On the discrete-time dynamics of the basic Hebbian neural network node.
In this paper, the dynamical behavior of the basic node used for constructing Hebbian artificial neural networks (NNs) is analyzed. Hebbian NNs are employed in communications and signal processing applications, among others. They have been traditionally studied on a continuous-time formulation whose validity is justified via some analytical procedures that presume, among other hypotheses, a specific asymptotic behavior of the learning gain. The main contribution of this paper is the study of a deterministic discrete-time (DDT) formulation that characterizes the average evolution of the node, preserving the discrete-time form of the original network and gathering a more realistic behavior of the learning gain. The new deterministic discrete-time model provides some unstability results (critical for the case of large similar variance signals) which are drastically different to the ones known for the continuous-time formulation. Simulation examples support the presented results, illustrating the practical limitations of the basic Hebbian model.


How to get rid of mitochondria: crosstalk and regulation of multiple mitophagy pathways.
Mitochondria are indispensable cellular organelles providing ATP and numerous other essential metabolites to ensure cell survival. Reactive oxygen species (ROS), which are formed as side reactions during oxidative phosphorylation or by external agents, induce molecular damage in mitochondrial proteins, lipids/membranes and DNA. To cope with this and other sorts of organellar stress, a multi-level quality control system exists to maintain cellular homeostasis. One critical level of mitochondrial quality control is the removal of damaged mitochondria by mitophagy. This process utilizes parts of the general autophagy machinery, e.g. for the formation of autophagosomes but also employs mitophagy-specific factors. Depending on the proteins utilized mitophagy is divided into receptor-mediated and ubiquitin-mediated mitophagy. So far, at least seven receptor proteins are known to be required for mitophagy under different experimental conditions. In contrast to receptor-mediated pathways, the Pink-Parkin-dependent pathway is currently the best characterized ubiquitin-mediated pathway. Recently two additional ubiquitin-mediated pathways with distinctive similarities and differences were unraveled. We will summarize the current state of knowledge about these multiple pathways, explain their mechanism, and describe the regulation and crosstalk between these pathways. Finally, we will review recent evidence for the evolutionary conservation of ubiquitin-mediated mitophagy pathways.


PEtab-Interoperable specification of parameter estimation problems in systems biology.
Reproducibility and reusability of the results of data-based modeling studies are essential. Yet, there has been-so far-no broadly supported format for the specification of parameter estimation problems in systems biology. Here, we introduce PEtab, a format which facilitates the specification of parameter estimation problems using Systems Biology Markup Language (SBML) models and a set of tab-separated value files describing the observation model and experimental data as well as parameters to be estimated. We already implemented PEtab support into eight well-established model simulation and parameter estimation toolboxes with hundreds of users in total. We provide a Python library for validation and modification of a PEtab problem and currently 20 example parameter estimation problems based on recent studies.


Can Clean Delivery Kits Prevent Infections? Lessons from Traditional Birth Attendants in Nigeria.
In resource-poor settings, perinatal infections contribute significantly to maternal and neonatal deaths, and the use of clean delivery kits (CDKs) has been proposed as a tool to reduce the risk of infection-related deaths. This study aims to assess the acceptability and effectiveness of CDKs in preventing infections in deliveries attended by traditional birth attendants (TBAs) in Abeokuta, Nigeria. The study was a cluster-randomized trial with 67 birth centres/clusters, 453 births/mothers, and 457 babies randomized to intervention or control arms; intervention involved supplementation of delivery with JANMA CDKs. Interviews were conducted at the birth homes, and the primary outcomes were neonatal infection and puerperal fever. The association between infection and perinatal risk factors was tested using the Chi-square and Fisher's exact tests. CDKs were well accepted by TBAs. The incidence of puerperal fever and neonatal infection was 1.1% and 11.2%, respectively. Concurrent infection was found in 1 (0.22%) of the mother-neonate pair. There was no significant association between any of the sociodemographic factors and infection for both mothers and neonates. PROM and prolonged labour were significantly associated with puerperal infection. All mothers with puerperal fever were from the control group. Compared to the control group, the relative risk of puerperal infection and neonatal infection in the intervention group was 0.08 (0.004 -1.35, p = 0.079) and 0.64 (0.37 to 1.1, p = 0.10), respectively. CDKs hold promising results in attenuating maternal infections in resource-poor settings. Larger studies with greater statistical power are required to establish statistically reliable information.


Ultrastructural and functional features of the developing mammalian heart: a brief overview.
The heart undergoes marked ultrastructural alterations during fetal and postnatal development. Early in fetal development, cardiac myocytes contain abundant pools of glycogen, scattered mitochondria and sparse, peripheral myofibrils. Transverse tubules are absent, and sarcoplasmic reticulum and intercalated discs are poorly developed. During late fetal and early postnatal development, myofibrils extend into the myocyte interior and attain a mature appearance, and the glycogen pools are reduced in size. In addition, transverse tubules develop and the morphological appearance of the sarcoplasmic reticulum and intercalated disc becomes increasingly complex. Experimental studies in sheep, corroborated by clinical studies in humans, also point to marked functional changes during development. In the fetus, the right ventricle is the dominant pumping chamber because right ventricular output exceeds left ventricular output, while pulmonary arterial and aortic pressures are similar. This functional difference is reflected in myocardial blood flow patterns, with blood flow to the right ventricle exceeding that to the left ventricle. The ventricular outputs equalize after birth, but a functional left ventricular dominance rapidly emerges following a postnatal increase in systemic vascular resistance and a decrease in pulmonary vascular resistance. This postnatal switchover in functional dominance is accompanied by a corresponding alteration in the relative level of ventricular myocardial blood flows. Consistent with right ventricular dominance in utero, myocytes in the right ventricle of the fetal sheep are larger and contain more myofibrillar material than those in the left ventricle. Left ventricular myocytes become larger than right ventricular myocytes after birth, but this adaptation to altered postnatal haemodynamics requires some weeks to become fully established.


Sativex® effects on promoter methylation and on CNR1/CNR2 expression in peripheral blood mononuclear cells of progressive multiple sclerosis patients.
Multiple sclerosis (MS) is a chronic demyelinating central nervous system (CNS) disease that involve oligodendrocyte loss and failure to remyelinate damaged brain areas causing a progressive neurological disability. Studies in MS mouse model suggest that cannabinoids ameliorate symptoms as spasticity, tremor and pain reducing inflammation via cannabinoid-mediated system. The aim of our study is to investigate the changes in cannabinoid type 1 (CNR1) and 2 (CNR2) receptors mRNA expression levels and promoter methylation in peripheral blood mononuclear cells (PBMCs) of MS secondary progressive (MSS-SP) patients treated with Sativex®. Our cohort included MSS-SP patients, that at the time of Sativex® treatment, are treated (n=7), not treated (n=11) or that had terminated interferon-β-1b (IFN-β-1b) therapy (n=12). By Methylation Sensitive High Resolution Melting (MS-HRM), we characterized the methylation profile of CNR1 and CNR2 promoter region, while the relative mRNA transcript levels of these two genes were evaluated in the same samples by Quantitative Real-Time PCR (qRT-PCR) analysis. We did not find different pattern of cytosine-phosphate-guanine (CpG) methylation in the CNR1/CNR2 promoter region of all MSS-SP patients treated with Sativex®. In addition, CNR1 and CNR2 expression did not significantly differ in MSS-SP patients not treated with IFN-β-1b vs. them that have suspended, while in MSS-SP patients treated with IFN-β-1b during Sativex® therapy we found a specific decrease of the CNR2 expression levels. These results suggest that the different expression of cannabinoid receptors by Sativex® treatment in leukocytes might be regulated through a molecular mechanism that involve interferon modulation.


Evolutionary diversification of the flowers in angiosperms.
Angiosperms and their flowers have greatly diversified into an overwhelming array of forms in the past 135 million years. Diversification was shaped by changes in climate and the biological environment (vegetation, interaction with other organisms) and by internal structural constraints and potentials. This review focuses on the development and structural diversity of flowers and structural constraints. It traces floral diversification in the different organs and organ complexes (perianth, androecium, gynoecium) through the major clades of extant angiosperms. The continuously improved results of molecular phylogenetics provide the framework for this endeavor, which is necessary for the understanding of the biology of the angiosperms and their flowers. Diversification appears to work with innovations and modifications of form. Many structural innovations originated in several clades and in special cases could become key innovations, which likely were hot spots of diversification. Synorganization between organs was an important process to reach new structural levels, from which new diversifications originated. Complexity of synorganization reached peaks in Orchidaceae and Apocynaceae with the independent evolution of pollinaria. Such a review throughout the major clades of angiosperms also shows how superficial and fragmentary our knowledge on floral structure in many clades is. Fresh studies and a multidisciplinary approach are needed.


Dried blood spot analysis for elements of nutritional concern as demonstrated in studies of Galápagos land iguanas (Conolophus species).
Dried blood spot (DBS) technology is valuable in providing simple means of storing blood samples from wildlife with small blood volumes. Methods designed for heavy metal analysis on DBS become more useful if extended to elements of nutritional significance. (1) Development of procedures for measuring Mn, Fe, Co, Cu, Zn, Se and Mo in DBS; (2) use the designed methods in health assessments of Galápagos land iguanas (Conolophus species). Elements were measured by inductively coupled plasma/mass spectrometry (ICP-MS) following acid digestion of whole blood or DBS from the same animal for direct comparison. Study animals comprised free-ranging iguanas from separate islands in the Galápagos archipelago. DBS spikes (Mn, Fe, Co, Cu, Zn, Se and Mo) demonstrated accuracy to ∼100 ppb; reporting limits were set there except for Fe and Zn which were set at 1000 ppb. Plasma samples - generally preferable for nutritional element diagnostics - were submitted from Galápagos land iguanas along with DBS as part of a large-scale health assessment. In plasma versus DBS concentration comparisons, Fe, Cu, Se and Mn correlated well with R^2 values of 0.799, 0.818, 0.896 and 0.899, respectively, and slopes ranging 0.88 - 1.3. Co and Zn showed greater scatter. Mo had insufficient points above its reporting limit and offered advantages for toxicity assessments. Bland-Altman diagrams showed flat scatter between 2x standard deviation boundaries with no undue trends except for Mn which had few points above its reporting limit. Bias, defined as the average difference [DBS - plasma] divided by the average value, was relatively low throughout, with values of - 19.3 % (Fe), - 48.7 % (Co), - 19.6 % (Cu), - 6.9 % (Zn), - 21.4 % (Se) and + 40.7 % (Mn). Normal distribution assessment of iguana Cu, Zn, Se and Fe plasma values showed unanticipated divergences between two species. The DBS approach for nutritional element analysis offers a suitable methodology for determining crucial elements Mn, Fe, Co, Cu, Zn, Se, and Mo in veterinary samples. Analyses of samples from Conolophus species revealed interesting divergences particularly for Cu, Zn, Se and Fe, elements generally associated with defense against oxidative stress.


