Manganese modifies Neurotrophin-3 (NT3) and its tropomyosin receptor kinase C (TrkC) in the cortex: Implications for manganese-induced neurotoxicity.
Manganese (Mn), an essential micronutrient, has the potential to induce apoptosis. The NT3/TrkC ligand/receptor pair known as part of the classic neurotrophic theory plays a critical role in neuronal survival. However, whether the NT3/TrkC-mediated signaling pathways are involved in Mn-induced apoptosis of cortical neurons remains unknown. The present study was designed to investigate the interactions between NT3/TrkC-mediated signaling pathways and Mn-induced apoptosis in cortical neurons. This study showed that subacute Mn exposure significantly increased the levels of pro-apoptotic Bax while decreasing the levels of anti-apoptotic Bcl 2 in the cortex compared with the corresponding control. Markedly reduced NT3 and TrkC levels along with decreased Ras/MAPK and PI3/Akt signaling in the cortex were observed following subacute Mn exposure. We further found increased levels of Bax, cleaved caspase-3, and the total apoptosis rate, and decreased levels of Bcl 2, NT3, TrkC, and Ras/MAPK and PI3/Akt signaling in Mn-treated primary cortical neurons. Pretreatment with hNT3 or Z-VAD-FAM ameliorated Mn-induced apoptosis by increasing the levels of NT3 and TrkC and its Ras/MAPK and PI3/Akt signaling pathways. Taken together, our findings clearly indicate that NT3/TrkC and mediated Ras/MAPK and PI3/Akt signaling pathways play a crucial role in Mn-induced neurotoxicity.


The Stress and Resilience Town Hall: A systems response to support the health workforce during COVID-19 and beyond.
The COVID-19 pandemic is a traumatic stressor resulting in anxiety, depression, post-traumatic stress, and burnout among healthcare workers. We describe an intervention to support the health workforce and summarize results from its 40-week implementation in a large, tri-state health system during the COVID-19 pandemic. We conducted 121 virtual and interactive Stress and Resilience Town Halls attended by 3555 healthcare workers. Town hall participants generated 1627 stressors and resilience strategies that we coded and analyzed using rigorous qualitative methods (Kappa = 0.85). We identify six types of stressors and eight types of resilience strategies reported by healthcare workers, how these changed over time, and how town halls were responsive to emerging health workforce needs. We show that town halls dedicated to groups working together yielded 84% higher mean attendance and more sharing of stressors and resilience strategies than those offered generally across the health system, and that specific stressors and strategies are reported consistently while others vary markedly over time. The virtual and interactive Stress and Resilience Town Hall is an accessible, scalable, and sustainable intervention to build mutual support, wellness, and resilience among healthcare workers and within hospitals and health systems responding to emerging crises, pandemics, and disasters.


Towards an automatic early stress recognition system for office environments based on multimodal measurements: A review.
Stress is a major problem of our society, as it is the cause of many health problems and huge economic losses in companies. Continuous high mental workloads and non-stop technological development, which leads to constant change and need for adaptation, makes the problem increasingly serious for office workers. To prevent stress from becoming chronic and provoking irreversible damages, it is necessary to detect it in its early stages. Unfortunately, an automatic, continuous and unobtrusive early stress detection method does not exist yet. The multimodal nature of stress and the research conducted in this area suggest that the developed method will depend on several modalities. Thus, this work reviews and brings together the recent works carried out in the automatic stress detection looking over the measurements executed along the three main modalities, namely, psychological, physiological and behavioural modalities, along with contextual measurements, in order to give hints about the most appropriate techniques to be used and thereby, to facilitate the development of such a holistic system.


Stretch-induced activation of ERK in myocytes is p38 and calcineurin-dependent.
Activation of specific mitogen-activated protein kinases (MAPKs) has been suggested to be involved in phenotype modulation of cells subjected to mechanical strain, which may be common to different mechano-sensitive cell types. We have submitted C2C12 myocytes to a static stretch and examined its effect upon the activation of ERK. Stretch induced a rapid but transient activation of ERK. This activation was however prevented when cells were pre-treated with inhibitors of p38 and calcineurin. The dependence of strain-induced ERK activation upon p38 suggests a cross-talk between these two pathways when mediating a response to a mechanical stimulus in this cell type. This suggests that cross relationships between these MAP kinases may be of crucial importance for myocyte phenotype modulation and differentiation in response to a mechanical stimulus.


Height on the draw-a-person: comparison of figure drawings of black and white male drug abusers.
The Draw-A-Person Test was administered to 30 black and 30 white male drug abusers, in a residential therapeutic community. Mean height and mean area of the female figures drawn by the black subjects were greater than the mean height and mean area of the female figures drawn by the white subjects. More black subjects drew the female figure taller than the male figure. Data were discussed in regard to family background and socio-cultural considerations.


Actin gene sequence from Euglena gracilis.
The full length coding sequence of the Euglena gracilis actin gene was determined by RT-PCR of Euglena gracilis mRNA. Conserved regions in the actin amino acid sequence were used as guides for the synthesis of degenerate primers. Sequence was obtained for 1,238 nucleotides, of which 1,131 were coding for 377 amino acids. Sequence comparisons showed a similarity with other actins of 56% to 80%. Even though most of the actin amino acid sequence was conserved, some regions showed high divergence, i.e. the DNase I-binding loop at the N-terminal region. The construction of a phylogenetic tree based on actin sequences from different organisms placed Euglena gracilis in a cluster with Trypanosoma brucei and Leishmania major.


Characteristics of single fallers and recurrent fallers among hospital in-patients.
Recurrent fallers constitute a minority of patients who fall but contribute considerably to the total number of falls recorded. To study the characteristics of recurrent fallers in a hospital setting. In a prospective observational study we investigated the characteristics of 1,025 patients admitted to a geriatric non-acute hospital. Patients were followed until discharge and were classified as non-fallers, single fallers or recurrent fallers. We identified 824 non-fallers, 136 single fallers and 65 recurrent fallers contributing 175 falls. Compared to non-fallers, recurrent fallers were more likely to have pre-admission falls (p = 0.004), confusion (p < 0.0001), an unsafe gait (p = 0.0001) and be on tranquillisers (p = 0.018) and antidepressants (p = 0.006). They had longer stays in hospital (p < 0.0001) and more nursing home discharges (p = 0.0001). There was considerable overlap with risk factors for single fallers but compared to this group they were more likely to be confused (p = 0.027), and on antidepressant medication (p = 0.009). They also had a longer length of stay (p < 0.001) and more nursing home discharges (p = 0.03). Confusion (p = 0.0001), unsafe gait (p = 0.0006) and antidepressants (p = 0.018) were independently associated with recurrent falls. It is important to recognise the risk factors that prospectively identify a recurrent faller because of the significant contribution to total falls by a relatively small number of patients. This may be useful not only in trying to reduce total falls but also in trying to reduce injury.


Effects of Subchronic Aluminum Exposure on Learning, Memory, and Neurotrophic Factors in Rats.
Aluminum (Al) is a neurotoxin that gradually accumulates in the brain in human life, resulting in oxidative brain injury related to Alzheimer's disease (AD) and other diseases. In this study, the learning and memory of rats exposed to different aluminum concentrations (0.0 g/L, 2.0 g/L, 4.0 g/L, and 8.0 g/L) were studied, and the learning and memory of rats were observed by shuttle box experiment. With hematoxylin and eosin staining, Western blot, immunofluorescence, and RT-PCR, the morphology of nerve cells in the hippocampus of rat brain were observed, and the levels of activator protein-1 (AP-1) gene and protein, nerve growth factor (NGF), neurotrophin-3 (NT3), glial cell line-derived neurotrophic factor (GDNF), and brain-derived neurotrophic factor (BDNF) gene and protein level, etc. The experimental results showed that subchronic aluminum exposure damaged learning and memory in rats. The cognitive function damage in rats was more evident after increasing the aluminum intake dose. The more aluminum intake, the more pronounced the histological changes in the hippocampus will be. The expression level and protein content of neurotrophic factors in the hippocampus of rats showed a negative correlation with aluminum intake. In this experiment, we explored the mechanism of aluminum exposure in learning and memory disorders, and provided some data reference for further elucidation of the damage mechanism of aluminum on the nervous system and subsequent preventive measures.


Oligodendrocyte PTEN is required for myelin and axonal integrity, not remyelination.
Repair of myelin injury in multiple sclerosis may fail, resulting in chronic demyelination, axonal loss, and disease progression. As cellular pathways regulated by phosphatase and tensin homologue deleted on chromosome 10 (PTEN; eg, phosphatidylinositol-3-kinase [PI-3K]) have been reported to enhance axon regeneration and oligodendrocyte maturation, we investigated potentially beneficial effects of Pten loss of function in the oligodendrocyte lineage on remyelination. We characterized oligodendrocyte numbers and myelin sheath thickness in mice with conditional inactivation of Pten in oligodendrocytes, Olig2-cre, Pten(fl/fl) mice. Using a model of central nervous system demyelination, lysolecithin injection into the spinal cord white matter, we performed short- and long-term lesioning experiments and quantified oligodendrocyte maturation and myelin sheath thickness in remyelinating lesions. During development, we observed dramatic hypermyelination in the corpus callosum and spinal cord. Following white matter injury, however, there was no detectable improvement in remyelination. Moreover, we observed progressive myelin sheath abnormalities and massive axon degeneration in the fasciculus gracilis of mutant animals, as indicated by ultrastructure and expression of SMI-32, amyloid precursor protein, and caspase 6. These studies indicate adverse effects of chronic Pten inactivation (and by extension, activation PI-3K signaling) on myelinating oligodendrocytes and their axonal targets. We conclude that PTEN function in oligodendrocytes is required to regulate myelin thickness and preserve axon integrity. In contrast, PTEN is dispensable during myelin repair, and its inactivation confers no detectable benefit.


