for stone disease is 20 to 40 years, although stones are seen in all age groups. There is a male to female ratio of 3:2. We conducted a systematic review and aimed to answer the following clinical questions: What are the effects of interventions for stone removal in people with asymptomatic kidney stones? What are the effects of interventions for the removal of symptomatic renal stones? What are the effects of interventions to remove symptomatic ureteric stones? What are the effects of interventions for the management of acute renal colic? We searched: Medline, Embase, The Cochrane Library, and other important databases up to May 2008 (Clinical Evidence reviews are updated periodically; please check our website for the most up-to-date version of this review). We included harms alerts from relevant organisations such as the US Food and Drug Administration (FDA) and the UK Medicines and Healthcare products Regulatory Agency (MHRA). We found 21 systematic reviews, RCTs, or observational studies that met our inclusion criteria. We performed a GRADE evaluation of the quality of evidence for interventions. In this systematic review, we present information relating to the effectiveness and safety of the following interventions: antispasmodic drugs; extracorporeal shockwave lithotripsy; intravenous fluids; non-steroidal anti-infammatory drugs (NSAIDs); opioids; oral fluids; percutaneous nephrolithotomy; and ureteroscopy.


Computer Vision Technology in the Differential Diagnosis of Cushing's Syndrome.
Cushing's syndrome is a rare disease characterized by clinical features that show morphological similarity with the metabolic syndrome. Distinguishing these diseases in clinical practice is challenging. We have previously shown that computer vision technology can be a potentially useful diagnostic tool in Cushing's syndrome. In this follow-up study, we addressed the described problem by increasing the sample size and including controls matched by body mass index. We enrolled 82 patients (22 male, 60 female) and 98 control subjects (32 male, 66 female) matched by age, gender and body-mass-index. The control group consisted of patients with initially suspected, but biochemically excluded Cushing's syndrome. Standardized frontal and profile facial digital photographs were acquired. The images were analyzed using specialized computer vision and classification software. A grid of nodes was semi-automatically placed on disease-relevant facial structures for analysis of texture and geometry. Classification accuracy was calculated using a leave-one-out cross-validation procedure with a maximum likelihood classifier. The overall correct classification rates were 10/22 (45.5%) for male patients and 26/32 (81.3%) for male controls, and 34/60 (56.7%) for female patients and 43/66 (65.2%) for female controls. In subgroup analyses, correct classification rates were higher for iatrogenic than for endogenous Cushing's syndrome. Regarding the advanced problem of detecting Cushing's syndrome within a study sample matched by body mass index, we found moderate classification accuracy by facial image analysis. Classification accuracy is most likely higher in a larger sample with healthy control subjects. Further studies might pursue a more advanced analysis and classification algorithm.


Total sleep deprivation on endogenous depression.
Ten endogenous depressive patients were deprived of sleep for two whole nights according to the following schedule: sleep/sleep deprivation/sleep/sleep deprivation/sleep. No drugs were administered. Experimental conditions were as neutral as possible. Blind and nonblind ratings were taken. The patients were generally rated as improved after sleep deprivation, but a substantial effect, though temporary with rapid relapse, occurred in only two cases. After subsequent sleep, relapse followed as a rule. The net antidepressive effect of the total procedure was slightly more than nil.


Nonparametric collective spectral density estimation with an application to clustering the brain signals.
In this paper, we develop a method for the simultaneous estimation of spectral density functions (SDFs) for a collection of stationary time series that share some common features. Due to the similarities among the SDFs, the log-SDF can be represented using a common set of basis functions. The basis shared by the collection of the log-SDFs is estimated as a low-dimensional manifold of a large space spanned by a prespecified rich basis. A collective estimation approach pools information and borrows strength across the SDFs to achieve better estimation efficiency. Moreover, each estimated spectral density has a concise representation using the coefficients of the basis expansion, and these coefficients can be used for visualization, clustering, and classification purposes. The Whittle pseudo-maximum likelihood approach is used to fit the model and an alternating blockwise Newton-type algorithm is developed for the computation. A web-based shiny App found at "https://ncsde.shinyapps.io/NCSDE" is developed for visualization, training, and learning the SDFs collectively using the proposed technique. Finally, we apply our method to cluster similar brain signals recorded by the for identifying synchronized brain regions according to their spectral densities.


New oral anticoagulants in atrial fibrillation and acute coronary syndromes: ESC Working Group on Thrombosis-Task Force on Anticoagulants in Heart Disease position paper.
Until recently, vitamin K antagonists were the only available oral anticoagulants, but with numerous limitations that prompted the introduction of new oral anticoagulants targeting the single coagulation enzymes thrombin (dabigatran) or factor Xa (apixaban, rivaroxaban, and edoxaban) and given in fixed doses without coagulation monitoring. Here we review the pharmacology and the results of clinical trials with these new agents in stroke prevention in atrial fibrillation and secondary prevention after acute coronary syndromes, providing perspectives on their future incorporation into clinical practice. In phase III trials in atrial fibrillation, compared with warfarin, dabigatran etexilate 150 mg B.I.D. reduced the rates of stroke/systemic embolism without any difference in major bleeding; dabigatran etexilate 110 mg B.I.D. had similar efficacy with decreased bleeding; apixaban 5 mg B.I.D. reduced stroke, systemic embolism, and mortality as well as major bleeding; and rivaroxaban 20 mg Q.D. was noninferior to warfarin for stroke and systemic embolism without a difference in major bleeding. All these agents reduced intracranial hemorrhage. Edoxaban is currently being evaluated in a further large phase III trial. Apixaban and rivaroxaban were evaluated in phase III trials for prevention of recurrent ischemia in patients with acute coronary syndromes who were mostly receiving dual antiplatelet therapy, with conflicting results on efficacy but consistent results for increased major bleeding. Overall, the new oral anticoagulants are poised to replace vitamin K antagonists for many patients with atrial fibrillation and may have a role after acute coronary syndromes. Although convenient to administer and manage, they present challenges that need to be addressed.


Rare Purulent Cardiac Tamponade Caused by Streptococcus Constellatus in a Young Immunocompetent Patient: Case Report and Review of the Literature.
BACKGROUND Purulent pericardial tamponade is a very rare occurrence in the current era of widespread antibiotic use. It is even rarer when caused by Streptococcus constellatus: a microorganism usually classified among the normal flora of the human body. It is occasionally diagnosed with certain predisposing factors. CASE REPORT We present the third case of Streptococcus constellatus cardiac tamponade reported in the current medical literature, occurring in a previously healthy young man who was initially admitted and treated for possible community-acquired pneumonia. The patient required immediate subxyphoid pericardiocentesis. He was also treated successfully with a lengthy course of both intravenous and oral antibiotics. Two months post-hospitalization, he was confirmed clinically stable with complete resolution of his purulent effusion. We also conducted a review of the literature for all Streptococcus milleri group purulent pericardial infections between 1984 and 2015. CONCLUSIONS Purulent cardiac tamponade caused by Streptococcus constellatus is extremely rare. It can be life threatening, however. Early appropriate diagnosis and therapeutic intervention are critical for a good outcome.


Regional Heterogeneity in Gene Expression, Regulation, and Coherence in the Frontal Cortex and Hippocampus across Development and Schizophrenia.
The hippocampus formation, although prominently implicated in schizophrenia pathogenesis, has been overlooked in large-scale genomics efforts in the schizophrenic brain. We performed RNA-seq in hippocampi and dorsolateral prefrontal cortices (DLPFCs) from 551 individuals (286 with schizophrenia). We identified substantial regional differences in gene expression and found widespread developmental differences that were independent of cellular composition. We identified 48 and 245 differentially expressed genes (DEGs) associated with schizophrenia within the hippocampus and DLPFC, with little overlap between the brain regions. 124 of 163 (76.6%) of schizophrenia GWAS risk loci contained eQTLs in any region. Transcriptome-wide association studies in each region identified many novel schizophrenia risk features that were brain region-specific. Last, we identified potential molecular correlates of in vivo evidence of altered prefrontal-hippocampal functional coherence in schizophrenia. These results underscore the complexity and regional heterogeneity of the transcriptional correlates of schizophrenia and offer new insights into potentially causative biology.


