Metrics of Diabetes Risk Are Only Minimally Improved by Exercise Training in Postmenopausal Breast Cancer Survivors.
Insulin resistance is a risk factor for breast cancer recurrence. How exercise training changes fasting and postglucose insulin resistance in breast cancer survivors is unknown. To evaluate exercise-induced changes in postglucose ingestion insulin concentrations, insulin resistance, and their associations with cancer-relevant biomarkers in breast cancer survivors. The University of Massachusetts Kinesiology Department. 15 postmenopausal breast cancer survivors not meeting the physical activity guidelines (150 min/week of exercise). A supervised 12-week aerobic exercise program (60 min/day, 3-4 days/week). Postglucose ingestion insulin was determined by peak insulin and area under the insulin curve (iAUC) during a 5-sample oral glucose tolerance test. Insulin sensitivity was estimated from the Matsuda composite insulin sensitivity index (C-ISI). Changes in fitness and body composition were determined from submaximal VO2peak and dual energy X-ray absorptiometry. Participants averaged 156.8 ± 16.6 min/week of supervised exercise. Estimated VO2peak significantly increased (+2.8 ± 1.4 mL/kg/min, P < .05) and body weight significantly decreased (-1.1 ± 0.8 kg, P < .05) following the intervention. There were no differences in fasting insulin, iAUC, C-ISI, or peak insulin following the intervention. Insulin was only significantly lower 120 min following glucose consumption (68.8 ± 34.5 vs 56.2 ± 31.9 uU/mL, P < .05), and there was a significant interaction with past/present aromatase inhibitor (AI) use for peak insulin (-11.99 non-AI vs +13.91 AI uU/mL) and iAUC (-24.03 non-AI vs +32.73 AI uU/mL). Exercise training had limited overall benefits on insulin concentrations following glucose ingestion in breast cancer survivors but was strongly influenced by AI use.


Interactions between carbon nanodots with human serum albumin and γ-globulins: The effects on the transportation function.
Carbon nanodots (C-dots) have attracted great attention as a new class of luminescent nanomaterials due to their superior physical and chemical properties. In order to better understand the basic behavior of C-dots in biological systems, a series of photophysical measurements were applied to study the interactions of C-dots with human serum albumin (HSA) and γ-globulins. The fluorescence of proteins was quenched by the dynamic mechanism rather than the formation of a protein/C-dots complex. The apparent dissociation constants of the C-dots bound to HSA and γ-globulins were of the same order of magnitude. Furthermore, it is proven that C-dots showed little influence on the conformation of HSA and γ-globulins. In addition, Fourier transform infrared and fluorescence spectroscopic studies demonstrated that the interaction between C-dots and two kinds of serum proteins was driven by hydrophobic and van der waals forces. Since the bioavailability of drugs can be modulated by their interactions with proteins, the variations of binding constants of three drugs with HSA and γ-globulins in the presence of different concentrations of C-dots (0-84 μmol L(-1)) have also been analyzed in this work, to reflect the effect of C-dots on the transportation function of HSA and γ-globulins.


Unilateral vocal fold immobility: a tertiary hospital's experience over 5 years.
The objective of the study was to determine the etiology and subsequent management of patients with unilateral vocal fold immobility (UVFI) and compare our results with other such studies. This was a retrospective case series of all patients that were treated for UVFI at one single tertiary referral centre between 2010 and 2014. The medical records of 161 patients over a 5-year period diagnosed with UVFI were analyzed. We looked at the patient demographics, side of immobility, etiology, management and voice assessment. A total of 21 patients were excluded due to varying reasons including second presentation and incomplete data. Our results demonstrated 37.1% of cases to be due to non-thyroid surgery (mainly vascular or anterior cervical spine surgery) compared to thyroid or parathyroid (18.6%). Carotid endarterectomy was the commonest cause followed by cervical spine discectomy or fusion. Other iatrogenic causes included thoracic surgery either involving the lung or not. Our results are very much in keeping with those seen by our colleagues in North America. A better appreciation of the causes of UVFI especially in cases not performed by otolaryngologists and head and neck surgeons should be highlighted and the necessary steps should be taken to prevent this iatrogenic complication.


Challenges facing workstyle reform for Japanese obstetricians and gynecologists revealed from time studies.
We aimed to grasp the actual working hours of Japanese obstetricians and gynecologists (OB/GYN doctors) as accurately as possible, using the same method of the Ministry of Health, Labour, and Welfare (MHLW). The time study targeted OB/GYN doctors working at 10 universities nationwide including Niigata University and 21 institutions which take a role of perinatal care in Niigata prefecture. Working hours per week were calculated based on the following categories: regular and overtime work inside the hospital, work outside the hospital, self-improvement, education, research, and others. Data on weekly working hours were converted to yearly data for analyses. A time study of 10 universities nationwide revealed that 30% of doctors work overtime for more than 1860 h even if they do not include on-call shifts in their working hours. In 21 institutions in Niigata, physicians in Niigata University worked more overtime than other hospitals. It became clear that community health care was supported by dispatching physicians working at university. Furthermore, the results of simulations predicted the pessimistic situation of perinatal medical care in Niigata. Our study showed the possibility to exist much more OB/GYN doctors who work more than 1860 h of overtime work per year than the data presented by the MHLW based on nation-wide survey in 2019. The fact that the working hours at the side jobs had a great influence on the increase in overtime work of physicians in University was the same result as the report of MHLW published in 2021.


On the importance of modeling fMRI transients when estimating effective connectivity: A dynamic causal modeling study using ASL data.
Effective connectivity is commonly assessed using blood oxygenation level-dependent (BOLD) signals. In (Havlicek et al., 2015), we presented a novel, physiologically informed dynamic causal model (P-DCM) that extends current generative models. We demonstrated the improvements afforded by P-DCM in terms of the ability to model commonly observed neuronal and vascular transients in single regions. Here, we assess the ability of the novel and previous DCM variants to estimate effective connectivity among a network of five ROIs driven by a visuo-motor task. We demonstrate that connectivity estimates depend sensitively on the DCM used, due to differences in the modeling of hemodynamic response transients; such as the post-stimulus undershoot or adaptation during stimulation. In addition, using a novel DCM for arterial spin labeling (ASL) fMRI that measures BOLD and CBF signals simultaneously, we confirmed our findings (by using the BOLD data alone and in conjunction with CBF). We show that P-DCM provides better estimates of effective connectivity, regardless of whether it is applied to BOLD data alone or to ASL time-series, and that all new aspects of P-DCM (i.e. neuronal, neurovascular, hemodynamic components) constitute an improvement compared to those in the previous DCM variants. In summary, (i) accurate modeling of fMRI response transients is crucial to obtain valid effective connectivity estimates and (ii) any additional hemodynamic data, such as provided by ASL, increases the ability to disambiguate neuronal and vascular effects present in the BOLD signal.


Evaluation of the inhibitory effects and the mechanism of terpenoids on Toxoplasma gondii tachyzoites.
Currently, symptomatic toxoplasmosis is treated with a combination of sulfadiazine and pyrimethamine. However, significant adverse effects and drug resistance have been reported. Terpenoids are widely found in nature, with numerous studies demonstrating that they have effective inhibitory effects on a variety of parasites. In this study, we employed intermediates or derivatives of lindenane sesquiterpenoids to evaluate their intracellular and extracellular inhibitory effects on Toxoplasma gondii tachyzoites and their cytotoxicity on macrophages. We demonstrated that two of these target terpenoids could effectively reduce the number of extracellular tachyzoites, probably by inducing tachyzoite apoptosis through altering tachyzoites mitochondrial membrane potential and calcium homeostasis. In addition, the two target terpenoids were able to promote intracellular tachyzoites elimination, possibly by enhancing macrophage activities. Two derivatives of lindenane sesquiterpenoids with low cytotoxicity to macrophages demonstrated direct and indirect antitoxoplasma effects.


Evidence for the participation of peripheral α5 subunit-containing GABAA receptors in GABAA agonists-induced nociception in rats.
The activation of GABAA receptor by γ-amino butyric acid (GABA) in primary afferent fibers produces depolarization. In normal conditions this depolarization causes a reduction in the release of neurotransmitters. Therefore, this depolarization remains inhibitory. However, previous studies have suggested that in inflammatory pain, GABA shifts its signaling from inhibition to excitation by an increased GABA-induced depolarization. The contribution of peripheral α5 subunit-containing GABAA receptors to the inflammatory pain is unknown. The purpose of this study was to investigate the possible pronociceptive role of peripheral α5 subunit-containing GABAA receptors in the formalin test. Formalin (0.5%) injection into the dorsum of the right hind paw produced flinching behavior in rats. Ipsilateral local peripheral pre-treatment (-10min) with exogenous GABA (0.003-0.03µg/paw) or common GABAA receptor agonists muscimol (0.003-0.03µg/paw), diazepam (0.017-0.056µg/paw) or phenobarbital (1-100µg/paw) significantly increased 0.5% formalin-induced nociceptive behavior. The pronociceptive effects of GABA (0.03µg/paw), muscimol (0.03µg/paw), diazepam (0.056µg/paw) and phenobarbital (100µg/paw) were prevented by either the GABAA receptor antagonist bicuculline (0.01-0.1µg/paw) or selective α5 subunit-containing GABAA receptor inverse agonist L-655,708 (0.017-0.17µg/paw). The α5 subunit-containing GABAA receptor protein was expressed in dorsal root ganglion (DRG) and dorsal spinal cord of naïve rats. The formalin injection did not modify α5 subunit-containing GABAA receptor expression. Overall, these results suggest that peripheral α5 subunit-containing GABAA receptors play a pronociceptive role in the rat formalin test.


Cross-sectional associations between trunk muscle composition, back pain, and physical function in the health, aging and body composition study.
Associations between trunk muscle composition and physical function have not been examined previously in older adults. We hypothesized that lower trunk muscle area and attenuation (higher fat infiltration) are associated with decreased functional capacity. The study sample consisted of a biracial cohort of well functioning men (739) and women (788) aged 70-79 from the Pittsburgh site of the Health, Aging and Body Composition (Health ABC) study. Computed tomography was used to measure trunk muscle area (cm2) and muscle attenuation (Hounsfield Unit [HU]) of the following muscle groups: lumbar paraspinals, lateral abdominals, and rectus abdominis. An average score was calculated for both trunk area and attenuation. The Health ABC Physical Performance Battery (PPB) and its individual components (usual and narrow walk, chair stands, and standing balance) were used to measure functional capacity. Linear regression analyses adjusting for demographic factors, height, body fat, thigh muscle composition, disease status, and low back pain (LBP) found that average trunk muscle area was not associated with any element of functional capacity (p >.10), whereas average trunk muscle attenuation was positively associated with the Health ABC Physical Performance Battery (p <.05) and chair stands (p <.001). Participants reporting higher LBP severity during the past year had lower muscle attenuation (p <.001 for trend), but there was no difference in average trunk muscle area according to LBP status. Findings suggest a link between trunk muscle composition and history of LBP as well as reduced functional capacity in older adults. Improving trunk muscle quality may lead to reduced LBP severity and improved functional status.