Involvement of mitochondrial K+ release and cellular efflux in ischemic and apoptotic neuronal death.
We measured and manipulated intracellular potassium (K+) fluxes in cultured hippocampal neurons in an effort to understand the involvement of K+ in neuronal death under conditions of ischemia and exposure to apoptotic stimuli. Measurements of the intracellular K+ concentration using the fluorescent probe 1,3-benzenedicarboxylic acid, 4,4'-[1,4,10,13-tetraoxa-7,16-diazacyclooctadecane-7,16-diylbis(5-methoxy-6,2-benzofurandiyl)]bis-, tetrakis [(acetyloxy) methyl] ester (PBFI) revealed that exposure of neurons to cyanide (chemical hypoxia), glutamate (excitotoxic insult) or staurosporine (apoptotic stimulus) results in efflux of K+ and cell death. Treatment of neurons with 5-hydroxydecanoate (5HD), an inhibitor of mitochondrial K+ channels, reduced K+ fluxes in neurons exposed to each insult and increased the resistance of the cells to death. K+ efflux was attenuated, levels of oxyradicals were decreased, mitochondrial membrane potential was stabilized and release of cytochrome c from mitochondria was attenuated in neurons treated with 5HD. K+ was rapidly released into the cytosol from mitochondria when neurons were exposed to the K+ channel opener, diazoxide, or to the mitochondrial uncoupler, carbonyl cyanide 4(trifluoromethoxy)phenylhydrazone (FCCP), demonstrating that the intramitochondrial K+ concentration is greater than the cytosolic K+ concentration. The release of K+ from mitochondria was followed by efflux through plasma membrane K+ channels. In vivo studies showed that 5HD reduces ischemic brain damage without affecting cerebral blood flow in a mouse model of focal ischemic stroke. These findings suggest that intracellular K+ fluxes play a key role in modulating neuronal oxyradical production and cell survival under ischemic conditions, and that agents that modify K+ fluxes may have therapeutic benefit in stroke and related neurodegenerative conditions.


Histologic findings in motor neuron disease. Relation to clinically determined activity, duration, and severity of disease.
Correlation of 18 histologic variables with age and sex of 24 patients with motor neuron disease (MND), and the duration, severity, and activity of their disease, showed that high density of atrophic fibers correlated with degree of muscle weakness and the worst prognosis and that type I grouping correlated with the best prognosis. Although both type I and type II fibers are involved in the majority of patients with MND, the data suggest that involvement of type I fibers is more important in relation to activity of the disease.


Comparative Effectiveness of Amiodarone and Lidocaine for the Treatment of In-Hospital Cardiac Arrest.
American Heart Association Advanced Cardiac Life Support (ACLS) guidelines support the use of either amiodarone or lidocaine for cardiac arrest caused by ventricular tachycardia or ventricular fibrillation (VT/VF) based on studies of out-of-hospital cardiac arrest. Studies comparing amiodarone and lidocaine in adult populations with in-hospital VT/VF arrest are lacking. Does treatment with amiodarone vs lidocaine therapy have differential associations with outcomes among adult patients with in-hospital cardiac arrest from VT/VF? This retrospective cohort study of adult patients receiving amiodarone or lidocaine for VT/VF in-hospital cardiac arrest refractory to CPR and defibrillation between January 1, 2000, and December 31, 2014, was conducted within American Heart Association Get With the Guidelines-Resuscitation (GWTG-R) participating hospitals. The primary outcome was return of spontaneous circulation (ROSC). Secondary outcomes were 24 h survival, survival to hospital discharge, and favorable neurologic outcome. Among 14,630 patients with in-hospital VT/VF arrest, 68.7% (n = 10,058) were treated with amiodarone and 31.3% (n = 4,572) with lidocaine. When all covariates were statistically controlled, compared with amiodarone, lidocaine was associated with statistically significantly higher odds of the following: (1) ROSC (adjusted OR [AOR], 1.15, P = .01; average marginal effect [AME], 2.3; 95% CI, 0.5 to 4.2); (2) 24 h survival (AOR, 1.16; P = 004; AME, 3.0; 95% CI, 0.9 to 5.1); (3) survival to discharge (AOR, 1.19; P < .001; AME, 3.3; 95% CI, 1.5 to 5.2); and (4) favorable neurologic outcome at hospital discharge (AOR, 1.18; P < .001; AME, 3.1; 95% CI, 1.3 to 4.9). Results using propensity score methods were similar to those from multivariable logistic regression analyses. Compared with amiodarone, lidocaine therapy among adult patients with in-hospital cardiac arrest from VT/VF was associated with statistically significantly higher rates of ROSC, 24 h survival, survival to hospital discharge, and favorable neurologic outcome.


The interrelationship of substance abuse and posttraumatic stress disorder. Epidemiological and clinical considerations.
This chapter reviews the data available on the relationship of substance abuse and posttraumatic stress disorder (PTSD). Delimiting the review to those studies of Vietnam veterans, we found that levels of combat exposure seemed to be positively related to subsequent alcohol use, although not all studies confirmed this relationship. When studies of patients seeking treatment for PTSD were examined, we learned that 60-80% of these patients had concurrent diagnoses of substance abuse, alcohol abuse, or dependence. Methodological limitations of all the studies are discussed and conclusions regarding the status of the PTSD-substance abuse relationship are drawn cautiously. Alternative suggestions for treatment are presented and discussed.


Expanding Reprogramming to Cardiovascular Progenitors.
Methods to generate expandable cardiac progenitor populations are desirable for developing cellular therapies for heart failure and for studying cardiac specification. Now in Cell Stem Cell, Lalit et al. (2016) and Zhang et al. (2016) report two different strategies for reprogramming adult mouse fibroblasts into highly expandable cardiovascular progenitor cells.


Incorporation of 5-bromodeoxycytidine in the adenovirus 2 replication origin interferes with nuclear factor 1 binding.
We have studied the binding of nuclear factor 1 (NFI), a human sequence-specific DNA-binding protein, to a DNA fragment substituted in vitro with 5-bromodeoxycytidine (5-BrdC). Even at low substitution grades binding of NFI to its recognition sequence was considerably lower than with the unsubstituted control fragment. We developed a procedure to cleave substituted DNA specifically at a BrdC residue and searched for contacts between NFI and 5-BrdC residues by an interference assay. Surprisingly, no specific contacts were found in or near the recognition sequence. It appeared instead that interference was inversely related to the distance of a 5-BrdC residue from the NFI binding site. Models to explain these results, including a possible sliding mechanism, are discussed.


Applicability to foraging simulation of a reinforcement schedule controlling the response energy of pigeons.
In optimal foraging theory (OFT), energy expenditure is an important variable for predicting foraging behavior. However, early studies, including operant simulations of foraging, did not measure energy expenditure. In the present study, an adjusting energy (AE) schedule was developed to control energy expenditure. Interresponse energy (IRE), a measure of the energy expenditure during a response, was calculated by dividing the square of the elapsed time between two consecutive responses by the square of the straight-line distance between the locations of the same two responses. An adjusting procedure was employed to estimate the indifference point between the requirements of the AE schedule and a fixed ratio (FR) schedule, which has been used in many operant simulations. In the adjusting procedure, pigeons adjusted the requirement of the AE schedule to that of the FR schedule. The results showed a systematic relationship between the requirements of the AE and FR schedules. Moreover, the total IRE per reinforcement systematically increased with the AE requirement. Thus, the present study demonstrates the utility of the AE schedule as a procedure for testing the validity of OFT.


Long-term treadmill exercise upregulated hippocampal learning-related genes without improving cognitive behaviour in socially isolated rats.
Some environment enrichments such as exercise has been reported to improve the diminished cognitive functions and related gene expression. Therefore, we aimed to investigate the effects of prolonged treadmill exercise on long-term learning and hippocampal gene expression, which involves learning and plasticity. Male Wistar rats (n = 32) randomly assigned into four groups: control (C), social isolation (SI), exercised (E), social isolation + exercise (SE) during postnatal days (PNDs) 21-34. Social isolation protocol was applied during 14 days by placing the rat alone in a cage. Rats were exercised daily, 5 days per week, for overall 4 weeks. Finally, learning performance was evaluated by the novel object recognition test. At the end of learning test, the rats were decapitated to isolate hippocampus tissues for learning related gene expression such as N-methyl-d-aspartate receptor (NMDAR) subunit genes (Grin1, Grin2a, Grin2b) and cyclin dependent kinase 5 (Cdk5), Cdk5 regulatory subunit p35 (Cdk5r), activity-regulated, cytoskeletal-associated protein (Arc), the immediate early gene (c-Fos, a marker of neuronal activation), doublecortin (DCX), achaetescute homolog 1 (ASCL1), brain-derived neurotrophic factor (BDNF) by real-time polymerase chain reaction (RT-PCR). Grin1, NMDAR subunit gene expression was increased significantly in E group compared to other groups. Grin2b, NMDAR subunit gene expression was increased in E group compared to the SI group. Cdk5 level increased in E group compared to the SE group. The ASCL1 gene expression increased in E group compare to the SE group. The DCX gene expression increasing in C group compared to SI and SE groups. Taken together these findings may point out that long-term social isolation down-regulated learning-related genes. However, treadmill exercise together with social isolation did not restore this down-regulation although treadmill exercise increased learning-related genes without improving cognitive behaviour.


Reduction of vertebral height with fragility vertebral fractures can induce variety of neurological deterioration.
The presence of vertebral fractures affect variations in the termination level of conus medullaris (TLCM) and alter neurological findings. However, few studies have examined association between vertebral fractures, TLCM, and neurological findings. Thus, we herein studied the number and severity of vertebral fractures, TLCM, and neurological findings to clarify the mechanism of neurological deterioration in patients with vertebral fractures. A total of 411 patients who underwent computed tomographic myelography were classified into those with (group F, n = 73) and those without vertebral fractures (group C, n = 338). We assessed correlations between TLCM and age, height, and gender in group C, differences in TLCM between groups F and C, and correlations between TLCM, and the number and severity score of fractures. Neurological evaluations were performed for the patellar tendon reflex (PTR), muscle weakness, sensory disturbance, and bladder contraction disorders. TLCM was most commonly located at the L1 vertebral body in group C and did not significantly differ with age, height, or gender. TLCM was most commonly located at L2 vertebral body in group F. TLCM was more caudally located in group F (P < 0.01). Additionally, there was a significant difference between TLCM and number of fractures, and the severity score of fractures (both P < 0.01). Twenty-three patients showed neurological deterioration by vertebral fractures. Some patients with T12 vertebral fracture showed hyperreflexia of PTR. Serious bladder contraction disorders were seen in patients with compression at close range of TLCM. We confirmed that vertebral fractures altered location of the TLCM, thus altering potential neurological symptoms. Moreover, there were correlations of the TLCM with the number and severity score of vertebral fractures. Spine surgeons should be cognizant of the relationship between TLCM, level of compressive lesion, and neurological findings to avoid the wrong level in spine surgery and unexpected neurological deteriorations after surgery.