A multidisciplinary approach for the identification of novel HIV-1 non-nucleoside reverse transcriptase inhibitors: S-DABOCs and DAVPs.
Among the FDA approved drugs for the treatment of AIDS, non-nucleoside reverse transcriptase inhibitors (NNRTIs) are essential components of first-line anti-HIV-1 therapy because of the less-severe adverse effects associated with NNRTIs administration in comparison to therapies based on other anti-HIV-1 agents. In this contest, 3,4-dihydro-2-alkoxy-6-benzyl-4-oxypyrimidines (DABOs) have been the object of many studies aimed at identifying novel analogues endowed with potent inhibitory activity towards HIV-1 wild type and especially drug-resistant mutants. Accordingly, based on the encouraging results obtained from the biological screening of our internal collection of S-DABO derivatives, we started with the systematic functionalization of the pyrimidine scaffold to identify the minimal required structural features for RT inhibition. Herein, we describe how the combination of synthetic, biological, and molecular modeling studies led to the identification of two novel subclasses of S-DABO analogues: S-DABO cytosine analogues (S-DABOCs) and 4-dimethyamino-6-vinylpyrimidines (DAVPs).


In Situ Microparticles Loaded with S-Nitrosoglutathione Protect from Stroke.
Treatment of stroke, especially during the first hours or days, is still lacking. S-nitrosoglutathione (GSNO), a cerebroprotective agent with short life time, may help if administered early with a sustain delivery while avoiding intensive reduction in blood pressure. We developed in situ forming implants (biocompatible biodegradable copolymer) and microparticles (same polymer and solvent emulsified with an external oily phase) of GSNO to lengthen its effects and allow cerebroprotection after a single subcutaneous administration to Wistar rats. Arterial pressure was recorded for 3 days (telemetry, n = 14), whole-blood platelet aggregation up to 13 days (aggregometry, n = 58), and neurological score, cerebral infarct size and edema volume for 2 days after obstruction of the middle cerebral artery by autologous blood clots (n = 30). GSNO-loaded formulations (30 mg/kg) induced a slighter and longer hypotension (-10 vs. -56 ± 6 mmHg mean arterial pressure, 18 h vs. 40 min) than free GSNO at the same dose. The change in pulse pressure (-50%) lasted even up to 42 h for microparticles. GSNO-loaded formulations (30 mg/kg) prevented the transient 24 h hyper-aggregability observed with free GSNO and 7.5 mg/kg-loaded formulations. When injected 2 h after stroke, GSNO-loaded microparticles (30 mg/kg) reduced neurological score at 24 (-62%) and 48 h (-75%) vs. empty microparticles and free GSNO 7.5 mg/kg and, compared to free GSNO, divided infarct size by 10 and edema volume by 8 at 48 h. Corresponding implants reduced infarct size and edema volume by 2.5 to 3 times. The longer (at least 2 days) but slight effects on arterial pressures show sustained delivery of GSNO-loaded formulations (30 mg/kg), which prevent transient platelet hyper-responsiveness and afford cerebroprotection against the consequences of stroke. In conclusion, in situ GSNO-loaded formulations are promising candidates for the treatment of stroke.


Corynespora cassiicola f. sp. schinii, a Potential Biocontrol Agent for the Weed Schinus terebinthifolius in the United States.
Schinus terebinthifolius (Anacardiaceae), Brazilian peppertree (BP), is a major environmental weed in many tropical and subtropical areas of the globe, including Florida, Hawai'i, and Australia. This plant has been the target of a classical biocontrol project in the United States involving pathogens collected in Brazil for several years. A fungus was found in the Brazilian state of Espírito Santo causing leaf spots and severe foliage blight on BP. Examination of the morphology and internal transcribed spacer sequence analysis confirmed that the fungus is a strain of Corynespora cassiicola. Preliminary host-range tests involving 24 species, including 11 species in the family Anacardiaceae, were conducted with the fungus, and specificity toward BP was confirmed. Plants of Brazilian pepper tree from populations in Florida and Hawai'i included in the tests became severely diseased. Therefore, the recognition of a new forma specialis-Corynespora cassiicola f. sp. schinii-is proposed. The specificity of this forma specialis and the severity of the disease it caused in the field and under controlled conditions indicate that it has the potential for use as a biocontrol agent for BP in areas where it is an exotic invasive species.


Polymeric Nanoparticles of Brazilian Red Propolis Extract: Preparation, Characterization, Antioxidant and Leishmanicidal Activity.
The ever-increasing demand for natural products and biotechnology derived from bees and ultra-modernization of various analytical devices has facilitated the rational and planned development of biotechnology products with a focus on human health to treat chronic and neglected diseases. The aim of the present study was to prepare and characterize polymeric nanoparticles loaded with Brazilian red propolis extract and evaluate the cytotoxic activity of "multiple-constituent extract in co-delivery system" for antileishmanial therapies. The polymeric nanoparticles loaded with red propolis extract were prepared with a combination of poly-ε-caprolactone and pluronic using nanoprecipitation method and characterized by different analytical techniques, antioxidant and leishmanicidal assay. The red propolis nanoparticles in aqueous medium presented particle size (200-280 nm) in nanometric scale and zeta analysis (-20 to -26 mV) revealed stability of the nanoparticles without aggregation phenomenon during 1 month. After freeze-drying method using cryoprotectant (sodium starch glycolate), it was possible to observe particles with smooth and spherical shape and apparent size of 200 to 400 nm. Attenuated total reflectance Fourier transform infrared spectroscopy (ATR-FTIR) and thermal analysis revealed the encapsulation of the flavonoids from the red propolis extract into the polymeric matrix. Ultra performance liquid chromatography coupled with diode array detector (UPLC-DAD) identified the flavonoids liquiritigenin, pinobanksin, isoliquiritigenin, formononetin and biochanin A in ethanolic extract of propolis (EEP) and nanoparticles of red propolis extract (NRPE). The efficiency of encapsulation was determinate, and median values (75.0 %) were calculated using UPLC-DAD. 2,2-Diphenyl-1-picryhydrazyl method showed antioxidant activity to EEP and red propolis nanoparticles. Compared to negative control, EEP and NRPE exhibited leishmanicidal activity with an IC50 value of ≅38.0 μg/mL and 31.3 μg/mL, 47.2 μg/mL, 154.2μg/mL and 193.2 μg/mL for NRPE A1, NRPE A2, NRPE A3 and NRPE A4, respectively. Nanoparticles loaded with red propolis extract in co-delivery system and EEP presented cytotoxic activity on Leishmania (V.) braziliensis. Red propolis extract loaded in nanoparticles has shown to be potential candidates as intermediate products for preparation of various pharmaceutical dosage forms containing red propolis extract in the therapy against negligible diseases such as leishmaniasis. Graphical Abstract Some biochemical mechanisms of cellular debridement of Leishmania (V.) braziliensis species by the flavonoids of red propolis extract (EEP) or NRPE loaded with red propolis extract.


512-Channel and 13-Region Simultaneous Recordings Coupled with Optogenetic Manipulation in Freely Behaving Mice.
The development of technologies capable of recording both single-unit activity and local field potentials (LFPs) over a wide range of brain circuits in freely behaving animals is the key to constructing brain activity maps. Although mice are the most popular mammalian genetic model, in vivo neural recording has been traditionally limited to smaller channel count and fewer brain structures because of the mouse's small size and thin skull. Here, we describe a 512-channel tetrode system that allows us to record simultaneously over a dozen cortical and subcortical structures in behaving mice. This new technique offers two major advantages - namely, the ultra-low cost and the do-it-yourself flexibility for targeting any combination of many brain areas. We show the successful recordings of both single units and LFPs from 13 distinct neural circuits of the mouse brain, including subregions of the anterior cingulate cortices, retrosplenial cortices, somatosensory cortices, secondary auditory cortex, hippocampal CA1, dentate gyrus, subiculum, lateral entorhinal cortex, perirhinal cortex, and prelimbic cortex. This 512-channel system can also be combined with Cre-lox neurogenetics and optogenetics to further examine interactions between genes, cell types, and circuit dynamics across a wide range of brain structures. Finally, we demonstrate that complex stimuli - such as an earthquake and fear-inducing foot-shock - trigger firing changes in all of the 13 brain regions recorded, supporting the notion that neural code is highly distributed. In addition, we show that localized optogenetic manipulation in any given brain region could disrupt network oscillations and caused changes in single-unit firing patterns in a brain-wide manner, thereby raising the cautionary note of the interpretation of optogenetically manipulated behaviors.