Aspergillus endocarditis and myocarditis in a patient with the acquired immunodeficiency syndrome (AIDS). A review of the literature.
We report the post-mortem findings of the case of a 31-year-old male who, through sexual contacts with a female drug addict, was found to be HIV-positive and developed the acquired immunodeficiency syndrome (AIDS) 2 years later. He was treated for various opportunistic infections over the next 7 years when he presented with cardiac abnormalities and multiple cerebral lesions which were responsible for his death. The results revealed Aspergillus fumigatus endocarditis and myocarditis with mycotic thromboembolic extension to the brain, spleen, kidney and pancreas. We review the literature of Aspergillus infection in patients with AIDS and more specifically cardiac involvement with this pathogen.


Atrial fibrillation in endomyocardial fibrosis is a marker of worse prognosis.
We studied the incidence of AF in patients with endomyocardial fibrosis (EMF) and its influence on prognosis and associated clinical events. One hundred and sixty consecutive patients with EMF were followed for a mean period of 4 years. Their mean age was 39.7 years. There were 114 women. During follow-up there were 56 deaths. Eighty-eight patients (55%) were submitted to surgical intervention. AF was observed in 58 cases (36.2%). The presence of AF was associated with a greater prevalence of dyspnea, peripheral edema, hepatomegaly, lower left ventricular ejection fraction, lower right ventricular systolic pressure (37.8 vs 45.6 mmHg, P=0.0392), and greater incidence of tricuspid regurgitation (86.0 vs 63.2%, P=0.004). AF was more frequent among patients in whom the disease involved the right ventricle, particularly those with intense fibrosis. Overall, patients with AF had a higher mortality rate than those who did not have AF (43.1 vs 30.3%, P=0.0195), but among those submitted to surgery, AF did not have an impact on survival. In conclusion, AF is frequent among patients with EMF. It is more prevalent among patients with right ventricular involvement and its presence is associated with a greater incidence of heart failure. AF is associated with worse prognosis, but surgery potentially reverses this bad evolution.


A dialysis rate method for the measurement of free iodothyronine and steroid hormones in blood.
From the dialysis rate of a small amount of tracer hormone from a serum sample through a semipermeable membrane towards an identical serum sample, the free hormone fraction can be calculated. This method is free from a number of artefacts that may occur in equilibrium dialysis.


Patient priorities for research: A focus group study of UK medical cannabis patients.
There has yet to be an evaluation of medical cannabis patient preferences with respect to future research. As such, prioritisation of research agendas has been largely driven by academia and industry. The primary aim of this study was to elicit priorities for research from medical cannabis patients in the United Kingdom (UK). Patients undergoing active treatment for health conditions with medical cannabis in the UK were invited to take part in focus groups from December 2021 to February 2022. An inductive thematic analysis of responses was performed. Participants also completed a ranking exercise whereby they assigned ten counters (each equivalent to £1 million GBP) to competing research priorities. 30 medical cannabis patients participated across 3 focus groups. The following themes were identified as research priorities: adverse events, comparison between cannabis-based medicinal products, health conditions, pharmacology of cannabis, types of study, healthcare professionals' attitudes, social environment, agriculture and manufacturing, and the cannabis plant. Participants assigned the highest proportion of research funding to 'assessment of effect on specific symptoms' (26 counters; 8.7%). This study highlighted specific themes within which to focus future research on medical cannabis. Clinically, there was a directive towards ensuring that research is condition- or symptom-specific. Participants also emphasised themes on the social impact of medical cannabis, such as knowledge of medical cannabis among healthcare professionals, stigma, and effects on driving and in the workplace. These findings can guide both research funders and researchers into effectively conducting research which fits within a more patient-centric model.


Association between time from cessation of oxytocin infusion for labor to delivery and intraoperative severe blood loss during cesarean section: a retrospective cohort study.
Aim: Prolonged oxytocin infusion during labor results in receptor desensitization and increases the risk of obstetric hemorrhage. We aimed to examine the association between recovery time (RT) and intraoperative severe blood loss in women who underwent a cesarean section after oxytocin pretreatment.Methods: We retrospectively assessed 103 Japanese women who underwent cesarean section after oxytocin pretreatment. RT (time from cessation of oxytocin infusion during labor to delivery) and intraoperative severe blood loss (active bleeding exceeding 1000 mL) were measured. Confounding factors were controlled, and RT cut-off value associated with severe blood loss and association between RT and intraoperative severe blood loss were assessed.Results: The mean maternal age was 34 years, and 100 (97.1%) women delivered at term. Mean RT was 121.6 min. The overall incidence of intraoperative severe blood loss was 22.3% (23/103). The cut-off point to predict intraoperative severe blood loss was 96 min (sensitivity, 65.2%; specificity, 81.3%). On multivariate analysis, an RT of ≤96 min [adjusted odds ratio (OR), 11.9; 95% confidence interval (CI), 3.32-42.7] and macrosomia (adjusted OR, 3.91; 95% CI, 1.10-13.8) were associated with intraoperative severe blood loss.Conclusions: Consideration of RT is helpful in the management of women undergoing cesarean section after oxytocin pretreatment.


Computed tomographic demonstration of inferior vena cava invasion in a case of hepatocellular carcinoma.
A case of hepatocellular carcinoma with neoplastic invasion of the inferior vena cava is presented. Computed tomograms before and after injection of contrast medium revealed an unusual density enhancement of the wall of the invaded portion of the vena cava. This sign is correlated with an angiographic sign of venous invasion present in our case.


High-speed parameter search of dynamic biological pathways from time-course transcriptomic profiles using high-level Petri net.
Dynamic simulation promises a deeper understanding of complex molecular mechanisms of biological pathways. How to determine the reaction kinetic parameters which govern the simulation results is still an open question in the field of systems biology. (1) Background: To execute simulation experiments, it is an essential first step to search effective values of model parameters. The complexity of biological systems and the experimental measurement technology severely limit the acquirement of accurate kinetic parameters. Previously proposed genomic data assimilation (GDA) approach enables users to handle parameter estimation using time-course information. However, it highly depends on successive time points and costs massive computational resource; (2) Methods: To address this problem, we present a new high-speed parameter search method for estimating the kinetic parameters of quantitative biological pathways using time-course transcriptomic profiles. The key idea of our method is to interactively prune the search space by introducing Probabilistic Linear-time Temporal Logic (PLTL) based model checking into GDA. (3) Results and conclusion: We demonstrated the effectiveness of our method by comparing with GDA on Mus musculus transcription circuits modelled by hybrid functional Petri net with extension. As a result, our method works faster and more accurate than GDA for both time-course datasets with dense and sparse observed values.


Associations of the ages at menarche and menopause with blood pressure and hypertension among middle-aged and older Chinese women: a cross-sectional analysis of the baseline data of the China Health and Retirement Longitudinal Study.
We evaluated the associations of the ages at menarche and menopause with blood pressure (BP) and hypertension using the baseline data of 7893 women from the China Health and Retirement Longitudinal Study, a nationally representative survey among Chinese adults aged ≥45 years. Multivariate linear and logistic regression analyses were performed to evaluate the associations of the ages at menarche and men`opause with BP and hypertension, respectively. Nonlinear associations were evaluated using spline analyses. After controlling for age, education, marital status, living areas, smoking, drinking, and medication use if necessary, an early onset of menarche by 1 year was associated with a 6% (95% confidence interval [CI]: 3-9%) higher odds of hypertension and 0.82 mm Hg (P < 0.001) and 0.41 mm Hg (P < 0.001) higher systolic and diastolic BP, respectively. When further controlling for the body mass index (BMI), blood glucose, and lipids, the associations were still significant. Spline analyses did not support U-shaped relationships between menarche age and hypertension risk (P = 0.35), systolic BP (P = 0.60), or diastolic BP (P = 0.70). When stratified by location of residence, menarche age was only associated with BP and hypertension among women living in rural areas. The age of menopause was positively associated with hypertension (odds ratio [OR] = 1.02 per year delay of menopause, 95% CI: 1.01-1.03). However, when further controlling for BMI, such an association no longer existed (OR = 1.01, P = 0.32). These findings indicated that the associations of menarche age with BP and hypertension may be modified by factors related to the area of residence in China, and the association between menopause age and hypertension was driven by BMI.


Optical modeling of random anti-reflective meta-surfaces for laser systems applications.
Optical performance of anti-reflective random meta-surfaces are studied numerically for coherent beam propagation systems, such as lasers. A methodology for the modeling of such optics performance is developed and applied to study the reflectivity and laser beam quality degradation. These quantitative metrics and design considerations highlight that reducing the size of the meta-surface period much below the light wavelength is not necessarily required.


The transfer of specific mitochondrial lipids and proteins to lipid droplets contributes to proteostasis upon stress and aging in the eukaryotic model system Saccharomyces cerevisiae.
Originally Lipid droplets (LDs) were considered as being droplets for lipid storage only. Increasing evidence, however, demonstrates that LDs fulfill a pleiotropy of additional functions. Among them is the modulation of protein as well as lipid homeostasis. Under unfavorable pro-oxidative conditions, proteins can form aggregates which may exceed the overall proteolytic capacity of the proteasome. After stress termination LDs can adjust and support the removal of these aggregates. Additionally, LDs interact with mitochondria, specifically take over certain proteins and thus prevent apoptosis. LDs, which are loaded with these harmful proteins, are subsequently eliminated via lipophagy. Recently it was demonstrated that this autophagic process is a modulator of longevity. LDs do not only eliminate potentially dangerous proteins, but they are also able to prevent lipotoxicity by storing specific lipids. In the present study we used the model organism Saccharomyces cerevisiae to compare the proteome as well as lipidome of mitochondria and LDs under different conditions: replicative aging, stress and apoptosis. In this context we found an accumulation of proteins at LDs, supporting the role of LDs in proteostasis. Additionally, the composition of main lipid classes such as phosphatidylcholines, phosphatidylethanolamines, phosphatidylinositols, phosphatidylglycerols, triacylglycerols, ceramides, phosphatidic acids and ergosterol of LDs and mitochondria changed during stress conditions and aging.