Immunohistochemical prediction of lapatinib efficacy in advanced HER2-positive breast cancer patients.
Molecular mechanisms of lapatinib resistance in breast cancer are not well understood. The aim of this study was to correlate expression of selected proteins involved in ErbB family signaling pathways with clinical efficacy of lapatinib. Study group included 270 HER2-positive advanced breast cancer patients treated with lapatinib and capecitabine. Immunohistochemical expression of phosphorylated adenosine monophosphate-activated protein (p-AMPK), mitogen-activated protein kinase (p-MAPK), phospho (p)-p70S6K, cyclin E, phosphatase and tensin homolog were analyzed in primary breast cancer samples. The best discriminative value for progression-free survival (PFS) was established for each biomarker and subjected to multivariate analysis. At least one biomarker was determined in 199 patients. Expression of p-p70S6K was independently associated with longer (HR 0.45; 95% CI: 0.25-0.81; p = 0.009), and cyclin E with shorter PFS (HR 1.83; 95% CI: 1.06-3.14; p = 0.029). Expression of p-MAPK (HR 1.61; 95% CI 1.13-2.29; p = 0.009) and cyclin E (HR 2.99; 95% CI: 1.29-6.94; p = 0.011) was correlated with shorter, and expression of estrogen receptor (HR 0.65; 95% CI 0.43-0.98; p = 0.041) with longer overall survival. Expression of p-AMPK negatively impacted response to treatment (HR 3.31; 95% CI 1.48-7.44; p = 0.004) and disease control (HR 3.07; 95% CI 1.25-7.58; p = 0.015). the efficacy of lapatinib seems to be associated with the activity of downstream signaling pathways - AMPK/mTOR and Ras/Raf/MAPK. Further research is warranted to assess the clinical utility of these data and to determine a potential role of combining lapatinib with MAPK pathway inhibitors.


Establishment of human Ig producing heterohybridomas by fusion of mouse myeloma cells with human lymphocytes derived from peripheral blood, bone marrow, spleen, lymph node, and synovial fluid. Effect of polyclonal prestimulation and cryopreservation.
50 fusion experiments were carried out to analyse heterohybridization efficiencies on mouse myeloma cells of the P3 X63 Ag8/653 line with human lymphocytes derived from peripheral blood, bone marrow, lymph node, spleen or synovial fluid. We found higher yields of growing and human Ig-producing hybridoma lines when lymphocytes from spleen or lymph node were fused. Although primary hybridomas could be established from fusions with bone marrow-derived cells, only in nine out of 1616 initially seeded wells was Ig production registered. Four fusions using immune cells from synovial fluid were made without success. Independently of the source of lymphocytes pokeweed mitogen (PWM) prestimulation had no enhancing effect on the percentage of wells with cell growth and this did not alter the IgM:IgG ratio in primary hybridomas (9:1), although cells from all compartments used here (with the exception of bone marrow cells) could be stimulated with PWM to produce both IgG and IgM in cultures. Cryopreserved lymphocytes from different sources could be used for fusions with comparable results registered for the fresh material.


Laminectomy and extension of instrumented fusion improves 2-year pain, disability, and quality of life in patients with adjacent segment disease: defining the long-term effectiveness of surgery.
Adjacent segment disease (ASD) may occur as a long-term consequence of spinal fusion and is associated with significant back and leg pain. Surgical management of symptomatic ASD consists of neural decompression and extension of fusion. However, conflicting results have been reported with respect to the long-term clinical effectiveness of revision surgery in this setting. We set out to comprehensively assess the long-term clinical outcome after revision surgery and determine its effectiveness in the treatment of adjacent segment disease. Fifty patients undergoing revision surgery for ASD-associated back and leg pain were included in this study. Baseline and 2-year Visual Analog Scale-Back Pain (VAS-BP), Visual Analog Scale-Leg Pain (VAS-LP), Oswestry Disability Index, physical and mental quality of life (Short Form-12 [SF-12] physical and mental component score [PCS and MCS]) and health-state utility (EuroQol [EQ-5D]) were assessed. A sustained improvement in VAS-BP (8.72 ± 1.85 vs. 3.92 ± 2.84, P = 0.001), VAS-LP (6.30 ± 3.90 vs. 3.02 ± 3.03, P = 0.001), Oswestry Disability Index (28.72 ± 9.64 vs. 18.48 ± 11.31, P = 0.001), SF-12 PCS (26.89 ± 8.85 vs. 35.58 ± 11.97, P = 0.001) and SF-12 MCS (44.66 ± 12.85 vs. 53.16 ± 9.46, P = 0.001) was observed 2 years after revision surgery, with a cumulative mean 2-year gain of 0.76 quality-adjusted life-years (EQ-5D). Median (interquartile range) time to narcotic independence and return to work was 1.7 (1.0-8.0) months and 2.0 (1.0-4.75) months, respectively. Patients undergoing decompression and extension of fusion for adjacent segment disease-associated back and leg pain reported long-term improvement in pain, disability, and both physical and mental quality of life, suggesting that revision surgery is a highly effective treatment strategy in this patient population.


Mitophagy: A crucial modulator in the pathogenesis of chronic diseases.
Mitophagy is an autophagic process through which damaged or dysfunctional mitochondria are specifically degraded to maintain cellular homeostasis. It is highly regulated by various signaling pathways such as the PTEN-induced putative kinase 1 (PINK1)/Parkin and NIP3-like protein X (NIX)/BNIP3 pathways. Additionally, it plays a crucial role in inducing some pathological processes. Notably, some evidence suggesting the association of mitophagy with the occurrence of chronic diseases such as Parkinson's disease (PD), cancer, diabetes, atherosclerosis (AS), and myocardial ischemia reperfusion (MIR) injury is available. Particularly, it has been reported that mitophagy could hinder the development of PD by activating the PINK1/Parkin pathway and acting as a defense mechanism against the induction of diabetes. Conversely, the induction of mitophagy plays dual roles in driving the process of cancer, AS, and MIR injury. In this review, we have explained the role and regulatory mechanisms through which mitophagy plays a role in these chronic pathologies. Importantly, the pharmacological targeting of mitophagy might prove to be a potential alternative for the treatment of these chronic diseases.


Genetic systems for monitoring interactions of transmembrane domains in bacterial membranes.
In recent years several systems have been developed to study interactions of TM domains within the inner membrane of the Gram-negative bacterium Escherichia coli. Mostly, a transmembrane domain of interest is fused to a soluble DNA-binding domain, which dimerizes in E. coli cytoplasm after interactions of the transmembrane domains. The dimeric DNA-binding domain subsequently binds to a promoter/operator region and thereby activates or represses a reporter gene. In 1996 the first bacterial system has been introduced to measure interactions of TM helices within a bacterial membrane, which is based on fusion of a transmembrane helix of interest to the DNA-binding domain of the Vibrio cholerae ToxR protein. Interaction of a transmembrane helix of interest within the membrane environment results in dimerization of the DNA-binding domain in the bacterial cytoplasm, and the dimeric DNA-binding domain then binds to the DNA and activates a reporter gene. Subsequently, systems with improved features, such as the TOXCAT- or POSSYCCAT system, which allow screening of TM domain libraries, or the GALLEX system, which allows measuring heterotypic interactions of TM helices, have been developed and successfully applied. Here we briefly introduce the currently most applied systems and discuss their advantages together with their limitations.


Neuromodulation Therapies in Pre-Clinical Models of Traumatic Brain Injury: Systematic Review and Translational Applications.
Traumatic brain injury (TBI) has been associated with several lasting impairments that affect quality of life. Pre-clinical models of TBI have been studied to further our understanding of the underlying short-term and long-term symptomatology. Neuromodulation techniques have become of great interest in recent years as potential rehabilitative therapies after injury because of their capacity to alter neuronal activity and neural circuits in targeted brain regions. This systematic review aims to provide an overlook of the behavioral and neurochemical effects of transcranial direct current stimulation (tDCS), transcranial magnetic stimulation (TMS), deep brain stimulation (DBS), and vagus nerve stimulation (VNS) in pre-clinical TBI models. After screening 629 abstracts, 30 articles were pooled for review. These studies showed that tDCS, TMS, DBS, or VNS delivered to rodents restored TBI-induced deficits in coordination, balance, locomotor activity and improved cognitive impairments in memory, learning, and impulsivity. Potential mechanisms for these effects included neuroprotection, a decrease in apoptosis, neuroplasticity, and the restoration of neural circuit abnormalities. The translational value, potential applicability, and the interpretation of these findings in light of outcome data from clinical trials in patients with TBI are discussed.


Nociceptive afferents selectively modulate the cardiac component of the peripheral chemoreceptor reflex via actions within the solitary tract nucleus.
Our previous findings showed that the nucleus of the solitary tract (NTS) mediated part of the tachycardia evoked during somatic noxious stimulation. Here, we investigated the interaction between somatic nociceptor- and peripheral chemoreceptor-evoked cardiac changes. We sought to determine whether this interaction occurred within the NTS, the primary site of termination of chemoreceptor afferents. In a working heart-brainstem preparation of rat, mechanical noxious activation of a forelimb evoked a tachycardia of 17.5+/-3 (mean+/-S.E.M.) b.p.m., whereas sodium cyanide (7-30 microg) stimulation of peripheral chemoreceptors produced a sub-maximal bradycardia of -140+/-15 b.p.m. During nociceptor stimulation the sodium cyanide-evoked bradycardia was attenuated to -42.6+/-12 b.p.m. but could be prevented by a multiple bilateral NTS microinjection of bicuculline (i.e. -173+/-18 b.p.m.). Furthermore, the activity of NTS neurones responding to peripheral chemoreceptor stimulation increased from 2.8+/-1.3 to 9.4+/-1.9 Hz during sodium cyanide injection (n=7; P<0.01). The latter response was attenuated reversibly to 2.9+/-0.9 Hz during simultaneous stimulation of the brachial nerve. Pressure ejection of bicuculline abolished this inhibitory action of brachial-nerve stimulation on the chemoreceptor-evoked excitatory synaptic response. We conclude that somatic noxious stimulation attenuates the chemoreceptor reflex-evoked bradycardia via a GABA(A)ergic mechanism in the NTS.


Fascial sling to correct male neurogenic sphincter incompetence: the McGuire/Raz approach.
We report the surgical approach used in 4 male myelomeningocele patients suffering from severe urinary incontinence owing to poor bladder compliance and sphincter incompetence. While bladder compliance was corrected by enlargement cystoplasty the sphincteric incompetence was treated by an autologous fascial sling around the bladder neck with excellent results. We consider this operation as an alternative to the artificial urinary sphincter in cases of male neurogenic sphincter incontinence.