Aberrant gene expression and sexually incompatible genomic imprinting in oocytes derived from XY mouse embryonic stem cells in vitro.
Mouse embryonic stem cells (ESCs) have the potential to differentiate into germ cells (GCs) in vivo and in vitro. Interestingly, XY ESCs can give rise to both male and female GCs in culture, irrespective of the genetic sex. Recent studies showed that ESC-derived primordial GCs contributed to functional gametogenesis in vivo; however, in vitro differentiation techniques have never succeeded in generating mature oocytes from ESCs due to cryptogenic growth arrest during the preantral follicle stages of development. To address this issue, a mouse ESC line, capable of producing follicle-like structures (FLSs) efficiently, was established to investigate their properties using conventional molecular biological methods. The results revealed that the ESC-derived FLSs were morphologically similar to ovarian primary-to-secondary follicles but never formed an antrum; instead, the FLSs eventually underwent abnormal development or cell death in culture, or formed teratomas when transplanted under the kidney capsule in mice. Gene expression analyses demonstrated that the FLSs lacked transcripts for genes essential to late folliculogenesis, including gonadotropin receptors and steroidogenic enzymes, whereas some other genes were overexpressed in FLSs compared to the adult ovary. The E-Cadherin protein, which is involved in cell-to-cell interactions, was also expressed ectopically. Remarkably, it was seen that oocyte-like cells in the FLSs exhibited androgenetic genomic imprinting, which is ordinarily indicative of male GCs. Although the FLSs did not express male GC marker genes, the DNA methyltransferase, Dnmt3L, was expressed at an abnormally high level. Furthermore, the expression of sex determination factors was ambiguous in FLSs as both male and female determinants were expressed weakly. These data suggest that the developmental dysfunction of the ESC-derived FLSs may be attributable to aberrant gene expression and genomic imprinting, possibly associated with uncertain sex determination in culture.


Relation of cardiovascular disease to potassium and magnesium deficiencies.
Potassium and magnesium deficiencies, particularly those induced by conventional loop and thiazide diuretic therapy, have been linked in clinical studies to an increased frequency in serious arrhythmias and mortality in acute myocardial infarction. Magnesium repletion has been shown not only to increase magnesium levels, but also to increase muscle potassium and to decrease the frequency of ventricular ectopic beats. Potassium replenishment alone may have a detrimental effect in magnesium-depleted patients. The potassium-sparing diuretic spironolactone (Aldactone) has been shown to spare both potassium and magnesium, and may therefore be a more appropriate diuretic therapy in patients at cardiovascular risk.


Concurrent validity of the Fitbit for assessing sedentary behavior and moderate-to-vigorous physical activity.
Recent advances in sensor technologies have promoted the use of consumer-based accelerometers such as Fitbit Flex in epidemiological and clinical research; however, the validity of the Fitbit Flex in measuring sedentary behavior (SED) and physical activity (PA) has not been fully determined against previously validated research-grade accelerometers such as ActiGraph GT3X+. Therefore, the purpose of this study was to examine the concurrent validity of the Fitbit Flex against ActiGraph GT3X+ in a free-living condition. A total of 65 participants (age: M = 42, SD = 14 years, female: 72%) each wore a Fitbit Flex and GT3X+ for seven consecutive days. After excluding sleep and non-wear time, time spent (min/day) in SED and moderate-to-vigorous PA (MVPA) were estimated using various cut-points for GT3X+ and brand-specific algorithms for Fitbit, respectively. Repeated measures one-way ANOVA and mean absolute percent errors (MAPE) served to examine differences and measurement errors in SED and MVPA estimates between Fitbit Flex and GT3X+, respectively. Pearson and Spearman correlations and Bland-Altman (BA) plots were used to evaluate the association and potential systematic bias between Fitbit Flex and GT3X+. PROC MIXED procedure in SAS was used to examine the equivalence (i.e., the 90% confidence interval with ±10% equivalence zone) between the devices. Fitbit Flex produced similar SED and low MAPE (mean difference [MD] = 37 min/day, P = .21, MAPE = 6.8%), but significantly higher MVPA and relatively large MAPE (MD = 59-77 min/day, P < .0001, MAPE = 56.6-74.3%) compared with the estimates from GT3X+ using three different cut-points. The correlations between Fitbit Flex and GT3X+ were consistently higher for SED (r = 0.90, ρ = 0.86, P < .01), but weaker for MVPA (r = 0.65-0.76, ρ = 0.69-0.79, P < .01). BA plots revealed that there is no apparent bias in estimating SED. In comparison with the GT3X+ accelerometer, the Fitbit Flex provided comparatively accurate estimates of SED, but the Fitbit Flex overestimated MVPA under free-living conditions. Future investigations using the Fitbit Flex should be aware of present findings.


Subcutaneous administration of genistein prior to lethal irradiation supports multilineage, hematopoietic progenitor cell recovery and survival.
Genistein, a non-toxic isoflavone from soybeans, has immunomodulating and radioprotective properties. In this study we investigated the mechanism for genistein-induced radioprotection by evaluating the recovery of bone marrow cells and peripheral blood hematology in lethally irradiated mice. CD2F1 male mice received a single subcutaneous injection of genistein (200 mg/kg) 24 h prior to a lethal, total body irradiation dose (8.75 Gy) of cobalt-60 gamma radiation. Survival and hematopoietic reconstitution were evaluated over nine weeks post-irradiation. Hematopoietic progenitor colony-forming cell assays were used to assess the reconstitution of bone marrow after radiation-induced myelosuppression. A total of 97% of genistein-treated mice survived after 30 days while 31% of vehicle-treated and 0% of untreated mice survived. The improvement in survival was related to accelerated neutrophil and platelet recovery, resulting from earlier and more pronounced multilineage, hematopoietic progenitor cell reconstitution in the femoral marrow compartment. Myeloid and erythroid progenitor cell numbers at day 15 post-irradiation were 6-fold to 20-fold higher in genistein-treated animals than in control animals. These results demonstrate that a single subcutaneous administration of genistein 24 h before irradiation provides significant radioprotection to the hematopoietic progenitor cell compartment.


Duration of viral shedding and culture positivity with postvaccination SARS-CoV-2 delta variant infections.
Isolation guidelines for severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) are largely derived from data collected prior to the emergence of the delta variant. We followed a cohort of ambulatory patients with postvaccination breakthrough SARS-CoV-2 infections with longitudinal collection of nasal swabs for SARS-CoV-2 viral load quantification, whole-genome sequencing, and viral culture. All delta variant infections in our cohort were symptomatic, compared with 64% of non-delta variant infections. Symptomatic delta variant breakthrough infections were characterized by higher initial viral load, longer duration of virologic shedding by PCR, greater likelihood of replication-competent virus at early stages of infection, and longer duration of culturable virus compared with non-delta variants. The duration of time since vaccination was also correlated with both duration of PCR positivity and duration of detection of replication-competent virus. Nonetheless, no individuals with symptomatic delta variant infections had replication-competent virus by day 10 after symptom onset or 24 hours after resolution of symptoms. These data support US CDC isolation guidelines as of November 2021, which recommend isolation for 10 days or until symptom resolution and reinforce the importance of prompt testing and isolation among symptomatic individuals with delta breakthrough infections. Additional data are needed to evaluate these relationships among asymptomatic and more severe delta variant breakthrough infections.


[Primary prevention of HIV infection].
Primary prophylaxis by a change of behaviour in the population continues to be the only effective possibility to prevent a further spread of the aids epidemic. The most important presupposition for this is the precise knowledge of the ways of transmission and the corresponding protective measures. Informations referring to this must turn up repeatedly and above all free of contradictions at all levels of the preventive campaign. The desired changes of behaviour suppose also a climate of confidence and solidarity. The physician's task of prevention is part of the comprehensive strategy of HIV-prevention in Switzerland (effect in width and depth on population- and target groups and on the individual level). It is naturally the physician's domain to look after and counsel individually. Most variable possibilities of beginning a conversation about Aids prevention offer themselves to him. He should put the indication for this as generous as possible and deal extensively with a possible risky behaviour which might not be conscious to both the patient and the physician. The HIV antibody test is indicated in certain situations, however the pre- and posttest counselling is in any case much more decisive than the result of the test. The preventive measures recommended up to now for the most important ways of transmission (sexual contact, intravenous drug use, perinatal transmission, blood/blood products) are still valid without modification.