Conventional and magnetization transfer MRI predictors of clinical multiple sclerosis evolution: a medium-term follow-up study.
The correlation between conventional MRI lesion load accumulation and multiple sclerosis clinical evolution is only modest. The assessment of brain parenchymal volume and of its changes over time may provide adjunctive MRI markers reflecting the more disabling aspects of multiple sclerosis pathology. Magnetization transfer (MT) MRI is sensitive to 'occult' multiple sclerosis-related brain damage and might also contribute to overcome the clinical/MRI paradox. In this study, we assessed the value of conventional and MT MRI-derived metrics in predicting the medium-term clinical evolution of patients with different multiple sclerosis phenotypes. We studied 73 patients, with relapsing-remitting multiple sclerosis (n = 34), secondary progressive multiple sclerosis (n = 19) and clinically isolated syndromes suggestive of multiple sclerosis (n = 20), and 16 healthy subjects. Brain dual-echo, T1-weighted (only in patients) and MT MRI scans were obtained at baseline and after 12 months. T2-hyperintense and T1-hypointense lesion volumes, normalized brain volume and average lesion MT ratio (MTR) were measured. MTR histograms from the whole brain tissue were also obtained. Clinical multiple sclerosis evolution and neurological disability were re-assessed in all patients after a median follow-up of 4.5 years. A multivariate analysis was performed to establish which clinical and MRI-derived variables were significant predictors of neurological deterioration at the end of the study period. At the end of follow-up, 34 patients showed significant neurological deterioration. The final multivariable model included average brain MTR percentage change after one year [P = 0.02, odds ratio (OR) = 0.86] and baseline T2-hyperintense lesion volume (P = 0.04, OR=1.04) as independent predictors of medium-term disability accumulation (r2 = 0.23). In this cohort of patients, abnormal values of average brain MTR changes showed a relatively high specificity (76.9%) and positive predictive value (59.1%) for Expanded Disability Status Scale score deterioration in individual cases. In patients with multiple sclerosis, a comprehensive estimation of the short-term changes of both conventional and MT MRI-detectable lesion burden might provide useful prognostic information for the medium-term clinical disease evolution.


Hormone therapy in epithelial ovarian cancer.
Although epidemiologic studies, animal experiments and receptor studies have shown that not only normal ovaries but also many malignant ovarian tumors can be considered as endocrine related and hormone dependent, the place of hormonal therapy in the management of patients with ovarian cancer remains unsettled. Most trials of hormonal treatment in ovarian cancer have been retrospective, involved only limited numbers of patients, and lacked important patient-related data and information pertaining to tumor characteristics. In addition, a variety of hormonal preparations with different degrees of potency and in different dosages were included in these studies. A literature review shows that response to hormonal therapy even in a preterminal setting, is modest, with about 8% objective response but almost no side effects. In a similar patient setting, more toxic therapeutic agents do not yield a better response. The place of hormonal therapy in the management of patients with epithelial ovarian cancer needs more thorough evaluation in well-designed randomized trials.


The ribosome-inactivating protein restrictocin deters insect feeding on Aspergillus restrictus.
The fungus-feeding beetle, Carpophilus freemani, consumed equal quantities of young mycelia, fewer phialides bearing mature spores and much fewer phialides bearing developing spores of Aspergillus restrictus compared to those of Aspergillus nidulans when tested in diet choice assays. The degree to which specific fungal structures were consumed was inversely related to the localization of high levels of restrictocin, a ribosome-inactivating protein, to those structures. Pure restrictocin added to the insect diet at 1000 p.p.m. killed 38.5% of C. freemani larvae and 62.5% of Spodoptera frugiperda larvae in 48 h, but did not affect C. freemani adults or Helicoverpa zea larvae over the same interval. In diet choice assays, 1000 p.p.m. of restrictocin deterred feeding by adult C. freemani and Sitophilus zeamais compared to control diets. Thus, restrictocin production and localization may have a natural defensive role against insect feeding at times critical to spore formation by A. restrictus, and may have potential as an insect control agent.


Symptoms of depression and anxiety among elite high school student-athletes in Sweden during the COVID-19 pandemic: A repeated cross-sectional study.
The COVID-19 pandemic precipitated numerous changes in daily life, including the cancellation and restriction of sports globally. Because sports participation contributes positively to the development of student-athletes, restricting these activities may have led to long-term mental health changes in this population. Using a repeated cross-sectional study design, we measured rates of depression using the Patient Health Questionnaire-2 and anxiety using the Generalized Anxiety Disorder-2 scale in student-athletes attending elite sport high schools in Sweden during the second wave of the pandemic (February 2021; n = 7021) and after all restrictions were lifted (February 2022; n = 6228). Depression among student-athletes decreased from 19.8% in 2021 to 17.8% in 2022 (p = .008, V = .026), while anxiety screening did not change significantly (17.4% to 18.4%, p > .05). Comparisons between classes across years revealed older students exhibited decreases in depressive symptoms, while younger cohorts experienced increases in symptoms of anxiety from 2021 to 2022. Logistic regressions revealed that being female, reporting poorer mental health due to COVID-19, and excessive worry over one's career in sports were significant predictors of both depression and anxiety screenings in 2022. Compared to times when sports participation was limited, the lifting of restrictions was associated with overall reduced levels of depression, but not anxiety.


Alcoholic Beverage Preference and Dietary Habits: A Systematic Literature Review.
The aim of this review is to systematically and critically evaluate the existing literature into the association between alcoholic beverage preference and dietary habits in adults. A literature search was conducted in the databases of Medline (PubMed), ISI Web of Knowledge, and PsycINFO for studies published up to March 2013. From a total of 4,345 unique hits, 16 articles were included in this systematic review. Two independent reviewers extracted relevant data for each study and assessed study quality. 14 cross-sectional and two ecological studies from the United States and several European countries were included. Across different study populations and countries, persons with a beer preference displayed in general less healthy dietary habits. A preference for wine was strongly associated with healthier dietary habits in Western study populations, whereas studies in Mediterranean populations did not observe this. Dietary habits of persons with another preference or who were abstinent were less reported. This review has shown that the preference for a specific alcoholic beverage is associated with diet. Thus, it might not be the alcoholic beverage but the underlying dietary patterns that are related to health outcomes.


Donor specific anti-HLA antibodies in hematopoietic stem cell transplantation. Single Center prospective evaluation and desensitization strategies employed.
In the setting of mismatched-hematopoietic stem cells transplantation, the detection of antibodies directed against donor-specific HLA allele(s) or antigen(s) (DSA) represents a barrier for engraftment. It is thus necessary to plan an immunosuppressive strategy, or to select an alternative donor. This prospective study aimed at evaluating the efficacy of our strategy for testing DSAs and the efficacy of the desensitization strategy (DS) employed between November 2017 and November 2020. The anti-HLA antibody search was performed using the Luminex bead assays (Lifecode ID and LSA I/II-Immucor) and expressed as mean fluorescence intensity (MFI >1,000 positive). If the patient had DSAs and no alternative donors, a DS was employed with rituximab (day -15), 2 single volume plasmaphereses (PP; days -9 and -8), intravenous immunoglobulins (day -7) and infusion of HLA selected platelets, if persistent DSAs were directed against class I HLA. DS was scheduled with or without PP, according to the DSA MFI (>1,000 or <5,000) and FCXM (flow cytometry crossmatch). Twenty-two out of 126 patients (17.46%) showed anti-HLA antibodies, 5 of them DSAs (3.97% of total); 3 patients underwent DS obtaining engraftment. Female gender (p=0.033) and a history of previous pregnancies or miscarriages (p=0.009) showed a statistically significant impact on alloimmunization. Factors associated with a delayed neutrophil engraftment were patient's female gender (p=0.039), stem cell source (p=0.025), and a high HSCT-specific comorbidity index (p=0.028). None of the analyzed variables, including the DSA detection, influenced engraftment. Our study confirms the importance to test DSAs in mismatched-hematopoietic stem cells transplantation The DS used proved successful in removing DSAs. Prospective multicenter studies are needed to better define and validate consensus strategies on DSA management in HSCT.


The influence of personal communities on the self-management of medication taking: A wider exploration of medicine work.
There is a lack of focus on the broader social context, networks and influences on medicine-taking as part of illness work. This work adopts a social network approach and seeks to explicate the nature of medicine-taking work that people with multiple long-term conditions (LTCs) and their social network members (SNMs) do in attempting to take their medications on a daily basis, the division of labour amongst these members and when and why SNMs become involved in that work. Semi-structured interviews were conducted with 20 people who had multiple LTCs. Medication networks were constructed and the division of labour in relation to medication-work was explored. Four types of medication-work emerged: medication articulation, surveillance, emotional and informational. Involvement of SNMs in medication-work was selective, performed primarily by family members, within the home. Involvement reflected network composition and/or an individual's conceptualisation/presentation of self. Our findings support and extend the conceptualisation of routine medicine-taking as a type of work. Furthermore, we illustrate the involvement of SNMs in aspects of medicine-work. Health professionals should explore and support the role of SNMs in medicine-taking where possible. Future research should explore the implications of network types and compositions on medicine-taking and associated work.


7-Nitroindazole attenuates vasodilation from cerebellar parallel fiber stimulation but not acetylcholine.
We used the relatively selective inhibitor of neuronal nitric oxide synthase 7-nitroindazole (7-NI) to test the hypothesis that the increases in local cerebellar blood flow (BFcrb) elicited by activation of the cerebellar parallel fibers (PF) are mediated by neuronal production of nitric oxide. In halothane-anesthetized rats, the cerebellar cortex was exposed and superfused with Ringer solution (37 degrees C; pH 7.3-7.4). The PF were stimulated electrically (100 microA, 30 Hz, 40 s), while BFcrb was monitored at the site of stimulation by a laser-Doppler flow probe. In vehicle-treated rats (n = 5), PF stimulation increased BFcrb by 61 +/- 5% (P < 0.05; analysis of variance and Tukey's test). 7-NI attenuated the increase in BFcrb dose dependently (10-100 mg/kg i.p.; n = 5 animals/dose) and by 55 +/- 7% at 100 mg/kg (P < 0.05). The attenuation of the response to PF stimulation was correlated with the degree of inhibition of calcium-dependent brain nitric oxide synthase activity, measured ex vivo by the citrulline assay (n = 21). 7-NI also attenuated the cerebrovasodilation elicited by hypercapnia (PCO2 = 50-60 mmHg) but did not affect the vasodilation evoked by acetylcholine (10 microM; n = 4; P > 0.05; t-test), a response mediated by endothelial nitric oxide synthase. 7-NI did not attenuate the BFcrb increase evoked by the nitric oxide donor S-nitroso-N-acetylpenicillamine (1 mM; n = 5; P > 0.05; t-test). Similarly, 7-NI did not affect resting systemic arterial pressure. These observations suggest that selective inhibition of neuronal nitric oxide synthase by 7-NI attenuates the increases in BFcrb evoked by PF stimulation. The findings provide additional support to the hypothesis that the increase in BFcrb evoked by PF stimulation is mediated, in part, by glutamate-induced activation of neuronal nitric oxide synthase.


