Self-Directed Weight Loss Strategies: Energy Expenditure Due to Physical Activity Is Not Increased to Achieve Intended Weight Loss.
Reduced physical activity and almost unlimited availability of food are major contributors to the development of obesity. With the decline of strenuous work, energy expenditure due to spontaneous physical activity has attracted increasing attention. Our aim was to assess changes in energy expenditure, physical activity patterns and nutritional habits in obese subjects aiming at self-directed weight loss. Energy expenditure and physical activity patterns were measured with a portable armband device. Nutritional habits were assessed with a food frequency questionnaire. Data on weight development, energy expenditure, physical activity patterns and nutritional habits were obtained for 105 patients over a six-month period from an initial cohort of 160 outpatients aiming at weight loss. Mean weight loss was -1.5 ± 7.0 kg (p = 0.028). Patients with weight maintenance (n = 75), with substantial weight loss (>5% body weight, n = 20) and with substantial weight gain (>5% body weight, n = 10) did not differ in regard to changes of body weight adjusted energy expenditure components (total energy expenditure: -0.2 kcal/kg/day; non-exercise activity thermogenesis: -0.3 kcal/kg/day; exercise-related activity thermogenesis (EAT): -0.2 kcal/kg/day) or patterns of physical activity (duration of EAT: -2 min/day; steps/day: -156; metabolic equivalent unchanged) measured objectively with a portable armband device. Self-reported consumption frequency of unfavorable food decreased significantly (p = 0.019) over the six-month period. An increase in energy expenditure or changes of physical activity patterns (objectively assessed with a portable armband device) are not employed by obese subjects to achieve self-directed weight loss. However, modified nutritional habits could be detected with the use of a food frequency questionnaire.


The Effect of Class Imbalance on Precision-Recall Curves.
In this note, I study how the precision of a binary classifier depends on the ratio r of positive to negative cases in the test set, as well as the classifier's true and false-positive rates. This relationship allows prediction of how the precision-recall curve will change with r, which seems not to be well known. It also allows prediction of how Fβ and the precision gain and recall gain measures of Flach and Kull (2015) vary with r.


Information oncologic tools produced by the U.S. National Cancer Institute: how much are they known and used? Results of an enquiry in Italy.
A survey was conducted among specialized users in order to establish the level of knowledge and diffusion of four informative tools: Cancerlit, PDQ, Clinprot, Cancergram. A questionnaire was distributed to 975 graduate researchers working in four national cancer research institutes in the north (2), center (1) and south (1) of Italy, as well as to the students of the Postgraduate School of Oncology (Oncology 2nd) of the "La Sapienza" University of Rome. Respondents were 486 (49.8%): only 167 (34.4%) declared to know Cancerlit, 56 (11.5%) PDQ, 10 (2%) Clinprot and 39 (8%) Cancergrams. Moreover, there is a time gap of nearly 6 years between the availability on-line of these tools and the information of its existence by the users. Most of the respondents were in favor of inserting courses on the use of information tools into the University curriculum and of training in the use of the tools.


Building memories on prior knowledge: behavioral and fMRI evidence of impairment in early Alzheimer's disease.
Impaired memory is a hallmark of prodromal Alzheimer's disease (AD). Prior knowledge associated with the memoranda improves memory in healthy individuals, but we ignore whether the same occurs in early AD. We used functional MRI to investigate whether prior knowledge enhances memory encoding in early AD, and whether the nature of this prior knowledge matters. Patients with early AD and Controls underwent a task-based fMRI experiment where they learned face-scene associations. Famous faces carried pre-experimental knowledge (PEK), while unknown faces with which participants were familiarized prior to learning carried experimental knowledge (EK). Surprisingly, PEK strongly enhanced subsequent memory in healthy controls, but importantly not in patients. Partly nonoverlapping brain networks supported PEK vs. EK associative encoding in healthy controls. No such networks were identified in patients. In addition, patients displayed impaired activation in a right sub hippocampal region where activity predicted successful associative memory formation for PEK stimuli. Despite the limited sample sizes of this study, these findings suggest that the role prior knowledge in new learning might have been so far overlooked and underestimated in AD patients. Prior knowledge may drive critical differences in the way healthy elderly and early AD patients learn novel associations.


The serodiagnosis of tuberculosis in children: an evaluation of an ELISA test using IgG antibodies to M. tuberculosis, strain H37 RV.
An enzyme-linked immunosorbent assay (ELISA) for detecting serum IgG antibodies to an autoclaved suspension of Mycobacterium tuberculosis H37 RV was evaluated as a diagnostic tool in 132 clinical cases of childhood tuberculosis. The mean (SD) optical density value in these patients was 0.115 (0.122) compared with 0.022 (0.017) in the control group of patients who had non-tuberculous acute respiratory tract infections. Using as a cut-off level the mean of the control plus 2 standard deviations, the test sensitivity was 62% and the specificity was 98% in all the patients with a clinical diagnosis of tuberculosis. In the culture-positive group (n = 35), the sensitivity was 69%. A positive correlation was shown between the optical density levels and increasing age and chronicity of infection but not with respect to tuberculin skin reactivity, nutritional status and the duration of prior therapy. In addition, BCG vaccination (presence of a scar) did not affect the ELISA result. We conclude that this ELISA test is a useful diagnostic test in children.


Differences in invitro responses of the hypothalamo-pituitary-gonadal hormonal axis between low- and high-egg-producing turkey hens.
Low-egg-producing hens (LEPH) ovulate less frequently than high-egg-producing hens (HEPH) and exhibit differences in mRNA levels for components of the hypothalamo-pituitary-gonadal (HPG) axis, suggesting differential responsiveness to trophic stimulation. Ovulation frequency is governed by the production of the pituitary gonadotropins and feedback of the ovarian follicle steroid hormones, which are regulated by HPG axis stimulation and inhibition at the hypothalamic level. The pituitary and follicle cells from LEPH and HEPH were subjected to in vitro hormonal treatments to stimulate or inhibit the HPG axis, followed by expression analysis of mRNA levels for HPG axis genes and radioimmunoassays for steroid hormone production. Statistical analysis was performed using the mixed models procedure of SAS. The pituitary cells from HEPH showed upregulation of genes associated with ovulation stimulation, whereas cells from LEPH showed upregulation of genes associated with inhibition of ovulation. High-egg-producing hens' follicle cells displayed a higher sensitivity and responsiveness to gonadotropin treatment. Level of egg production impacted ovulation-related gene expression in the pituitary cells as well as steroid hormone production in the follicle cells, with HEPH displaying a greater positive response to stimulation. These findings indicate that differences in egg production among turkey hens likely involve differential responsiveness of the cells within the HPG axis.