Structural brain change in Attention Deficit Hyperactivity Disorder identified by meta-analysis.
The authors sought to map gray matter changes in Attention Deficit Hyperactivity Disorder (ADHD) using a novel technique incorporating neuro-imaging and genetic meta-analysis methods. A systematic search was conducted for voxel-based structural magnetic resonance imaging studies of patients with ADHD (or with related disorders) in relation to comparison groups. The authors carried out meta-analyses of the co-ordinates of gray matter differences. For the meta-analyses they hybridised the standard method of Activation Likelihood Estimation (ALE) with the rank approach used in Genome Scan Meta-Analysis (GSMA). This system detects three-dimensional conjunctions of co-ordinates from multiple studies and permits the weighting of studies in relation to sample size. For gray matter decreases, there were 7 studies including a total of 114 patients with ADHD (or related disorders) and 143 comparison subjects. Meta-analysis of these studies identified a significant regional gray matter reduction in ADHD in the right putamen/globus pallidus region. Four studies reported gray matter increases in ADHD but no regional increase was identified by meta-analysis. In ADHD there is gray matter reduction in the right putamen/globus pallidus region. This may be an anatomical marker for dysfunction in frontostriatal circuits mediating cognitive control. Right putamen lesions have been specifically associated with ADHD symptoms after closed head injuries in children.


Förster resonant energy transfer in orthogonally arranged chromophores.
We investigate the ultrafast resonant energy transfer of a perylene bisimide dyad by pump-probe spectroscopy, chemical variation, and calculations. This dyad undergoes transfer with near-unit quantum efficiency, although the transition dipole moments of the donor and acceptor are in a perfectly orthogonal arrangement to each other in the equilibrium geometry. According to the point dipole approximation used in Förster theory, no energy transfer should occur. Experimentally we do, however, find an ultrafast transfer time of 9.4 ps. With the transition density cube approach we show that in the orthogonal arrangement the Coulombic interactions do not contribute to the electronic coupling. Through the change of the spacer in both length and chemical character, we can clearly exclude any Dexter-type energy transfer. The temperature effects on the Förster resonant energy transfer rate demonstrate that energy transfer is enabled through low-frequency ground-state vibrations, which break the orthogonal arrangement of the transition dipole moments. The dyads presented here therefore are a first example that shows with extreme clarity the decisive role vibrational motion plays in energy transfer processes.


Development and validation of a maleimide-based enzyme-linked immunosorbent assay for the detection of tetrodotoxin in oysters and mussels.
The recent detection of tetrodotoxins (TTXs) in puffer fish and shellfish in Europe highlights the necessity to monitor the levels of TTXs in seafood by rapid, specific, sensitive and reliable methods in order to protect human consumers. A previous immunoassay for TTX detection in puffer fish, based on the use of self-assembled monolayers (SAMs) for the immobilization of TTX on maleimide plates (mELISA), has been modified and adapted to the analysis of oyster and mussel samples. Changing dithiol for cysteamine-based SAMs enabled reductions in the assay time and cost, while maintaining the sensitivity of the assay. The mELISA showed high selectivity for TTX since the antibody did not cross-react with co-occurring paralytic shellfish poisoning (PSP) toxins and no interferences were observed from arginine (Arg). Moreover, TTX-coated maleimide plates stored for 3 months at -20°C and 4°C were stable, thus when pre-prepared, the time to perform the assay is reduced. When analyzing shellfish samples, matrix effects and toxin recovery values strongly depended on the shellfish type and the sample treatment. Blank oyster extracts could be directly analyzed without solid-phase extraction (SPE) clean-up, whereas blank mussel extracts showed strong matrix effects and SPE and subsequent solvent evaporation were required for removal. However, the SPE clean-up and evaporation resulted in toxin loss. Toxin recovery values were taken as correction factors (CFs) and were applied to the quantification of TTX contents in the analysis of naturally-contaminated shellfish samples by mELISA. The lowest effective limits of detection (eLODs) were about 20 and 50µg/kg for oyster extracts without and with SPE clean-up, respectively, and about 30µg/kg for mussel extracts with both protocols, all of them substantially below the eLOD attained in the previous mELISA for puffer fish (230µg/kg). Analysis of naturally-contaminated samples by mELISA and comparison with LC-MS/MS quantifications demonstrated the viability of the approach. This mELISA is a selective and sensitive tool for the rapid detection of TTX in oyster and mussel samples showing promise to be implemented in routine monitoring programs to protect human health.


Phase-locking of two self-seeded tapered amplifier lasers.
We report on the phase-locking of two diode lasers based on self-seeded tapered amplifiers. In these lasers, a reduction of linewidth is achieved using narrow-band high-transmission interference filters for frequency selection. The lasers combine a compact design with a Lorentzian linewidth below 200 kHz at an output power of 300 mW for a wavelength of 780 nm. We characterize the phase noise of the phase-locked laser system and study its potential for coherent beam-splitting in atom interferometers.


Small-molecule screen identifies modulators of EWS/FLI1 target gene expression and cell survival in Ewing's sarcoma.
Ewing's sarcoma family of tumors (EFT) is characterized by the presence of chromosomal translocations leading to the expression of oncogenic transcription factors such as, in the majority of cases, EWS/FLI1. Because of its key role in Ewing's sarcoma development and maintenance, EWS/FLI1 represents an attractive therapeutic target. Here, we characterize PHLDA1 as a novel direct target gene whose expression is repressed by EWS/FLI1. Using this gene and additional specific well-characterized target genes such as NROB1, NKX2.2 and CAV1, all activated by EWS/FLI1, as a read-out system, we screened a small-molecule compound library enriched for FDA-approved drugs that modulated the expression of EWS/FLI1 target genes. Among a hit-list of nine well-known drugs such as camptothecin, fenretinide, etoposide and doxorubicin, we also identified the kinase inhibitor midostaurin (PKC412). Subsequent experiments demonstrated that midostaurin is able to induce apoptosis in a panel of six Ewing's sarcoma cell lines in vitro and can significantly suppress xenograft tumor growth in vivo. These results suggest that midostaurin might be a novel drug that is active against Ewing's cells, which might act by modulating the expression of EWS/FLI1 target genes.


Acid sphingomyelinase: No potential as a biomarker for multiple sclerosis.
Multiple sclerosis (MS) lacks reliable biomarkers that reflect disease activity. Recent evidence suggests that an altered sphingolipid metabolism is associated with MS pathogenesis. To explore acid sphingomyelinase (ASM) activity and altered sphingolipid metabolism as potential biomarkers in serum of MS patients, to predict active and progressive disease, and response to disease modifying therapy (DMT). Levels of serum ASM activity were longitudinally analyzed in 40 clinically isolated syndrome, 64 relapsing remitting (RR) and 10 primary progressive MS patients, and 22 healthy controls (HC). ASM activity and sphingolipid levels were measured in a different sample of 61 RRMS patients using DMT. A significant difference in ASM activity levels was observed between MS patients and HC (p < 0.001). There was no correlation between ASM activity levels and disease activity, progression or response to DMT. Ceramide (Cer)-C16:0 , Cer-C24:0 and sphingomyelin (SM)-C20:0, SM-C22:0, SM-C24:0 and SM-C24:1 showed a significant increase during fingolimod use. Although higher levels in MS patients were found, ASM activity levels do not show potential as a biomarker for predicting disease activity, progression or response to DMT. Two ceramides and four types of sphingomyelin require further investigation as potential markers for treatment response.


Impossible shadows and lightness constancy.
The intersection between an illumination and a reflectance edge is characterised by the 'ratio-invariant' property, that is the luminance ratio of the regions under different illumination remains the same. In a CRT experiment, we shaped two areas, one surrounding the other, and simulated an illumination edge dividing them in two frames of illumination. The portion of the illumination edge standing on the surrounding area (labelled contextual background) was the contextual edge, while the portion standing on the enclosed area (labelled mediating background) was the mediating edge. On the mediating background, there were two patches, one per illumination frame. Observers were asked to adjust the luminance of the patch in bright illumination to equate the lightness of the other. We compared conditions in which the luminance ratio at the contextual edge could be (i) equal (possible shadow), or (ii) larger (impossible shadow) than that at the mediating edge. In addition, we manipulated the reflectance of the backgrounds. It could be higher for the contextual than for the mediating background; or, vice versa, lower for the contextual than for the mediating background. Results reveal that lightness constancy significantly increases when: (i) the luminance ratio at the contextual edge is larger than that at the mediating edge creating an impossible shadow, and (ii) the reflectance of the contextual background is lower than that of the mediating one. We interpret our results according to the albedo hypothesis, and suggest that the scission process is facilitated when the luminance ratio at the contextual edge is larger than that at the mediating edge and/or the reflectance of the including area is lower than that of the included one. This occurs even if the ratio-invariant property is violated.