The Michael Mason prize: early rheumatoid arthritis--the window narrows.
RA is a chronic disease in which synovitis drives joint destruction. Immunomodulatory therapy in the established phase of disease limits synovitis, and slows the rate of joint destruction, but is not curative. Increasing evidence suggests that the very early phase of RA, within the first few months after the onset of symptoms, represents a pathologically distinct and temporally transient window during which outcomes can be more effectively modulated by therapy. Furthermore, recent data show that we can accurately predict the development of RA in patients with very early synovitis, using clinical and serological measures. This makes very early targeted treatment a realistic possibility. However, it remains the case that the majority of patients with very early synovitis delay for prolonged periods before seeking medical help. Effective public engagement, to reduce this delay, is the key to translate advances in the fields of pathology, prognostication and therapy into benefit for patients with new onset RA.


Quality improvement in health care--a fundamental analysis and South African perspective.
This article is based on the opening address that was delivered at a national conference with the launching of the South African Society for Quality in Health Care (SASQuaH). The quality improvement process and principles are analyzed, as well as an exposition of the South African situation. The concepts regarding quality, quality improvement and quality assurance are clarified, the different stakeholders in formalised quality improvement explained, as well as the motives for and principles of quality improvement in health services. The different external and internal mechanisms in quality improvement are analyzed, including the principle of community ownership and international collaboration. Continuous development and capacity building regarding quality improvement is recommended at all levels of health service delivery.


Vaginal estrogen and mammogram results: case series and review of literature on treatment of genitourinary syndrome of menopause (GSM) in breast cancer survivors.
To examine mammographic density before and after at least 1 year of vaginal estrogen use in a small cohort of healthy postmenopausal women and women with a personal history of breast cancer. We extracted data via chart review of patients from a single practitioner's menopause specialty clinic in Baltimore, MD. Mammographic change was primarily determined via the Bi-RADS scoring system, including the Bi-RADS density score. In addition, we conduct a narrative review of the current literature on the usage of local estrogen therapy, and systemic and local alternatives in the treatment of genitourinary syndrome of menopause (GSM) in breast cancer survivors. Twenty healthy postmenopausal women and three breast cancer survivors fit our inclusion criteria. Amongst these two groups, we did not find an increase in mammographic density after at least 1 year and up to 18 years of local vaginal estrogen. Ospemifene use in one patient did not appear to be associated with any change in Bi-RADS score. Our narrative review found little data on the effects of vaginal estrogen therapy or newer alternative systemic therapies such as ospemifene on mammographic density. Low-dose vaginal estrogen use for 1 or more years in a small cohort of women with GSM did not appear to be associated with any changes in breast density or Bi-RADS breast cancer risk scores in the majority of study participants, including three breast cancer survivors. Larger long-term controlled clinical trials should be conducted to examine the effects of low-dose vaginal estrogen on mammographic density in women with and without a personal history of breast cancer. Furthermore, relative efficacy and risk of vaginal estrogen compared with other forms of treatment for GSM should also be studied in long-term trials.


Early determinants of H2O2-induced endothelial dysfunction.
Reactive oxygen species (ROS) can stimulate nitric oxide (NO(*)) production from the endothelium by transient activation of endothelial nitric oxide synthase (eNOS). With continued or repeated exposure, NO(*) production is reduced, however. We investigated the early determinants of this decrease in NO(*) production. Following an initial H(2)O(2) exposure, endothelial cells responded by increasing NO(*) production measured electrochemically. NO(*) concentrations peaked by 10 min with a slow reduction over 30 min. The decrease in NO(*) at 30 min was associated with a 2.7-fold increase in O(2)(*-) production (p < 0.05) and a 14-fold reduction of the eNOS cofactor, tetrahydrobiopterin (BH(4), p < 0.05). Used as a probe for endothelial dysfunction, the integrated NO(*) production over 30 min upon repeated H(2)O(2) exposure was attenuated by 2.1-fold (p = 0.03). Endothelial dysfunction could be prevented by BH(4) cofactor supplementation, by scavenging O(2)(*-) or peroxynitrite (ONOO(-)), or by inhibiting the NADPH oxidase. Hydroxyl radical (()OH) scavenging did not have an effect. In summary, early H(2)O(2)-induced endothelial dysfunction was associated with a decreased BH(4) level and increased O(2)(*-) production. Dysfunction required O(2)(*-), ONOO(-), or a functional NADPH oxidase. Repeated activation of the NADPH oxidase by ROS may act as a feed forward system to promote endothelial dysfunction.


NONLINEAR AUTOREGRESSIVE LATENT TRAJECTORY MODELS.
Autoregressive latent trajectory (ALT) models combine features of latent growth curve models and autoregressive models into a single modeling framework. The development of ALT models has focused primarily on models with linear growth components, but some social processes follow nonlinear trajectories. Although it is straightforward to extend ALT models to allow for some forms of nonlinear trajectories, the identification status of such models, approaches to comparing them with alternative models, and the interpretation of parameters have not been systematically assessed. In this paper we focus on two forms of nonlinear autoregressive latent trajectory (NLALT) models. The first form allows for a quadratic growth trajectory, a popular form of nonlinear latent growth curve models. The second form derives from latent basis models, or freed loading models, that allow for arbitrary growth processes. We discuss details concerning parameterization, model identification, estimation, and testing for the two forms of NLALT models. We include a simulation study that illustrates potential biases that may arise from fitting alternative models to data derived from an autoregressive process and individual-specific nonlinear trajectories. In addition, we include an extended empirical example modeling growth trajectories of weight from birth through age 2.


A species-specific DNA probe obtained from Streptococcus salivarius subsp. thermophilus detects strain restriction polymorphism.
Genomic polymorphism in Streptococcus salivarius subsp. thermophilus was revealed by DNA restriction pattern analysis. A 4.2-kb variable DNA fragment was cloned from strain NST7 and hybridised with the DNA of 25 strains allowing an easy detection of intraspecific RFLP. Strong and weak hybridisation signals were observed and the latter were specifically revealed by a 2.1-kb fragment of the probe. Probe specificity was demonstrated by the absence of homology with DNA of strains belonging to 10 other species, with the exception of S. salivarius subsp. salivarius, confirming a close relationship between S. salivarius and S. thermophilus.


Ultrashort pulse polarization control in silicon waveguides.
The nonlinear polarization dynamics of ultrashort optical pulses propagating in a low birefringent silicon waveguide is theocratically and numerically studied, with a static electric field applied across the waveguide. It is shown that the pulse shape and polarization evolution can be efficiently controlled by adjusting the magnitude of the applied dc field. It is also demonstrated that the polarization instability regime can be achieved in such waveguides - despite the presence of strong linear losses - by appropriately engineering the spatial distribution of the control field along the waveguide. The simulations indicate that short silicon waveguides can serve as a viable platform for developing re-configurable all-optical and/or optically assisted electro-optic devices in the spectral range spanning from near- to mid-infrared.


Current Status and Future of Gene Engineering in Livestock.
The application of gene engineering in livestock is necessary for various reasons, such as increasing productivity and producing disease resistance and biomedicine models. Overall, gene engineering provides benefits to the agricultural and research aspects, and humans. In particular, productivity can be increased by producing livestock with enhanced growth and improved feed conversion efficiency. In addition, the application of the disease resistance models prevents the spread of infectious diseases, which reduces the need for treatment, such as the use of antibiotics; consequently, it promotes the overall health of the herd and reduces unexpected economic losses. The application of biomedicine could be a valuable tool for understanding specific livestock diseases and improving human welfare through the development and testing of new vaccines, research on human physiology, such as human metabolism or immune response, and research and development of xenotransplantation models. Gene engineering technology has been evolving, from random, time-consuming, and laborious methods to specific, time-saving, convenient, and stable methods. This paper reviews the overall trend of genetic engineering technologies development and their application for efficient production of genetically engineered livestock, and provides examples of technologies approved by the United States (US) Food and Drug Administration (FDA) for application in humans.


The perceptual learning of time-compressed speech: A comparison of training protocols with different levels of difficulty.
Speech perception can improve substantially with practice (perceptual learning) even in adults. Here we compared the effects of four training protocols that differed in whether and how task difficulty was changed during a training session, in terms of the gains attained and the ability to apply (transfer) these gains to previously un-encountered items (tokens) and to different talkers. Participants trained in judging the semantic plausibility of sentences presented as time-compressed speech and were tested on their ability to reproduce, in writing, the target sentences; trail-by-trial feedback was afforded in all training conditions. In two conditions task difficulty (low or high compression) was kept constant throughout the training session, whereas in the other two conditions task difficulty was changed in an adaptive manner (incrementally from easy to difficult, or using a staircase procedure). Compared to a control group (no training), all four protocols resulted in significant post-training improvement in the ability to reproduce the trained sentences accurately. However, training in the constant-high-compression protocol elicited the smallest gains in deciphering and reproducing trained items and in reproducing novel, untrained, items after training. Overall, these results suggest that training procedures that start off with relatively little signal distortion ("easy" items, not far removed from standard speech) may be advantageous compared to conditions wherein severe distortions are presented to participants from the very beginning of the training session.


Activation of pyruvate dehydrogenase by dichloroacetate has the potential to induce epigenetic remodeling in the heart.
Dichloroacetate (DCA) promotes pyruvate entry into the Krebs cycle by inhibiting pyruvate dehydrogenase (PDH) kinase and thereby maintaining PDH in the active dephosphorylated state. DCA has recently gained attention as a potential metabolic-targeting therapy for heart failure but the molecular basis of the therapeutic effect of DCA in the heart remains a mystery. Once-daily oral administration of DCA alleviates pressure overload-induced left ventricular remodeling. We examined changes in the metabolic fate of pyruvate carbon (derived from glucose) entering the Krebs cycle by metabolic interventions of DCA. (13)C6-glucose pathway tracing analysis revealed that instead of being completely oxidized in the mitochondria for ATP production, DCA-mediated PDH dephosphorylation results in an increased acetyl-CoA pool both in control and pressure-overloaded hearts. DCA induces hyperacetylation of histone H3K9 and H4 in a dose-dependent manner in parallel to the dephosphorylation of PDH in cultured cardiomyocytes. DCA administration increases histone H3K9 acetylation in in vivo mouse heart. Interestingly, DCA-dependent histone acetylation was associated with an up-regulation of 2.3% of genes (545 out of 23,474 examined). Gene ontology analysis revealed that these genes are highly enriched in transcription-related categories. This evidence suggests that sustained activation of PDH by DCA results in an overproduction of acetyl-CoA, which exceeds oxidation in the Krebs cycle and results in histone acetylation. We propose that DCA-mediated PDH activation has the potential to induce epigenetic remodeling in the heart, which, at least in part, forms the molecular basis for the therapeutic effect of DCA in the heart.