Taurine, hypotaurine, and GABA uptake by cultured neuroblastoma cells.
Uptake of [3H]taurine, [35S]hypotaurine, and [3H] gamma-aminobutyric acid (GABA) was studied in neuroblastoma C1300 cells in Krebs-Ringer-Hepes-glucose medium (pH 7.4). The uptakes consisted of nonsaturable penetration (taurine and hypotaurine) and two saturable transport components: high affinity for taurine, hypotaurine, and GABA and low affinity for hypotaurine and GABA. The affinity of the high-affinity uptake was highest for hypotaurine but the transport capacity was greatest for taurine. GABA uptake was almost abolished by taurine and hypotaurine. Hypotaurine also strongly inhibited taurine uptake, whereas GABA had only a moderate inhibitory effect on taurine and hypotaurine uptakes. The mutual inhibition suggests that these amino acids use the same transport sites when entering the cells.


[Autoimmune response following administration of a protein preparation from the caudate nuclei of cattle brains to rabbits].
Plasma membranes of bovine caudate nuclei were solubilized by means of Triton X-100. Detergent-extracted membrane proteins were separated by chromatography on DEAE-cellulose and the affinity adsorbent proposed for the isolation of nicotinic cholinergic receptor protein (N-ChR) from Torpedo electrical organ. Half of the rabbits immunized with purified protein preparation developed a syndrome of myasthenia type. This effect resembled the autoimmune response to N-ChR from fish electrical organs. It is assumed that the N-ChR present in the caudate nucleus has antigen determinants similar to N-ChRs of the electrical organs and skeletal muscles.


Significance of stromal markers in atypical polypoid adenomyoma.
Atypical polypoid adenomyoma (APA) may progress to endometrioid carcinoma and may mimic myoinvasive carcinoma on biopsy specimens. Here, we present a case of an APA of the uterine cervix hysteroscopically treated, which recurred two years after and progressed to endometrioid carcinoma. In all biopsy specimens and in the hysterectomy specimen, the benign APA component showed an unusual immunohistochemical stromal pattern (periglandular fringe-like CD10 pattern, diffuse h-caldesmon positivity, p16 negativity), which is typical of myoinvasive carcinoma. Interestingly, the other three cases of cancerized APA assessed for h-caldesmon in the literature showed diffuse stromal positivity also in the benign APA component. Our case shows that the stromal markers used for differentiating between APA and myoinvasive carcinoma may be misleading even when their pattern seems unequivocal. Furthermore, our case suggests that h-caldesmon positivity might be a prognostic marker for progression of APA to carcinoma. Further studies are encouraged in this regard.


Association Between Thyroid Hormone Status and Trace Elements in Serum of Patients with Nodular Goiter.
The present study investigated the status of calcium and magnesium as well as essential trace elements including iodine, selenium, copper, iron, and zinc in adults residing in the Zhytomyr region of Ukraine. In addition, the relative risk of goiter occurrence was evaluated. In this comparative study, 40 adults without goiter (control group) and 16 adults with diagnosed nodular goiter (NG) were examined. Inductively coupled plasma optical emission spectrometry (ICP-OES) was used for the measurements of Mg, Ca, Se, Zn, Cu, and Fe in serum of patients with NG and control group. Patients with nodular goiter had lower serum values of Ca, Mg, Se, Cu, Fe, and Zn than those in the control group. The presence of mild iodine deficiency was evident in both groups with the median urinary iodine excretion (UIE) 80.5 μg/L in the control group and 64.5 μg/L in goiter group. There was a positive association between goiter presence and low concentration of Ca in serum (odds ratio (OR) = 2.29 (1.26-3.55), p < 0.05) in the NG group. High relative risk of goiter was observed at low concentrations of magnesium (OR = 3.33 (1.39-7.62), p < 0.05) and selenium (OR = 1.63, (1.16-1.78), p < 0.05) in comparison with OR values in the control group. Low concentrations of Ca, Mg, Zn, and Se in serum combined with reduced UIE resulted in the highest risk of goiter (OR = 12.5, (2.15-79.42), p < 0.01). This study proved that Thyroglobulin concentration in serum is the reliable indicator of nodular goiter. We also suggest that a combination of low concentrations of Ca, Mg, Zn, Cu, and Se in blood serum, and reduced iodine concentration in urine resulted in the highest risk of nodular goiter development.


Comparison of comet-based approaches to assess base excision repair.
DNA repair plays an essential role in maintaining genomic stability, and can be assessed by various comet assay-based approaches, including the cellular repair assay and the in vitro repair assay. In the cellular repair assay, cells are challenged with a DNA-damaging compound and DNA damage removal over time is assessed. In the in vitro repair assay, an early step in the repair process is assessed as the ability of a cellular extract to recognize and incise damaged DNA in substrate nucleoids from cells treated with a DNA-damaging compound. Our direct comparison of both assays in eight cell lines and human peripheral blood lymphocytes indicated no significant relationship between these DNA repair assays (R2 = 0.084, P = 0.52). The DNA incision activity of test cells measured with the in vitro repair assay correlated with the background level of DNA damage in the untreated test cells (R2 = 0.621, P = 0.012). When extracts were prepared from cells exposed to DNA-damaging agents (10 mM KBrO3 or 1 µM Ro 19-8022 plus light), the incision activity was significantly increased, which is in line with the notion that base excision repair is inducible. The data presented suggest that the two assays do not measure the same endpoint of DNA repair and should be considered as complementary.


Truncation of the cytoplasmic tail of the lutropin/choriogonadotropin receptor prevents agonist-induced uncoupling.
An agonist-induced change in the functional properties of a constant number of receptors seems to be a ubiquitous phenomenon involved in the regulation of cell surface receptors. Although the mechanisms responsible for this phenomenon (called uncoupling or desensitization) have been studied in detail using beta 2-adrenergic receptors it is unclear if the models derived from these studies are applicable to other members of the family of G protein-coupled receptors. Since it has been shown previously that truncation of the C-terminal cytoplasmic tail of the beta 2-adrenergic receptor results in a delay in the onset of agonist-induced uncoupling (Bouvier, M., Hausdorff, W.P., De Blasi, A., O'Dowd, B.F., Kobilka, B.K., Caron , M.G., and Lefkowitz, R.J. (1988) Nature 333, 370-373), we now present experiments designed to test the effects of a similar truncation of the lutropin/choriogonadotropin (LH/CG) receptor on its functional properties. The results presented herein show that (i) clonal lines of human embryonic kidney cells stably transfected with cDNAs encoding for the wild-type (rLHR-wt) or a mutant receptor truncated at amino acid residue 631 (rLHR-t631) express functional LH/CG receptors as judged by their ability to bind hCG and to respond to it with increased cAMP accumulation; (ii) a preincubation of the cells expressing rLHR-wt with hCG leads to a reduction in the ability of hCG to activate adenylylcyclase; and (iii) this reduction is severely blunted in cells expressing rLHR-t631. These results demonstrate that the C-terminal cytoplasmic tail of the LH/CG receptor is necessary for agonist-induced uncoupling.


Toward manipulating serotonin signaling via the microbiota-gut-brain axis.
It is now well established in humans that there is a bidirectional pathway of communication between the central and enteric nervous systems in which members of the microbiome participate. This microbiota-gut-brain axis (MGBA) is crucial for normal development and physiology, and its dysregulation has been implicated in a range of neurological and intestinal disorders. Investigations into the mechanistic underpinnings of the MGBA have identified serotonin as a molecule of particular interest. In this review, we highlight recent advances toward understanding the role of endogenous serotonin in microbial communities, how microbial communities bidirectionally interact with host serotonin, and potential future engineering opportunities to leverage these novel mechanisms for biomedical applications.


Relationship Between Control Over Nursing Practice and Job Satisfaction and Quality of Patient Care.
Control over nursing practice (CONP) in the work setting is associated with several positive consequences such as increased job satisfaction, support of teamwork, decreased patient mortality, and improved quality of care. The purpose of this study was to examine the level of perceived CONP among Jordanian registered nurses (RNs) and determine its relationship with their job satisfaction and quality of patient care. A descriptive, cross-sectional correlational design was used. A convenience sample of 230 RNs was recruited from 4 hospitals. The RNs had a moderate level of perceived CONP. This control was positively correlated with their work satisfaction and perception of the quality of patient care that they delivered. Nurse managers should be encouraged to take into consideration nurses' perceived CONP to improve working conditions for nurses.


Use of a cDNA library for the study of mRNA changes during muscle differentiation.
A cDNA library was used to measure changes in many individual mRNAs during muscle differentiation in culture. A library of 1000 clones was constructed from total myofiber poly(A) RNA. About 23% of these clones gave a detectable colony hybridization signal using end-labeled myofiber mRNA, the remainder containing muscle sequences too rare to be detected with this assay. The 230 positive clones were grouped into four classes based on relative visual intensity. Reconstruction experiments using pure globin mRNA enable us to determine the approximate percentage of total RNA made up by each mRNA hybridizing to a cDNA clone. Those clones containing sequences complementary to developmentally regulated mRNAs were identified by a differential hybridization procedure. The cDNA library was screened with end-labeled mRNA from both undifferentiated myoblasts and differentiated myofibers. Although the bulk of the clones hybridized essentially the same with both RNA populations, several dozen were found which hybridized differentially. Some clones contained sequences which were not present at all in myoblasts and present in very high quantities in myofibers. Others contained sequences found in both myoblasts and myofibers but in increased quantities in the differentiated cells. Still others contained sequences which decreased in quantity during muscle differentiation. The clones in the first group were chosen for immediate analysis since they likely contain contractile protein mRNA sequences. However, all the characterized cDNA clones can now be used as probes to study the chromosomal organization and developmental expression of genes active during muscle differentiation.