Quantitative 99mTc-pertechnetate thyroid scintigraphy in normal beagles.
This study was done to investigate the validity of published canine thyroid/salivary (T/S) ratios of approximately 1 in normal dogs and to determine thyroid uptake of 99mTc-pertechnetate (pertechnetate) measured as percent uptake of injected dose. These parameters were evaluated in 13 Beagle dogs over a 4 hour period. Mean +/- standard deviation (SD) and median T/S ratios of 1.2 +/- 0.3 and 1.1 were essentially the same at twenty minutes and 1 hour. T/S values ranged from 0.9 to 2.2 at 20 minutes and from 0.8 to 2.4 at 1 hour. T/S ratio values progressively declined over the subsequent time intervals with mean +/- SD and median values of 0.6 +/- 0.2 and 0.6, respectively, measured at 4 hours. The mean +/- SD maximum percent dose uptake of pertechnetate within the thyroid gland was 0.55% +/- 0.15% with a range of 0.28% to 0.90%. The mean +/- SD time interval from injection of pertechnetate to maximum uptake within the thyroid gland was 160 +/- 55 minutes with a range 31-240 minutes. The data derived from this study of normal dogs may be useful in the evaluation of dogs with thyroidal diseases including hypothyroidism.


Collaboration between ZEBET, FRAME, and ECVAM: FRAME's contribution to establishing the Three Rs in Europe.
FRAME has more than any other institution contributed to the implementation of the Three Rs concept in Europe. The first step was achieved by establishing the European Research Group on Alternatives to Toxicity Testing (ERGATT), in which individual scientists in several European countries began to collaborate to promote the development and validation of in vitro toxicity tests in order to refine, reduce and replace the use of animals in toxicity testing. The first successful project was the start of the INVITOX in vitro toxicology database, which is today managed by ECVAM. Major milestones were the establishment in 1989 of ZEBET, the first national centre for alternatives, in Germany, and of ECVAM in 1991. In 1990, ERGATT and CAAT organised a workshop in Amden, Switzerland, at which European and US scientists developed the concept of experimental validation of toxicity tests, which today remains the basis for the independent, scientific validation process. ECVAM has applied this validation concept in all of its successful validation studies, which have provided in vitro toxicity tests that have been accepted for regulatory purposes by the EU Commission and by the OECD. By re-launching the ATLA journal in 1983, FRAME provided another powerful scientific tool for promoting the Three Rs in Europe and around the world. The close cooperation of scientists from FRAME, ZEBET and ECVAM has most effectively promoted and established the Three Rs as the basic scientific, ethical and legal concept for refining, reducing and replacing the use of experimental animals.


Population-based interventions for the prevention of fall-related injuries in older people.
Fall-related injuries are a significant cause of morbidity and mortality in the older population. Summary information about counter-measures that successfully address the risk factors for fall-related injuries in research settings has been widely disseminated. However, less available is evidence-based information about successful roll-out of these counter-measures in public health programmes in the wider community. Population-based interventions in the form of multi-strategy, multi-focused programmes are hypothesised to result in a reduction in population-wide injury rates. This review tests this hypothesis with regards to fall-related injuries among older people. To assess the effectiveness of population-based interventions, defined as coordinated, community-wide, multi-strategy initiatives, for reducing fall-related injuries among older people. The search strategy was based on electronic searches, handsearches of selected journals and snowballing from reference lists of selected publications. Studies were independently screened for inclusion by two reviewers. Included studies were those that reported changes in medically treated fall-related injuries among older people following the implementation of a controlled population-based intervention. Data were independently extracted by two reviewers. Meta-analysis was not appropriate, due to the heterogeneity of the included studies. Out of 23 identified studies, five met the criteria for inclusion. There were no randomised controlled trials. Significant decreases or downward trends in fall-related injuries were reported in all five of the included studies with the relative reduction in fall-related injuries ranging from 6 to 33%. Despite methodological limitations of the evaluation studies reviewed, the consistency of reported reductions in fall-related injuries across all programmes support the preliminary claim that the population-based approach to the prevention of fall-related injury is effective and can form the basis of public health practice. Randomised, multiple community trials of population-based interventions are indicated to increase the level of evidence in support of the population-based approach. Research is also required to elucidate the barriers and facilitators in population-based interventions that influence the extent to which population programmes are effective.


The Rutgers Alcohol Problem Index for Adolescent Alcohol and Drug Problems: A Comprehensive Modern Psychometric Study.
The Rutgers Alcohol Problem Index (RAPI) has been used extensively as a measure of alcohol-related problems experienced by adolescents and young adults. The present study aimed to comprehensively examine the psychometric profile and criterion-related validity of an 18-item RAPI adapted to measure negative consequences resulting from alcohol and other drug use in an at-risk adolescent population. Categorical confirmatory factor analysis and the graded response model were used for evaluations of the latent factor structure, item properties, test information, and item invariance across gender, age groups (middle vs. late adolescence), and drug use profiles (readily available drugs vs. other illicit drugs), as well as computation of the item response theory scale scores for each of 617 alternative high school students (44% female). A reduced set of 16 RAPI items provided plausible evidence of unidimensionality and good measurement precision at a relatively wide range of the latent trait continuum. All but two items were invariant across the drug use profiles. The other illicit drug group had higher levels of the latent substance-related problems than the readily available drug group. Criterion-related validity was supported using a measure of past-year polysubstance use. The RAPI can be effectively modified to measure negative consequences resulting from alcohol and other drug use and reflects a sufficiently wide range of the latent continuum of problem use.


The prosensory function of Sox2 in the chicken inner ear relies on the direct regulation of Atoh1.
The proneural gene Atoh1 is crucial for the development of inner ear hair cells and it requires the function of the transcription factor Sox2 through yet unknown mechanisms. In the present work, we used the chicken embryo and HEK293T cells to explore the regulation of Atoh1 by Sox2. The results show that hair cells derive from Sox2-positive otic progenitors and that Sox2 directly activates Atoh1 through a transcriptional activator function that requires the integrity of Sox2 DNA binding domain. Atoh1 activation depends on Sox transcription factor binding sites (SoxTFBS) present in the Atoh1 3' enhancer where Sox2 directly binds, as shown by site directed mutagenesis and chromatin immunoprecipitation (ChIP). In the inner ear, Atoh1 enhancer activity is detected in the neurosensory domain and it depends on Sox2. Dominant negative competition (Sox2HMG-Engrailed) and mutation of the SoxTFBS abolish the reporter activity in vivo. Moreover, ChIP assay in isolated otic vesicles shows that Sox2 is bound to the Atoh1 enhancer in vivo. However, besides activating Atoh1, Sox2 also promotes the expression of Atoh1 negative regulators and the temporal profile of Atoh1 activation by Sox2 is transient suggesting that Sox2 triggers an incoherent feed-forward loop. These results provide a mechanism for the prosensory function of Sox2 in the inner ear. We suggest that sensory competence is established early in otic development through the activation of Atoh1 by Sox2, however, hair cell differentiation is prevented until later stages by the parallel activation of negative regulators of Atoh1 function.


Small molecules bound to unique sites in the target protein binding cleft of calcium-bound S100B as characterized by nuclear magnetic resonance and X-ray crystallography.
Structural studies are part of a rational drug design program aimed at inhibiting the S100B-p53 interaction and restoring wild-type p53 function in malignant melanoma. To this end, structures of three compounds (SBi132, SBi1279, and SBi523) bound to Ca(2+)-S100B were determined by X-ray crystallography at 2.10 A (R(free) = 0.257), 1.98 A (R(free) = 0.281), and 1.90 A (R(free) = 0.228) resolution, respectively. Upon comparison, SBi132, SBi279, and SBi523 were found to bind in distinct locations and orientations within the hydrophobic target binding pocket of Ca(2+)-S100B with minimal structural changes observed for the protein upon complex formation with each compound. Specifically, SBi132 binds nearby residues in loop 2 (His-42, Phe-43, and Leu-44) and helix 4 (Phe-76, Met-79, Ile-80, Ala-83, Cys-84, Phe-87, and Phe-88), whereas SBi523 interacts with a separate site defined by residues within loop 2 (Ser-41, His-42, Phe-43, Leu-44, Glu-45, and Glu-46) and one residue on helix 4 (Phe-87). The SBi279 binding site on Ca(2+)-S100B overlaps the SBi132 and SBi523 sites and contacts residues in both loop 2 (Ser-41, His-42, Phe-43, Leu-44, and Glu-45) and helix 4 (Ile-80, Ala-83, Cys-84, Phe-87, and Phe-88). NMR data, including saturation transfer difference (STD) and (15)N backbone and (13)C side chain chemical shift perturbations, were consistent with the X-ray crystal structures and demonstrated the relevance of all three small molecule-S100B complexes in solution. The discovery that SBi132, SBi279, and SBi523 bind to proximal sites on Ca(2+)-S100B could be useful for the development of a new class of molecule(s) that interacts with one or more of these binding sites simultaneously, thereby yielding novel tight binding inhibitors specific for blocking protein-protein interactions involving S100B.