Inactivation of arf-bp1 induces p53 activation and diabetic phenotypes in mice.
It is well accepted that the Mdm2 ubiquitin ligase acts as a major factor in controlling p53 stability and activity in vivo. Although several E3 ligases have been reported to be involved in Mdm2-independent p53 degradation, the roles of these ligases in p53 regulation in vivo remain largely unknown. To elucidate the physiological role of the ubiquitin ligase ARF-BP1, we generated arf-bp1 mutant mice. We found that inactivation of arf-bp1 during embryonic development in mice resulted in p53 activation and embryonic lethality, but the mice with arf-bp1 deletion specifically in the pancreatic β-cells (arf-bp1(FL/Y)/RIP-cre) were viable and displayed no obvious abnormality after birth. Interestingly, these mice showed dramatic loss of β-cells as mice aged, and >50% of these mice died of severe diabetic symptoms before reaching 1 year of age. Notably, the diabetic phenotype of these mice was largely reversed by concomitant deletion of p53, and the life span of the mice was significantly extended (p53(LFL/FL)/arf-bp1(FL/Y)/RIP-cre). These findings underscore an important role of ARF-BP1 in maintaining β-cell homeostasis in aging mice and reveal that the stability of p53 is critically regulated by ARF-BP1 in vivo.


Anharmonic Densities of States for Vibrationally Excited I-(H2O), (H2O)2, and I-(H2O)2.
Monte Carlo sampling calculations were performed to determine the anharmonic sum of states, Nanh( E), for I-(H2O), (H2O)2, and I-(H2O)2 versus internal energy up to their dissociation energies. The anharmonic density of states, ρanh( E), is found from the energy derivative of Nanh( E). Analytic potential energy functions are used for the calculations, consisting of TIP4P for H2O···H2O interactions and an accurate two-body potential for the I-···H2O fit to quantum chemical calculations. The extensive Monte Carlo samplings are computationally demanding, and the use of computationally efficient potentials was essential for the calculations. Particular emphasis is directed toward I-(H2O)2, and distributions of its structures versus internal energy are consistent with experimental studies of the temperature-dependent vibrational spectra. At their dissociation thresholds, the anharmonic to harmonic density of states ratio, ρanh( E)/ρh( E), is ∼2, ∼ 3, and ∼260 for I-(H2O), (H2O)2, and I-(H2O)2, respectively. The large ratio for I-(H2O)2 results from the I-(H2O)2 → I-(H2O) + H2O dissociation energy being more than 2 times larger than the (H2O)2 → 2H2O dissociation energy, giving rise to highly mobile H2O molecules near the I-(H2O)2 dissociation threshold. This work illustrates the importance of treating anharmonicity correctly in unimolecular rate constant calculations.


Potential complementarity between forest carbon sequestration incentives and biomass energy expansion.
There is a growing literature on the potential contributions the global forest sector could make toward long-term climate action goals through increased carbon sequestration and the provision of biomass for energy generation. However, little work to date has explored possible interactions between carbon sequestration incentives and bioenergy expansion policies in forestry. This study develops a simple conceptual model for evaluating whether carbon sequestration and biomass energy policies are carbon complements or substitutes. Then, we apply a dynamic structural model of the global forest sector to assess terrestrial carbon changes under different combinations of carbon sequestration price incentives and forest bioenergy expansion. Our results show that forest bioenergy expansion can complement carbon sequestration policies in the near- and medium-term, reducing marginal abatement costs and increasing mitigation potential. By the end of the century these policies become substitutes, with forest bioenergy expansion increasing the costs of carbon sequestration. This switch is driven by relatively high demand and price growth for pulpwood under scenarios with forest bioenergy expansion, which incentivizes management changes in the near- and medium-term that are carbon beneficial (e.g., afforestation and intensive margin shifts), but requires sustained increases in pulpwood harvest levels over the long-term.


A reference equation for maximal aerobic power for treadmill and cycle ergometer exercise testing: Analysis from the FRIEND registry.
Background Maximal oxygen uptake (VO2max) is a powerful predictor of health outcomes. Valid and portable reference values are integral to interpreting measured VO2max; however, available reference standards lack validation and are specific to exercise mode. This study was undertaken to develop and validate a single equation for normal standards for VO2max for the treadmill or cycle ergometer in men and women. Methods Healthy individuals ( N = 10,881; 67.8% men, 20-85 years) who performed a maximal cardiopulmonary exercise test on either a treadmill or a cycle ergometer were studied. Of these, 7617 and 3264 individuals were randomly selected for development and validation of the equation, respectively. A Brazilian sample (1619 individuals) constituted a second validation cohort. The prediction equation was determined using multiple regression analysis, and comparisons were made with the widely-used Wasserman and European equations. Results Age, sex, weight, height and exercise mode were significant predictors of VO2max. The regression equation was: VO2max (ml kg-1 min-1) = 45.2 - 0.35*Age - 10.9*Sex (male = 1; female = 2) - 0.15*Weight (pounds) + 0.68*Height (inches) - 0.46*Exercise Mode (treadmill = 1; bike = 2) ( R = 0.79, R2 = 0.62, standard error of the estimate = 6.6 ml kg-1 min-1). Percentage predicted VO2max for the US and Brazilian validation cohorts were 102.8% and 95.8%, respectively. The new equation performed better than traditional equations, particularly among women and individuals ≥60 years old. Conclusion A combined equation was developed for normal standards for VO2max for different exercise modes derived from a US national registry. The equation provided a lower average error between measured and predicted VO2max than traditional equations even when applied to an independent cohort. Additional studies are needed to determine its portability.


Resting state functional connectivity correlates of emotional awareness.
Multiple neuroimaging studies have now linked emotional awareness (EA), as measured by the Levels of Emotional Awareness Scale (LEAS), with activation in regions of neural networks associated with both conceptualization (i.e., default mode network [DMN] regions) and interoception (i.e., salience network [SN] regions) - consistent with the definition of EA as one's ability to appropriately recognize, conceptualize, and articulate the emotions of self and other in fine-grained, differentiated ways. However, no study has yet tested the hypothesis that greater LEAS scores are associated with greater resting state functional connectivity (FC) within these networks. Twenty-six adults (13 female) underwent resting state functional magnetic resonance imaging, and also completed the LEAS. Using pre-defined functional ROIs from the DMN and SN, we observed that LEAS scores were significantly positively correlated with FC between several regions of both of these networks, even when controlling for differences in general intelligence (IQ). These results suggest that higher EA may be associated with more efficient information exchange between brain regions involved in both interoception- and conceptualization-based processing, which could plausibly contribute to more differentiated bodily feelings and more fine-grained conceptualization of those feelings.


Somatostatin receptor subtypes in human type 2 diabetic islets.
Somatostatin inhibits hormone release through 5 G protein-coupled somatostatin receptors (sst1-sst5). However, the role of somatostatin in islet physiology is not fully known. The immunoreactivity to sst1 to sst5 in normal human endocrine pancreas has been described. The present study reports the expression of sst1 to sst5 in human pancreatic islets with type 2 diabetes mellitus. Pancreatic autopsy specimens from individuals with type 2 diabetes mellitus and matched controls were double immunostained to demonstrate sst1 to sst5 in the major islet cell types. Most apparent differences in type 2 diabetic islets were the lack of sst1 and sst4 in glucagon cells and sst1-3 and 4 in somatostatin cells, whereas minor changes were demonstrated in insulin cells. The pancreatic polypeptide cells showed a reversed staining pattern in diabetic islets compared with the controls. In type 2 diabetes mellitus, the sst pattern differed from that of the controls in somatostatin, pancreatic polypeptide, and glucagon cells, to a minor extent in insulin cells. It is unclear whether the changes in sst patterns are primarily due to the diabetes or secondary to metabolic disturbances. However, this study may be the basis for further functional studies to evaluate the role of sst1 to sst5 in the diabetic state.


New-onset of postoperative atrial fibrillation is likely to recur in the absence of other triggers.
Incident atrial fibrillation (AF) is reported in 10%-65% of patients without previous AF diagnosis after open heart surgery. The risk of late AF recurrence after a postoperative AF onset is unclear, and it is controversial whether AF limited to the postoperative period should elicit oral anticoagulation (OAC) therapy. The primary objective of this study was to evaluate the long-term recurrence of AF in patients developing new-onset peri-procedural AF. Patients (n=189) with available baseline and follow-up data included in Left Atrial Appendage Closure with Surgery trial were coded for known AF at baseline and for postoperative first-time AF diagnosis. AF occurrence was classified as follows: peri-procedural ≤7 days postoperatively, early >7 days but ≤3 months and late >3 months. Patients with no AF recurrence registered during follow-up were invited to undergo Holter monitoring. A total of 163 (86.2%) patients had no history of AF. Among these, 80 (49.1%) developed new-onset peri-procedural AF. After a mean follow-up of 3.7±1.6 years, late AF occurred in 35 of the 80 (43.8%) patients who developed peri-procedural AF and in 6 additional patients (7.2%) who remained in sinus rhythm until discharge (hazard ratio [HR] 9.3, 95% CI 3.8-22.4, p<0.001). Patients with peri-procedural AF and early AF had 12.24 times higher risk of late AF (95% CI 4.76-31.45, p<0.001) as compared to the group with no postoperative AF. New-onset of AF after open heart surgery has a high rate of recurrence and should not be regarded as a self-limiting phenomenon secondary to surgery.