Intra-articular Osteoid Osteoma of the Olecranon Fossa.
Osteoid osteomas are benign primary bone lesions characterized by a central nidus with surrounding reactive sclerosis, classically presenting as worsening nocturnal pain relieved by non-steroidal anti-inflammatory medications (NSAIDs). These most commonly occur in intracortical bone and the diaphysis of long bones. As a rare entity, intra-articular osteoid osteomas present unusually, often resulting in a delayed or incorrect diagnosis. We present a case of an intra-articular osteoid osteoma, emphasizing the importance of MRI in aiding diagnosis in this atypical location.


Nrf2 Inhibits the Progression of Parkinson's Disease by Upregulating AABR07032261.5 to Repress Pyroptosis.
Parkinson's disease (PD) is associated with dysregulated neural cell death, such as pyroptosis, but its regulatory mechanisms are poorly understood. This study investigated roles of nuclear factor E2-related factor 2 (Nrf2) in regulating pyroptosis and PD development. Cellular and rat PD models established by 6-OHDA exposure were subjected to Nrf2 overexpression. Neurobehavioral functions were assessed by the traction test, Morris Water Maze, and open field test. Cell proliferation was analyzed by MTS assay, while flow cytometry was applied to quantify levels of reactive oxygen species (ROS) and apoptosis. Nissl bodies in rat brains were detected by Nissl staining, and cell apoptosis in brain tissues was assessed by terminal deoxynucleotidyl transferase dUTP nick-end labeling. Differential expression of lncRNA and mRNA was characterized by deep sequencing. A cellular PD model was successfully established by inducing PC12 cell differentiation with nerve growth factor-β and exposing differentiated cells to 6-OHDA. Cells exhibited significantly increased ROS levels, enhanced pyroptosis, and inhibited Nrf2 phosphorylation. The rat PD model exhibited impaired muscle strength, increased pyroptosis, and repressed Nrf2 phosphorylation. Nrf2 overexpression effectively repressed pyroptosis in both cellular and rat PD models. Marked alterations of lncRNA and mRNA profiles were induced by Nrf2 overexpression in the cellular PD model, which involved multiple signaling pathways. Silencing of the lncRNA AABR07032261.5 significantly promoted pyroptosis in the cellular PD model. Nrf2 suppressed PD pathogenesis in cellular and animal models by promoting AABR07032261.5, which repressed pyroptosis.


Radiation induced hypopharyngeal stenosis masquerading as the larynx: a case report.
Radiation to the head and neck is commonly used in the treatment of cancers. A side effect in some patients is the development of pharyngeal and or esophageal strictures. Hypopharyngeal strictures can resemble edematous larynges. If mistakenly so identified, tracheal tubes placed through these structures are unlikely to result in tracheal intubation and more likely to cause obstruction, esophageal intubation or mediastinal damage. This report points out the development of hypopharyngeal stenosis following radiation. The location and appearance of hypopharyngeal stenosis during laryngoscopy are illustrated. The report points out the deficiencies of newer supraglotic laryngoscopes in this situation and underscores the benefits of flexible fiberoptic devices.


Hospitalizations for chronic pancreatitis in Allegheny County, Pennsylvania, USA.
Population-based estimates for chronic pancreatitis (CP) are scarce. We determined incident CP hospitalization rates and the risk of pancreatitis-related readmissions in Allegheny County, Pennsylvania, USA. We used Pennsylvania Health Care Cost Containment Council (PHC4) dataset to identify all unique White and Black Allegheny County residents with incident hospitalization for CP from years 1996-2005. We noted presence of alcoholism codes (from one year before index hospitalization until last contact) and pancreatitis-related readmissions until the third quarter of 2007. Age-, sex-, and race-adjusted (to US 2000 population) rates/100,000 were calculated. 988 unique County residents with incident hospitalization for CP were identified. Of these, 37.6% also received alcoholism codes. Overall hospitalization rate was 7.75/100,000 (95% CI 7.26-8.24), which remained stable throughout the study period. Patients with alcoholism codes were significantly younger (47.2 vs. 58.0 years), more likely to be male (71.4 vs. 36.6%), and Black (38.5 vs. 17.7%). Hospitalization rates were significantly higher (2.4-fold) in Blacks (vs. Whites), particularly for those with alcoholism codes. During follow-up (median 45 months), pancreatitis-related readmissions were common, significantly more so for patients with alcoholism codes. CP hospitalization rates over a one-decade period were stable. Readmissions were highest among patients with a diagnosis of alcoholism. and IAP.


Importance of Aedes albopictus in veterinary medicine.
To assess the role of Aedes albopictus in transmission of filarial nematodes of veterinary importance, researches were carried out in different geographical areas. In Italy a first research was performed to study the susceptibility of Ae. albopictus to Dirofilaria repens, D. immitis and Setaria labiatopapillosa. The development of L3 larvae was longer than in other species of mosquitoes but Ae. albopictus could be a suitable vector of filariae. To understand the role of Ae. albopictus in the natural transmission of Dirofilaria and to assess the risk for animal and human health, in 2000, 2001 and 2002 another study was carried out in the town of Padua. A total of 2,534 Ae. albopictus were caught on human-attracted mosquitoes. Specific primers and sequencing identified filarial DNA as D. immitis; Ae. albopictus was proved a natural vector of D. immitis. Similar results were confirmed in Central Italy also for D. repens. The presence of Ae. albopictus increased the probability of transmission of canine and human dirofilariosis in urban environment and it could change the epidemiology of dirofilariosis, in particular for what concern the time of biting and the risk season. These aspects must be considered to outline a correct prophylaxis.


Identification of a cis-acting dendritic targeting element in the mRNA encoding the alpha subunit of Ca2+/calmodulin-dependent protein kinase II.
In mammalian neurons a selected group of mRNAs, including the transcript encoding the alpha subunit of Ca2+/calmodulin-dependent protein kinase II, is found in dendrites. The molecular mechanisms underlying extrasomatic RNA trafficking are not well described. It is thought that dendritic transcripts contain cis-acting elements that direct their selective subcellular sorting. Here we report the identification of an extrasomatic targeting element in the 3' untranslated region of the mRNA encoding the alpha subunit of Ca2+/calmodulin-dependent protein kinase II. In primary hippocampal neurons, this 1200-nucleotide-spanning, cis-acting element is sufficient to mediate dendritic localization of chimeric reporter transcripts. The trafficking signal does not share any striking sequence similarity with a previously characterized dendritic targeting element in transcripts encoding the microtubule-associated protein 2. In dendrites of transfected primary neurons, recombinant RNAs form granules with an average diameter of 0.45 microm that may represent preferential RNA docking sites or multimolecular transport units. These findings imply that extrasomatic sorting of individual dendritic mRNAs involves at least partially distinct molecular mechanisms, as well as large trafficking complexes.