Inflammation gene variants and susceptibility to albuminuria in the U.S. population: analysis in the Third National Health and Nutrition Examination Survey (NHANES III), 1991-1994.
Albuminuria, a common marker of kidney damage, serves as an important predictive factor for the progression of kidney disease and for the development of cardiovascular disease. While the underlying etiology is unclear, chronic, low-grade inflammation is a suspected key factor. Genetic variants within genes involved in inflammatory processes may, therefore, contribute to the development of albuminuria. We evaluated 60 polymorphisms within 27 inflammatory response genes in participants from the second phase (1991-1994) of the Third National Health and Nutrition Examination Survey (NHANES III), a population-based and nationally representative survey of the United States. Albuminuria was evaluated as logarithm-transformed albumin-to-creatinine ratio (ACR), as ACR ≥ 30 mg/g, and as ACR above sex-specific thresholds. Multivariable linear regression and haplotype trend analyses were conducted to test for genetic associations in 5321 participants aged 20 years or older. Differences in allele and genotype distributions among non-Hispanic whites, non-Hispanic blacks, and Mexican Americans were tested in additive and codominant genetic models. Variants in several genes were found to be marginally associated (uncorrected P value < 0.05) with log(ACR) in at least one race/ethnic group, but none remained significant in crude or fully-adjusted models when correcting for the false-discovery rate (FDR). In analyses of sex-specific albuminuria, IL1B (rs1143623) among Mexican Americans remained significantly associated with increased odds, while IL1B (rs1143623), CRP (rs1800947) and NOS3 (rs2070744) were significantly associated with ACR ≥ 30 mg/g in this population (additive models, FDR-P < 0.05). In contrast, no variants were found to be associated with albuminuria among non-Hispanic blacks after adjustment for multiple testing. The only variant among non-Hispanic whites significantly associated with any outcome was TNF rs1800750, which failed the test for Hardy-Weinberg proportions in this population. Haplotypes within MBL2, CRP, ADRB2, IL4R, NOS3, and VDR were significantly associated (FDR-P < 0.05) with log(ACR) or albuminuria in at least one race/ethnic group. Our findings suggest a small role for genetic variation within inflammation-related genes to the susceptibility to albuminuria. Additional studies are needed to further assess whether genetic variation in these, and untested, inflammation genes alter the susceptibility to kidney damage.


Adolescent health experience after abortion or delivery (AHEAD) trial: formative protocol for intervention development to prevent rapid, repeat pregnancy.
There is a high unmet need for modern contraception among adolescents, and adolescent girls who have already been pregnant are especially vulnerable to a rapid, repeat pregnancy (defined as a subsequent pregnancy within two years). The Adolescent Health Experience after Abortion or Delivery (AHEAD) trial will design, pilot, finalize, and ultimately evaluate an intervention targeted at reducing rapid repeat pregnancy. This protocol presents the methods for the first phase--formative research to identify key determinants of contraceptive use and rapid, repeat unintended pregnancy among adolescents. The determinants of adolescent pregnancy are known to vary by context; therefore, a dissimilar set of three countries will be selected to enable evaluation of the intervention in diverse cultural, political and economic environment, and to allow the intervention to be tested with a fuller range of ever-pregnant adolescents, including those who have chosen to terminate their pregnancy as well as those who are mothers. We will also consider marital status in settings where it is common for adolescents to marry. Focus group discussions (FGDs) will be conducted to examine barriers and facilitators to using contraception; preferred methods of overcoming these barriers; and perceptions of the services and information received. Key informant (KI) interviews will take place with various cadres of healthcare providers, health and education officials, and members of key youth and health organizations that work with adolescents. These interviews will focus on perceptions of pregnant adolescents; perceived information, skills, and motivations required for adolescent uptake of contraception; and experiences, challenges, and attitudes encountered during interactions. The findings from this first formative phase will be used to develop an intervention for preventing rapid, repeat unintended pregnancy among adolescents. This intervention will be piloted in a second phase of the AHEAD trial.


Combined treatment for acromegaly with long-acting somatostatin analogs and pegvisomant: long-term safety for up to 4.5 years (median 2.2 years) of follow-up in 86 patients.
We previously reported on the efficacy, safety, and quality of life (QoL) of long-acting somatostatin analogs (SSA) and (twice) weekly pegvisomant (PEG-V) in acromegaly and improvement after the addition of PEG-V to long-acting SSA. To assess the long-term safety in a larger group of acromegalic patients over a larger period of time: 29.2 (1.2-57.4) months (mean (range)). Pegvisomant was added to SSA monotherapy in 86 subjects (37 females), to normalize serum IGF1 concentrations (n=63) or to increase the QoL. The median dosage was 60.0 (20-200) mg weekly. After a mean treatment period of 29.2 months, 23 patients showed dose-independent PEG-V related transient liver enzyme elevations (TLEE). TLEE occurred only once during the continuation of combination therapy, but discontinuation and re-challenge induced a second episode of TLEE. Ten of these patients with TLEE also suffered from diabetes mellitus (DM). In our present series, DM had a 2.28 odds ratio (CI 1.16-9.22; p=0.03) higher risk for developing TLEE. During the combined therapy, a clinical significant decrease in tumor size by more than 20% was observed in 14 patients. Two of these patients were previously treated by pituitary surgery, 1 with additional radiotherapy and all other patients received primary medical treatment. Long-term combined treatment with SSA and twice weekly PEG-V up to more than 4 years seems to be safe. Patients with both acromegaly and DM have a 2.28 higher risk of developing TLEE. Clinical significant tumor shrinkage was observed in 14 patients during combined treatment.


Analysis of the relationship between nurse influences over flexible working and commitment to nursing.
The paper explores the theoretical and practical bases of both commitment and control within the context of temporal aspects of flexible working in nursing. The aims of the paper are to examine the relationships between nurses' shift patterns, influence over shift pattern and realization of shift preference and commitment to nursing. Data were collected through a postal questionnaire completed by 2987 British nurses employed in hospitals, care homes and hospices. Principal components analysis was used to identify common factors among responses to a series of 33 statements about working life. Data were analysed using ANOVA and multiple regression techniques. Permanent night shift nurses reported lower levels of commitment to nursing. As predicted, influence over shift patterns and realization of preferred shift pattern were positively associated with commitment to nursing, although the relationship was weak. Positive perceptions of career development opportunities were a stronger predictor of commitment to nursing. Results are discussed in light of previous ethnographic research on nurses' shift patterns. The opportunity to explore quantitatively the effects of shift-related decisions on commitment using a large sample is useful. While influence and shift type were predictors of commitment, the positive impact they were expected to have was smaller than expected. Similarly, the negative effects of not having influence or of working permanent night shifts were smaller than expected and the statistical significance of such small effects relies heavily on the large sample obtained. Other variables, particularly career development prospects, outweigh the influence of shift-related variables on commitment. The research gives a clear message to human resource managers involved with nurse management: so long as nurses have a strong perception of career development potential the otherwise negative influences of shift impact can be minimized.


Detecting couplings between interacting oscillators with time-varying basic frequencies: instantaneous wavelet bispectrum and information theoretic approach.
In the natural world, the properties of interacting oscillatory systems are not constant, but evolve or fluctuating continuously in time. Thus, the basic frequencies of the interacting oscillators are time varying, which makes the system analysis complex. For studying their interactions we propose a complementary approach combining wavelet bispectral analysis and information theory. We show how these methods uncover the interacting properties and reveal the nature, strength, and direction of coupling. Wavelet bispectral analysis is generalized as a technique for detecting instantaneous phase-time dependence for the case of two or more coupled nonlinear oscillators whereas the information theory approach can uncover the directionality of coupling and extract driver-response relationships in complex systems. We generate bivariate time-series numerically to mimic typical situations that occur in real measured data, apply both methods to the same time-series and discuss the results. The approach is applicable quite generally to any system of coupled nonlinear oscillators.