Simultaneous optimization of limited sampling points for pharmacokinetic analysis of amrubicin and amrubicinol in cancer patients.
Limited sampling points for both amrubicin (AMR) and its active metabolite amrubicinol (AMR-OH) were simultaneously optimized using Akaike's information criterion (AIC) calculated by pharmacokinetic modeling. In this pharmacokinetic study, 40 mg/m(2) of AMR was administered as a 5-min infusion on three consecutive days to 21 Japanese lung cancer patients. Blood samples were taken at 0, 0.08, 0.25, 0.5, 1, 2, 4, 8 and 24 h after drug infusion, and AMR and AMR-OH concentrations in plasma were quantitated using a high-performance liquid chromatography. The pharmacokinetic profile of AMR was characterized using a three-compartment model and that of AMR-OH using a one-compartment model following a first-order absorption process. These pharmacokinetic profiles were then integrated into one pharmacokinetic model for simultaneous fitting of AMR and AMR-OH. After fitting to the pharmacokinetic model, 65 combinations of four sampling points from the concentration profiles were evaluated for their AICs. Stepwise regression analysis was applied to select the sampling points for AMR and AMR-OH to predict the area under the concentration-time curves (AUCs) at best. Of the three combinations that yielded favorable AIC values, 0.25, 2, 4 and 8 h yielded the best AUC prediction for both AMR (R(2) = 0.977) and AMR-OH (R(2) = 0.886). The prediction error for AUC was less than 15%. The optimal limited sampling points of AMR and AMR-OH after AMR infusion were found to be 0.25, 2, 4 and 8 h, enabling less frequent blood sampling in further expanded pharmacokinetic studies for both AMR and AMR-OH.


A typical postcesarean epithelioid trophoblastic lesion with placenta increta: Case report and literature review.
We report an extremely rare case of unusual atypical epithelioid trophoblastic lesion from placenta increta. A 25-year-old Chinese woman with a history of 1 cesarean delivery was admitted to the hospital at 37 weeks' gestation. Magnetic resonance imaging (MRI) showed the feature of placenta increta. An elective primary cesarean section was scheduled and a healthy baby was born. The patient was suggestive of placenta increta and implantation site was resected. Histology indicated a lesion consisting of epithelioid trophoblastic cells with an intermediate pattern between a classical placental site nodule and an epithelioid trophoblastic tumor. This may be linked to her previous cesarean delivery, which supports the relationship between atypical epithelioid trophoblastic lesion formation and surgical interventions. Her serum human chorionic gonadotropin (hCG) was 352.4 IU/L after delivery and gradually decreased to normal level.


Early detection of Fasciola gigantica infection in buffaloes by enzyme-linked immunosorbent assay and dot enzyme-linked immunosorbent assay.
In an attempt to develop a suitable serological test for early detection of Fasciola gigantica infection in buffaloes, a group of proteins were isolated from the somatic antigen of the parasite by immunoaffinity chromatography. The process of isolation of the proteins has been standardized and significant level of repeatability was achieved. To test the diagnostic potentiality of the antigens, two serological tests, viz., enzyme-linked immunosorbent assay (ELISA) and dot enzyme-linked immunosorbent assay, were standardized using the sera from experimentally noninfected (group A) and infected (group B) animals. Further, the sensitivity and the specificity of the tests were evaluated employing the field sera from animals of different parasitic load viz., F. gigantica positive (group C), F. gigantica and Gastrothylax crumenifer positive (group D), F. gigantica and Gigantocotyle explanatum positive (group E), a group of sera without F. gigantica but other trematode infection (group F), only G. crumenifer positive (group G), only G. explanatum positive (group H), G. crumenifer and G. explanatum positive (group I), and PM negative (group J) collected from slaughterhouses of Bareilly (Uttar Pradesh, India) and Patna (Bihar, India). In plate ELISA, the sensitivity of the antigen and the test was 75.75% while the specificity was 97%, 95%, and 98%, respectively, against G. crumenifer, G. explanatum, and mixed infection of G. crumenifer and G. explanatum, respectively. In the case of dot ELISA the sensitivity was 86.5% and specificity was 92.3%, 94.7%, and 90%, respectively, against G. crumenifer, G. explanatum, and mixed infection of G. crumenifer and G. explanatum, respectively. The potentiality of the antigen in the diagnosis of field infection is discussed.


Effect of Chêneau style braces on vertebral wedging amongst individuals with adolescent idiopathic scoliosis.
It is generally accepted that braces can stop curve progression but little evidence exists regarding structural improvement in the spine using spinal bracing. Our study aimed to investigate the possible structural improvements of vertebral wedging with high correction bracing. The aim of our study was to assess whether spinal brace treatment may influence vertebral wedging in adolescent idiopathic scoliosis (AIS). We reviewed our database according to the following inclusion criteria: girls with a diagnosis of AIS, Risser 0-2, age 10-14 years with Cobb angles greater than 35°. Our study cohort consisted of 27 patients fulfilling the inclusion criteria with an average brace wearing time of 16.6 h per day and Cobb angles between 36° and 79°. The target value for our study was the apical vertebra wedging, measured twice before brace treatment commenced and twice after the average follow-up period of 20.5 months of treatment. The average apex wedging noted before brace wearing started was 9.8° (median: 9) and after a period of 20.5 months of brace wearing, it had reduced to an average of 5.8° (median: 4.9), (p < 0.001). This would indicate a structural correction of 44%. Our study supports the hypothesis that spinal high correction braces improve the degree of vertebral wedging in skeletally immature girls with AIS. Structural corrections of the apical vertebra seem possible when high correction asymmetric braces are used in the treatment of patients with AIS.


UCP1 -3826 A>G polymorphism affects weight, fat mass, and risk of type 2 diabetes mellitus in grade III obese patients.
We investigated whether or not the UCP1 -3826 A>G polymorphism is associated with obesity and related metabolic disorders in grade III obese patients. 150 obese patients (body mass index ≥35 kg/m(2)) who were candidates for bariatric surgery were studied. Weight (kg), body mass index (kg/m(2)); fat free mass (kg), fat mass (kg), energy intake (kcal), level of physical activity, plasma levels of glucose, total cholesterol, low-density lipoprotein, high-density lipoprotein (HDL), triacylglycerols, and the prevalence of comorbidities associated with obesity were collected from medical records. Polymorphism rs1800592 genotyping was performed through allelic discrimination method in real time polymerase chain reaction using the TaqMan predesigned SNP Genotyping Assays kits. The t test was done to determine if genotypes of each polymorphism are associated with anthropometric and body composition variables. Linear regression models were used for age, sex, height, physical activity, and energy intake in weight and body composition variations (P < 0.05). Among these 150 individuals (47.2 ± 10.5 y, 80% women) the distribution of AA, AG, and GG was 41.3%, 45.3%, and 13.4%, respectively. Weight and body fat were lower in individuals who were carriers of a mutated allele G. It was observed that mutated homozygotes (GG) had a lower frequency of type 2 diabetes mellitus compared with those of wild allele (AA+AG). UCP1 -3826 A>G polymorphism is associated with weight, body fat mass, and risk of type 2 diabetes mellitus in obese individuals candidates for bariatric surgery.


The Fusarium oxysporum sti35 gene functions in thiamine biosynthesis and oxidative stress response.
The sti35 gene of the vascular wilt fungus Fusarium oxysporum was originally identified based on induced expression under stress conditions. In this study, the transcriptional regulation and biological function of sti35 were examined in the tomato pathogen F. oxysporum f.sp. lycopersici. Expression of sti35 was repressed by thiamine and induced by high temperatures. Sti35 transcripts were detected both during early and late stages of infection of tomato plants by F. oxysporum. Heterologous expression of the sti35 cDNA restored thiamine prototrophy in a Saccharomyces cerevisiae thi4 mutant and increased UV tolerance in a uvr(-) mutant of Escherichia coli. Targeted Deltasti35 knockout mutants of F. oxysporum exhibited a thiamine auxotrophic phenotype and reduced tolerance to the superoxide-generating agent menadione, indicating that Sti35 has a dual role in thiamine biosynthesis and oxidative stress response. RT-PCR analysis revealed the presence of differential RNA splicing of the second 5'-UTR intron, suggesting that thiamine may regulate sti35 expression via a post-transcriptional mechanism. F. oxysporum transformants carrying a transcriptional fusion of the sti35 promoter to the lacZ reporter gene produced high levels of beta-galactosidase activity when grown in the absence, but not in the presence of thiamine. Thus, the sti35 promoter represents a useful tool for the controlled expression of genes of interest in F. oxysporum.


Autophagy is a survival force via suppression of necrotic cell death.
Macroautophagy or autophagy is a self-digesting mechanism that the cellular contents are engulfed by autophagosomes and delivered to lysosomes for degradation. Although it has been well established that autophagy is an important protective mechanism for cells under stress such as starvation via provision of nutrients and removal of protein aggregates and damaged mitochondria, there is a very complex relation between autophagy and cell death. At present, the molecular cross-talk between autophagy and apoptosis has been well discussed, while the relationship between autophagy and programmed necrotic cell death is less understood. In this review we focus on the role of autophagy in necrotic cell death by detailed discussion on two important forms of necrotic cell death: (i) necroptosis and (ii) poly-(ADP-ribose) polymerase (PARP)-mediated cell death. It is believed that one important aspect of the pro-survival function of autophagy is achieved via its ability to block various forms of necrotic cell death.


Environmental, energy, and economic impact assessment of sludge management alternatives based on incineration.
In this study, different management strategies for sewage sludge disposal were evaluated associated with environmental, energy, and economic impact, using life cycle assessment (LCA), cumulative energy demand (CED) and life cycle costing (LCC) approaches. Four scenarios, including mono-incineration, co-incineration in municipal solid wastes (MSW) incineration plant, co-incineration in coal-fired power plant and co-incineration in cement kiln, were assessed. The environmental burdens generated from the sludge incineration contributed primarily to the global warming, followed by eutrophication, marine aquatic ecotoxicity, and human toxicity potential across the four scenarios. Furthermore, mono-incineration scenario appeared to be the most environmentally unfriendly, energy and economy intensive alternative, with the LCA, CED and LCC value of 5.41E-09, 1736 MJ and 1.84 million CNY, respectively. By contrast, co-incineration in cement kiln exhibited the lowest CED (368 MJ), LCC (0.59 million CNY), and environmental burdens (1.02E-09). In addition, the sensitivity analysis indicated that four scenarios were sensitive to the changes in the electricity efficiency and the moisture content contained in sewage sludge, suggesting that it was of great significance to enhance the efficiency of sludge dewatering and thermal drying The findings of this study can provide scientific reference for selecting the optimal strategies for the most environmentally and economically friendly sewage sludge management with optimum energy efficiency.