Flame atomic absorption spectrometric determination of cadmium(II) and lead(II) after their solid phase extraction as dibenzyldithiocarbamate chelates on Dowex Optipore V-493.
An enrichment procedure for cadmium and lead after their solid phase extraction as dibenzyldithiocarbamate chelates on Dowex Optipore V-493 has been established prior to their flame atomic absorption spectrometric determinations. The analytical parameters including pH, amounts of dibenzyldithiocarbamate, sample volume, etc., were investigated. The effects of alkaline and earth alkaline ions and some metal ions on the retentions of analytes on Dowex Optipore V-493 resin were examined. Under the optimized conditions, the detection limits (3s, n=21) for cadmium and lead were 0.43 microg L(-1) and 0.65 microg L(-1), respectively. The relative standard deviation (R.S.D.), and the recoveries of standard addition for this method were lower than 5% (n=11) and 95-102%, respectively. Three standard reference samples (LGC 6010 Hard drinking water, NIST SRM 2711 Montana soil and GBW 07605 Tea) were introduced for accuracy and precision of analytical data. The proposed solid phase extraction system was successfully applied to the analysis of environmental samples.


Influence of methylphenidate on eating in obese men.
Rapid synaptic dopamine transport or reduced brain dopamine receptor signaling may influence energy intake. Methylphenidate, a dopamine reuptake inhibitor, increases brain synaptic dopamine and produces anorexia, suggesting that it may reduce energy intake. We investigated the effects of two doses of short-acting methylphenidate on energy intake over one meal in obese adult males. Nine obese males (>85th BMI percentile) ingested a placebo or a moderate dose (0.5 mg/kg) or a high dose (1.0 mg/kg) of methylphenidate in a within-subject double-blind acute laboratory study. One hour after ingestion, pizza consumption was measured in a naturalistic laboratory setting. Participants reduced energy intake by 23% for the moderate dose vs. the placebo (p < 0.02), but there was no significant difference for the high dose vs. the moderate dose (p > 0.05). Participants consumed 34% fewer kilocalories after ingesting the lowest effective dose of methylphenidate compared with placebo (725.7 +/- 404.5 vs.1095 +/- 271.1 kcal, p < 0.01). Seven of nine subjects responded to the moderate dose. The increase in perceived drug effect above placebo was correlated with the reduction in energy intake for both the moderate (r = -0.85, p = 0.004) and the high (r = -0.75 p = 0.021) doses. Hunger scores were not different across drug doses or placebo before drug administration. Methylphenidate reduced energy intake of a highly palatable food over one meal by one-third in obese adult males. Dopamine transport inhibition may be an effective component of a comprehensive treatment for obesity.


Electrocardiography is Unreliable to Detect Potential Lethal Hyperkalemia in Patients with Non-dialysis Chronic Kidney Disease.
Hemodialysis patients with hypercalcemia are less likely to manifest the usual electrocardiographic changes associated with hyperkalemia than in those with normal renal function. This study was conducted to determine whether electrocardiography (ECG) is a reliable indicator to detect severe life-threatening hyperkalemia in non-dialysis CKD patients. The study was conducted at three referral university hospitals between July 2017 and June 2018. Severe hyperkalemia was defined as serum potassium concentration ≥ 8.0 mEq/L. Serum potassium, sodium, bicarbonate, calcium, and creatinine concentrations were measured and simultaneous 12-lead ECG was obtained. Patients with end-stage renal disease receiving renal replacement therapy were excluded. Also excluded were patients with the usual ECG abnormalities to hyperkalemia. Of the 438 patients screened, 10 (2.3%) aged 2-14 years with severe hyperkalemia and normal ECG findings were identified. Median serum potassium level was 8.6 mEq/L (range 8.2-9.0). All had regular sinus rhythm. P, QRS, ST segment, T morphology, PR and QT interval, and QRS duration were all normal. Hyperkalemia was associated with CKD, metabolic acidosis, and hypercalcemia in all cases. Therapy with intravenous 0.9% saline, sodium bicarbonate, glucose, insulin, calcium, and salbutamol corrected the hyperkalemia in 7 patients. The remaining three patients evinced arrhythmias requiring hemodialysis. Although rare, non-dialysis CKD patients with hypercalcemia may not manifest the usual electrographic abnormalities associated with hyperkalemia. Thus, a normal ECG finding in non-dialysis CKD patients should be interpreted with caution.


Adsorption and controlled release performances of flavor compounds by rice bran insoluble dietary fiber improved through steam explosion method.
In this study, steam explosion was employed as a modification process for rice bran insoluble dietary fiber (RBIDF) to improve the flavor adsorption and controlled release capacities of RBIDF. Results showed that the flavor adsorption ability of RBIDF was effectively improved due to the unfolding structure, increased specific surface area and pore volume and exposure of more functional groups after steam explosion treatment. The mechanism of the flavor adsorption behavior of modified RBIDF was preliminarily explored using adsorption kinetics and isotherms combined with SEM and DSC analysis. Results showed that the Langmuir isotherm model and pseudo-second-order kinetic model yielded the best fit to the adsorption data, indicating monolayer adsorption of flavor onto the modified RBIDF, and the adsorption was mainly driven by chemisorption process. The flavor release profile of modified RBIDF was investigated by HS-SPME/GC-MS and E-nose. After long-time storage, the flavor compounds were retained at a higher concentration in the modified RBIDF compared with the untreated RBIDF, indicating that the steam explosion treatment prolonged the retention time and enhanced the retention and controlled release capacities of RBIDF for flavor compounds. This study provides indications for potential applications of steam explosion-modified RBIDF as a novel flavor delivery system and functional ingredient.


Biological monitoring for ochratoxin A and citrinin and their metabolites in urine samples of infants and children in Bangladesh.
Ochratoxin A (OTA) and citrinin (CIT) are nephrotoxic mycotoxins, found in various foodstuffs and in animal feed, and may cause adverse effects on animal and human health. Previous biomonitoring data indicate a frequent co-exposure of Bangladeshi adults to these mycotoxins. However, since such data are not yet available for young children, a vulnerable part of the population, we conducted this study to assess their exposure to OTA and CIT and compare it with that of adults in Bangladesh. In total, 154 urine samples were collected from infants and children in Rajshahi (n = 88) and Dhaka (n = 66) district of Bangladesh. OTA, CIT, and their metabolites were analyzed by a sensitive HPLC-FLD or LC-MS/MS method, respectively. Overall, OTA and CIT biomarkers were detectable in 72.7% and 54.9% of urines, respectively. The mean OTA and OTα levels in urines were higher in children (0.13 ng/mL and 0.28 ng/mL, respectively) than in infants (0.08 ng/mL and 0.05 ng/mL, respectively). Regarding region, the mean level of OTA was higher in samples from Rajshahi district (0.13 ng/mL) than from Dhaka district (0.09 ng/mL), while the mean OTα level was 2-fold higher in the Dhaka. The total CIT biomarker concentration was significantly higher in children (2.16 ng/mL) than in infant (0.70 ng/mL) urines (p < 0.05), and the mean concentration of HO-CIT was about 6-fold higher than that of parent compound CIT. A provisional daily intake for CIT was calculated and exceeded a preliminary value set by EFSA (0.2 μg/kg bw) in 23.3% and 11.9% of children and infants, respectively. OTA and CIT biomarker concentrations in the young children cohorts are higher than those found in Bangladeshi adults in summer, but lower than in winter season. The new results indicate frequent co-exposure to nephrotoxic mycotoxins that varies between the cohorts and regions in Bangladesh.


Synthetic oxytocin and breastfeeding: reasons for testing an hypothesis.
Synthetic oxytocin is widely used in developed countries and in emerging countries as well. It is the most common medical intervention in childbirth. A great variety of side effects are plausible. There are in particular theoretical reasons to test the hypothesis that the increasing incidence of breastfeeding difficulties and the frequent earlier than desired cessation of breastfeeding are related to the use of synthetic oxytocin during labour. There have already been some studies that tend to support this hypothesis. Four hundred of the 7465 children born in 2006 at the Carlos Haya University Hospital (Malaga, Spain) were randomly selected. By interviewing the mothers, information about feeding type and duration was obtained in 2011 for 316 children. Among the 189 children who were born after labours induced or augmented with synthetic oxytocin, the odds ratio for bottle-feeding was 1.451 and the odds ratio for withdrawal at 3 months was 2.294. In addition, the Battelle Developmental Inventory was used to assess at age five 148 children (84 born with synthetic oxytocin): the odds ratio for neuropsychological development disorders after use of oxytocin was 1.46. The main limitation of such a preliminary study is that in the context of a tertiary Spanish hospital the possible effects of synthetic oxytocin on the quality and duration of breastfeeding cannot be easily dissociated from the effects of other components of pharmacological assistance during labour, particularly epidural fentanyl (a synthetic opioid analgesic). This comment is valid for all studies exploring the side effects of synthetic oxytocin in obstetric units of developed countries, including explorations through videotapes of the effects of synthetic oxytocin on primitive neonatal reflexes. It is also valid for studies exploring the side effects of obstetric analgesia without taking into account the use of synthetic oxytocin. This is why we underline the importance of conducting such studies in emerging countries (e.g. China and Brazil) where synthetic oxytocin is widely used while there are no epidural services. Studies of oxytocin given electively at the onset of third stage of labour (after the birth of the neonate) should help to interpret possible effects on the quality and duration of breastfeeding.


Design, synthesis, and biological evaluations of substituted pyrazoles as pyrrolomycin analogues against staphylococcal biofilm.
The formation of biofilm enables Staphylococcus aureus to resist antibiotics and causes chronic infections. Several compounds of pyrrolomycins are potent antibacterial agents which display inhibition upon staphylococcal biofilms. We designed and synthesized two series of substituted pyrazoles as pyrrolomycin analogues. Compounds 17a, 17d and 17h displayed potent antibacterial activity against various vancomycin-resistant Enterococcus faecalis (VRE) and methicillin-resistant Staphylococcus aureus (MRSA), and 17d showed the most potent activity against MRSA (MIC = 0.0625 μg/mL), vancomycin-intermediate Staphylococcus aureus (VISA) (MIC = 0.0313 μg/mL). Further study indicated that compound 17h could significantly reduce the biofilm formation of MRSA and exhibited promising selectivity. In vitro liver microsomal stability was also evaluated and the results manifested that 17h was metabolically stable in human liver microsomes.


Vitreolysis.
Pars plana vitreolysis appears to be a promising procedure and should be investigated more thoroughly during the years ahead. 22 cases are reported, which showed maintenance of the visual acuity in 20 eyes over a follow-up period averaging 13 months. The technique and indications have been discussed for this new procedure.