Breast cancer and the brain: a neurodevelopmental hypothesis to explain the opposing effects of caloric deprivation during the Dutch famine of 1944-1945 on breast cancer and its risk factors.
Most studies on calorie deprivation and cancer risk in rodents show reductions in tumor occurrence. However, the few human studies on calorie restriction are conflicting. An overview is given of results in the DOM (diagnostic onderzoek mammacarcinoom) cohorts among women exposed to the Dutch Famine of 1944-1945. Opposing effects were found on risk factors (shortening of leg length, later menarche, and earlier menopause), whereas urinary estrogens and plasma insulin-like growth factor (IGF)-1 and IGF binding protein-3 were increased, as was breast cancer itself. Exposure between 2 and 10 y old was an unexpected window of susceptibility to the effects of calorie deprivation. The effects of famine exposure were most clearly seen in women who never gave birth. These opposing observations can be explained by a neurodevelopmental hypothesis on set-point shifts at the level of the diencephalons/hypothalamus, either directly or from rebound effects. Such a mechanism reflects old evolutionary adaptation systems in lower and higher organisms to cope with periods of stress and famine by adjusting, for example, reproductive functions. These effects in exposed women may later also affect their unexposed offspring. This hypothesis provides several testable, hormone-mediated corollaries on the relationships between the role of calories in a Westernized lifestyle and human cancer risk. The underlying developmental perspective, as opposed to a risk factor approach, can explain why certain ages, even before breast development, are especially sensitive to effects of large fluctuations in calories. The observations presented may have implications for preventive strategies such as promoting moderation of calorie intake to curb cancer risks.


Person-Centred, Culturally Appropriate Music Intervention to Improve Psychological Wellbeing of Residents with Advanced Dementia Living in Australian Rural Residential Aged Care Homes.
This quasi-experimental, nonrandomized intervention study reports the effect of person-centred, culturally appropriate music on psychological wellbeing of residents with advanced dementia in five rural residential aged care homes in Australia. Seventy-four residents attended in person-centred music sessions and culturally appropriate group sessions. Interest, response, initiation, involvement, enjoyment, and general reactions of the residents were assessed using the Music in Dementia Assessment Scale (MiDAS), and interviews and focus groups were conducted with aged care staff and musicians. The overall effect of person-centred sessions at two-time points were: during the intervention-351.2 (SD 93.5); and two-hours post intervention-315.1 (SD 98.5). The residents presented a moderate to high level of interest, response, initiation, involvement, and enjoyment during the session and at post-intervention. However, the MiDAS sub-categories' mean scores differed between the time-points: interest (t59 = 2.8, p = 0.001); response (t59 = 2.9, p = 0.005); initiation (t59 = 2.4, p = 0.019); and involvement (t59 = 2.8, p = 0.007), indicating a significant decline in the effect of person-centred music over time. Interestingly, during the period of time, most of the residents were observed with no exhibitions of agitation (87.5%), low in mood (87.5%), and anxiousness (70.3%), and with a presentation of relaxation (75.5%), attentiveness (56.5%), and smiling (56.9%). Themes from qualitative data collected regarding culturally appropriate group music sessions were behavioural change, meaningful interaction, being initiative, increased participation, and contentment. The findings suggest that the integration of music into care plans may reduce the residents' agitation and improve their emotional wellbeing in rural aged care homes.


Efficient distribution estimation for data with unobserved sub-population identifiers.
We study efficient nonparametric estimation of distribution functions of several scientifically meaningful sub-populations from data consisting of mixed samples where the sub-population identifiers are missing. Only probabilities of each observation belonging to a sub-population are available. The problem arises from several biomedical studies such as quantitative trait locus (QTL) analysis and genetic studies with ungenotyped relatives where the scientific interest lies in estimating the cumulative distribution function of a trait given a specific genotype. However, in these studies subjects' genotypes may not be directly observed. The distribution of the trait outcome is therefore a mixture of several genotype-specific distributions. We characterize the complete class of consistent estimators which includes members such as one type of nonparametric maximum likelihood estimator (NPMLE) and least squares or weighted least squares estimators. We identify the efficient estimator in the class that reaches the semiparametric efficiency bound, and we implement it using a simple procedure that remains consistent even if several components of the estimator are mis-specified. In addition, our close inspections on two commonly used NPMLEs in these problems show the surprising results that the NPMLE in one form is highly inefficient, while in the other form is inconsistent. We provide simulation procedures to illustrate the theoretical results and demonstrate the proposed methods through two real data examples.


[The symptomatic treatment of amyotrophic lateral sclerosis].
Patients with amyotrophic lateral sclerosis (ALS) have symptoms of progressive muscle weakness, of disturbed speech and swallowing, and in the terminal phase those of respiratory weakness. Treatment options, in particular those for excessive weight loss and respiratory weakness, should be introduced to the patients and their families when the patient is emotionally capable and before dysarthria severely hampers communication. Special equipment for keeping the patient as mobile as possible should be made available much earlier than in the case of other diseases of the muscles as in ALS progression is much faster. Cramps, pathological crying or laughter, spasms, and spasticity can all be treated by medication. When speech can no longer be understood, adaptive strategies such as sign language, mime, posture and communication apparatus varying from a note pad to advanced computer systems can be used. Sialorrhoea, caused by difficulty swallowing with its accompanying danger of aspiration can be halted by the use of medication, by radiotherapy and by the injection into the salivary glands of botulin A toxin. Weight loss, also a result of dysphagia, can be avoided by eating frequent small meals or if necessary performing a percutaneous endoscopic or radiological gastroscopy. Excess mucus in the respiratory tract can be treated with anticholinergics. Difficulty in coughing up thick and sticky mucus cannot always be adequately helped. Respiratory weakness is treatable by external respiratory supportive therapy using a nasal mask, as well as invasive respiratory support via a trachcostoma and by treating the symptoms of respiratory weakness. The latter form of treatment is palliative and forms part of terminal care. During the terminal phase restlessness, anxiety, pain, and dyspnoea require the most attention. Treatment requires careful multidisciplinary cooperation.


Photoactivated Refractive Index Anisotropy in Fluorescent Thiophene Derivatives.
The optical control of anisotropy in materials is highly advantageous for many technological applications, including the real-time modulation of another light signal in photonic switches and sensors. Here, we introduce three thiophene derivatives with a donor-acceptor structure, which feature different positions of an electron-acceptor nitrile group, and both photoalignment and luminescence properties. Quantum chemical calculations highlight the presence of trans-forms stable at room temperature and metastable cis-isomers. Besides photoluminescence peaked at 440-460 nm and 0.4 ns lifetime, the three nonlinear optical chromophores exhibit photoinduced anisotropy of the refractive index closely depending on the specific molecular structure, with higher values of birefringence at lower driving signal being obtained for ortho substitution of the nitrile group. All-optical modulation of an external light beam at rates of hundreds of hertz is demonstrated in the fluorescent systems. This finding opens an interesting route to multispectral photonic switches embedded in the active layers of light-emitting devices.


Differences between sexes in thermoregulatory responses and exercise time during endurance exercise in a hot environment following pre-cooling with ice slurry ingestion.
This study aimed to examine differences between sexes in thermoregulatory responses and exercise time after ice slurry ingestion in a hot environment. Twenty-four healthy adults (male n = 12, body weight (BW) = 65.8 ± 10.3; female n = 12, BW = 58.2 ± 10.0) ingested 7.5 g/kg of either ice slurry at -1 °C (ICE) or control water at 20 °C (CON) before cycling at 55%VO2 max in a hot environment (controlled at 38 °C, 40% relative humidity). Rectal (Tre) and skin (Tsk) temperature, heart rate, sweat rate, respiratory gases, ratings of thermal sensation (TS), thermal comfort (TC), and rating of perceived exertion (RPE) were measured. Ice slurry did not improve exercise time in both sexes despite Tre was significantly lower in ICE than CON in both sexes. Tre, Tsk, HR, sweat rate and TS did not differ between sexes. TC and RPE in ICE were significantly higher during exercise in males than in females. In conclusion, there were no sex differences in the effects of pre-cooling with ice slurry ingestion; however, pre-cooling with ice slurry may be more effective in mitigating ratings of TC and RPE in females than males.


Methylation Status of SP1 Sites within miR-23a-27a-24-2 Promoter Region Influences Laryngeal Cancer Cell Proliferation and Apoptosis.
DNA methylation plays critical roles in regulation of microRNA expression and function. miR-23a-27a-24-2 cluster has various functions and aberrant expression of the cluster is a common event in many cancers. However, whether DNA methylation influences the cluster expression and function is not reported. Here we found a CG-rich region spanning two SP1 sites in the cluster promoter region. The SP1 sites in the cluster were demethylated and methylated in Hep2 cells and HEK293 cells, respectively. Meanwhile, the cluster was significantly upregulated and downregulated in Hep2 cells and HEK293 cells, respectively. The SP1 sites were remethylated and the cluster was significantly downregulated in Hep2 cells into which methyl donor, S-adenosyl-L-methionine, was introduced. Moreover, S-adenosyl-L-methionine significantly increased Hep2 cell viability and repressed Hep2 cell early apoptosis. We also found that construct with two SP1 sites had highest luciferase activity and SP1 specifically bound the gene cluster promoter in vitro. We conclude that demethylated SP1 sites in miR-23a-27a-24-2 cluster upregulate the cluster expression, leading to proliferation promotion and early apoptosis inhibition in laryngeal cancer cells.


Autoradiographic localization of beta-1 and beta-2 adrenoceptors in guinea pig atrium and regions of the conducting system.
[125I]Cyanopindolol binding to slide mounted guinea pig atrial sections was time-dependent (Ki = 2.4 X 10(8) M-1 min-1, K-1 = 1.2 X 10(-3) min-1) saturable (24-32 pM) and stereoselective with respect to the isomers of propranolol. Competition binding curves with the beta-1 selective antagonist CGP 20712A and beta-2 selective antagonist ICI 118,551 revealed the presence of beta-1 and beta-2 adrenoceptors in the proportions of 85 to 15%. X-ray film exposed to sections of guinea pig heart incubated previously with [125I]cyanopindolol with or without ICI 118,551 (70 nM) or CGP 20712A (100 nM) or (-)-propranolol (1 microM) showed a high density and even distribution of beta-1 adrenoceptors and a lower density and even distribution of beta-2 adrenoceptors over the myocardium. Beta-2 adrenoceptors also were located in the epicardium, the adventitia of the pulmonary artery, aorta, superior and inferior vena cava and the endothelium of the superior and inferior vena cava. Development of nuclear emulsion-coated coverslips confirmed these observations and, in addition, localized beta-2 adrenoceptors to nerve tissue and the aortic valve. In the papillary muscle and surrounding Purkinje cells there was a high density and even distribution of beta-1 and a much lower density of beta-2 adrenoceptors. The atrioventricular node, atrioventricular bundle and ventricular Purkinje system also contained both beta-1 and beta-2 adrenoceptors and in comparison with the surrounding myocardium, these tissues had a slightly greater density of beta-2 adrenoceptors.