Interaction of silver nanoparticles with plasma transport proteins: A systematic study on impacts of particle size, shape and surface functionalization.
Metallic nanoparticles are an important and widely used materials in development of nano-enabled medicine. For that reason, their interaction with biological molecules has to be systematically examined, as use of nanoparticles can lead to altered biological functions. In this study, we evaluated the interaction between silver nanoparticles (AgNPs) and two important plasma transport proteins - albumin and α-1-acid glycoprotein. To investigate comprehensively how different physico-chemical properties impact interaction of proteins with nanosurface, AgNPs of different size, shape and surface coating was prepared. The study was conducted using UV-Vis absorption, fluorescence, inductively coupled plasma mass spectrometry, circular dichroism spectroscopy, transmission electron microscopy, dynamic and electrophoretic light scattering techniques. The results showed significant complexities of the nano-bio interface and binding affinities of proteins onto surface of different AgNPs, which were affected by both AgNPs and protein properties. The most significant role on AgNPs-protein interaction had the coating agents used for AgNPs surface stabilization. Our findings should improve safe-by-design approach to development of the metallic nanomaterials for medical use.


Biosynthesis of gold nanoparticles and inhibition of various stages of bacterial biofilms formed by drug-resistant Aeromonas hydrophila, Escherichia coli, and Klebsiella pneumoniae.
Biofilm development by bacteria is a serious issue in many different sectors, including the food, pharmaceutical, water, petroleum, and others. Because biofilms are difficult to remove due to their capacity to survive in bad environments, attempts were undertaken throughout this work to biologically manufacture gold nanoparticles with possible antibiofilm capabilities. During this study, 19 morphologically distinct fungal cultures were isolated from soil samples and used to synthesize gold nanoparticles. Among 19 fungi, 7 were producing gold nanoparticles and these particles were subjected to antimicrobial activity. Among 7 gold nanoparticles, nanoparticles synthesized from S2-10 isolate of fungi exhibited high antimicrobial activity toward biofilm-forming drug-resistant bacteria including Klebsiella pneumoniae (14.6 ± 0.57 mm), Aeromonas hydrophila (13.3 ± 0.57 mm) and Escherichia coli (10.0 ± 1.0 mm), and was used for further studies. The chosen nanoparticle also shown strong anti-biofilm action against test microorganisms on a variety of material substrates, including plastic, metal, and glass. These nanoparticles also exhibited inhibition of microcolony/initial biofilm, mature and preformed biofilm formed by Klebsiella pneumoniae (53.42 ± 4.45%, 62.14 ± 7.45%, and 24.12 ± 9.87%, respectively), Aeromonas hydrophila (43.89 ± 0.95%, 59.79 ± 1.19%, and 54.85 ± 6.50%, respectively) and Escherichia coli (45.28 ± 6.88%, 45.17 ± 3.94%, and 32.39 ± 4.16%, respectively). The physicochemical characterization of potential nanoparticles was conducted using UV-visible Spectrophotometer, X-ray diffractionSpectroscopy (XRD), Energy Dispersive X-ray (EDX) spectroscopy, Fourier Transform Infrared Spectroscopy (FTIR), and Scanning Electron Microscopy (SEM). These physicochemical analyses confirmed the formation of gold nanoparticles which were found to be spherical in shape with the size between 30 and 40 nm. Furthermore, the potential fungal isolate S2-10 was characterized as Aspergillus niger isolate CJ14 with the help of morphological and molecular techniques and deposited at National Center for Biotechnology Information (NCBI) (Accession number MN519438.1). Hence, it can be concluded that the gold nanoparticles produced using A. niger isolate CJ14 contain significant antibiofilm properties and can be used for developing antibiofilm products for future applications.


Oxytocin reduces background anxiety in a fear-potentiated startle paradigm.
Oxytocin reportedly decreases anxious feelings in humans and may therefore have therapeutic value for anxiety disorders, such as post-traumatic stress disorder (PTSD). As PTSD patients have exaggerated startle responses, a fear-potentiated startle paradigm in rats may have face validity as an animal model to examine the efficacy of oxytocin in treating these symptoms. Oxytocin (0, 0.01, 0.1, or 1.0 μg, subcutaneously) was given either 30 min before fear conditioning, immediately after fear conditioning, or 30 min before fear-potentiated startle testing to assess its effects on acquisition, consolidation, and expression of conditioned fear, respectively. Startle both in the presence and absence of the fear-conditioned light was significantly diminished by oxytocin when administered at acquisition, consolidation, or expression. There was no specific effect of oxytocin on light fear-potentiated startle. In an additional experiment, oxytocin had no effects on acoustic startle without previous fear conditioning. Further, in a context-conditioned test, previous light-shock fear conditioning did not increase acoustic startle during testing when the fear-conditioned light was not presented. The data suggest that oxytocin did not diminish cue-specific conditioned nor contextually conditioned fear, but reduced background anxiety. This suggests that oxytocin has unique effects of decreasing background anxiety without affecting learning and memory of a specific traumatic event. Oxytocin may have antianxiety properties that are particularly germane to the hyper-vigilance and exaggerated startle typically seen in PTSD patients.


The specificity of H-2-restricted cytotoxic T lymphocytes directed to AKR/Gross leukemia virus-induced tumors. I. Isolation of a selectively resistant variant tumor subclone.
The AKR.H-2bSL1 tumor cell line is susceptible to H-2Kb-restricted cytotoxic T lymphocytes (CTL) directed against the subclass of AKR endogenous leukemia virus-induced tumors that express the Gross cell surface antigen (anti-AKR/Gross virus CTL). A variant subclone (cl.18-5) of AKR.H-2bSL1 was isolated, whose susceptibility to lysis by conventional or cloned lines of anti-AKR/Gross virus CTL was approximately 5% or less than that of the parental tumor. The cl.18-5 variant was also ineffective when used as an in vivo priming cell or an in vitro stimulator cell in the generation of anti-AKR/Gross virus CTL or as an unlabeled target cell in competitive inhibition assays. These results implied that the failure of cl.18-5 to be lysed was due to a lack of recognition by the CTL. In contrast, cl.18-5 was able to be lysed by and stimulate the generation of predominantly H-2Db-restricted CTL with apparent specificity for AKR minor histocompatibility antigens. The variant line was also about as susceptible as the parental AKR.H-2bSL1 line to both allogeneic CTL and to H-2Kb-restricted, TNP-specific CTL. Thus, the lack of recognition of cl.18-5 by anti-AKR/Gross virus CTL did not appear to be due to a failure to express functional H-2 products or to a generalized insusceptibility to H-2-restricted CTL. Rather, cl.18-5 appeared to be a selective variant and a useful probe for studying the specificity of anti-AKR/Gross virus CTL.


Prospective randomized comparison of two different sized percutaneous endoscopically placed gastrostomy tubes.
We performed a prospective randomised study of two different sized percutaneous endoscopic gastrostomy (PEG) tubes to determine if tube size influenced the incidence of PEG-related complications. Patients were given prophylactic cefuroxime, if not already on antibiotics at the time of PEG insertion. Fifty-two PEGs were successfully placed, 26 in each group. Most patients who required a PEG had suffered a cerebrovascular event (82.7%). There were no procedure-related deaths. The mean ages (standard deviation) for the 12 and 20 French Gauge (FG) groups were 78.7 (8.9) and 73.9 (14.4) years, respectively, with no statistical difference. There were no significant differences in mortality (9 deaths in the 12 FG and 11 deaths in the 20 FG groups), number of peristomal infections (8 infections in the 12 FG and 12 infections in the 20 FG groups), episodes of leakage (12 leakages in the 12 FG and 17 leakages in the 20 FG groups) or tube blockage (2 blockage episodes in the 12 FG and 1 blockage episode in the 20 FG groups) between the two groups over a follow-up period of 190 days. The incidence of insertion- and feeding-related complications was thus not influenced by tube size. As the smaller PEG tubes were easier and less traumatic to insert we conclude that there are grounds for considering the more widespread use of the narrower diameter 12 FG PEG tubes.


Effect of cyclosporine A on male reproduction in rats.
We examined the effects of the immunosuppressive drug cyclosporine A on male reproduction in sexually mature rats. The drug was administered subcutaneously in 3 doses (10, 20 and 40 mg. per kg. daily) for 14 days. Cyclosporine A administration resulted in a dose-dependent decline in body and reproductive organ weights, histology of the testis showed definite degenerative changes, and sperm counts and motility decreased. Spermatozoa from treated rats retained the cytoplasmic droplet and they were decapitated. Consequently, sterility occurred in rats treated with high doses of cyclosporine A. A reduction in circulating testosterone and an increase in gonadotropins were noted. The possibility existed that the cyclosporine A-induced alterations in the male reproductive system were secondary to the hepatic or nephrotoxic effects of the drug. Therefore, we assessed liver and kidney function by assay of the serum levels of bilirubin, alkaline phosphatase, serum glutamic oxaloacetic transaminase, serum glutamic pyruvic transaminase and creatinine. Except for an increase in bilirubin, none of the other liver function studies was altered, thereby eliminating hepatocellular damage as the etiology of impaired reproductive function in cyclosporine A-treated rats. We observed an increase in creatinine levels in animals treated with higher doses of cyclosporine A, consistent with mild renal failure. However, the fact that a decline in reproductive function was seen even in the group treated with 10 mg. per kg. cyclosporine A daily, which had normal serum creatinine levels, suggests that the alterations in male reproduction seen with this therapy are not entirely the result of nephrotoxicity.


Arrhythmogenic effect of a crude extract from sea anemone Condylactis gigantea: possible involvement of rErg1 channels.
Sea anemones possess a number of peptide toxins that target ion channels which provide powerful tools to study the molecular basis of diverse signaling pathways. It is also acknowledged that currents through Erg1 K(+) channels in cardiac myocytes are important for electrical stability of the heart and alterations in its activity has been linked to the onset of a potentially life-threatening heart condition named long QT syndrome type 2. Here, we report that a crude extract from sea anemone Condylactis gigantea significantly increases the QT interval and has arrhythmogenic effects in the rat heart. Furthermore, a bioassay-guided purification procedure allowed the isolation of a chromatographic fraction containing a major component with a molecular mass of 4478 Da from the crude extract, which causes a significant inhibition of whole-cell patch-clamp currents through recombinant Erg1 channels, responsible of the rapid delayed rectifying current crucial for electrical activity in the heart. Further studies could provide relevant information on the molecular mechanism of C. gigantea peptide toxins which represent promising tools in studying the physiology of diverse ion channels.