Neonatal lipopolysaccharide exposure impairs sexual development and reproductive success in the Wistar rat.
We investigated, in rats, whether neonatal exposure to bacterial lipopolysaccharide (LPS) impairs sexual development, sexual decline, and reproductive behaviour in later life. Rats were administered either LPS (Salmonella enterica, serotype enteritidis, 0.05 mg/kg, ip) or saline (equivolume) on days 3 and 5 postpartum. The immediate and long-term effect of treatment on HPA and HPG hormones, testicular morphology, and mating behaviour was assessed. Neonatal LPS exposure induced a significant increase in corticosterone compared to controls, as well as reduced testosterone and LH in males and LH in females immediately following neonatal drug exposure. Neonatal LPS exposure disrupted the normal weight-to-age ratio of puberty onset in males and females, and impaired sexual performance in adulthood. Reproductive function was reflected in significantly diminished sperm presence in rats that had received neonatal LPS. LPS-treated females exhibited LH suppression during puberty, and males demonstrated testosterone suppression in late adulthood. Testosterone and LH surges during mating were significantly reduced in adult offspring treated with LPS as neonates. Furthermore, animals exposed to neonatal LPS and subsequent stress in adulthood, exhibited significantly blunted corticosterone responses. Morphometric assessment of testes taken from neonates revealed reduced gonocyte genesis immediately following LPS exposure and increased seminiferous disorganisation of the epithelium in these animals in adulthood. This research demonstrates the long-term impact of neonatal bacterial exposure on reproductive success given that early life exposure to bacteria disrupted puberty onset and sexual performance. Associated changes in neuroendocrine functioning suggest a possible mechanism through which a subfertile phenotype may arise.


Physical mechanisms in neuroelectromagnetic therapies.
Physical parameters that are used to characterize different types of electromagnetic devices used in neurotherapy can include power, frequency, carrier frequency, current, magnetic field intensity, and whether an application is primarily electric or primarily magnetic. Currents can range from tens of microamperes to hundreds of milliamperes, magnetic fields from tens of microtesla to more than one tesla, and frequencies from a few Hz to more than 50 GHz. A division into three device categories is proposed, based on the current applied and the specificity of the therapeutic signal. Two research areas have great potential for new neuroelectromagnetic strategies. Studies of endogenous neural oscillatory states suggest using external fields to reinforce or inhibit such states. Also, various independent groups have reported that weak magnetic fields, in particular ion cyclotron resonance fields, are capable of sharply altering behavior in rats.


Rice-based electrolyte drinks more effective than water in replacing sweat losses during hot weather training and operations.
Heat-related injury presents significant threats to the health and operational effectiveness of Soldiers and military operations. In 2012, active component, U.S. Armed Forces experienced 365 incident cases of heat stroke and 2,257 incident cases of ?other heat injury.? Most of these occurred among recruit and enlisted personnel and most were under the age of 30. In conditioned military personnel, a rice-based oral rehydration solution was superior to water alone at maintaining body weight and, by inference, enabled Soldiers to better maintain their the state of hydration during prolonged exercise in high ambient temperatures. In view of the health risks associated with dehydration and their effects on training and operations, this study suggests that the consumption of beverages containing electrolytes and a rice-based carbohydrate is superior to the consumption of water alone in preventing dehydration and heat related illness.


Risk undermined in the bilateral pharmaceutical regulatory system in Taiwan.
The concept of Pharmacovigilance Planning and Risk Minimization Planning (PVP/RMP), initiated by the International Conference on Harmonization (ICH), addressed an important conceptual change from monitoring the safety of individual medicine to proactively conducting risk prevention for the minimization of medication error. However, the implementation of PVP/RMP is a challenge in societies like Taiwan where irrational medication and co-medication is prevalent. It is even more difficult in Taiwan where two regulatory bodies are governing pharmaceutical affairs, namely Taiwan Food and Drug Administration (TFDA) in charge of Western Medicine (WM) and the Department of Chinese Medicine and Pharmacy (DCMP) in charge of Traditional Chinese Medicine (TCM). There are thus dual-tract drug approval panels, two GMP controls and two independent adverse drug event reporting systems. This rendered irrational co-medication of WM and TCM undetectable and the standard tools for monitoring pharmacovigilance inapplicable. The bilateral regulatory system is conceptually unscientific in accordance with PVP/RMP and unethical from humanity point of view. The first part of this review delivers (1) social aspects of polypharmacy in Taiwan; (2) regulatory aspects of pharmaceutical administration; (3) risks undermined in the bilateral regulatory system and (4) pharmacoepidemiology in relation to the risk of polypharmacy. As evidence-based medicine (EBM) forms the fundamental risk-benefit assessment on medication, the second part of this review delivers (1) the scientific aspects of the beauty and the odds of biological system that governs host-xenobiotics interaction; (2) conceptual evolution from product management (pharmacovigilance) to risk management (PVP/RMP); (3) non-biased due process is essential for risk-benefit assessment on medicinal products and (4) the opinion of the authors on system building for safe medication.


Voltage-dependent ion channels in the mouse RPE: comparison with Norrie disease mice.
We studied electrophysiological properties of cultured retinal pigment epithelial (RPE) cells from mouse and a mouse model for Norrie disease. Wild-type RPE cells revealed the expression of ion channels known from other species: delayed-rectifier K(+) channels composed of Kv1.3 subunits, inward rectifier K(+) channels, Ca(V)1.3 L-type Ca(2+) channels and outwardly rectifying Cl(-) channels. Expression pattern and the ion channel characteristics current density, blocker sensitivity, kinetics and voltage-dependence were compared in cells from wild-type and Norrie mice. Although no significant differences were observed, our study provides a base for future studies on ion channel function and dysfunction in transgenic mouse models.


Increased proton pump inhibitor and NSAID exposure in irritable bowel syndrome: results from a case-control study.
Patients with irritable bowel syndrome (IBS) seen by a gastroenterologist often utilize medications that may alter intestinal homeostasis. The question arises whether exposure to these drugs is associated with the development of IBS symptoms. Aim of this study was therefore to assess the use of PPIs and NSAIDs in patients with IBS versus controls. Cases of IBS from the last 5 years were reviewed. All patients having had at least one prescription for a particular drug (PPIs, NSAIDs, SSRIs, diuretics, ACE inhibitors) in the 6 months prior to the time of initial symptom onset were considered exposed. The control group consisted of individuals randomly selected from the general population. 287 cases of IBS were retrieved for analysis together with 287 age and sex-matched controls. Exposure to PPIs and NSAIDs was significantly higher in IBS patients, whereas no association between ACE inhibitor use and IBS was found. PPIs were not significantly associated when excluding patients with gastrointestinal reflux disease or functional dyspepsia. Exposure to SSRIs was also positively associated with IBS, but only when patients with psychiatric comorbidity were included in the analyses. Medications that may alter intestinal homeostasis such as NSAIDs and PPIs were more frequently used in IBS patients compared to controls. This association might be relevant for everyday clinical practice, but it is remains to be elucidated whether this association is of etiological nature.


Transient ischemic dilation ratio in 82Rb PET myocardial perfusion imaging: normal values and significance as a diagnostic and prognostic marker.
In myocardial perfusion SPECT, transient ischemic dilation ratio (TID) is a well-established marker of severe ischemia and adverse outcome. However, its role in the setting of (82)Rb PET is less well defined. We analyzed 265 subjects who underwent clinical rest-dipyridamole (82)Rb PET/CT. Sixty-two subjects without a prior history of cardiac disease and with a normal myocardial perfusion study had either a low or a very low pretest likelihood of coronary artery disease or negative CT angiography. These subjects were used to establish a reference range of TID. In the remaining 203 patients with an intermediate or high pretest likelihood, subgroups with normal and abnormal TID were established and compared with respect to clinical variables, perfusion defect scores, left ventricular function, and absolute myocardial flow reserve. Follow-up was obtained for 969 ± 328 d to determine mortality by review of the social security death index. In the reference group, TID ratio was 0.98 ± 0.06. Accordingly, a threshold for abnormal TID was set at greater than 1.13 (0.98 + 2.5 SDs). In the study group, 19 of 203 patients (9%) had an elevated TID ratio. Significant differences between subgroups with normal and abnormal TID ratio were observed for ejection fraction reserve (5.0 ± 6.4 vs. 1.8 ± 7.9; P < 0.05), difference between end-systolic volume (ESV) at rest and stress (ΔESV[stress-rest]; 1.8 ± 7.4 vs. 12.3 ± 13.0 mL; P < 0.0001), difference between end-diastolic volume (EDV) at rest and stress (ΔEDV[stress-rest]; 10.8 ± 11.5 vs. 23.8 ± 14.6 mL; P < 0.0001), summed rest score (1.8 ± 3.8 vs. 3.8 ± 7.6; P < 0.05), summed stress score (3.0 ± 5.4 vs. 7.5 ± 9.8; P < 0.002), summed difference score (1.3 ± 2.6 vs. 3.7 ± 5.3; P < 0.02), and global myocardial flow reserve (2.1 ± 0.8 vs. 1.7 ± 0.6; P < 0.02). Additionally, TID-positive patients had a significantly lower overall survival probability (P < 0.05). In a subgroup analysis of patients without regional perfusion abnormalities, TID-positive patients' overall survival probability was significantly smaller (P < 0.03), and TID was an independent predictor (exponentiation of the B coefficients [Exp(b)] = 6.22; P < 0.009) together with an ejection fraction below 45% (Exp[b] = 6.16; P < 0.002). The present study suggests a reference range of TID for (82)Rb PET myocardial perfusion imaging that is in the range of previously established values for SPECT. Abnormal TID in (82)Rb PET is associated with more extensive left ventricular dysfunction, ischemic compromise, and reduced global flow reserve. Preliminary outcome analysis suggests that TID-positive subjects have a lower overall survival probability.