Pregnancy weight gain and breast cancer risk.
BACKGROUND: Elevated pregnancy estrogen levels are associated with increased risk of developing breast cancer in mothers. We studied whether pregnancy weight gain that has been linked to high circulating estrogen levels, affects a mother's breast cancer risk. METHODS: Our cohort consisted of women who were pregnant between 1954-1963 in Helsinki, Finland, 2,089 of which were eligible for the study. Pregnancy data were collected from patient records of maternity centers. 123 subsequent breast cancer cases were identified through a record linkage to the Finnish Cancer Registry, and the mean age at diagnosis was 56 years (range 35 - 74). A sample of 979 women (123 cases, 856 controls) from the cohort was linked to the Hospital Inpatient Registry to obtain information on the women's stay in hospitals. RESULTS: Mothers in the upper tertile of pregnancy weight gain (>15 kg) had a 1.62-fold (95% CI 1.03-2.53) higher breast cancer risk than mothers who gained the recommended amount (the middle tertile, mean: 12.9 kg, range 11-15 kg), after adjusting for mother's age at menarche, age at first birth, age at index pregnancy, parity at the index birth, and body mass index (BMI) before the index pregnancy. In a separate nested case-control study (n = 65 cases and 431 controls), adjustment for BMI at the time of breast cancer diagnosis did not modify the findings. CONCLUSIONS: Our study suggests that high pregnancy weight gain increases later breast cancer risk, independently from body weight at the time of diagnosis.


Current and future directions of pharmacokinetic and pharmacokinetic-pharmacodynamic modelling and simulation: population approach group in Europe 19th annual meeting.
The Population Approach Group in Europe meeting is the annual meeting of a group of scientists interested in quantitative pharmacology approaches. This meeting provides insights on new modelling approaches and applications thereof in different therapeutic areas and in different phases of drug development, in which modelling and simulation is set 'at work'. These meeting highlights mainly focus on the contributions pertinent to this journal, such as physiology-based pharmacokinetic modelling, modelling of pharmacokinetic-pharmacodynamic relationships (including binary/categorical end points) in non-clinical studies and also include presentations of junior scientists in the annual Lewis Sheiner Student Session.


A Novel Vaccine Candidate: Recombinant Toxoplasma gondii Perforin-Like Protein 2 Stimulates Partial Protective Immunity Against Toxoplasmosis.
Toxoplasma gondii is an apicomplexan pathogen infecting 2 billion people and numerous livestock, causing a major threat to economies and human health. Passive-active immunoprophylaxis is an efficient approach to provide protection against toxoplasmosis. T. gondii perforin-like protein 2 (TgPLP2) contains a membrane attack complex/perforin (MACPF) domain, making it a potential vaccine candidate. Here, we aimed to assess the protection efficacy of TgPLP2 using Bagg albino/c (BALB/c) mice model. The Escherichia coli system was used to obtain the recombinant TgPLP2 (rTgPLP2). Mice challenged by anti-rTgPLP2 polyclonal antibodies (PcAb) pretreated tachyzoites showed obviously increased survival outcomes. In addition, mice that passively received anti-rTgPLP2 PcAb following a lethal dose of tachyzoites infection had longer survival time compared with phosphate-buffered saline (PBS) controls. Furthermore, we demonstrated that immunization with rTgPLP2 could prolong survival in RH strain infected mice and resulted in the lowest brain cysts size and number of Prugniaud (PRU) genotype II strain infected mice. High levels of Toxoplasma-specific IgG, IgG1, IgG2a, and cytokines (IFN-γ and IL-10) were produced after two immunizations with rTgPLP2. Together these results indicated that TgPLP2 can induce both humoral and cellular immune responses to protect host against infection and thus is a potential candidate for T. gondii vaccines.


[A review of 86 fractures of the lower end of the femur treated by internal fixation using A.O. blade-plates (author's transl)].
The authors have treated 86 fractures of the lower end of the femur by internal fixation with an A.O. blade-plate. An adequate follow up was made in 71 cases. A classification of the fractures is given. Half of the fractures were either complex or involved the joint. The technique that was used is described. The authors consider that an immediate cancellous autogenous bone graft is worthwhile. 8 cases of non-union, 7 stiff knees and 31 malunions were seen; 70 per cent of cases achieved satisfactory function.


Gastro-intestinal stromal tumor (GIST): case report.
Gastro-intestinal stromal tumors (GISTs), as currently defined, represent the largest category of primary non epithelial neoplasms of the gastrointestinal tract. They arise from mesenchymal cells located in the wall of the organ and show a remarkable variability in their differentiation pathways. For this reason there is relevant degree of confusion in their interpretation. On the basis of immunohistochemical and ultrastructural studies these neoplasms are divided into several categories: leiomyomas, schwannomas and less differentiated tumors referred as GIST. In the small bowel GIST are uncommon. Usually asymptomatic, they could be the cause of surgical emergencies like massive bleeding, obstruction, intussusception or perforation. Generally benign, an higher percentage of malignant cases are described in the small bowel. The Authors report a case of malignant GIST of the small intestine presented with bowel obstruction by ileal invagination. In this case, as usually it happens in malignant GIST, the final diagnosis was obtained by an abdominal surgical exploration.


Charge transport in molecular junctions: From tunneling to hopping with the probe technique.
We demonstrate that a simple phenomenological approach can be used to simulate electronic conduction in molecular wires under thermal effects induced by the surrounding environment. This "Landauer-Büttiker's probe technique" can properly replicate different transport mechanisms, phase coherent nonresonant tunneling, ballistic behavior, and hopping conduction. Specifically, our simulations with the probe method recover the following central characteristics of charge transfer in molecular wires: (i) the electrical conductance of short wires falls off exponentially with molecular length, a manifestation of the tunneling (superexchange) mechanism. Hopping dynamics overtakes superexchange in long wires demonstrating an ohmic-like behavior. (ii) In off-resonance situations, weak dephasing effects facilitate charge transfer, but under large dephasing, the electrical conductance is suppressed. (iii) At high enough temperatures, kBT/ϵB > 1/25, with ϵB as the molecular-barrier height, the current is enhanced by a thermal activation (Arrhenius) factor. However, this enhancement takes place for both coherent and incoherent electrons and it does not readily indicate on the underlying mechanism. (iv) At finite-bias, dephasing effects may impede conduction in resonant situations. We further show that memory (non-Markovian) effects can be implemented within the Landauer-Büttiker's probe technique to model the interaction of electrons with a structured environment. Finally, we examine experimental results of electron transfer in conjugated molecular wires and show that our computational approach can reasonably reproduce reported values to provide mechanistic information.