Efficiently quantifying DNA methylation for bulk- and single-cell bisulfite data.
DNA CpG methylation (CpGm) has proven to be a crucial epigenetic factor in the mammalian gene regulatory system. Assessment of DNA CpG methylation values via whole-genome bisulfite sequencing (WGBS) is, however, computationally extremely demanding. We present FAst MEthylation calling (FAME), the first approach to quantify CpGm values directly from bulk or single-cell WGBS reads without intermediate output files. FAME is very fast but as accurate as standard methods, which first produce BS alignment files before computing CpGm values. We present experiments on bulk and single-cell bisulfite datasets in which we show that data analysis can be significantly sped-up and help addressing the current WGBS analysis bottleneck for large-scale datasets without compromising accuracy. An implementation of FAME is open source and licensed under GPL-3.0 at https://github.com/FischerJo/FAME.


Peripheral extracorporeal membrane oxygenation (ECMO) in patients with posttransplant cardiac graft failure.
We sought to report the usefulness of extracorporeal membrane oxygenation (ECMO) in heart transplant patients. Between March 2002 and August 2004, 14 heart transplant patients (11 men and three women, 36 +/- 15 years old, range = 12 to 50) with primary graft failure underwent peripheral ECMO implantation. Three patients had pulmonary hypertension and three had been transplanted with hearts from marginal donors. At the time of implantation, all were in severe cardiogenic shock despite maximal inotropic support. In six patients, the ECMO was implanted in the operating room since cardiopulmonary bypass could not be weaned. In the eight remaining patients, ECMO was implanted in the intensive care unit, during the first 48 hours in seven cases. In one patient, implantation was performed during external resuscitation. In all cases, femoral vessels were canulated using the Seldinger technique after anterior wall exposure. Distal arterial perfusion of the lower limb was systematically used. Pump outflow was high enough in all the cases (mean: 2.6 +/- 0.2 L/min/m(2)). Three patients died on circulatory support. One patient was implanted with a total artificial heart after a few hours and another one underwent unsuccessful emergent retransplantation. Nine patients were weaned from ECMO after a mean duration of 5 +/- 2.5 days. Among them, one died of infection at 10 days after weaning and seven others were discharged to rehabilitation centers. Fast operating room or bedside implantation of a peripheral ECMO allows the physician to stabilize the hemodynamic status of patients with cardiac graft failure, potentially leading toward myocardial recovery.


Time dependent and other possible changes of the intracellular D-glucose and Na+ concentration in the cortical tissue of rabbit kidney perfused "in vitro".
The mean intracellular D-glucose concentration (Gi), the ratio between Gi and the serosal extracellular concentration (Gi/Ge) and the cell Na+ (Nai+) were calculated on cortex slices at increasing times after their excision from two groups of kidneys perfused with blood containing 18.1 +/- 0.4 and 23.1 +/- 2.0 mmol.l-1 plasma glucose concentration. The distribution ratios Gi/Ge, always lower than 1, were substantially unchanged up to at least 50s after removal of the tissue from the organ. Between these time limits the calculated Gi and Gi/Ge does not seem to be affected by a rapid glucose breakdown in the isolated tissue. Neither such low values depend on a degrading of the intraextracellular Na+ concentration gradient, since this gradient seems to remain unchanged throughout the same time. In another group of experiments, in which phloretin was added to the perfusing blood (mean 0.6 X 10(-3) mol.l-1) at different glucose levels, the mean values of Gi/Ge in the cortical tissue were higher (in many cases greater than 1) than those obtained in a group of control kidneys perfused in absence of phloretin. According to the presented data, the mean Gi and Gi/Ge values, as calculated by us, seem to correspond to those existing in the functioning tissue just before its excision. The already suggested view that the low values of Gi and Gi/Ge could depend on the possible existence of a mechanism actively extruding glucose from the basolateral membrane of tubular cells seems to be reinforced.


Association between Payer-Class and Patient Satisfaction Scores in an Academic Urology Outpatient Clinic.
As physician reimbursement models transition from fee-for-service to value-based purchasing, patient satisfaction is becoming an important payment modifier. Studies have shown correlation between nonmodifiable patient attributes and satisfaction scores. This study aims to investigate potential patient bias in survey scores associated with payer type in an academic urology outpatient clinic setting. The survey results of 20,944 patients seen in an academic urology clinic were analyzed, and the weighted mean±SD scores of patient experience in 6 major payer categories were compared: 1) commercial insurance, 2) Medicare, 3) Medicaid, 4) other government plans, 5) workers' comp and 6) self-pay/uncompensated. ANOVA and Tukey-Kramer Honestly Significant Difference were utilized to assess statistical significance. Medicare patients reported the highest satisfaction scores (93.03), followed by other government coverage (91.49), commercial insurance (91.32), workers' comp (90.72), Medicaid (89.21) and self-pay/uncompensated (89.00). ANOVA analysis resulted in an F value of 40.198 (p <0.001). Tukey-Kramer demonstrated statistical significance in the difference between the means of the following groups: commercial vs Medicaid (p <0.001), commercial vs Medicare (p <0.001), commercial vs self-pay/uncompensated (p <0.01), other government vs Medicaid (p <0.001), other government vs Medicare (p <0.01), Medicaid vs Medicare (p <0.001), Medicare vs self-pay/uncompensated (p <0.001). Our data suggest that patient experience scores are impacted by the type of coverage the patient carries and not only by the quality of care provided. Urology clinicians should be aware of these nuances when selecting MIPS quality reporting metrics. Currently, these biased scores may have an impact on physicians' reimbursement, and policymakers should consider adjusting reimbursement according to payer-mix.


Development of a real-time reverse transcription PCR assay for detection of a novel nidovirus associated with a neurological disease of the Australian brushtail possum (Trichosurus vulpecula).
To develop a quantitative reverse transcription PCR (RT-qPCR) assay for detection of the putative wobbly possum disease (WPD) virus and to apply this test to investigate the viral load in archival tissues from past WPD transmission studies. The real-time assay was developed as a two-step RT-qPCR in a SYBR green format and validated using serial dilutions of a linearised plasmid containing target DNA. The copy number values were normalised to the amount of RNA in each reverse transcription reaction and presented as the number of viral copies per μg of total [corrected] RNA. The viral load was determined in archival samples from animals that had received inoculations of infectious WPD tissue suspensions. Thirty samples originating from 22 possums, comprising five samples from three healthy possums and 25 samples from 19 possums that had received inoculations of infectious WPD tissue suspensions were tested. The assay was linear (R(2) > 0.99) within the tested range from 1 to 10(7) target copies/µL, with an efficiency of >90%. The intra-assay variability CV values ranged from 0.8 to 4.5% for different standards, with the inter-assay variability CV values ranging from 0.4 to 2.5%, indicating good precision and reproducibility of the assay. The novel nidovirus was detected in all 25 samples from WPD-affected possums. Tissues from three control possums and from one experimentally infected rabbit were negative for WPD RNA. The viral load in WPD-positive tissues differed between individual possums and between tissue types, ranging from 2.2 to 359,980 copies/pg RNA. The highest viral load was detected in liver, followed by brain, spleen, kidney and urine. There was a more than four log difference in the viral load between pools of tissues originating from two outbreaks of WPD in different geographical regions. Detection of viral RNA in a variety of tissues from WPD affected possums, including brain, is consistent with the multi-organ distribution of histopathological lesions observed in WPD. Our data suggest that liver may constitute the sample of choice for diagnostic testing. Differences in the viral load in tissues from possums inoculated with infectious WPD tissue suspensions from Rotorua or Invermay origin suggest that WPD viruses with different biological properties may exist. We have developed a RT-qPCR assay for detection of the putative WPD virus. The test showed good sensitivity and reproducibility over the wide dynamic range of template concentrations. It provides a tool for future diagnostic and research purposes.


Oral Fabrol (oral N-acetyl-cysteine) in chronic bronchitis.
Five hundred and twenty-six patients suffering from chronic bronchitis were randomized to receive either N-acetylcysteine (NAC) or placebo during a 6-month period. The aim was to compare the number of acute exacerbations in the two groups. General practitioners were asked to enter patients with a diagnosis of chronic bronchitis, based on the MRC criteria. We failed to find any statistically significant difference in the number of exacerbations between the two treatment groups although there was a slight trend towards improvement in the NAC group during the first 3 months of the trial. The tolerability was similar for both treatments. Patients taking NAC showed a reduction in number of days on which they were incapacitated and this result was statistically significant.


Lipid fingerprinting in women with early-onset preeclampsia: a first look.
The aim of this preliminary study was to characterize the plasma lipid profiling of women with preeclampsia. Plasma samples of 8 pregnant women with early-onset preeclampsia and 8 normal pregnant women were evaluated. Lipids were extracted from plasma using the Bligh-Dyer protocol. The extracts were subjected to MALDI-MS. Data matrix was exported for partial least squares discriminant analysis (PLS-DA) and a parameter VIP was employed to reflect the variable importance in the discriminant analysis. The major discriminant variables were selected and underwent to Mann-Whitney U test. A total of 1290 ions were initially identified and twelve m/z signals were highlighted as the most important lipids for the discrimination of patients with preeclampsia. The identification of these differential lipids was carried out through Lipid Database Search. The main classes identified were glycerophosphocholines [GP01], glycerophosphoserines [GP03], glycerophosphoglycerols [GP04], glycosyldiradylglycerols [GL05] and glycerophosphates [GP10].


Meal-stimulated cholecystokinin release and exocrine pancreatic secretion in dogs.
To confirm the role of cholecystokinin (CCK) and secretin in digestion, exocrine pancreatic secretion, plasma CCK, and secretin were measured simultaneously in six dogs prepared with gastric and pancreatic fistulas after feeding a solid meal. Plasma CCK concentration determined by radioimmunoassay increased significantly from the basal level, reached a peak 35 min after meal ingestion, and after a dip it further increased toward the end of the 3-h observation. Pancreatic protein output increased significantly, peaked at the fifth 10-min period, and then declined progressively. Plasma CCK concentration and pancreatic protein output correlated significantly during the first postprandial hour. Plasma secretion demonstrated significant elevation at 15 min and a peak at 25 min after a meal. Plasma secretin correlated significantly with both bicarbonate output and flow rate during the 3 h. Simultaneous measurements of plasma CCK and secretin and of pancreatic secretion suggested that postprandial pancreatic secretion is primarily mediated by releases of CCK and secretin, but these hormones do not seem to be the only factors responsible for the secretion.