Thickness prediction in metal alloys using nuclear techniques and artificial neural network: Modelling.
Metal alloys are widely used in the aerospace, biological, civil and automotive industries, thus being very important to develop techniques to identify these alloys. Nuclear technique based on gamma densitometry is a non-invasive technique that is able to identify metal alloys using a radiation source and a scintillator detector. The measurement geometry and the dataset for training an artificial neural network were developed using the MCNP6 code. Therefore, this study proposes the thickness prediction of five aluminum alloys (2024-O, 2090-T83, 3003, 5086-O and 7075-O), a titanium alloy, and two steel carbon alloys (stainless 302 and stainless 316) ranging from 2 to 50 mm for four different gamma-ray radiation energies using gamma transmission and artificial neural network. A study to evaluate the reliability of the results was performed by analyzing the uncertainties in the data from the simulation with the MCNP6 code and the data predicted by ANN. The results indicate that it was possible to predict all the alloys thicknesses using the energy of 137Cs radiation source, in which more than 96% of the cases with 5% of relative error, even for a group of alloys with very close densities values as aluminum alloys.


A rare case of spontaneous esophageal rupture (Boerhaave's syndrome) associated with pulmonary rupture.
Spontaneous esophageal rupture (Boerhaave's Syndrome) is a life-threatening condition that requires early diagnosis and effective treatment. The authors describe a rare case of spontaneous rupture of the esophagus associated with pulmonary rupture in a 54-year-old man. The anatomical basis for the occurrence of a spontaneous esophageal rupture associated with pulmonary rupture is discussed.


Association Between Nighttime Discharge from the Intensive Care Unit and Hospital Mortality: A Multi-Center Retrospective Cohort Study.
We aimed to determine the impact of nighttime discharge from the intensive care unit (ICU) to the ward on hospital mortality and readmission rates in consecutive critically ill patients admitted to five Canadian ICUs. We hypothesized that hospital mortality and readmission rates would be higher for patients discharged after hours compared with discharge during the day. A multi-center retrospective cohort study was carried out at five hospitals in Edmonton, Canada, between July 2002 and December 2009. Nighttime discharge was defined as discharge from the ICU occurring between 07:00 pm and 07:59 am. Logistic regression analysis was used to explore the associations between nighttime discharge and outcomes. Of 19,622 patients discharged alive from the ICU, 3,505 (17.9%) discharges occurred during nighttime. Nighttime discharge occurred more commonly among medical than surgical patients (19.9% vs. 13.8%, P < 0.001) and among those with more comorbid conditions, compared with daytime discharged patients. Crude hospital mortality (11.8% versus 8.8%, P < 0.001) was greater for nighttime discharged as compared to daytime discharged patients. In a multivariable analysis, after adjustment for comorbidities, diagnosis and source of admission, nighttime discharge remains associated with higher mortality (odds ratio [OR] 1.29; 95% CI, 1.14 to 1.46, P < 0.001). This finding was robust in two sensitivity analyses examining discharges occurring between 00:00 am and 04:59 am (OR 1.28; 1.12-1.47; P < 0.001) and for those who died within 48 h of ICU discharge without readmission (OR 1.24; 1.07-1.42, P = 0.002). There was no difference in ICU readmission for nighttime compared with daytime discharges (7.4% vs. 6.9 %, p = 0.26). However, rates were higher for nighttime discharges in community compared with tertiary hospitals (7.7% vs. 5.7%, P = 0.023). In a large integrated health region, 1 in 5 ICU patients are discharged at nighttime, a factor with increasing occurrence during our study and shown to be independently associated with higher hospital mortality.


First-Aid Hydrogel Wound Dressing with Reliable Hemostatic and Antibacterial Capability for Traumatic Injuries.
First-aid for severe traumatic injuries in the battlefield or pre-hospital environment, especially for skin defects or visceral rupture, remains a substantial medical challenge even in the context of the rapidly evolving modern medical technology. Hydrogel-based biomaterials are highly anticipated for excellent biocompatibility and bio-functional designability. Yet, inadequate mechanical and bio-adhesion properties limit their clinical application. To address these challenges, a kind of multifunctional hydrogel wound dressing is developed with the collective multi-crosslinking advantages of dynamic covalent bonds, metal-catechol chelation, and hydrogen bonds. The mussel-inspired design and zinc oxide-enhanced cohesion strategy collaboratively reinforce the hydrogel's bio-adhesion in bloody or humoral environments. The pH-sensitive coordinate Zn2+ -catechol bond and dynamic Schiff base with reversible breakage and reformation equip the hydrogel dressing with excellent self-healing and on-demand removal properties. In vivo evaluation in a rat ventricular perforation model and Methicillin-resistant Staphylococcus aureus (MRSA)-infected full-thickness skin defect model reveal excellent hemostatic, antibacterial and pro-healing effectiveness of the hydrogel dressing, demonstrating its great potential in dealing with severe bleeding and infected full-thickness skin wounds.


Reversible control of magnetism in FeRh thin films.
The multilayer of approximate structure MgO(100)/[nFe51Rh49(63 Å)/57Fe51Rh49(46 Å)]10 deposited at 200 °C is primarily of paramagnetic A1 phase and is fully converted to the magnetic B2 phase by annealing at 300 °C for 60 min. Subsequent irradiation by 120 keV Ne+ ions turns the thin film completely to the paramagnetic A1 phase. Repeated annealing at 300 °C for 60 min results in 100% magnetic B2 phase, i.e. a process that appears to be reversible at least twice. The A1 → B2 transformation takes place without any plane-perpendicular diffusion while Ne+ irradiation results in significant interlayer mixing.


Development of a novel approach for secondary modelling in predictive microbiology: incorporation of microbiological knowledge in black box polynomial modelling.
This research deals with the development of a novel secondary modelling procedure within the framework of predictive microbiology. The procedure consists of three steps: (i) careful formulation of the available microbiological information, both from literature and from the experimental case study at hand, (ii) translation of these requirements in mathematical terms under the form of partial derivatives throughout the complete interpolation region of the experimental design, and (iii) determination of parameter values with suitable optimisation techniques for a flexible black box modelling approach, e.g., a polynomial model or an artificial neural network model. As a vehicle for this procedure, the description of the maximum specific growth rate of Lactobacillus sakei in modified BHI-broth as influenced by suboptimal temperature, water activity, sodium lactate and dissolved carbon dioxide concentration is under study. The procedure results in a constrained polynomial model with excellent descriptive and interpolating features in comparison with an extended Ratkowsky-type model and classical polynomial model, by combining specific properties of both model types. The developed procedure is illustrated on the description of the lag phase as well. It is stressed how the confrontation with experimental data is very important to appreciate the descriptive and interpolating capacities of new or existing models, which is nowadays not always carefully performed. Alternatively, the first two steps of the novel procedure can be used as a tool to demonstrate clearly (possible) interpolative shortcomings of an existing model with straightforward spreadsheet calculations.


The aldosterone index could be used to diagnose the dominant gland in primary aldosteronism - a retrospective study.
Failed cannulation in the right adrenal vein, which makes the sampling results in the contralateral vein and inferior vena cava (IVC) nonsense, is the main obstacle of using adrenal vein sampling (AVS) in the lateralisation diagnosis in primary aldosteronism (PA). We performed a retrospective study to evaluate the specificity and sensitivity of using the aldosterone index (AI) in PA lateralisation diagnosis. We enrolled 116 patients who were diagnosed with PA and then underwent AVS in the West China Hospital of Sichuan University from April 2015 to April 2017. The AI, calculated by dividing the aldosterone concentration of the failed side by the aldosterone concentration of IVC, was used for lateralisation diagnosis if the cannulation was judged to be failed by traditional method. Patients with dominant adrenal gland based on successful AVS were included in subgroup 2 (n = 75), while the patients diagnosed with a dominant gland using AI method were enrolled in subgroup 1 (n = 41). No significant difference of clinical and biochemical findings between the two groups was detected (p value after operation > 0.05). ROC analysis was performed to test the specificity and sensitivity based on the AI in subgroup 2. The AUC for dominant gland detection was 0.76, which resulted in 91.3% sensitivity and 67.53% specificity. The positive and negative likelihood ratios were 2.81. Our data suggested that the modified strategy using AI to diagnose the dominant gland in PA is an efficient method when cannulation has failed in the right side.


Examining resurgence and repetition with the evolutionary theory of behavior dynamics.
Recently, Redner et al. (2022) examined the nature of resurgence across repeated iterations of the traditional three-phase resurgence procedure with four pigeons. Although extant research findings in this area are mixed, Redner et al. found that resurgence generally increased in magnitude with repetition. These findings provide a baseline against which future research examining resurgence using this three-phase procedure can be compared and contrasted. The purpose of the present investigation was to examine resurgence via concurrent schedule arrangements similar to those described by Redner et al. with 30 artificial organisms (AOs) animated by the Evolutionary Theory of Behavior Dynamics (McDowell, 2004). We quantified the prevalence of resurgence across iterations and found that resurgence occurred in 86.7 % (156 of 180) iterations across all 30 AOs. This is strikingly similar to the resurgence prevalence estimates of 87.5 % reported by both Redner et al. (2022). However, we also found that the magnitude of target responding generally did not change significantly with repetition. This finding is inconsistent with Redner et al. but is consistent with the predictions of prominent quantitative models of behavioral persistence and a number of relevant studies (Volkert et al., 2009; Gratz et al., 2019). We also conducted exploratory analyses to examine how several variables (e.g., sensitivity to reinforcement, reinforcer magnitude, number of sessions of exposure to various phases) affect the prevalence and magnitude of resurgence among AOs.


Field measurements of key parameters associated with nocturnal OBT formation in vegetables grown under Canadian conditions.
The objective of this study was to provide the parameter values required to model OBT formation in the edible parts of plants following a hypothetical accidental tritium release to the atmosphere at night. The parameters considered were leaf area index, stomatal resistance, photosynthesis rate, the photosynthetic production rate of starch, the nocturnal hydrolysis rate of starch, the fraction of starch produced daily by photosynthesis that appears in the fruits, and the mass of the fruit. Values of these parameters were obtained in the summer of 2002 for lettuce, radishes and tomatoes grown under typical Canadian environmental conditions. Based on the maximum observed photosynthetic rate and growth rate, the fraction of starch translocated to the fruit was calculated to be 17% for tomato fruit and 14% for radish root.


Beliefs and attitudes towards the consumption of sugar-free products in a sample of Italian adolescents.
The objective of this study was to investigate beliefs and attitudes towards sugar-free products in a sample of Italian teenagers. A total of 10 secondary public schools in the county of Rome were randomly selected. The probability of being extracted was set proportional to the number of students. A random sample of teenagers was selected from the schools for participating in a food consumption survey. The sample included 233 adolescents, aged 15-21 y. A total of 224 adolescents agreed to participate in the study aimed at assessing beliefs and attitudes. Two questionnaires were completed at the first day of the last period of the 4-day food survey. The first questionnaire contained questions on attitudes and beliefs toward a healthy diet. The second questionnaire, aimed at assessing adolescents' attitudes towards the consumption of sugar-free products, was based in part on the social-psychological model of the Theory of Planned Behaviour. In general, the adolescents of this sample seemed to be quite uncertain in their responses. On average, they did not express an emphatic point of view with regard to the act of eating sugar-free products. The results of the assessment of relationships between the variables of the theoretical model revealed the lack of causal links among beliefs, attitudes, intention and consumption of sugar-free products. Perceived behavioural control was not a significant predictor of intention. The target of further studies aimed at investigating attitudes towards sugar-free products should be tailored according to the quantity of consumption of such products in order to represent a sample of frequent and high consumers of sugar-free products. European Commission Quality of Life and Management of Living Resources Fifth framework Programme. QLRT-1999-00155.