Thiourea as a template for photodimerization of azastilbenes.
In this study we have explored the potential of thiourea (TU) as a template to preorient stilbazoles and bispyridylethylenes (azastilbenes) in the crystalline state. TU is able to preorient eleven azastilbenes toward dimerization in the crystalline state. While cocrystals of these eleven olefins photodimerized to a single dimer expected based on crystal packing, pure crystals of these olefins either were nonreactive or gave a mixture of dimers. The differential photobehavior of the pure crystals and cocrystals highlights the importance of TU in templating the olefins in a photoreactive orientation in the crystalline state. X-ray crystallographic and photochemical studies have identified a few azastilbenes that photodimerize in spite of not being arranged in an ideal orientation in the crystalline state. These as well as a few examples already reported in the literature suggest that it is important to recognize that molecules could experience large amplitude motions in the crystalline state, especially when energized by light. Short-term lattice instability caused by photoexcitation can be effective in driving a photochemical reaction. Thus one should view the crystalline arrangement of molecules upon light exposure as "dynamic" rather than "static" as determined from X-ray structure analysis.


Cadmium-induced hepatotoxicity in rats and the protective effect of naringenin.
This experiment pertains to the protective role of naringenin against cadmium (Cd)-induced oxidative stress in the liver of rats. Cadmium is a major environmental pollutant and is known for its wide toxic manifestations. Naringenin is a naturally occurring citrus flavonone which has been reported to have a wide range of pharmacological properties. In the present investigation cadmium (5mg/kg) was administered orally for 4 weeks to induce hepatotoxicity. Liver damage induced by cadmium was clearly shown by the increased activities of serum hepatic marker enzymes namely aspartate transaminase (AST), alanine transaminase (ALT), alkaline phosphatase (ALP), lactate dehydrogenase (LDH), gamma glutamyl transferase (GGT) and serum total bilirubin (TB) along with the increased level of lipid peroxidation indices (thiobarbituric acid reactive substances (TBARS) and lipid hydroperoxides) and protein carbonyl contents in liver. The toxic effect of cadmium was also indicated by significantly decreased levels of enzymatic antioxidants (superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx) and glutathione S-transferase (GST)) and non-enzymatic antioxidants (reduced glutathione (GSH), vitamin C and vitamin E). Administration of naringenin at a dose of (50mg/kg) significantly reversed the activities of serum hepatic marker enzymes to their near-normal levels when compared to Cd-treated rats. In addition, naringenin significantly reduced lipid peroxidation and restored the levels of antioxidant defense in the liver. The histopathological studies in the liver of rats also showed that naringenin (50mg/kg) markedly reduced the toxicity of cadmium and preserved the normal histological architecture of the tissue. The present study suggested that naringenin may be beneficial in ameliorating the cadmium-induced oxidative damage in the liver of rats.


Expression of human A4V mutant Cu,Zn superoxide dismutase in Schizosaccharomyces pombe: investigations of its toxic properties.
Cu,Zn superoxide dismutase (SOD1) is an antioxidant enzyme that catalyzes the removal of superoxide radicals generated in various biological oxidations. Amyotrophic lateral sclerosis (ALS) is one of the most common neurodegenerative disorders, occurring in families (FALS) and sporadically (SALS). FALS and SALS are distinguishable genetically but not clinically. More than 100 point mutations in the human SOD 1 gene have been identified that cause FALS. In order to determine the effects of mutant SOD protein, we first cloned wild-type and A4V mutant human SOD1 into Schizosaccharomyces pombe. This study shows viabilities and some antioxidant properties including SOD, catalase, proteasomal activity, and protein carbonyl levels of transformants in SOD1 deleted strain (MN415); and its parental strain (JY741) at different stress conditions. There was no more oxidative damage in the human mutant SOD carrying the transformant strain compared with other strains. These results may help to explain whether ALS progresses as a consequence of cellular oxidative damage.


Measurement of quality of life: current state of the art.
In recent years several investigators have developed procedures for measuring the quality of life of individuals. This paper focuses on an empirical approach to defining the main determinants of quality of life and the development and application of procedures for surveying the importance of each component in representative samples of 3 age groups of Americans. These age groups included those persons 30, 50, and 70 years of age. The survey also included reports by these persons concerning the extent to which their needs and wants were met with respect to each of these components. By using the critical incident technique it was found that there were 15 components that included all of the 6,500 activities and experiences reported by a wide variety of Americans from various geographic areas as contributing either positively or negatively to their quality of life. In the survey of 500 men and 500 women at each of the 3 age levels it was found that the 5 dimensions they most frequently described as important included: health, children, understanding yourself, work, and spouse. The 5 dimensions for which they reported their needs and wants were least well met were: participating in government, active recreation, learning and education, creative expression, and helping others. The 5 dimensions that correlated most highly with their reports of overall quality of life at present were: material comforts, work, health, active recreation, and learning and education. Suggestions are made for revising these procedures for use with persons with a disability to improve their quality of life.


Characteristics of Patients With Sertoli and Leydig Cell Testis Neoplasms From a National Population-Based Registry.
Sertoli and Leydig cell tumors (SCT and LCT) are uncommon testis neoplasms. Data regarding patient demographics and outcomes are limited to small series. We further characterized these tumors using a large cancer database. The Surveillance, Epidemiology, and End Results (SEER) database was queried from 2004 to 2012. International Classification of Diseases for Oncology (ICD-O) codes identified SCT and LCT. Common germ cell tumors (GCT) provided a reference group. Age, race, histology, tumor size, stage, and cancer-specific mortality (CSM) were compared. Thirty-one men had SCT, 76 had LCT, and 17,998 had GCTs. Median follow-up for SCT, LCT, and GCTs was 46, 38, and 50 months, respectively. Median ages for SCT and LCT were 39 and 47, respectively, which was older than those with GCT (34 years; P < .001). African American race was more common in SCT (23%) and LCT (24%) patients compared to GCT (3%, P < .001). LCT most commonly presented with stage I disease (98.5%), while patients with SCT presented at higher stages (35% with stage II/III). CSM was highest in patients with SCT (32% vs. 2% LCT and 7% GCT, P < .001). Median survival of those with CSM was similar between SCT, LCT, and GCTs (15, 12, and 14 months, respectively). Compared to GCT, SCT and LCT present at older ages and are more common in African Americans. Metastasic disease at presentation and CSM rates are higher in SCT compared to LCT and GCT, suggesting a clinically relevant distinction between these histologies. Better characterization of these rare neoplasms will continue to inform patient counseling and management.


Advancements in nano-enabled therapeutics for neuroHIV management.
This viewpoint is a global call to promote fundamental and applied research aiming toward designing smart nanocarriers of desired properties, novel noninvasive strategies to open the blood-brain barrier (BBB), delivery/release of single/multiple therapeutic agents across the BBB to eradicate neurohuman immunodeficiency virus (HIV), strategies for on-demand site-specific release of antiretroviral therapy, developing novel nanoformulations capable to recognize and eradicate latently infected HIV reservoirs, and developing novel smart analytical diagnostic tools to detect and monitor HIV infection. Thus, investigation of novel nanoformulations, methodologies for site-specific delivery/release, analytical methods, and diagnostic tools would be of high significance to eradicate and monitor neuroacquired immunodeficiency syndrome. Overall, these developments will certainly help to develop personalized nanomedicines to cure HIV and to develop smart HIV-monitoring analytical systems for disease management.