Effects of antimicrobial peptides on membrane dynamics: A comparison of fluorescence and NMR experiments.
Antimicrobial peptides (AMPs) represent a promising class of compounds to fight resistant infections. They are commonly thought to kill bacteria by perturbing the permeability of their cell membranes. However, bacterial killing requires a high coverage of the cell surface by bound peptides, at least in the case of cationic and amphipathic AMPs. Therefore, it is conceivable that peptide accumulation on the bacterial membranes might interfere with vital cellular functions also by perturbing bilayer dynamics, a hypothesis that has been termed "sand in the gearbox". Here we performed a systematic study of such possible effects, for two representative peptides (the cationic cathelicidin PMAP-23 and the peptaibol alamethicin), employing fluorescence and NMR spectroscopies. These approaches are commonly applied to characterize lipid order and dynamics, but sample different time-scales and could thus report on different membrane properties. In our case, fluorescence anisotropy measurements on liposomes labelled with probes localized at different depths in the bilayer showed that both peptides perturb membrane fluidity and order. Pyrene excimer-formation experiments showed a peptide-induced reduction in lipid lateral mobility. Finally, laurdan fluorescence indicated that peptide binding reduces water penetration below the headgroups region. Comparable effects were observed also in fluorescence experiments performed directly on live bacterial cells. By contrast, the fatty acyl chain order parameters detected by deuterium NMR spectroscopy remained virtually unaffected by addition of the peptides. The apparent discrepancy between the two techniques confirms previous sporadic observations and is discussed in terms of the different characteristic times of the two approaches. The perturbation of membrane dynamics in the ns timescale, indicated by the multiple fluorescence approaches reported here, could contribute to the antimicrobial activity of AMPs, by affecting the function of membrane proteins, which is strongly dependent on the physicochemical properties of the bilayer.


Algorithm for vector autoregressive model parameter estimation using an orthogonalization procedure.
We review the derivation of the fast orthogonal search algorithm, first proposed by Korenberg, with emphasis on its application to the problem of estimating coefficient matrices of vector autoregressive models. New aspects of the algorithm not previously considered are examined. One of these is the application of the algorithm to estimate coefficient matrices of a vector autoregressive process with time-varying coefficients when multiple realizations of the said process are available. Computer simulations were also performed to characterize the statistical properties of the estimates. The results show that even for shorter time series the algorithm works well and obtains good estimates of the time-varying parameters. Statistical characterization indicates that the standard deviation of the estimates decreases as 1 square root N (N being the length of the time series), a typical behavior of least-squares estimators. Another key aspect of the approach, which has previously been considered, is its direct extension to the parameter estimation of vector nonlinear autoregressive models. Nonlinear terms can be added to the model and the same algorithm can be applied to effectively estimate their associated parameters. Using chaotic time series generated from the Lorenz equations, the algorithm produces a model that captures the nonlinear structure of the data and exhibits the same chaotic attractor as that of the original system.


LOX-1/LOXIN: the yin/yang of atheroscleorosis.
Atherosclerosis is the first cause of death in industrialized countries. Together with traditional risk factors (male gender, hypercholesterolemia, hypertension, diabetes, smoking and age), non-traditional risk factors have also been described as predisposing to this disease. Among these, oxidized low density lipoproteins (OxLDL) have been described in correlation to many proatherogenic processes. Many of the effects of OxLDL are mediated by the lectin like oxidized low density lipoprotein receptor 1 (LOX-1), expressed on endothelial cells, macrophages, SMCs and platelets. LOX-1 is encoded by the lectin like oxidized low density lipoprotein receptor 1 (OLR1) gene, located in the p12.3-p13.2 region of human chromosome 12. Variations on this gene have been studied extensively both at the functional and epidemiological level. Despite the fact that functional roles for two variants have been demonstrated, the epidemiological studies have provided inconsistent and inconclusive results. Of particular interest, it has been demonstrated that a linkage disequilibirum block of SNPs located in the intronic sequence of the OLR1 gene modulates the alternative splicing of OLR1 mRNA, leading to different ratios of LOX-1 full receptor and LOXIN, an isoform lacking part of the functional domain. As demonstrated, LOXIN acts by blocking the negative effective of LOX-1 activation. Here we review the state of the art regarding LOX-1, LOXIN, and the functional effects that are associated with the interaction of these molecules.


The effect of vanadium on essential element uptake of Setaria viridis' seedlings.
High concentrations of vanadium, a ubiquitous element in the environment, in growing media leads to deformation of root structure and leaf chlorosis and necrosis, consequently affecting the translocations of nutrients and essential elements. However, the effects of vanadium on essential element uptake, and the interactions of essential elements in the presence of vanadium, remain incompletely understood. To elucidate the effects of different concentrations of vanadium on major and trace essential elements and plant growth, a native plant species growing in a vanadium mining area, Setaria viridis (dog tail's grass), was incubated in solutions containing 0-55.8 mg/L vanadium. The shoot accumulation of four major essential elements and five trace essential elements was detected, and the root length and stem height were measured. The results showed that vanadium in soil solution enhanced the accumulation of all major essential elements in shoot. Vanadium concentrations lower than 47.4 mg/L showed an obvious positive (p < 0.05) effect on P accumulation and translocation. In the case of trace essential elements, there were threshold values for solution vanadium stimulation of element uptake. The threshold values for Cu and Zn, Fe, and Mo uptake were 4.3, 16.3, and 40.6 mg/L, respectively. When vanadium levels surpassed these values, accumulation was suppressed and the solution vanadium concentrations attenuated the solution-to-shoot translocation of most of the essential elements. Among the trace essential elements, translocation of Fe was obviously enhanced (p < 0.05) by vanadium. Solution vanadium also enhanced plant growth at lower concentrations and inhibited it at higher levels. The threshold values for stem height and root length were 36.8 and 16.3 mg/L, respectively. Concentrations of 40 and 55.8 mg/L vanadium in soil solution caused a 50% decrease in root length and stem height, respectively, showing that root length of Setaria viridis is more susceptible to vanadium toxicity than stem growth.


Hypertonic sodium chloride induction of cyclooxygenase-2 occurs independently of NF-kappaB and is inhibited by the glucocorticoid receptor in A549 cells.
Cellular response to a hypertonic environment is important for fluid clearance in the lung. Hypertonicity modulates prostaglandin synthesis by influencing cyclooxygenase-2 (COX-2) expression in tissues such as liver and kidney via a mitogen-activated protein kinase (MAPK)-dependent pathway. However, little is known about COX-2 expression in response to hypertonicity in the lung. COX-2 mRNA accumulation induced by hypertonic NaCl was detected after 1 h of treatment, and COX-2 mRNA continued to accumulate until 18 h, the longest time point examined, in human alveolar epithelial A549 cells. This induction was a transcriptional event that occurred in the absence of the protein synthesis inhibitor cycloheximide and was the result of enhanced promoter activity, as examined with the use of full-length COX-2 promoter-driven reporter plasmids. The induction of COX-2 expression by hypertonic NaCl did not require the activation of NF-kappaB. The p38 MAPK inhibitor, SB203580, or MEK1/2 inhibitor, U0126, inhibited hypertonic induction of COX-2 expression. We examined whether the hypertonic induction of COX-2 was under the influence of glucocorticoid; we found that COX-2 promoter activity and mRNA and protein levels were depressed by dexamethasone and antagonized by the glucocorticoid receptor (GR) antagonist RU486. Our data demonstrate that the induction of COX-2 expression by hypertonic NaCl occurs independently of NF-kappaB and is inhibited by the GR in A549 cells.


Preparation of gluten free bread enriched with green mussel (Perna canaliculus) protein hydrolysates and characterization of peptides responsible for mussel flavour.
Green mussel protein hydrolysates (GMPH) utilization for the enrichment of gluten-free bread followed by characterization of flavour peptides using chromatography and electronic nose techniques have been done. The degree of hydrolysis was carried out in each protease digest, and the higher degree of hydrolysis was observed in pepsin digestion. Gluten-free (GF) bread was formulated by using buckwheat flour (BWF), rice flour (RF) and chickpea flour (CPF) (70:20:10) and GMPH were added in the range of 0-20% in the GF bread for enrichment with GMPH. Radar plot of the electronic nose analysis showed that the sensors P30/2, T30/1 and T70/2 had a higher response to the GF bread and GMPH. Consequently, the peptide sequence was obtained manually by ESI-MS spectra of GMPH (KGYSSYICDK) and F-II (SSYCIVKICDK). Flavour quality was 97% discriminately comparable to the GMPH and F-II fractions. Mussel flavoured GF bread can be included in the celiac diet.


Pollen limitation may be a common Allee effect in marine hydrophilous plants: implications for decline and recovery in seagrasses.
Pollen limitation may be an important factor in accelerated decline of sparse or fragmented populations. Little is known whether hydrophilous plants (pollen transport by water) suffer from an Allee effect due to pollen limitation or not. Hydrophilous pollination is a typical trait of marine angiosperms or seagrasses. Although seagrass flowers usually have high pollen production, floral densities are highly variable. We evaluated pollen limitation for intertidal populations of the seagrass Zostera noltei in The Netherlands and found a significant positive relation between flowering spathe density and fruit-set, which was suboptimal at <1200 flowering spathes m(-2) (corresponding to <600 reproductive shoots m(-2)). A fragmented population had ≈35 % lower fruit-set at similar reproductive density than a continuous population. 75 % of all European populations studied over a large latitudinal gradient had flowering spathe densities below that required for optimal fruit-set, particularly in Southern countries. Literature review of the reproductive output of hydrophilous pollinated plants revealed that seed- or fruit-set of marine hydrophilous plants is generally low, as compared to hydrophilous freshwater and wind-pollinated plants. We conclude that pollen limitation as found in Z. noltei may be a common Allee effect for seagrasses, potentially accelerating decline and impairing recovery even after environmental conditions have improved substantially.


Mineralization of 4-fluorocinnamic acid by a Rhodococcus strain.
A bacterial strain capable of aerobic degradation of 4-fluorocinnamic acid (4-FCA) as the sole source of carbon and energy was isolated from a biofilm reactor operating for the treatment of 2-fluorophenol. The organism, designated as strain S2, was identified by 16S rRNA gene analysis as a member of the genus Rhodococcus. Strain S2 was able to mineralize 4-FCA as sole carbon and energy source. In the presence of a conventional carbon source (sodium acetate [SA]), growth rate of strain S2 was enhanced from 0.04 to 0.14 h(-1) when the culture medium was fed with 0.5 mM of 4-FCA, and the time for complete removal of 4-FCA decreased from 216 to 50 h. When grown in SA-supplemented medium, 4-FCA concentrations up to 1 mM did not affect the length of the lag phase, and for 4-FCA concentrations up to 3 mM, strain S2 was able to completely remove the target fluorinated compound. 4-Fluorobenzoate (4-FBA) was transiently formed in the culture medium, reaching concentrations up to 1.7 mM when the cultures were supplemented with 3.5 mM of 4-FCA. Trans,trans-muconate was also transiently formed as a metabolic intermediate. Compounds with molecular mass compatible with 3-carboxymuconate and 3-oxoadipate were also detected in the culture medium. Strain S2 was able to mineralize a range of other haloorganic compounds, including 2-fluorophenol, to which the biofilm reactor had been exposed. To our knowledge, this is the first time that mineralization of 4-FCA as the sole carbon source by a single bacterial culture is reported.