Physiologic and innate immunity responses to bacterial lipopolysaccharide administration in beef heifers supplemented with OmniGen-AF.
Nutritional alternatives to strengthen animal immunocompetence are critical for welfare and productivity in livestock systems, such as beef cattle operations. This experiment evaluated physiological and innate immunity effects of supplementing an immunomodulatory feed ingredient (Omnigen-AF; Phibro Animal Health, Teaneck, NJ, USA) to beef heifers administered bacterial lipopolysaccharide (LPS). In total, 8 non-pregnant, non-lactating nulliparous Angus×Hereford heifers (676±4 days of age) were ranked by BW (473±8 kg), and assigned to crossover design containing two periods of 34 days each. Heifers were housed in individual pens and had ad libitum access to meadow foxtail (Alopecurus pratensis L.) hay, water and a granulated commercial vitamin+mineral mix. Within each period, heifers received (as-fed basis) 227 g/day of dried distillers grains including (OMN) or not (CON) 56 g of Omnigen-AF for 34 days. On day 28 of each period (0800 h), heifers received an intravenous bolus dose (0.5 μg/kg of BW, diluted in 5 ml of 0.9% sterile saline) of bacterial LPS (Escherichia coli 0111:B4). Hay DM intake was recorded daily from day 0 to 34. Blood was collected at -1, 0, 0.5, 1, 1.5, 2, 2.5, 3, 4, 5, 6, 24, 48, 72, 96, 120 and 144 h relative to LPS administration. Heifer intravaginal temperature was recorded every 10 min from -0.5 to 10 h relative to LPS administration. No treatment effect was detected (P=0.35) for hay DM intake during the experiment. No treatment effects were detected (P⩾0.64) for intravaginal temperature and plasma concentrations of tumor necrosis-α, cortisol and haptoglobin, which increased (time effect, P<0.01) for OMN and CON heifers and peaked at 4.5, 2, 4 and 48 h, respectively, after LPS administration. No treatment effects were detected (P⩾0.35) for whole blood mRNA expression of chemokine ligand 5, tumor necrosis-α, cyclooxygenase 2 and interleukin 8, which also increased (time effect, P<0.01) for OMN and CON heifers and peaked at 0.5, 1.5, 2 and 2.5 h, respectively, after LPS administration. Whole blood mRNA expression of interleukin 8 receptor and L-selectin were also similar (P⩾0.61) between OMN and CON heifers, and decreased (time effect, P<0.01) for both treatments reaching nadir levels at 1 and 2.5 h, respectively, after LPS administration. Collectively, OMN supplementation did not modulate the physiological and innate immunity responses of beef heifers to bacterial LPS administration.


Using theory to interpret beliefs in migrants diagnosed with latent TB.
Tuberculosis (TB) is a serious health threat to migrant farm workers in the Midwestern United States. This article describes characteristics of migrant culture and lifestyle, economic, and health challenges that may impact screening, diagnosis, and adherence with complex medication regimens associated with TB. A brief overview of TB discusses the historical perspective of the disease and describes the stages, transmission, and incidence among migrant populations. Several theoretical models, such as the Health Belief Model (HBM) and social cognitive theory, were considered by the authors to guide understanding of migrant beliefs about TB. A qualitative research study conducted with 23 Hispanic migrants with latent TB infection is presented. Discussion of the research findings describes environmental, cognitive, and social factors that were barriers to screening, diagnosis, and treatment. The article concludes with a description of recent migrant health clinic updates designed to improve the worker' health status and considerations for environmental and educational change.


Walking improvement in chronic incomplete spinal cord injury with exoskeleton robotic training (WISE): a randomized controlled trial.
Clinical trial. To demonstrate that a 12-week exoskeleton-based robotic gait training regimen can lead to a clinically meaningful improvement in independent gait speed, in community-dwelling participants with chronic incomplete spinal cord injury (iSCI). Outpatient rehabilitation or research institute. Multi-site (United States), randomized, controlled trial, comparing exoskeleton gait training (12 weeks, 36 sessions) with standard gait training or no gait training (2:2:1 randomization) in chronic iSCI (>1 year post injury, AIS-C, and D), with residual stepping ability. The primary outcome measure was change in robot-independent gait speed (10-meter walk test, 10MWT) post 12-week intervention. Secondary outcomes included: Timed-Up-and-Go (TUG), 6-min walk test (6MWT), Walking Index for Spinal Cord Injury (WISCI-II) (assistance and devices), and treating therapist NASA-Task Load Index. Twenty-five participants completed the assessments and training as assigned (9 Ekso, 10 Active Control, 6 Passive Control). Mean change in gait speed at the primary endpoint was not statistically significant. The proportion of participants with improvement in clinical ambulation category from home to community speed post-intervention was greatest in the Ekso group (>1/2 Ekso, 1/3 Active Control, 0 Passive Control, p < 0.05). Improvements in secondary outcome measures were not significant. Twelve weeks of exoskeleton robotic training in chronic SCI participants with independent stepping ability at baseline can improve clinical ambulatory status. Improvements in raw gait speed were not statistically significant at the group level, which may guide future trials for participant inclusion criteria. While generally safe and tolerable, larger gains in ambulation might be associated with higher risk for non-serious adverse events.


Oxidative stress and mitochondrial DNA damage in heart failure.
Recent experimental and clinical studies have suggested that oxidative stress is enhanced in heart failure. The production of oxygen radicals is increased in the failing heart while antioxidant enzyme activities are preserved. Mitochondrial electron transport is an enzymatic source of oxygen radical generation and also a target against oxidant-induced damage in the failing myocardium. Chronic increases in oxygen radical production in the mitochondria can lead to a catastrophic cycle of mitochondrial DNA (mtDNA) damage, as well as functional decline, further oxygen radical generation, and cellular injury. Reactive oxygen species induce myocyte hypertrophy, apoptosis, and interstitial fibrosis by activating matrix metalloproteinases. These cellular events play an important role in the development and progression of maladaptive cardiac remodeling and failure. Therefore, oxidative stress and mtDNA damage are good therapeutic targets. Overexpression of peroxiredoxin-3 (Prx-3), mitochondrial antioxidant, or mitochondrial transcription factor A (TFAM) could ameliorate the decline in mtDNA copy number in failing hearts. Consistent with alterations in mtDNA, the decrease in oxidative capacities is also prevented. Therefore, the activation of Prx-3 or TFAM expression could ameliorate the pathophysiological processes seen in myocardial failure. Inhibition of oxidative stress and mtDNA damage could be novel and potentially effective treatment strategies for heart failure.


Inhibition of human proteases: from target identification to therapy. 23-24 April 1998, Boston, MA, USA.
This two-day meeting, sponsored by the Cambridge Healthtech Institute, was attended by around 70 scientists, primarily from the pharmaceutical industry, with a small number from academia. Most of the industry representatives were from smaller companies engaged in drug development; virtually no multinational companies were represented. Four main topics were covered: target identification and pathophysiology; structural analysis and drug design; screening for proteinase inhibitors; and, clinical development. The target identification session was by far the strongest. Only two posters were shown and podium presentations predominated. Several speakers dealt in depth with the chemistry of their company's line of potential inhibitors; a detailed description of the numerous possible chemical modifications is beyond the scope of this review.


Hydrogen peroxide and nitric oxide mediated cold- and dehydration-induced myo-inositol phosphate synthase that confers multiple resistances to abiotic stresses.
myo-Inositol phosphate synthase (MIPS) is the key enzyme of myo-inositol synthesis, which is a central molecule required for cell metabolism and plant growth as a precursor to a large variety of compounds. A full-length fragment of MfMIPS1 cDNA was cloned from Medicago falcata that is more cold-tolerant than Medicago sativa. While MfMIPS1 transcript was induced in response to cold, dehydration and salt stress, MIPS transcript and myo-inositol were maintained longer and at a higher level in M. falcata than in M. sativa during cold acclimation at 5 °C. MfMIPS1 transcript was induced by hydrogen peroxide (H(2) O(2)) and nitric oxide (NO), but was not responsive to abscisic acid (ABA). Pharmacological experiments revealed that H(2) O(2) and NO are involved in the regulation of MfMIPS1 expression by cold and dehydration, but not by salt. Overexpression of MfMIPS1 in tobacco increased the MIPS activity and levels of myo-inositol, galactinol and raffinose, resulting in enhanced resistance to chilling, drought and salt stresses in transgenic tobacco plants. It is suggested that MfMIPS1 is induced by diverse environmental factors and confers resistance to various abiotic stresses.


Dummy molecularly imprinted microbeads as solid-phase extraction material for selective determination of phthalate esters in water.
The aim of this study was to investigate the usability of newly synthesized dummy molecularly imprinted microbeads (DMIMs) as a solid phase extraction (SPE) material to determine six phthalate esters (PEs) in water by GC-MS analysis. Diethyl phthalate (DEP) was used as a dummy template to prepare poly(ethylene glycol dimethacrylate N-methacryloyl-l-tryptophan methyl ester) [PEMATrp)] DMIMs by using suspension polymerization. The PEMATrp DMIMs were characterized by using Fourier transform infrared spectroscopy (FTIR), X-ray photoelectron spectroscopy (XPS) and scanning electron microscopy (SEM). Firstly, the adsorption capacities of the DMIMs prepared in different template molecule (DEP) to functional monomer (MATrp) ratios were investigated by using DEP solutions in the concentration range of 1-500mg/L at pH 3.0. Styrene and vanillic acid were used to evaluate the selectivity of the prepared DMIMs towards the template molecule (DEP). Then, the best analytical conditions were investigated for the simultaneous determination of dimethyl phthalate (DMP), diethyl phthalate (DEP), di-n-butyl phthalate (DBP), benzylbutyl phthalate (BBP), di-(2-ethylhexyl) phthalate (DEHP) and di-n-octyl phthalate (DnOP) in aqueous media by using the PEMATrp DMIMs as SPE material. Validation experiments showed that the PEMATrp DMIMs-SPE method had good linearity at 12.5-250.0μg/L (0.988-0.999), good precision (1.2-5.9%), and limits of detection in a range of 0.31-0.41μg/L.