Denervation techniques.
The bladder can be denervated at several levels: centrally, at the level of the sacral nerves; peripherally, on the pelvic nerves; or in the bladder, by cutting the relevant structures, by injecting substances toxic to nerves, or by hyperbaric bladder distension. These procedures target the sensory or motor nerves to weaken or to interrupt the detrusor reflex arc. Most of the procedures introduced previously, e.g. bladder trans-section by open operation, endoscopic or transvesical phenolization, hyperbaric bladder distension, and peripheral denervation of the bladder, have now been abandoned. Although some of these techniques had a high initial success rate in abolishing detrusor overactivity and in controlling incontinence, the relapse rate within 18 months approached 100%. In the early 1950s, much of the denervation surgery was performed on sacral roots and nerves. Nowadays, sacral de-afferentiation of the bladder by dorsal sacral root rhizotomy of S2-S5, using specialized techniques, either intradurally or at the conal level, has proved to be a very effective procedure for patients with spinal cord injuries and detrusor hyper-reflexia, functional low compliance and reflex incontinence, which cannot be managed by any other means. In conclusion, although in approximately 90% of patients the symptoms of an overactive bladder will be eliminated or improved by conservative methods of treatment, bladder denervation procedures still have a place in the management of these patients. In particular, with these procedures a low-pressure bladder system can be obtained and urinary continence restored in patients with complete suprasacral spinal cord lesions.


[Gestational diabetes under clinical conditions in aspired normoglycemia: investigation for correlation of blood glucose daily profiles and fetometric ultrasound parameters].
One of the therapeutic aims in gestational diabetes (GDM) is to prevent the development of fetal hypertrophy by adaptation of maternal glycemic control. Relating to this context, maternal blood glucose daily profiles and fetal biometry ultrasound parameters were analysed for a possible correlation. A special focus was given to the question as to whether a latency period exists for this possible connection. 152 pregnancies affected by GDM, without fetal malformations or aneuploidies, were enrolled. Altogether, 746 ultrasound examinations consisting of 7 fetometric parameters each and 1 288 blood glucose daily profiles originating from the 20 (th) to 40 (th) gestational week were systematically investigated for interrelation by correlation and multiple regression analysis. No robust, constant correlation between the analysed parameters could be observed. However, marked differences between latency periods were noticed. Blood glucose parameters, which revealed significant regressions with fetal abdominal circumference, had an average time lag of 6.2 +/- 2.5 weeks, whereas the latency period for head circumference averaged 2.4 +/- 1.2 weeks. The overall small-for-gestational-age (SGA) rate was 20 %, pregnant women with a body mass index > 30 kg / m (2) revealed the highest rate of 28 %. Therapeutic intervention depending on sonographically detected hypertrophy must be considered as being delayed. The currently valid therapeutic criteria including intended normoglycemia and regular fetometric ultrasound controls cannot prevent markedly high SGA rates, especially among obese women, in adequately treated GDM.


Tumor Lysis Syndrome After Bilobectomy for Typical Carcinoid Tumor of the Lung.
Tumor lysis syndrome is a life-threatening complication comprising hyperuricemia, hyperkalemia, hyperphosphatemia, and hypocalcemia attributed to release of intracellular contents. Although traditionally associated with leukemia and lymphoma after chemotherapy, it is known to occur in solid malignancies. We herein report a rare case of this syndrome after resection of bulky carcinoid tumor of the lung.


Multipotent mesenchymal stromal cell therapy and risk of malignancies.
Cell therapy with Multipotent Mesenchymal Stromal Cells (MSC) holds enormous promise for the treatment of a large number of degenerative and immune/inflammatory diseases. Their multilineage differentiation potential, immunoprivilege and capacity of promoting recovery of damaged tissues coupled with anti-inflammatory and immunosuppressive properties are the focus of a multitude of clinical studies currently underway. The recognized clinical potential of MSC repairing/immunomodulatory effects now encompasses graft-versus-host disease, hematologic malignancies, cardiovascular diseases, neurologic and inherited diseases, autoimmune diseases, organ transplantation, refractory wounds, and bone/cartilage defects among others. However, it has been suggested that both the need of extensive ex vivo culture for MSC clinical use, and their proangiogenic, anti-apoptotic and immunomodulatory properties may act together as tumor promoters, raising significant safety concerns. This paper will review the available data on in vitro MSC maldifferentiation and the ability of MSC to sustain tumor growth in vivo, with the aim to clarify whether MSC-based therapeutic approaches may carry actual risk of malignancies.


Splitting of the P3 component during dual-task processing in a patient with posterior callosal section.
When two concurrent sensorimotor tasks have to be performed at a short time interval, the second response is generally delayed at a central decision stage. However, in patients who have undergone full or partial transection of forebrain fibers connecting the two hemispheres (split-brain), independent structures subserving all processing stages should reside in each disconnected hemisphere, thus predicting parallel processing of dual tasks. Surprisingly, this prediction is usually not verified behaviorally. We reasoned that brain imaging with high-density recordings of event-related potentials (ERPs) could clarify the extent and limits of parallel processing in callosal patients. We studied a patient (AC) with posterior callosal section in a lateralized number-comparison task. Behaviorally, the split-brain patient showed robust dual-task interference, superficially similar to the psychological refractory period (PRP) effect in the control group of 14 healthy subjects, but significantly different in important aspects such as slowing of response times in the first task. Analysis of ERPs revealed that the parietal P3 component became split into distinct contralateral components in the patient, and was dramatically reduced for targets in his left visual field. In contrast to the control group, P3 latencies showed minimal to nonexistent postponement related to dual-task processing in the patient. In summary, our findings suggest that the left and right hemisphere networks normally involved in a single distributed "global neuronal workspace" that underlies the generation of the P3 component and serial processing, became strongly decoupled after a posterior callosal lesion.


Normal testosterone plasma levels in non-abstinent alcoholics.
Testosterone (T), prolactin (PRL), and luteinizing hormone (LH) levels were estimated in plasma of 32 male chronic alcoholics during a period of their usual alcohol consumption and compared to the values of 32 age-matched controls. T levels were not different from normals, while both PRL and LH were significantly elevated. The subgroup of patients with severe liver injury (n = 11) showed elevated PRL (2P less than 0.001), but normal LH and T, while patients with normal hepatic function (n = 21) showed normal PRL levels, elevated LH, and a tendency to elevated T levels.


Learning from community pharmacists' initial experiences of a workplace-based training program.
To prepare community pharmacists for the provision of clinical and patient-focused services, a novel postgraduate course for community pharmacists in the United Kingdom was developed. The program incorporated personal development planning against a personal development framework, workplace mentoring, employment of work-based assessment tools, activities that encouraged increased inter-professional working, reflection, and opportunities for peer support. Objectives were to identify course components that support development, describe the effect on practice, and explore the perceived advantages and disadvantages of this model of postgraduate education. Interviews were conducted with a purposive sample of 15 community pharmacists after they had completed approximately one year of the three-year course. A topic guide covering approaches to learning, rationale for course selection, course experiences to date, and impact on practice was used. Interview recordings were thematically analyzed. Two themes were identified. 'Support for learning' describes the components of the course that provided support for learning, including opportunities to learn with and from others, workplace mentoring, and facilitated access to general practitioners. 'Outcomes of learning' encompasses how the course was a way of effecting change within existing roles and the increase in confidence and motivation to change practice. The model has merit in supporting community pharmacists to develop the confidence and competence required for extended clinical and patient-focused roles. While this model of learning seems to provide educational value, further research is required to determine whether the additional resources required to provide workplace mentoring, use work-based assessment tools, and encourage inter-professional working are justified.


Mapping GPs' motivation — it's not all about the money: a nationwide cross-sectional survey study from Denmark.
Understanding physicians' motivation may be essential for policymakers if they are to design policies that cater to physicians' wellbeing, job retention, and quality of care. However, physicians' motivation remains an understudied area. To map GPs' work motivation. A cross-sectional analysis using registry and survey data from Denmark. Survey data were used to measure four types of motivation: extrinsic motivation, intrinsic motivation, user orientation, and public service motivation. These were combined with register data on the characteristics of the GP, practice, and area. Using latent profile analysis, the heterogeneity in GPs' motivation was explored; the associations between GPs' motivation and the GP, practice, and area characteristics were estimated using linear regression analyses. There was substantial heterogeneity in GPs' motivations. Five classes of GPs were identified with different work motivations: class 1 'it is less about the money' - probability of class membership 53.2%; class 2 'it is about everything' - 26.5%; class 3 'it is about helping others' - 8.6%; class 4 'it is about the work' - 8.2%; and class 5 'it is about the money and the patient' - 3.5%. Linear regression analyses showed that motivation was associated with GP, practice, and area characteristics to a limited extent only. GPs differ in their work motivations. The finding that, for many GPs, 'it is not all about the money' indicated that their different motivations should be considered when designing new policies and organisational structures to retain the workforce and ensure a high quality of care.


Tumor variation in the cancer family syndrome: ovarian cancer.
The Cancer Family Syndrome is a hereditary disorder (autosomal dominant), characterized by early onset cancer of the colon (particularly the proximal colon) and endometrium, with an excess of multiple primary cancers. Recent evidence reflects the possibility of an even broader tumor spectrum consisting of carcinoma of the stomach, the ovary, the kidney, and possibly other organs. A family with the cardinal features of the Cancer Family Syndrome is described, including two sisters and their mother who had ovarian carcinoma at early ages (38, 46, and 49 years). Two of these three women have shown unusual tolerance to cancer, despite invasion of the primary tumors. Implications for cancer surveillance and management are discussed.