ZZEF1 is a Histone Reader and Transcriptional Coregulator of Krüppel-Like Factors.
The ZZ-type zinc finger and EF-hand domain protein 1 (ZZEF1) is a multidomain-containing protein. Mutations of ZZEF1 has been implicated in several kinds of human diseases such as diabetes and cancers. However, the function of the ZZEF protein remains largely unknown. Here we show that ZZEF1 functions as a histone H3 reader. The second ZZ domain of ZZEF1 (ZZEF1ZZ2) binds to the N-terminus of histone H3 and is capable of accommodating common epigenetic marks on the H3 tail. The N-terminal amino acids, especially Ala1, of H3 and an acidic cavity of ZZEF1ZZ2 are critical for the ZZ-H3 interaction. RNA-seq analysis in human lung cancer cell line H1299 reveals that downregulated genes upon ZZEF1 depletion are specifically enriched in genes regulated by Krüppel-like factors. Indeed, ZZEF1 physically interacts with KLF9 and KLF6, and regulates a common set of target genes of these transcription factors. Together, our findings suggest a model in which ZZEF1 binds to histone H3 tail and promotes KLF9/6-mediated gene regulation.


[Damage to the isolated heart due to adrenaline perfused with a saline solution].
Perfusion of the isolated heart with Krebs solution containing 5 and 20 microgram/ml of adrenaline induced cardiocyte micronecrosis. Perfusion with 0.5 microgram/ml of adrenaline induced no micronecrosis. Dispersion analysis showed a statistically significant effect of adrenaline concentrations on the degree of the cardionecrotic effect. The fact of micronecrosis appearance in the isolated heart during its perfusion with saline solution requires revision of the hypothesis on the leading role of blood factor in the realization of the cardionecrotic effect of adrenaline. The appearance of micronecroses with the action of adrenaline in concentrations which activate the mechanism of amines uptake by the heart myocytes speaks in favour of the casual relationship between the accumulation of the biogenic amines by myocytes and the development of their necrosis.


Analysis of ligand binding curves in terms of species fractions.
The ligand binding curve for a macromolecular system presents the average number of ligand molecules bound per macromolecule as a function of the chemical potential or the logarithm of the ligand concentration. We show that various observable properties of this curve, for example its asymptotes and derivatives, are expressible in terms of linear combinations of the mole fractions alphai of macromolecules binding i molecules of ligand. Whenever enough such properties of the binding curve are known, the linear equations in alphai can be solved to give the mole fractions of each of the various macromolecular species. An application of these results is that a Hill plot for hemoglobin-ligand equilibrium where the asymptotes approach unit slope can be made to yield the four Adair constants by a simple algebraic method. A second use is that a knowledge of the first and second derivatives of the binding curve at points along the curve can yield the species fractions as functions of the degree of saturation without direct knowledge of the ligand binding constants. These methods are illustrated by some numerical examples.


The picture exchange communication system.
The Picture Exchange Communication System (PECS) was developed as a means to teach children with autism and related developmental disabilities a rapidly acquired, self-initiating, functional communication system. Its theoretical roots combine principles from applied behavior analysis and guidelines established within the field of alternative and augmentative communication. This approach has several potential advantages relative to imitation-based strategies (both vocal and gestural) and symbol selection strategies. The system begins with the exchange of simple icons but rapidly builds "sentence" structure. The system also emphasizes developing the request function prior to developing responding to simple questions and commenting. The development of requesting with a sentence structure also permits the rapid development of attributes more traditionally taught within a receptive mode. The relationship between the introduction of PECS and various other behavioral issues (i.e., social approach and behavior management) as well as its relationship to the codevelopment of speech are reviewed.


Multiproxy characterization of sedimentary facies in a submarine sulphide mine tailings dumping site and their environmental significance: The study case of Portmán Bay (SE Spain).
Mining activities are essential to our society, but ore extraction and treatment produce waste that must be stored in safe places without harm to the environment. For a long time, seafloor disposal has been viewed as a cheap option with barely visible impacts. In Portmán Bay, SE of Spain, large amounts of tailings from open pit sulphide mining were discharged directly into the coastal sea over 33 years, thus forming a massive deposit that completely infilled the bay and expanded seawards over the inner continental shelf. Here we present the first multiproxy physicochemical characterization of the submarine tailings in Portmán Bay, mostly by using non-destructive techniques, also including pre-dumping and post-dumping sediments. Eight distinct sedimentary facies, grouped in four stratigraphic units, have been thus identified in a set of up to 4.3 m long gravity cores totalling more than 60 m. Geogenic and anthropogenic geochemical proxies consistently allow differentiating pre-dumping sediments from tailings. Potentially toxic metals if made bioavailable can reach high concentrations in units including or formed exclusively by tailings (i.e. up to 3455, 2755 and 1007 mg kg-1 for Pb, As, and Zn, respectively). Some physical properties, such as magnetic susceptibility, are particularly useful as the tailings are rich in Fe-bearing minerals (>30% Fe in some layers). Estimated sedimentation rates show a strong gradient from proximal to distal locations, with rates in excess of 50 cm yr-1 to less than 1 cm yr-1. We ultimately document the history of the transformation of Portmán Bay from an almost natural state to a new condition after a long period of massive dumping of mine tailings. Our study provides guidance to further assessments in a context where the diversity of marine environments impacted by the disposal of mine waste is expected to grow in the near future.


Calcium oscillations and mammalian egg activation.
Fertilization in all species studied to date induces an increase in the intracellular concentration of free calcium ions ([Ca2+]i) within the egg. In mammals, this [Ca2+]i signal is delivered in the form of long-lasting [Ca2+]i oscillations that begin shortly after fusion of the gametes and persist beyond the time of completion of meiosis. While not fully elucidated, recent evidence supports the notion that the sperm delivers into the ooplasm a trigger of oscillations, the so-called sperm factor (SF). The recent discovery that mammalian sperm harbor a specific phospholipase C (PLC), PLCzeta has consolidated this view. The fertilizing sperm, and presumably PLCzeta promote Ca2+ release in eggs via the production of inositol 1,4,5-trisphosphate (IP3), which binds and gates its receptor, the type-1 IP3 receptor, located on the endoplasmic reticulum, the Ca2+ store of the cell. Repetitive Ca2+ release in this manner results in a positive cumulative effect on downstream signaling molecules that are responsible for the completion of all the events comprising egg activation. This review will discuss recent advances in our understanding of how [Ca2+]i oscillations are initiated and regulated in mammals, highlight areas of discrepancies, and emphasize the need to better characterize the downstream molecular cascades that are dependent on [Ca2+]i oscillations and that may impact embryo development.


VISTAL--a new 2D visualization tool of protein 3D structural alignments.
We offer a tool, denoted VISTAL, for two-dimensional visualization of protein structural alignments. VISTAL describes aligned structures as a series of matched secondary structure elements, colored according to the three-dimensional distance of their Calpha atoms. VISTAL can be downloaded from http://trantor.bioc.columbia.edu/~kolodny/software.html.


Effects of food and grapefruit juice on single-dose pharmacokinetics of blonanserin in healthy Chinese subjects.
The purpose of this study was to investigate the potential effects of a meal and grapefruit juice on the pharmacokinetics of blonanserin and its metabolite N-desethyl blonanserin in healthy Chinese volunteers. This was a single-centre, open-label, fixed-sequence study, where 12 healthy Chinese volunteers received a single dose of 8 mg blonanserin after an overnight fast in period 1 (reference), a high-fat meal during period 2 and with co-administration of 250 mL of grapefruit juice in period 3. The washout period was 7 days. Series of plasma samples were collected after each dose to determine concentrations of blonanserin and its metabolite N-desethyl blonanserin using liquid chromatography-tandem mass spectrometry. Pharmacokinetic parameters were estimated by non-compartmental analysis and compared between periods by standard average bioequivalence ANOVA. Adverse events were monitored throughout the study. All subjects completed the study. High-fat meals significantly increased blonanserin exposure (AUCt) 2.58-fold (90% CI 2.21, 3.02), relative to the reference period. Co-administration of blonanserin with grapefruit juice remarkably prolonged elimination half-life of blonanserin (from 9.7 to 21.4 h) and significantly increased exposures to blonanserin and N-desethyl blonanserin by 5.82-fold (90% CI 4.57, 7.42) and 1.81-fold (90% CI 1.65, 1.98), respectively. These results suggested that blonanserin was largely metabolised in the intestinal tract before becoming systemically available, and both food and grapefruit juice enhanced exposure to blonanserin and N-desethyl blonanserin. Grapefruit juice increased bioavailability and may have reduced systemic clearance of blonanserin. Further intestinal CYP3A4 and hepatic CYP3A4 might be postulated to explain the delayed elimination of blonanserin. Dose adjustment of blonanserin is needed on the basis of co-intake of known strong CYP3A4 inhibitor. Patients taking high-dose blonanserin also need to be cautious about the ingestion of grapefruit juice.


Gonadotropin gene targeting and biological implications.
Pituitary gonadotropins FSH and LH are heterodimeric glycoproteins consisting of a common alpha and a hormone-specific beta subunit that are non-covalently linked. These hormones orchestrate gonadal growth, differentiation, and function by regulating both steroid-ogenesis and gametogenesis. Advances in the past two decades in manipulating the mouse genome by site-specific mutagenesis have heralded a new dimension to our understanding of the biology of gonadotropins. Using these gene-targeting approaches, knockout mice lacking the hormone-specific gonadotropin subunits, and hence the functional dimeric hormones, have been generated. These individual gonadotropin-deficient mice are useful to delineate the distinct in vivo biological roles of FSH and LH. These mice also serve as valuable genetic tools to study the signaling mechanisms within the gonads and help a better understanding of some forms of human infertility.