Assessment of calcium homeostasis in the critically ill surgical patient. The diagnostic pitfalls of the McLean-Hastings nomogram.
Hypocalcemia is a common problem in critically ill surgical patients. We prospectively evaluated whether measurement of the total serum calcium (Ca) concentration or calculation of the serum ionized Ca level (by the McLean-Hastings nomogram) accurately reflects the measured serum ionized Ca level. Although 71% and 58% of 156 predominantly surgical intensive care unit (ICU) patients were hypocalcemic by the total serum Ca or calculated ionized Ca level, respectively, only 12% were hypocalcemic by directly measured serum ionized Ca measurement. The total serum Ca and calculated ionized Ca concentrations were sensitive (95% and 89%, respectively) but lacked specificity (32% and 46%, respectively) in predicting ionized hypocalcemia. Analyses of Ca binding to albumin in the serum of surgical ICU patients and normal subjects suggested that there is a circulating factor in critically ill patients that increases the binding of Ca to albumin. These observations may explain why the McLean-Hastings nomogram underestimates the protein-induced changes in serum Ca in critically ill surgical subjects. We conclude that: total serum Ca and calculated ionized Ca concentrations are poor indicators of the true serum ionized Ca status in critically ill surgical patients, and we recommend direct measurement of serum ionized Ca levels in these patients; and variability in the affinity of Ca for binding proteins in critical illness may explain the poor correlation between serum total and ionized Ca measurements.


Effectiveness of neuromotor task training for children with developmental coordination disorder: a pilot study.
The aim of this pilot study was to evaluate the effectiveness of a Neuromotor Task Training (NTT), recently developed for the treatment of children with Developmental Coordination Disorder (DCD) by pediatric physical therapists in the Netherlands. NTT is a task-oriented treatment program based upon recent insights from motor control and motor learning research. Ten children with DCD (intervention group) were tested before and after 9 and 18 treatment sessions on the Movement ABC and a dysgraphia scale in order to measure the effectiveness of treatment on gross and fine motor skills in general and handwriting in particular. Five children (no-treatment control group) were tested twice with a time lag of nine weeks on the Movement ABC in order to measure spontaneous improvement. No improvement was measured for the children in the no-treatment control group, whereas a significant improvement was found for children in the intervention group for both quality of handwriting and performance on the Movement ABC after 18 treatment sessions.


Cerenkov losses: a limit for bandgap determination and Kramers-Kronig analysis.
Measuring low energy losses in semiconductors and insulators with high spatial resolution becomes attractive with the increasing availability of modern transmission electron microscopes (TEMs) equipped with monochromators, C(s) correctors and energy filters. In this paper, we demonstrate that Cerenkov losses pose a limit for the interpretation of low energy loss spectra (EELS) in terms of interband transistions and bandgap determination for many materials. If the velocity of a charged particle in a medium exceeds the velocity of light, photons are emitted leading to a corresponding energy loss of a few electronvolt. Since these losses are strong for energies below the onset of interband transitions, they change the apparent loss function of semiconductors and insulators, with the risk of erroneous interpretation of spectra. We measured low energy losses of Si and GaAs with a monochromated TEM demonstrating the effect of sample thickness on Cerenkov losses. Angle resolved EELS and energy filtered diffraction patterns (taken without a monochromator) show the extremely narrow angular distribution of Cerenkov losses. The latter experiment provides a method that allows to decide whether Cerenkov radiation masks the very low loss signal in EELS.


Final report on the safety assessment of benzaldehyde.
Benzaldehyde is an aromatic aldehyde used in cosmetics as a denaturant, a flavoring agent, and as a fragrance. Currently used in only seven cosmetic products, its highest reported concentration of use was 0.5% in perfumes. Benzaldehyde is a generally regarded as safe (GRAS) food additive in the United States and is accepted as a flavoring substance in the European Union. Because Benzaldehyde rapidly metabolizes to Benzoic Acid in the skin, the available dermal irritation and sensitization data demonstrating no adverse reactions to Benzoic Acid were considered supportive of the safety of Benzaldehyde. Benzaldehyde is absorbed through skin and by the lungs, distributes to all well-perfused organs, but does not accumulate in any specific tissue type. After being metabolized to benzoic acid, conjugates are formed with glycine or glucuronic acid, and excreted in the urine. Little acute toxicity was seen. The oral LD(50) of Benzaldehyde in rats and mice ranged from 800 to 2850 mg/kg. The intraperitoneal LD(50) in white rats was 3265 mg/kg. In short-term oral studies, the no observed adverse effect level (NOAEL) was 400 mg/kg in rats and mice. In subchronic oral studies, the NOAEL was 400 mg/kg in rats and 600 mg/kg in mice. In a 16-week feeding study, rats given up to 10,000 ppm showed no signs of toxicity. Repeated inhalation of volatilized Benzaldehyde produced ocular and nasal irritation at 500 ppm and death in rabbits at 750 ppm. Undiluted Benzaldehyde was irritating to rabbit eyes, causing edema, erythema, and pain. Benzaldehyde was determined not to be a contact sensitizer, but did produce allergic reactions in a maximization test. Clinical reports of allergy to Benzaldehyde are rare. Benzoic Acid did not produce irritation or sensitization reactions in human clinical studies. Benzoic Acid also failed to produce reactions in phototoxicity and photosensitization tests. Neither Benzaldehyde, Benzoic Acid, nor Sodium Benzoate are reproductive or developmental toxicants at doses that are nontoxic to the mother. In a behavioral study, blood levels of 0.12 ng/ml Benzaldehyde produced a 44% reduction in motor activity in mice. Benzaldehyde did not produce mutations in bacterial assays, but did produce chromosomal abnormalities in Chinese hamster cells and increased mutations in a mouse lymphoma forward mutation assay. Benzaldehyde was evaluated by the National Toxicology Program, which found no evidence of carcinogenicity in rats, and some evidence of carcinogenicity in mice. Several studies have suggested that Benzaldehyde can have carcinostatic or antitumor properties. Overall, at the concentrations used in cosmetics, Benzaldehyde was not considered a carcinogenic risk to humans. Although there are limited irritation and sensitization data available for Benzaldehyde, the available dermal irritation and sensitization data and ultraviolet (UV) absorption and phototoxicity data demonstrating no adverse reactions to Benzoic Acid support the safety of Benzaldehyde as currently used in cosmetic products.


Comparison of the survival of different isolates of SARS-CoV-2 in evaporating aerosols.
Numerous variants of SARS-CoV-2 with increased transmissibility have emerged over the course of the pandemic. Potential explanations for the increased transmissibility of these variants include increased shedding from infected individuals, increased environmental stability, and/or a lower infectious dose. Upon exhalation of a respiratory particle into the environment, water present in the particle is rapidly lost through evaporation, resulting in a decrease in particle size. The aim of the present study was to compare the losses of infectivity of different isolates of SARS-CoV-2 during the rapid evaporation of aerosol particles that occurs immediately post-generation to assess if there are differences suggestive of increased survival, and ultimately greater transmissibility, for more recent variants. Losses of infectivity of several isolates of SARS-CoV-2 suspended in viral culture media were assessed following aerosolization and evaporation in a flowing chamber. The results demonstrate that losses of infectivity measured post-evaporation were similar for three different isolates of SARS-CoV-2, including isolates from the more recent Delta and Omicron lineages. The average loss in infectivity across all three isolates was 61 ± 15% (-0.46 ± 0.17 log10 TCID50/L-air) at a relative humidity <30%. These results, together with those from several previous studies, suggest that it is unlikely that an increase in environmental stability contributes to the observed increases in transmissibility observed with more recent variants of SARS-CoV-2.


Association of polymorphisms in GCKR and TRIB1 with nonalcoholic fatty liver disease and metabolic syndrome traits.
In several genome-wide association studies, nonalcoholic fatty liver disease and alanine aminotransferase susceptibility variants have been identified in several genes, including LYPLAL1, ZP4, GCKR, HSD17B13, PALLD, PPP1R3B, FDFT1, TRIB1, COL13A1, CPN1, ERLIN1, CWF19L1, EFCAB4B, PZP, and NCAN. To investigate the relationship between these genes and nonalcoholic fatty liver disease in the Japanese population, we genotyped 540 patients and 1012 control subjects for 18 variations. We performed logistic regression analyses to characterize the association between the tested variations and nonalcoholic fatty liver disease. Metabolic syndrome and histological traits were also analyzed by linear regression. We also examined GCKR rs780094, TRIB1 rs2954021, and PNPLA3 rs738409 for epistatic effects. The A-allele of rs780094 in GCKR (P = 0.0024) and the A-allele of rs2954021 TRIB1 (P = 4.5×10⁻⁵) were significantly associated with nonalcoholic fatty liver disease. GCKR rs780094 was also associated with decreased plasma glucose, and increased triglycerides in the patient and control groups. GCKR rs780094 was also associated with an increased ratio of visceral to subcutaneous fat area in the patients with nonalcoholic fatty liver disease. Variations in GCKR, TRIB1, and PNPLA3 independently influenced nonalcoholic fatty liver disease and had no epistatic effects. Our data suggest variations in GCKR and TRIB1 are involved in the development of nonalcoholic fatty liver disease.


Associations between internet sex seeking and STI associated risk behaviours among men who have sex with men.
This exploratory study identified associations between internet sex seeking and HIV associated risk behaviours among a high risk sample of men who have sex with men (MSM). A cross sectional survey of men attending a sex resort was conducted. Of 164 men asked to participate, 91% completed a self administered questionnaire. The questionnaire assessed demographic variables and (using a 3 month recall period) men's HIV associated sexual risk behaviours. Potential confounding variables were assessed and controlled, as needed, by multivariate analysis. Men currently resided in 14 states. One sixth reported being HIV positive. 57% of the men reported using the internet to seek sex. Differences in critical behaviours (unprotected anal sex and number of partners) were not found. However, compared to those not seeking sex by internet, men using the internet to meet sex partners were more likely to report fisting (adjusted odds ratio = 3.3, p = 0.04), having group sex (prevalence ratio (PR) = 1.2, p = 0.0001), using poppers during sex (PR = 1.94, p = 0.0001), and using ecstasy during sex (PR = 2.7, p = 0.04). Internet sex seeking men were also significantly more likely to report meeting sex partners in bathhouses (PR = 2.2, p = 0.0001), bars (PR = 1.5, p = 0.001), parks (PR = 3.2, p = 0.006), and circuit parties (PR = 8.9, p = 0.007). Among MSM attending a sex resort, those using the internet to seek sex partners may have modestly elevated risks for acquiring or transmitting sexually transmitted infections. Subsequent studies should investigate the utility of using the internet as forum for promoting safer sex behaviours among high risk MSM.