The use of self-learning modules to facilitate learning of basic science concepts in an integrated medical curriculum.
This study used qualitative and quantitative approaches to evaluate the effectiveness of self-learning modules (SLMs) developed to facilitate and individualize students' learning of basic medical sciences. Twenty physiology and nineteen microanatomy SLMs were designed with interactive images, animations, narrations, and self-assessments. Of 41 medical students, 40 students voluntarily completed a questionnaire with open-ended and closed-ended items to evaluate students' attitudes and perspectives on the learning value of SLMs. Closed-ended items were assessed on a five-point Likert scale (5 = high score) and the data were expressed as mean ± standard deviation. Open-ended questions further evaluated students' perspectives on the effectiveness of SLMs; student responses to open-ended questions were analyzed to identify shared patterns or themes in their experience using SLMs. The results of the midterm examination were also analyzed to compare student performance on items related to SLMs and traditional sessions. Students positively evaluated their experience using the SLMs with an overall mean score of 4.25 (SD ± 0.84). Most students (97%) indicated that the SLMs improved understanding and facilitated learning basic science concepts. SLMs were reported to allow learner control, to help in preparation for subsequent in-class discussion, and to improve understanding and retention. A significant difference in students' performance was observed when comparing SLM-related items with non-SLM items in the midterm examination (P < 0.05). In conclusion, the use of SLMs in an integrated basic science curriculum has the potential to individualize the teaching and improve the learning of basic sciences.


Basketball-Specific Small-Sided Games Training during Ramadan Intermitting Fasting: Do Changes in Body Composition, Sleep Habits, and Perceived Exertion Affect Technical Performance?
The objective of this study was to assess the effects of an additional small-sided games (SSGs) training program during Ramadan intermitting fasting (RIF) on technical performance depending on changes in body composition, sleep habits, and ratings of perceived exertion (RPE). Twenty-four professional male basketball players from the Tunisian first division participated in this study. The players were randomly assigned to an intervention group (INT; n = 12) or an active control group (CON; n = 12). Both groups completed a four-week SSG training program (three sessions per week). During the first and fourth weeks of the SSGs training, the two groups were evaluated to detect changes in technical performance, dietary intake, body composition, sleep quality index (PSQI) survey outcomes, RPE, heart rate (HR), and blood lactate concentration [La]. During the fourth week of the RIF period, body composition, dietary intake, sleep latency, sleep duration, and HR significantly decreased only for INT (p < 0.001). However, RPE significantly increased (p < 0.001), and technical performances were negatively affected (p < 0.01). MANCOVA (adjusted for the percentage of change in sleep duration, body mass, and RPE) showed no significant differences in either group. In conclusion, our results showed that the technical performance of professional basketball male players was significantly affected at the end of RIF independently of changes in RPE, sleep habits, and body composition.


Normal hematopoiesis is maintained by activated bone marrow CD4+ T cells.
CD4(+) T cells produce hematopoietic-related cytokines and are essential for hematopoiesis stimulation during infection and hematologic recovery after bone marrow transplantation. However, it remains unclear if T cells are necessary to maintain normal hematopoiesis. We report here that, in T-cell-deficient mice, terminal differentiation of myeloid progenitors is defective, resulting in very low levels of granulocytes in the periphery. Hematopoiesis is restored after thymus graft or reconstitution with CD4(+) T cells but not CD8(+) T cells. Bone marrow CD4(+) T cells have an activated phenotype and produce cytokines, apparently, in the absence of exogenous stimulation. Transgenic mice carrying T-cell receptor specific for an ovalbumin peptide presented in the context of a specific class II molecule (I-A(d)) (DO11.10 RAG(-/-)) show the same hematopoietic deficiency as athymic mice. Their bone marrow CD4(+) T cells are not activated, suggesting that hematopoiesis maintenance requires the presence of cognate antigen in order to activate bone marrow T-helper cells. In fact, priming of transgenic mice with ovalbumin restores normal hematopoiesis. The data show that the current concept of "normal hematopoiesis" does not reflect a basal bone marrow activity, but it is an antigen-induced state maintained by constant activation of bone marrow CD4(+) T cells.


Rehabilitation of native reptiles and amphibians in DuPage County, Illinois.
Between 1980 and 1994, 586 individuals of 20 native reptile and amphibian species were evaluated at a wildlife rehabilitation facility in DuPage County, Illinois (USA). On examination, 48% of reptiles and 52% of amphibians were injured, and 2% of each class were diseased; remaining animals were uninjured. The most frequent causes of injury to reptiles were automobile collisions (55%), ingestion of fishing tackle (19%), and domestic animal attacks (8%). Automobile collisions (38%), lawn and garden accidents (33%), and pet collection (19%) were the most frequent causes of injury to amphibians. We released 354 reptiles and 54 amphibians into appropriate habitat available in local forest preserves, representing case release prevalences of 84% and 68% for each class, respectively.


Tyrosine and threonine phosphorylation of an immunoaffinity-purified 44-kDa MAP kinase.
We have approached the functioning of a MAP kinase, which is thought to be a "switch kinase" in the phosphorylation cascade initiated from various receptor tyrosine kinases including the insulin receptor. To do so, antipeptide antibodies were raised against the C-terminal portion of ERK1 (extracellular signal-regulated kinase 1), a protein kinase belonging to the family of MAP kinases. With these antipeptide antibodies, we observed the following: (i) a 44-kDa protein can be specifically recognized both under native and denaturing conditions; (ii) a 44-kDa phosphoprotein can be revealed in 32P-labeled cells; its phosphorylation is stimulated by insulin, sodium orthovanadate, and okadaic acid; (iii) a MBP kinase activity can be precipitated, which phosphorylates MBP on threonine residues, and which is stimulated by insulin, sodium orthovanadate, okadaic acid, and fetal calf serum; (iv) this MBP kinase activity appears to be correlated with the in vivo induced phosphorylation of the 44-kDa protein. We next studied the in vitro phosphorylation of this 44-kDa/ERK1-immunoreactive protein. A time- and manganese-dependent phosphorylation was stimulated by the in vitro addition of sodium orthovanadate. Phosphoamino acid analysis of the in vitro phosphorylated 44-kDa protein revealed both threonine and tyrosine phosphorylation. Importantly, this in vitro phosphorylation of MAP kinase results in activation of phosphorylation of added MBP substrate. As a whole, our data indicate that the 44-kDa phosphoprotein identified by our antipeptide antibodies very likely corresponds to a MAP kinase.(ABSTRACT TRUNCATED AT 250 WORDS)


Factors mediating cancer risk in systemic lupus erythematosus.
Patients with systemic lupus erythematosus (SLE) are at an elevated risk for certain cancers compared to the population at large. Cancers seen at higher rates in the SLE population include hematologic malignancies, such as non-Hodgkin lymphoma, and cancers of the lung and thyroid. SLE patients also have a decreased risk for certain malignancies, such as breast cancer, melanoma, and prostate cancer. We review the literature on risk factors for malignancy in patients with SLE and discuss the exogenous and innate factors that are thought to contribute to the unique pattern of cancer risk observed in this patient population. These risk factors are important for providers of SLE patients to understand in order to maintain high clinical suspicion and detect malignancy as soon as possible. Further research is needed to determine the most effective guidelines on counseling patients on cancer screening and prevention.