Application of the ultrasonic technique for the pathophysiological analysis of the pericardial disease.
(1) In large pericardial effusion, echography showed a revolving motion of the heart. This characteristic motion was interfered by the pericardial adhesion. Real time cross-sectional scanning was useful for detecting the adhesion, especially the localized one. When excessive cardiac motion coexisted with tachycardia, alternation of the heart position accompanied by electrical alternans was observed. Based on the results, the mechanisms for the alternation of the heart position was discussed. (2) Echogram suggesting the pericardial thickening was found in about a half of patients with constrictive pericarditis. Another findings, which may reflect the anatomical and functional changes in constrictive pericarditis, were also presented. Although each of them is not pathognomonic, concurrence of them may suggest the presence of constrictive pericarditis. (3) Inflow velocity in jugular vein was markedly decreased in cardiac tamponade. The inspiratory increase in the inflow velocity was observed in mild cardiac tamponade, but not in severe one. Flow velocity pattern of jugular vein in constrictive pericarditis was characterized by the appearance of the reverse flow being coexistent with rapidly decelerating inflow in diastole.


Prevalence and predictors of bone health among perinatally HIV-infected adolescents.
Long-term complications of HIV including low bone mineral density remain a concern. We studied the prevalence and predictors of low bone mineral density among South African perinatally HIV-infected adolescents (PHIVA) on antiretroviral therapy (ART). Cross-sectional analysis. Bone health was evaluated by measuring the calcaneus stiffness index among PHIVA on ART. Low stiffness index was defined as z-score less than -2 SD compared with age-matched and sex-matched HIV-uninfected adolescents (HIV-). Overall, 407 PHIVA (median age: 14 years; 50.4% female; median age at ART initiation: 4.2 years) and 92 HIV- (median age: 13.7 years; 54.4% female) were included. Median duration on ART was 9.8 years (interquartile range 6.8-11.5) with 38% initiating ART at 2 years or less of age. Stiffness index was lower in PHIVA (-0.19 vs. 0.43, P ≤ 0.001), respectively. During puberty, mean stiffness index increased with Tanner Stage in both PHIVA and HIV- but these increases were larger among HIV-; Tanner Stage II-III (96 vs. 101, P = 0.009) and Tanner Stage IV-V (104 vs. 112, P = 0.001). Among PHIVA, 52 (13%) had low stiffness index. After adjusting for age, sex and Tanner Stage, use of lopinavir/ritonavir [odds ratio (OR) = 2.31, P = 0.012] and viral load more than 50 copies/ml (OR = 2.06, P = 0.023) were associated with increased risk of low stiffness index, while use of efavirenz (OR = 0.41, P = 0.009) was associated with decreased risk of low stiffness index. Stiffness index was a significantly lower in PHIVA than in HIV-, especially during puberty. Among PHIVA, detectable viral load and use of lopinavir/ritonavir were risk factors for low stiffness index. Further longitudinal studies are important to determine the clinical implications.


A comparison of balance-correcting responses induced with platform-translation and shoulder-pull perturbation methods.
The understanding of reactive balance control mechanisms in humans emanates from studies utilizing a variety of perturbation methods, instructions, and sensory conditions. The use of different perturbation methods may produce method-specific balance-correcting responses. This study evaluated balance-correcting responses induced with platform-translation and shoulder-pull methods with equilibrated perturbation intensities, and whether the absence of vision affects balance-correcting responses differently between perturbation methods. Fifteen healthy young males participated. Unexpected forward and backward platform-translation and shoulder-pull perturbations were induced with eyes-open and eyes-closed. Participants were asked to behave naturally. Forward stepping trials were analyzed. Margin of stability (MOS) was calculated from the position data of reflective markers placed strategically around the body. MOS was reported at step onset and at foot contact. MOS was smaller at step onset (0.01 ± 0.01 m) and at foot contact (0.09 ± 0.01 m) during platform-translation trials and at both time points during shoulder-pull trials (0.04 ± 0.01 m and 0.17 ± 0.01 m, respectively). The absence of vision did not affect MOS at step onset. At foot contact during platform-translation with eyes-closed MOS was larger (0.11 ± 0.01 m) than with eyes-open (0.08 ± 0.01 m), but not different between eyes-open (0.17 ± 0.01 m) and eyes-closed (0.18 ± 0.01 m) during shoulder-pull. Participants required a second step to recover balance in 14% of the platform-translation and 3% of the shoulder-pull trials. During platform-translation trials participants demonstrated smaller MOS which placed them in a less favorable circumstance for balance recovery. Platform-translation appears to be more challenging than shoulder-pull perturbation in terms of balance recovery. This study underscores that caution is required when interpreting results of studies utilizing different perturbation paradigms.


[Glutathione-S-transferase activity in the blood of scorpion fish (Scorpaena porcus) depending on sex, age and season].
Glutathione-S-transferase activity in the blood of scorpion fish inhabiting coastal areas of Sevastopol depending on sex, age and season were studied. The decrease of glutathione-S-transferase activities has been observed with fish ageing.


Common genetic variation in the glucokinase gene (GCK) is associated with type 2 diabetes and rates of carbohydrate oxidation and energy expenditure.
Glucokinase (GCK) plays a role in glucose metabolism and glucose-stimulated insulin secretion. Rare mutations in GCK cause MODY. We investigated whether common variation (minor allele frequency ≥0.01) in GCK is associated with metabolic traits and type 2 diabetes. Four exonic single-nucleotide polymorphisms (SNPs) and three SNPs predicted to cause loss of promoter function were identified in whole-genome sequence data from 234 Pima Indians. These seven tag SNPs and rs4607517, a type 2 diabetes variant established in other studies, were analysed in 415 full-heritage non-diabetic Pima Indians characterised for metabolic traits, and 7,667 American Indians who had data on type 2 diabetes and BMI. A novel 3' untranslated region (3'UTR) SNP, chr7:44184184-G/A, was associated with the rate of carbohydrate oxidation post-absorptively (β = 0.22 mg [kg estimated metabolic body size (EMBS)](-1) min(-1), p = 0.005) and during a hyperinsulinaemic-euglycaemic clamp (β = 0.24 mg [kg EMBS](-1) min(-1), p = 0.0002), the rate of carbohydrate oxidation in a respiratory chamber (β = 311 kJ/day, p = 0.03) and 24 h energy expenditure, which was attributable to the thermic effect of food (β = 520 kJ/day, p = 3.39 × 10(-6)). This 3'UTR SNP was also associated with diabetes (OR 1.36, 95% CI 1.11, 1.65, p = 0.002), where the A allele (allele frequency 0.05) was associated with a lower rate of carbohydrate oxidation, lower 24 h energy expenditure and higher risk for diabetes. In a Cox proportional hazards model, a rate of insulin-stimulated carbohydrate oxidation lower than the mean rate at baseline predicted a higher risk for developing diabetes than for those above the mean (hazard rate ratio 2.2, 95% CI 1.3, 3.6, p = 0.002). Common variation in GCK influences the rate of carbohydrate oxidation, 24 h energy expenditure and diabetes risk in Pima Indians.


Localization and neurochemical characteristics of the extrinsic sympathetic neurons projecting to the pylorus in the domestic pig.
The pylorus, an important part of the digestive tract controlling the flow of chyme between the stomach and the duodenum, is widely innervated by intrinsic and extrinsic nerves. To determine the locations of postganglionic sympathetic perikarya that innervate the pylorus of the domestic pig, a retrograde tracing method with application of Fast Blue tracer was used. All positive neuronal cell bodies (ca. 1750) were found in the celiac-cranial mesenteric ganglion complex (CSMG), however, the coeliac poles of this complex provided the major input to the pylorus. Afterwards, the immunohistochemical staining procedure was applied to determine biologically active substances expressed in the FB-labeled perikarya. Approximately 77% of the FB-positive cell bodies contained tyrosine hydroxylase (TH), 87% dopamine β-hydroxylase (DβH), 40% neuropeptide Y (NPY), 12% somatostatin (SOM) and 7% galanin (GAL). The presence of all these substances in the ganglion tissue was confirmed by RT-PCR technique. Double immunocytochemistry revealed that all of the TH-positive perikarya contained DβH, about 40% NPY, 12% SOM and 8% GAL. Additionally, all above-cited immunohistochemical markers as well as VIP, PACAP, ChAT, LEU, MET, SP and nNOS were observed within nerve fibers associated with the FB-positive perikarya. Immunocytochemical labeling of the pyloric wall tissue disclosed that TH+, DβH+ and NPY+ nerve fibers innervated ganglia of the myenteric and submucosal plexuses, blood vessels, both muscular layers and the muscularis mucosae; nerve fibers immunoreactive to GAL mostly innervated both muscular layers, while SOM+ nerve fibers were observed within the myenteric plexus. Presented study revealed sources of origin and immunohistochemical characteristics of the sympathetic postganglionic perikarya innervating the porcine pylorus.


Initial experience with use of hydrogel microcoils in embolization of pulmonary arteriovenous malformations.
The purpose of this study is to describe our initial experience with embolization of pulmonary arteriovenous malformations (PAVMs) using hydrogel microcoils. The technical and radiological outcomes were retrospectively reviewed in seven patients with nine simple-type PAVMs (median feeder size 4 mm, range 3-6 mm) who underwent embolization. Hydrogel microcoils were mainly used, and detachable bare microcoils were combined as needed to occlude the terminal feeding artery just before the sac. Of a total of 43 microcoils, 30 (69.8%) hydrogel microcoils were deployed in eight PAVMs with the median number 3.5 (range 2 to 6) per lesion. All hydrogel microcoils were successfully deployed without microcatheter stuck or malposition. In the remaining one small PAVM, only soft bare microcoils were used, however, resulting in recanalization requiring additional coils in the second session. The venous sac was substantially shrunk in all lesions treated with hydrogel microcoils with the median size reduction rate 95.0% (range 81.8% to 99.0%) during the median follow-up period 10 months (range 6 to 18 months). In conclusion, hydrogel microcoils were safely and effectively applied for occluding PAVMs with relatively small feeders.