Antitumor activities of JTP-74057 (GSK1120212), a novel MEK1/2 inhibitor, on colorectal cancer cell lines in vitro and in vivo.
The MAPK pathway is one of the most important pathways for novel anticancer drug development. We performed high-throughput screening for compounds that induce expression of p15INK4b, and identified JTP-74057 (GSK1120212), which is being evaluated in ongoing phase I, II and III clinical trials. We characterized its antitumor activities in vitro and in vivo. JTP-74057 strongly inhibited MEK1/2 kinase activities, but did not inhibit another 98 kinase activities. Treatment by JTP-74057 resulted in growth inhibition accompanied with upregulation of p15INK4b and/or p27KIP1 in most of the colorectal cancer cell lines tested. Daily oral administration of JTP-74057 for 14 days suppressed tumor growth of HT-29 and COLO205 xenografts in nude mice. Notably, tumor regression was observed only in COLO205 xenografts, and COLO205 was much more sensitive to JTP-74057-induced apoptosis than HT-29 in vitro. Treatment with an Akt inhibitor enhanced the JTP-74057-induced apoptosis in HT-29 cells. Finally, JTP-74057 exhibited an additive or a synergistic effect in combination with the standard-of-care agents, 5-fluorouracil, oxaliplatin or SN-38. JTP-74057, a highly specific and potent MEK1/2 inhibitor, exerts favorable antitumor activities in vitro and in vivo. Sensitivity to JTP-74057-induced apoptosis may be an important factor for the estimation of in vivo efficacy, and sensitivity was enhanced by an Akt inhibitor. These results suggest the usefulness of JTP-74057 in therapeutic applications for colorectal cancer patients.


Elucidating the role of pigment epithelium-derived factor (PEDF) in metabolic PCOS models.
PCOS is the most common endocrinopathy in women; associated with obesity and insulin resistance (IR). IR leads to accumulation of advanced-glycation-end-products (AGEs) and their receptor, RAGE. PCOS patients have increased levels of vascular endothelial growth factor (VEGF), interleukin 6/8 (IL-6/8) and anti-Mϋllerian-hormone (AMH). PEDF is a secreted-glycoprotein known for its anti-angiogenic and anti-inflammatory properties. We aimed to elucidate the role of PEDF in the pathogenesis and treatment of PCOS. We used a prenatal PCOS mouse model and fed the female offspring a high-fat diet, inducing metabolic PCOS (met.PCOS) characteristics. Female offspring were divided into three groups: control; met.PCOS; met.PCOS + recombinant PEDF (rPEDF). Met.PCOS mice gained more weight, had elevated serum IL-6 and higher mRNA levels of AMH, PEDF and RAGE in their granulosa cells (GCs) than met.PCOS + rPEDF mice. An in vitro Met.PCOS model in human GCs (KGN) line was induced by prolonged incubation with insulin/AGEs, causing development of IR. Under the same conditions, we observed an elevation of VEGF, IL-6/8 mRNAs, concomitantly with an increase in PEDF mRNA, intracellular protein levels, and an elevation of PEDF receptors (PEDF-Rs) mRNA and protein. Simultaneously, a reduction in the secretion of PEDF from GCs, was measured in the medium. The addition of rPEDF (5 nM) activated P38 signaling, implying that PEDF-Rs maintained functionality, and negated AGE-induced elevation of IL-6/8 and VEGF mRNAs. Decreased PEDF secretion may be a major contributor to hyperangiogenesis and chronic inflammation, which lie at the core of PCOS pathogenesis. rPEDF treatment may restore physiological angiogenesis inflammatory balance, thus suggesting a potential therapeutic role in PCOS.


Low-volume strength and endurance training prevent the decrease in exercise hyperemia induced by non-dominant forearm immobilization.
We examined the effect of 3-week upper limb immobilization on conduit artery cross-sectional area and peak hyperemia (BF(peak)) after exhaustive dynamic handgrip exercise (Ex(dyn)), and that of low-volume strength and endurance training during immobilization. Healthy volunteers (n = 21; mean age, 22 years) were divided into 3 groups: immobilization only (IMM; n = 7), immobilization with training (STR + END; n = 7), and control (no immobilization or training, CNT; n = 7). Endurance training comprised Ex(dyn) at 30% maximum voluntary contraction (MVC) (duration of each session, ~60 s; twice weekly). Strength training involved intermittent isometric handgrip exercise at 70% MVC (duration of each session, 40 s; twice weekly), repeated 10 times. We used ultrasound methods to measure the brachial artery cross-sectional area and the BF(peak) after Ex(dyn) for 5 min pre- and post-immobilization. We found a significant group by time interaction in BF(peak) (p < 0.05). A significant decrease was found in BF(peak) in the IMM (p < 0.05) between pre- and post-immobilization and a protective effect in the STR + END. The 3-week upper limb immobilization did not influence the baseline artery cross-sectional area. In conclusion, BF(peak) decreased after 3-week upper limb immobilization and a combination of strength training and endurance training preserved the blunted BF(peak).


Prognosis of symptomatic versus asymptomatic autoimmune hepatitis: a study of 68 patients.
Autoimmune hepatitis (AIH) is a chronic liver disorder of unknown etiology. Disease presentation ranges from asymptomatic to symptomatic onset, fulminant, acute, or chronic. To evaluate the prognosis of patients with asymptomatic versus symptomatic AIH and to determine the role of early treatment and prognostic factors in this subgroup of patients. Sixty-eight patients with AIH were retrospectively evaluated for clinical presentation, liver function tests, autoantibody profile, liver biopsy, treatment, and long-term prognosis. Twenty-three patients were classified as having asymptomatic AIH. They were compared with the 45 AIH patients with symptomatic onset. Patients with symptomatic and asymptomatic presentations were indistinguishable by age, but there was a relative male predominance in the asymptomatic group. Aminotransferase and immunoglobulin levels were lower in the asymptomatic group. In contrast, the histopathologic picture was indistinguishable between the two groups. Lobular hepatitis of a moderate to severe degree, portal fibrosis, and bridging fibrosis were detected in similar frequencies in both groups. Asymptomatic onset led to a delay in diagnosis and, consequently, in the initiation of treatment and was associated with use of relatively lower doses of corticosteroids. Response to treatment and long-term prognosis of asymptomatic patients was better than that observed in the symptomatic group. Asymptomatic onset of AIH is relatively common. The clinical presentation appears in correlation with liver functions tests but not with liver histology. This subgroup of AIH carries a better response to treatment and a favorable prognosis.


Finding space for participation: fisherfolk mobility and co-management of Lake Victoria fisheries.
The literature on fisheries co-management is almost silent on the issue of the movement of fisherfolk within fisheries, although such movement must have implications for the effectiveness of co-management. The introduction of co-management often involves the formation of new structures that should enable the participation of key stakeholder groups in decision-making and management, but such participation is challenging for migrating fishers. The article reports on a study on Lake Victoria, East Africa, which investigated the extent of movement around the lake and the implications of movement for how fishers participate and are represented in co-management, and the implications of the extent and nature of movement for co-management structures and processes. The analysis draws on the concept of space from the literature on participation in development and on a framework of representation in fisheries co-management in addressing these questions. The created space is on an 'invited' rather than open basis, reflecting the top-down nature of implementation and the desire to secure participation of different occupational groups, as well as women in a male-dominated sector. The more powerful boat owners dominate positions of power within the co-management system, particularly as the levels of co-management, from sub-district to national, are traversed. The limited power and resources of boat crew are exacerbated by the degree and nature of movement around the lake, making effective participation in co-management decision-making a challenge.


Celastrol-loaded bovine serum albumin nanoparticles target inflamed neutrophils for improved rheumatoid arthritis therapy.
Inflammatory neutrophils (INEs), motivated by cytokines, continue to migrate into the inflamed joints, driving the development of RA. Hence, inducing apoptosis of INEs to reduce recruitment at inflamed joints is an effective strategy for the treatment of RA. However, simply apoptotic INEs may trigger the release of neutrophil extracellular traps (NETs) and accelerate the inflammatory process. To overcome these drawbacks, an RGD-modified bovine serum albumin (BSA) nanoparticles (CBR NPs) was fabricated to selectively target INEs in situ for intracellular delivery of CLT. Studies have demonstrated that CBR NPs can selectively target circulating INEs and induce INEs apoptosis. Meanwhile, CBR NPs inhibited the activation of NETs via NF-κB pathway and the release of Cit-H3 thereby blocking the release process of NETs. In collagen-induced arthritis (CIA) mouse model, CBR NPs suppressed the inflammatory response, and reduced the toxic effects of CLT. In summary, this study shed light on an innovative approach to treat RA by inducing apoptosis of circulating INEs and inhibiting NETs. STATEMENT OF SIGNIFICANCE: RGD-modified bovine serum albumin (BSA) nanoparticles for delivering celastrol, abbreviated as CBR NPs, were constructed to inhibit the infiltration of circulating inflammatory neutrophils (INEs) into inflamed joints while inhibiting the release of NETs to alleviate tissue damage. CBR NPs were prepared for the first time to induce apoptosis of INEs; CBR NPs could inhibit the release of NETs while inducing apoptosis of INEs in vivo and vitro cellular experiments; CBR NPs had favorable anti-inflammatory effects and low toxicity side-effects in collagen-induced arthritis (CIA) mouse models. The application of nanotechnology to induce apoptosis of INEs while inhibiting the release of NETs was a promising approach for the treatment of RA.


TASK (TWIK-related acid-sensitive K+ channel) is expressed in glomerulosa cells of rat adrenal cortex and inhibited by angiotensin II.
The present study was conducted to explore the possible contribution of a recently described leak K+ channel, TASK (TWIK-related acid-sensitive K+ channel), to the high resting K+ conductance of adrenal glomerulosa cells. Northern blot analysis showed the strongest TASK message in adrenal glomerulosa (capsular) tissue among the examined tissues including heart and brain. Single-cell PCR demonstrated TASK expression in glomerulosa cells. In patch-clamp experiments performed on isolated glomerulosa cells the inward current at -100 mV in 30 mM [K+] (reflecting mainly potassium conductance) was pH sensitive (17+/-2% reduction when the pH changed from 7.4 to 6.7). In Xenopus oocytes injected with mRNA prepared from adrenal glomerulosa tissue the expressed K+ current at -100 mV was virtually insensitive to tetraethylammonium (3 mM) and 4-aminopyridine (3 mM). Ba2+ (300 microM) and Cs+ (3 mM) induced voltage-dependent block. Lidocaine (1 mM) and extracellular acidification from pH 7.5 to 6.7 inhibited the current (by 28% and 16%, respectively). This inhibitory profile is similar (although it is not identical) to that of TASK expressed by injecting its cRNA. In oocytes injected with adrenal glomerulosa mRNA, TASK antisense oligonucleotide reduced significantly the expression of K+ current at -100 mV, while the sense oligonucleotide failed to have inhibitory effect. Application of angiotensin II (10 nM) both in isolated glomerulosa cells and in oocytes injected with adrenal glomerulosa mRNA inhibited the K+ current at -100 mV. Similarly, in oocytes coexpressing TASK and ATla angiotensin II receptor, angiotensin II inhibited the TASK current. These data together indicate that TASK contributes to the generation of high resting potassium permeability of glomerulosa cells, and this background K+ channel may be a target of hormonal regulation.