The complete mitogenome of blue swimming crab Portunus pelagicus Linnaeus, 1766 (Crustacea: Decapoda: Portunidae).
The complete mitochondrial genome (mitogenome) of the blue swimming crab Portunus pelagicus Linnaeus, 1766 (Crustacea: Decapoda: Portunidae) was determined in this study. The full length mitogenome is 16 157 bp in size, and consists of 13 protein-coding genes, 22 tRNA genes, two rRNA genes, and a non-coding control region, with the base composition of 33.70% for A, 18.99% for C, 12.22% for G, and 35.09% for T. The gene order of P. pelagicus mainly retains as the pancrustacean ground pattern, except for a single translocation of tRNA(His) gene. The mitogenome data provide a basis for further studies on population genetics and phylogenetics.


Increased asthma medication use in association with ambient fine and ultrafine particles.
The association between particulate air pollution and asthma medication use and symptoms was assessed in a panel study of 53 adult asthmatics in Erfurt, Germany in winter 1996/1997. Number concentrations of ultrafine particles, 0.01-0.1 microm in diameter (NC(0.01-0.1), mean 17,300 x cm(-3), and mass concentrations of fine particles 0.01-2.5 microm in diameter (MC(0.01-2.5)), mean 30.3 microg x m(-3), were measured concurrently. They were not highly correlated (r=0.45). The associations between ambient particle concentrations and the prevalence of inhaled beta2-agonist, corticosteroid use and asthma symptoms, were analysed separately with logistic regression models, adjusting for trend, temperature, weekend, holidays, and first order autocorrelation of the error. Cumulative exposures over 14 days of ultrafine and fine particles were associated with corticosteroid use. Beta2-agonist use was associated with 5-day mean NC(0.01-0.1) and MC(0.01-2.5). The prevalence of asthma symptoms was associated with ambient particle concentrations. The results suggest that reported asthma medication use and symptoms increase in association with particulate air pollution and gaseous pollutants such as nitrogen dioxide.


Social regulation, medicalisation and the nurse's role: insights from an analysis of nursing documentation.
Medicine is recognised as a dominant source of governmentality and social regulation, and although nursing has been implicated in the same process, analytical work in this area has been sparse. The article aims to present an analysis of nursing records in order to understand the structural and social processes that mediate the texts. 45 sets of nursing records drawn from four clinical sites in Ireland were subjected to a discourse analysis. This article focuses on two main themes that were derived from data: (i) the manner in which nurses controlled, regulated and invigilated patients' activities of daily living and (ii) the way in which activities of daily living were mediated by a biomedical worldview in the clinical settings. Through the organising framework of Activities of Daily Living (ADLs), normative social practices relating to hygiene, eating and drinking, sleeping and so forth were surveyed and monitored within clinical settings. We construct qualitative categories around a range of ways that nurses assessed and judged patients' capacities at ADLs. Furthermore, it is argued that the framework of ADLs epitomises the medicalisation of normative social practices, whereupon the most mundane of normal functions become redefined as an actual or potential clinical pathology, legitimating nursing interventions. According to the nursing documentation, biochemical interventions in the form of various medications were the most dominant means through which nurses attempted to restore or improve the functional capacity of an ADL. We conclude by proposing that nurses' invigilation of patients' ADLs is not necessarily a repressive feature of nursing practice, but rather has the potential to be used to advocate on patients' behalf in certain circumstances.


[Influence of prostatilen on smooth muscle organs functional activity in surgical patients (clinical and experimental study)].
The action of prostatilen on contractile activity of smooth muscles of isolated line slices of urine bladder of Wistar rats (myography) and arterial vessels of cat kidneys (resistography) was studied. On the basis of clinical cases effectiveness of prostatilen was analysed as a treatment restorting urine bladder function in acute reflex urinary retention after operations in the area of rectal sphincter, as well as in treatment of patients with chronic prostatitis. It is shown, that prostatilen produces contractile action on smooth muscles of renal blood vessels in cats and urine bladder walls in rats and it raises contractile activity of smooth muscles of human urine bladder. The results of experimental and clinical investigations make it possible to recommend the application of this bioregulating preparation for treatment and prophylaxis of disturbances in urination.


Electrochemical isothermal-capacitance-transient spectroscopy: a new depth profiling method of deep levels.
The authors report on a new depth profiling method of deep levels, which we call electrochemical isothermal-capacitance-transient spectroscopy (EICTS). This is combined with electrochemical capacitance-voltage using the Schottky barrier of etchable electrolyte and isothermal-capacitance-transient spectroscopy using the capacitance-transient profile at a fixed temperature. We proved its validity by applying to the ZnSe:N epitaxial film of thickness of more than 1000 nm and comparing the characteristics of an obtained deep level with the results measured by conventional deep-level detection techniques. It is expected that EICTS is very effective to assess the deep levels of wide-bandgap semiconductors that suffer from various point defects and their complexes.


Calibration of some instruments for measuring peak expiratory flow.
The performance of instruments for measuring peak expiratory flow have been investigated. This evaluation was carried out with two designs of spirometer, two types of pneumotachograph and three versions of the Wright peak flow meter. The appropriate flow profiles were generated by means of a special type of flow calibrator. The errors produced by the instruments varied from zero to about 26%.


Characterization of Anion Effects on the Nitrate-Sensitive ATP-Dependent Proton Pumping Activity of Soybean (Glycine max L.) Seedling Root Microsomes.
The ATP-dependent proton-pumping activity of soybean (Glycine max L.) root microsomes is predominantly nitrate sensitive and presumably derived from the tonoplast. We used microsomes to characterize anion effects on proton pumping of the tonoplast vesicles using two distinctly different techniques.Preincubation of the vesicles with nitrate caused inhibition of proton pumping and ATPase activity, with similar concentration dependence. Fluoride, which preferentially inhibits the plasma membrane ATPase, inhibited ATPase activity strongly at concentrations which did not affect proton pumping activity.Addition of potassium salts, after a steady-state pH gradient is established in the absence of such salts, caused an increased pH gradient which was due to alleviation of Delta Psi and subsequent increased influx of H(+) into these vesicles. This anion-induced increase in the pH gradient could be used as a measure of the relative anion permeabilities, which were of the order Br(-) = NO(3) (-) > Cl(-) >> SO(4) (2-). Phosphate and fluoride caused no increase in the pH gradient. Since the concentration dependence of KCl- and KNO(3)-induced quenching exhibited a saturable component, and since H(+) uptake was increased by only certain anions, the data suggest that there may be a relatively specific anion channel associated with tonoplast-derived vesicles.