Application of DNA machine in amplified DNA detection.
A DNA machine finds its application to construct an enzyme-free, label-free, cost-effective and novel approach for amplified DNA detection. The present approach is endowed with high sensitivity and selectivity. A detection limit as low as 0.2 nM was obtained by practical measurement.


Apoptosis in skin cancer development and regression.
Non-melanoma skin cancers (NMSC) are the most common malignant tumors in white population and their incidence has been increasing worldwide. Molecular events regulating cell survival, apoptosis, growth arrest as well as cell differentiation, are important contributors to the overall kinetics of benign and malignant cell growth and play a role in their development, progression and regression. Failure of these pathways can result in the loss of control over proliferation and lead to tumor development through the inactivation of tumor suppressor genes or the activation of oncogenes. Also, immunological mechanisms have been implicated in a phenomenon of tumor progression as well as spontaneous tumor regression. We have tried to summarize the main events in etiopatogenesis, development, progression and in some cases skin cancer regression. Further studies are needed to elucidate completely the details of apoptotic control in normal skin and determine factors resulting in apoptotic disbalance and disease.


Effectiveness of theta burst versus high-frequency repetitive transcranial magnetic stimulation in patients with depression (THREE-D): a randomised non-inferiority trial.
Treatment-resistant major depressive disorder is common; repetitive transcranial magnetic stimulation (rTMS) by use of high-frequency (10 Hz) left-side dorsolateral prefrontal cortex stimulation is an evidence-based treatment for this disorder. Intermittent theta burst stimulation (iTBS) is a newer form of rTMS that can be delivered in 3 min, versus 37·5 min for a standard 10 Hz treatment session. We aimed to establish the clinical effectiveness, safety, and tolerability of iTBS compared with standard 10 Hz rTMS in adults with treatment-resistant depression. In this randomised, multicentre, non-inferiority clinical trial, we recruited patients who were referred to specialty neurostimulation centres based at three Canadian university hospitals (Centre for Addiction and Mental Health and Toronto Western Hospital, Toronto, ON, and University of British Columbia Hospital, Vancouver, BC). Participants were aged 18-65 years, were diagnosed with a current treatment-resistant major depressive episode or could not tolerate at least two antidepressants in the current episode, were receiving stable antidepressant medication doses for at least 4 weeks before baseline, and had an HRSD-17 score of at least 18. Participants were randomly allocated (1:1) to treatment groups (10 Hz rTMS or iTBS) by use of a random permuted block method, with stratification by site and number of adequate trials in which the antidepressants were unsuccessful. Treatment was delivered open-label but investigators and outcome assessors were masked to treatment groups. Participants were treated with 10 Hz rTMS or iTBS to the left dorsolateral prefrontal cortex, administered on 5 days a week for 4-6 weeks. The primary outcome measure was change in 17-item Hamilton Rating Scale for Depression (HRSD-17) score, with a non-inferiority margin of 2·25 points. For the primary outcome measure, we did a per-protocol analysis of all participants who were randomly allocated to groups and who attained the primary completion point of 4 weeks. This trial is registered with ClinicalTrials.gov, number NCT01887782. Between Sept 3, 2013, and Oct 3, 2016, we randomly allocated 205 participants to receive 10 Hz rTMS and 209 participants to receive iTBS. 192 (94%) participants in the 10 Hz rTMS group and 193 (92%) in the iTBS group were assessed for the primary outcome after 4-6 weeks of treatment. HRSD-17 scores improved from 23·5 (SD 4·4) to 13·4 (7·8) in the 10 Hz rTMS group and from 23·6 (4·3) to 13·4 (7·9) in the iTBS group (adjusted difference 0·103 [corrected], lower 95% CI -1·16; p=0·0011), which indicated non-inferiority of iTBS. Self-rated intensity of pain associated with treatment was greater in the iTBS group than in the 10 Hz rTMS group (mean score on verbal analogue scale 3·8 [SD 2·0] vs 3·4 [2·0] out of 10; p=0·011). Dropout rates did not differ between groups (10 Hz rTMS: 13 [6%] of 205 participants; iTBS: 16 [8%] of 209 participants); p=0·6004). The most common treatment-related adverse event was headache in both groups (10 Hz rTMS: 131 [64%] of 204; iTBS: 136 [65%] of 208). In patients with treatment-resistant depression, iTBS was non-inferior to 10 Hz rTMS for the treatment of depression. Both treatments had low numbers of dropouts and similar side-effects, safety, and tolerability profiles. By use of iTBS, the number of patients treated per day with current rTMS devices can be increased several times without compromising clinical effectiveness. Canadian Institutes of Health Research.


Chemical properties of femoral gland secretions in the desert iguana,Dipsosaurus dorsalis.
This study investigates the chemistry of femoral gland secretions in the desert iguana,Dipsosaurus dorsalis (Lacertilia: Iguanidae), and discusses their possible functional significance. Electrophoretic and proton NMR studies indicated that the secretions are composed of approximately 80 % protein and 20% lipid material. Individual differences in polyacrylamide gel banding patterns of femoral gland proteins were found. Reflectance spectroscopy revealed that the secretions strongly absorb longwave ultraviolet light, a feature that may be important in the localization of secretion deposits in the environment.


Primitive nervous systems: electrophysiology of the pharynx of the polyclad flatworm, Enchiridium punctatum.
1. Electrical activity accompanying motor activity can be recorded from the excised pharynx of Enchiridium punctatum. Multiple stimuli elicit behaviour which consists of an initial aperture closure followed by extension and then peristalsis. If the stimulus parameters are increased the preparation bends from side to side instead of proceeding through the behavioural sequence. Bending appears to inhibit other movements differentially. 2. The conduction involved with peristalsis is polarized and proceeds in a proximal direction. 3. With stimulus intensities greater than those needed to produce the behavioural response an initial muscle potential (IMP) is evoked. The IMP is frequency sensitive. Maximum facilitation occurs within 100 ms and drops to 50% of maximum within 250 ms. 4. Conduction velocities of the IMP range from 0-05 m s-1 to 1-9 m s-1. Conduction velocities appear to increase with facilitation.


A phase IIb study to determine the safety and efficacy of candidate INfluenza Vaccine MVA-NP+M1 in combination with licensed Ina CTivated infl Uenza vaccine in adult S aged 65 years and above (INVICTUS): a study protocol.
Seasonal influenza has a significant annual global impact. Current influenza vaccines work by inducing strain-specific antibodies against the highly polymorphic surface proteins of the influenza virus and need to be redesigned every year, increasing their cost and limiting availability. There is a demand for a more efficacious vaccine, particularly in older adults in which the current vaccines show poor efficacy. The aim is to investigate a novel vaccine, MVA-NP+M1, which targets T cell responses to the nucleoprotein and matrix 1 core proteins of the influenza virus A, which are highly conserved, and therefore may provide long protection against a broad range of influenza strains. INVICTUS is a phase IIb study to determine the safety and efficacy of candidate INfluenza Vaccine MVA-NP+M1 in combination with licensed Ina CTivated infl Uenza vaccine in adult S aged 65 years and above is a randomised, participant-blinded, placebo-controlled, multi-centre phase IIb efficacy study planned for 2030 volunteers aged 65 and over, in primary care. The primary objective is to assess the efficacy of MVA-NP+M1 co-administered with licensed inactivated quadrivalent influenza vaccine in adults ≥65 years. Participants complete daily diaries to record solicited and unsolicited events in the first four weeks post vaccination, and influenza-like illness (ILI) symptoms and severity throughout the influenza season. We hypothesise an improvement in the primary outcome, a reduction in the average number of days spent with moderate or severe influenza-like illness during periods of influenza circulation, in the group administered with MVA-NP+M1, compared to those in the control group. Registration: ClinicalTrials.gov identifier NCT03300362. Protocol version: INVICTUS Protocol v3.0, 08 June06 2018.