Stretch-induced regulation of angiotensinogen gene expression in cardiac myocytes and fibroblasts: opposing roles of JNK1/2 and p38alpha MAP kinases.
The cardiac renin-angiotensin system (RAS) has been implicated in mediating myocyte hypertrophy, remodeling, and fibroblast proliferation in the hemodynamically overloaded heart. However, the intracellular signaling mechanisms responsible for regulation of angiotensinogen (Ao), a substrate of the RAS system, are largely unknown. Here we report the identification of JNK1/2 as a negative, and p38alpha as a major positive regulator of Ao gene expression. Isolated neonatal rat ventricular myocytes (NRVM) and fibroblasts (NRFB) plated on deformable membranes coated with collagen IV, were exposed to 20% equiaxial static-stretch (0-24 h). Mechanical stretch initially depressed Ao gene expression (4 h), whereas after 8 h, Ao gene expression increased in a time-dependent manner. Blockade of JNK1/2 with SP600125 increased basal Ao gene expression in NRVM (10.52+/-1.98 fold, P<0.001) and NRFB (13.32+/-2.07 fold, P<0.001). Adenovirus-mediated expression of wild-type JNK1 significantly inhibited, whereas expression of dominant-negative JNK1 and JNK2 increased basal and stretch-mediated (24 h) Ao gene expression, showing both JNK1 and JNK2 to be negative regulators of Ao gene expression in NRVM and NRFB. Blockade of p38alpha/beta by SB202190 or p38alpha by SB203580 significantly inhibited stretch-induced (24 h) Ao gene expression, whereas expression of wild-type p38alpha increased stretch-induced Ao gene expression in both NRVM (8.41+/-1.50 fold, P<0.001) and NRFB (3.39+/-0.74 fold, P<0.001). Conversely, expression of dominant-negative p38alpha significantly inhibited stretch response. Moreover, expression of constitutively active MKK6b (E) significantly stimulated Ao gene expression in the absence of stretch, indicating that p38 activation alone is sufficient to induce Ao gene expression. Taken together p38alpha was demonstrated to be a positive regulator, whereas JNK1/2 was found to be a negative regulator of Ao gene expression. Prolonged stretch diminished JNK1/2 activation, which was accompanied by a reciprocal increase in p38 activation and Ao gene expression. This suggests that a balance in JNK1/2 and p38alpha activation determines the level of Ao gene expression in myocardial cells.


In situ mechanical analysis of myofibrillar perturbation and aging on soft, bilayered Drosophila myocardium.
Drosophila melanogaster is a genetically malleable organism with a short life span, making it a tractable system in which to study mechanical effects of genetic perturbation and aging on tissues, e.g., impaired heart function. However, Drosophila heart-tube studies can be hampered by its bilayered structure: a ventral muscle layer covers the contractile cardiomyocytes. Here we propose an atomic force microscopy-based analysis that uses a linearized-Hertz method to measure individual mechanical components of soft composite materials. The technique was verified using bilayered polydimethylsiloxane. We further demonstrated its biological utility via its ability to resolve stiffness changes due to RNA interference to reduce myofibrillar content or due to aging in Drosophila myocardial layers. This protocol provides a platform to assess the mechanics of soft biological composite systems and, to our knowledge, for the first time, permits direct measurement of how genetic perturbations, aging, and disease can impact cardiac function in situ.


Tacrine-6-ferulic acid, a novel multifunctional dimer, inhibits amyloid-β-mediated Alzheimer's disease-associated pathogenesis in vitro and in vivo.
We have previously synthesized a series of hybrid compounds by linking ferulic acid to tacrine as multifunctional agents based on the hypotheses that Alzheimer's disease (AD) generates cholinergic deficiency and oxidative stress. Interestingly, we found that they may have potential pharmacological activities for treating AD. Here we report for the first time that tacrine-6-ferulic acid (T6FA), one of these compounds, can prevent amyloid-β peptide (Aβ)-induced AD-associated pathological changes in vitro and in vivo. Our results showed that T6FA significantly inhibited auto- and acetylcholinesterase (AChE)-induced aggregation of Aβ(1-40)in vitro and blocked the cell death induced by Aβ(1-40) in PC12 cells. In an AD mouse model by the intracerebroventricular injection of Aβ(1-40), T6FA significantly improved the cognitive ability along with increasing choline acetyltransferase and superoxide dismutase activity, decreasing AChE activity and malondialdehyde level. Based on our findings, we conclude that T6FA may be a promising multifunctional drug candidate for AD.


Population insulin sensitivity from sparsely sampled oral glucose tolerance tests.
This work aimed to estimate population-level insulin sensitivity (SI) from 2-hour oral glucose tolerance tests (OGTT) with less than 7 samples. The current methodology combines the OGTT mathematical model developed by Dalla Man et al., with nonlinear multilevel (NLML) statistical model to estimate population-level insulin sensitivity (SI) from sparsely sampled datasets (3 or 4 samples per subject obtained in 120 min). To validate our novel methodology of population SI estimation, we simulated 50 virtual subjects. We simulated 10 observations per subject over 240 minutes. After estimating their SI using the OGTT model, the virtual subjects were split into two groups, subjects with SI above the average and ones with below average. Subsequently, the simulated data were analyzed using statistical software and employing a t-test. The mean estimates of population SI for the two groups of virtual subjects and their respective 95% CI were compared to the estimates obtained with our novel NLML group SI estimates obtained using the 3 and 4 time points per subject. To further validate the performance of the novel NLML model, a set of 34 prediabetic and 30 diabetic subjects with T2D was used. As outlined above for the in-silico subjects, differences between the prediabetic and T2D subjects in regard to SI was assessed using the classical two-stage approach (individual SI estimation followed by statistical comparison of the two groups). The average estimates obtained with the classical two-stage approach were compared to the group estimated obtained with the NLML approach using 3 (0, 60, and 120 minutes) points per subject obtained in 120 minutes. Unique and identifiable individual estimates of SI were obtained for all virtual subjects. In comparison to the subjects with above average SI (n=25), the subjects with simulated below average SI (n=25) exhibited significantly lower insulin sensitivity (P<0.001). Our novel NLML population model confirmed these findings (4-point OGTT: P<0.001; 3-point OGTT: P<0.001). In a similar fashion to the one outlined for the virtual subjects, the median insulin sensitivities estimated with the classical two-stage approach were different between the prediabetic (n=34) and T2D subjects (n=32, P=0.004). Using 3 points per subject, our novel NLML model confirmed these findings (P<0.001). The population estimates of SI from OGTT data is an effective tool to assess population insulin sensitivity and assess differences that may not be possible when calculating individual SI or when less than 7 samples are available.


Understanding Mendelian errors in SNP arrays data using a Gochu Asturcelta pig pedigree: genomic alterations, family size and calling errors.
Up to 478 Gochu Asturcelta pig parents-offspring trios (61 different families) were genotyped using the Axiom_PigHDv1 Array to identify the causes of Mendelian errors (ME). Up to 545,364 SNPs were retained. Up to 40,540 SNPs gathering 292,297 allelic mismatches were identified and were overlapped with SINEs and LINEs (Sscrofa genome 11.1). Copy number variations (CNV) were called using PennCNV. ME were classified into eight different classes according to the trio member ("Trio" meaning no assignment) and the allele on which ME was identified: TrioA/B, FatherA/B, MotherA/B, OffspringA/B. Most ME occurred due to systematic causes: (a) those assigned to the Father, Mother or Offspring occurred by null or partial null alleles characterized by heterozygote deficiency, varied with family size, involved a low number of loci (6506), and gathered most mismatches (228,145); (b) TrioB errors varied with family size, covaried with SINEs, LINEs and CNV, and involved most ME loci (33,483) and mismatches (65,682); and (c) TrioA errors were non-systematic ME with no sampling bias involving 1.2% of mismatches only and a low number of loci (1939). The influence of TrioB errors on the overall genotyping quality may be low and, since CNV vary among populations, their removal should be considered in each particular dataset. ME assignable to the Father, Mother or Offspring may be consistent within technological platforms and may bias severely linkage or association studies. Most ME caused by null or partial null alleles can be removed using heterozygote deficiency without affecting the size of the datasets.


Effect of neuroleptics on phospholipase A2 activity in the brain of rats.
The effect of neuroleptics on phospholipase A2 (PLA2) activity in rat brain plasma membranes was studied. Chlorpromazine (10 mg/kg), fluphenazine (5 mg/kg), thioridazine (5 mg/kg), trifluoperazine (5 mg/kg), haloperidol (2 mg/kg), and sulpiride (100 mg/kg) were administered to rats intraperitoneally as a single dose or long-term treatment (4 weeks). The PLA2 activity was determined 24, 48, and 72 h after the last injection of a drug. The enzyme activity was decreased after a single or 4-week administration of chlorpromazine, trifluoperazine, haloperidol, and sulpiride. Fluphenazine and thioridazine caused an increase of PLA2 activity in rat brain both after a single dose and long-term administration. For the first time it was shown that neuroleptics cannot only inhibit but also increase, PLA2 activity. Elucidation of this fact requires further studies.


Identification of disease activity and health status cut-off points for the symptom state acceptable to patients with rheumatoid arthritis.
Established thresholds for low levels of disease activity need to be examined from a patients' perspective. To identify new cut-off points for patients' perception of satisfactory condition (patient acceptable symptom state (PASS)) in composite indices and patient-reported outcomes, and to examine the agreement between the new PASS cut-off points for composite indices and existing thresholds for remission, low and moderate disease activity. Patients with rheumatoid arthritis from a treatment register (n = 1496, 72.1% women, mean (SD) age 53.9 (13.5) years, disease duration 7.6 (9.1) years, 28-joint Disease Activity Score (DAS28) 4.98 (1.36)) responded during follow-up (12, 24 and 52 weeks) to a global dichotomised question on satisfactory condition (PASS). New PASS cut-off points were identified with the 75th centile estimation and receiver operating characteristic analyses for a variety of outcome measures, and cut-off points for composite indices were examined for agreement with the low disease activity threshold (1.625) of the Patient Activity Scale (PAS) and thresholds for remission, low and moderate disease activity in DAS28, Simplified Disease Activity Index (SDAI) and Clinical Disease Activity Index (CDAI). New PASS cut-off points for DAS28, SDAI and CDAI were in the moderate range of disease activity, and the cut-off point was 3.56 for PAS. Agreement between thresholds for disease activity levels and the PASS cut-off points was best for low disease activity (accuracy 64.5-74.6), and better for moderate disease activity (accuracy 61.3-67.2) than for remission (accuracy 30.7-45.8). The current PASS concept seems to be in the range of moderate disease activity.


Mono- and polyclonal antibodies as probes to study vitellin processing in embryos of the stick insect Carausius morosus.
During embryonic development, insect vitellins (Vt) are degraded by limited proteolysis to yield a number of lower-molecular weight polypeptides. The aim of the present study was to identify these polypeptides in the embryo and to verify how they relate to Vt polypeptides deposited in the oocyte during vitellogenesis. To this end a panel of poly- and monoclonal antibodies (Pab, Mab) was raised against Vt polypeptides and employed by immunoelectrophoresis and immunoblotting on embryos belonging to different developmental stages. Through this approach three major staining patterns were observed. First, Mab 4 reacts with both polypeptides B1 and E20, suggesting that polypeptide B1 is gradually trimmed to yield polypeptide E20 in late embryos. Second, Mab 12 is specific for polypeptide A3 which is retained unchanged throughout embryogenesis. Third, Pab anti-A2 and Mab 13 show that polypeptide A2 is processed to yield polypeptide E9 through limited proteolysis. In conclusion, the staining patterns reported in this study show that Vt polypeptides in developing embryos of the stick insect Carausius morosus undergo at least two major processing events concerning polypeptides B1 and A2.