[Comparison between primary anastomosis after intraoperative colonic defecation and Hartmann procedure in patients with obstructive left colon cancer].
To compare primary anastomosis after intraoperative colonic defecation and Hartmann procedure for obstructive left colon cancer. Clinical data of 68 patients who underwent emergent laparotomy for left colon cancer with acute bowel obstruction between January 2000 and January 2008 were analyzed retrospectively. Primary resection and anastomosis with intraoperative defecation was performed in 43 patients and Hartmann's procedure in 25 cases. Patients in both groups were comparable in terms of age, gender, nutritional status, underlying diseases, tumor location and stage, etc. The morbidity and mortality in the two groups were 25.6% vs 28.0% (P=0.761) and 2.3% vs 4.0% (P=0.369), respectively, and the differences were not statistically significant. The length of hospital stay (including first resection operation and second admission for colostomy closure) was (16.6+/-7.8) d in the primary anastomosis group and (24.6+/-9.4) d in the Hartmann procedure group, and the difference was statistically significant (P=0.002). The costs of hospitalization in the two groups were CNY 50,192.8+/-39,727.4 and CNY 58,382.1+/-30,304.9 (P=0.020). Primary resection with intraoperative colonic defecation is safe and effective, and should be considered as an alternative to Hartmann procedure for obstructive left colon cancer in selected patients.


[Chromatin diminution at the border of the XX and XXI centuries].
The size of genomes in eukaryotic organisms is one of the greatest mysteries of biology. As known from the middle of the XX century, the level of organization of a particular organism, does not depend on its genome size, i. e. on DNA amount in the nucleus. We believe that an actual function of non-coding DNA stands behind the phenomenon of chromatin diminution, known already for 100 years. Diminution of chromatin normally takes place in cells involved in body building and never occurs in developmental precursors of germ cells. Apparently, the former are cells, in which non-coding DNA is functionally significant. We cloned a fraction of DNA eliminated during chromatin diminution of Cyclops kolensis (Cyclopoida, Crustascea) and sequenced 90 clones totally making 32 kb. Taken together, the provided evidence has demonstrated a high organization ordering of DNA sequences restricted to the germ line. Chromatin diminution never takes place in human cells and in cells of the majority of animals. These cells may isolate non-coding DNA in other ways, making it unreactable for most enzymes and thus functionally cut off. Thus, a certain part of genome with a particular size and structure may serve for genetic isolation of species as shellfish or junk DNA are vital components rather than pieces of garbage.


Comparative effects of pH and ionic strength on protein-protein interactions, unfolding, and aggregation for IgG1 antibodies.
Changes in protein-protein interactions, protein unfolding, and nonnative aggregation were assessed for a series of human IgG1 antibodies as a function of pH and solution ionic strength (I). Unfolding transitions were characterized with differential scanning calorimetry. Protein-protein interactions were characterized with the apparent second virial coefficient (A(2)) from light scattering. Aggregation pathways were assessed using size-exclusion chromatography and multi-angle laser light scattering, aggregation kinetics, and structural changes monitored by circular dichroism spectroscopy and thioflavine T (ThT) binding. Ionic strength had relatively minor qualitative effects on unfolding, while pH had large effects for all four antibodies. A(2) was sensitive to both pH and I, and indicated that electrostatic interactions and nonuniform surface-charge distributions were important near neutral pH. Depending on solution pH and I, distinct aggregation pathways were found for each antibody, and these shared similar patterns versus pH, I, and A(2). Main differences observed across different antibodies included thermal unfolding transitions in DSC and the effects of pH and I on aggregation kinetics and pathways. These correlated strongly with whether aggregates of a given antibody bound ThT, suggesting possible differences with respect to conformational changes and/or regions of the proteins that are structurally involved in stabilizing the aggregates.


Determination of immunological homology between cellulosome subunits and cloned endoglucanases and xylanase of Clostridium thermocellum.
Immuno-cross reactivity between the subunits of Clostridium thermocellum cellulosome and cloned endogucanases and xylanase from the same bacterium was studied using the polyclonal antibodies raised against cloned enzymes. Dot blot analysis showed that the cellulosome, S8 and S11 subunits cross-reacted strongly with the antibodies of all cloned enzymes tested except that raised against CelC. Western blot analysis revealed that S8 and S11 subunits cross-reacted with the antibodies of CelA, CelB, CelD, CelG, CelH and XynZ, but the antibodies of CelB and CelG were highly specific for S8 and S11 subunits, respectively. Similar analysis using dissociated cellulosome showed that the antibodies of all cloned enzymes tested cross-reacted with more than one subunit of the cellulosome. Antibodies of CelC showed a very low cross-reactivity against all subunits of the cellulosome. The results indicate that immunological cross-reactivity studies could be useful, not only for demonstrating the similarities between native and cloned enzymes, but also for identifying native enzymes using antibodies of cloned enzymes.


Effects of immune checkpoint blockade on antigen-specific CD8+ T cells for use in adoptive cellular therapy.
Adoptive T-cell therapies have been successfully used as prophylaxis or treatment for immunocompromised patients at risk of viral infections or advanced cancers. Unfortunately, for some refractory cancers, they have failed. To overcome this, checkpoint inhibitors are used to rescue immune antitumor responses. We hypothesized that in vitro checkpoint blockade during T-cell stimulation and expansion with messenger RNA (mRNA)-pulsed DCs may enhance the activity of antigen-specific T cells and improve the efficacy of adoptive cellular therapy platforms. Human peripheral blood mononuclear cells were isolated from cytomegalovirus (CMV)-seropositive donors to generate DCs. These were pulsed with CMV matrix phosphoprotein 65 (CMVpp65)-mRNA to educate T cells in coculture for 15 days. Three checkpoint blockade conditions were evaluated (anti-PD1, anti-Tim3, and anti-PD1 + Tim3). IL-2 and antibodies blockades were added every 3 days. Immunophenotyping was performed on Day 0 and Day 15. Polyfunctional antigen-specific responses were evaluated upon rechallenge with CMVpp65 peptides. CMVpp65-activated CD8+ T cells upregulate Lag3 and Tim3 (P ≤ 0.0001). Tim3 antibody blockade alone or in combination led to a significant upregulation of Lag3 expression on CD8+ pp65Tetramer+ central memory, effector memory, and terminal effector memory cells re-expressing RA (TEMRA) T cells. This latter T-cell subset uniquely maintains double-positive Tim3/Lag3 expression after checkpoint blockade. By contrast, PD1 blockade had minimal effects on Tim3 or Lag3 expression. In addition, IFN-γ secretion was reduced in T cells treated with Tim3 blockade in a dose-dependent manner (P = 0.004). In this study, we have identified a potential activating component of Tim3 and linkage between Tim3 and Lag3 signaling upon blocking the Tim3 axis during T-cell-antigen-presenting cell interactions that should be considered when targeting immune checkpoints for clinical use.


Somnogenic cytokines and models concerning their effects on sleep.
All the sleep-promoting substances currently identified also have other biological activities. Despite years of effort, a single specific central nervous system sleep center has not been described. These observations led us to propose a biochemical model of a sleep activational system in which the effects of several sleep factors are integrated into a regulatory scheme. These sleep factors interact by altering the metabolism, production, or activity of each other and thereby result in multiple feedback loops. This web of interactions leads to sleep stability in that minor challenges to the system will not greatly alter sleep. The system, however, is responsive to strong perturbations, such as sleep deprivation and infectious disease. The sleep-promoting effects of cytokines and their interactions with prostaglandins and the neuroendocrine system are used to illustrate the functioning of a part of the sleep activational system under normal conditions and during infectious disease. Although the actions of individuals sleep factors are not specific to sleep, their interactions at various levels of the neuraxis can mediate a specific sleep response. Such a system would also be responsive to the autonomic and environmental parameters that alter sleep.


FOXD1 facilitates pancreatic cancer cell proliferation, invasion, and metastasis by regulating GLUT1-mediated aerobic glycolysis.
Although FOXD1 has been found to be involved in the malignant processes of several types of cancers, its role in pancreatic cancer (PC) is not well understood. This study aimed to investigate the expression and function of FOXD1 in PC. We found that FOXD1 mRNA and protein expression were upregulated in PC tissues compared with non-tumor tissues, and high expression level of FOXD1 was associated with an adverse prognostic index of PC. The results of in vitro and in vivo assays indicate that overexpression of FOXD1 promotes aerobic glycolysis and the capacity of PC cells to proliferate, invade, and metastasize, whereas FOXD1 knockdown inhibits these functions. The results of mechanistic experiments suggest that FOXD1 can not only directly promote SLC2A1 transcription but also inhibit the degradation of SLC2A1 through the RNA-induced silencing complex. As a result, FOXD1 enhances GLUT1 expression and ultimately facilitates PC cell proliferation, invasion, and metastasis by regulating aerobic glycolysis. Taken together, FOXD1 is suggested to be a potential therapeutic target for PC.


SUMOylation regulates the intracellular fate of ZO-2.
The zonula occludens (ZO)-2 protein links tight junctional transmembrane proteins to the actin cytoskeleton and associates with splicing and transcription factors in the nucleus. Multiple posttranslational modifications control the intracellular distribution of ZO-2. Here, we report that ZO-2 is a target of the SUMOylation machinery and provide evidence on how this modification may affect its cellular distribution and function. We show that ZO-2 associates with the E2 SUMO-conjugating enzyme Ubc9 and with SUMO-deconjugating proteases SENP1 and SENP3. In line with this, modification of ZO-2 by endogenous SUMO1 was detectable. Ubc9 fusion-directed SUMOylation confirmed SUMOylation of ZO-2 and was inhibited in the presence of SENP1 but not by an enzymatic-dead SENP1 protein. Moreover, lysine 730 in human ZO-2 was identified as a potential modification site. Mutation of this site to arginine resulted in prolonged nuclear localization of ZO-2 in nuclear recruitment assays. In contrast, a construct mimicking constitutive SUMOylation of ZO-2 (SUMO1ΔGG-ZO-2) was preferentially localized in the cytoplasm. Based on previous findings the differential localization of these ZO-2 constructs may affect glycogen-synthase-kinase-3β (GSK3β) activity and β-catenin/TCF-4-mediated transcription. In this context we observed that ZO-2 directly binds to GSK3β and SUMO1ΔGG-ZO-2 modulates its kinase activity. Moreover, we show that ZO-2 forms a complex with β-catenin. Wild-type ZO-2 and ZO-2-K730R inhibited transcriptional activity in reporter gene assays, whereas the cytosolic SUMO1ΔGG-ZO-2 did not. From these data we conclude that SUMOylation affects the intracellular localization of ZO-2 and its regulatory role on GSK3β and β-catenin signaling activity.


Improving the care of mentally ill patients in a tertiary emergency department: development of a psychiatric assessment and planning unit.
The aim was to describe the Psychiatric Assessment and Planning Unit (PAPU), established at the Royal Melbourne Hospital (RMH) to improve access to psychiatric care for patients presenting via the Emergency Department (ED). PAPU was opened in January 2007. Data was recorded from RMH ED databases to compare ED lengths of stay (LOS) prior to and after establishing PAPU. Interventions including requirement for one-to-one nursing (i.e. one nurse dedicated to the care and observation of one patient) and mechanical restraint and unarmed security threats are routinely documented and were compared before and after PAPU opened. Demographic data and inpatient LOS information was collected by clinicians in PAPU. During the 12 months of initial evaluation we found that PAPU assisted with reducing the number of patients waiting excessive LOS in the RMH ED essentially to zero, due to both expedient transfers from ED and increased direct admissions from the community. We were also able to demonstrate a reduction in mechanical restraint, security codes for unarmed threats and one-to-one nursing in the ED. The establishment of the PAPU at RMH has resulted in significant improvement in the care of patients with mental illness in the least restrictive environment, based on decreased LOS and need for interventions.


PD-L1 Positivity Associated With Presence of Tertiary Lymphoid Structures and High-Stage Disease in Upper Tract Urothelial Carcinoma.
Persistent antigen exposure leads to the accumulation of lymphocytes and subsequent tertiary lymphoid structures (TLS). We investigated the relationship of tumor microenvironment (TME) with respect to programmed death ligand 1 (PD-L1), its receptor programmed death 1 (PD-1), and TLS in upper tract urothelial carcinoma (UTUC) cases and compared them with UTUC associated with urothelial bladder carcinoma (UTUC-BCa). We retrospectively identified 72 patients with UTUC. Representative slides were reviewed, and TLS were counted. Immunohistochemical stains for PD-1 and PD-L1 were performed. PD-1-positive lymphocytes were counted and H-score for PD-L1-positive membranous staining was determined. PD-L1 expression in the tumor was present in 55.1% of the UTUC cases. Higher stage was associated with increased PD-L1 expression (P = .035). TLS were present in 33.3% and their presence was significantly associated with PD-L1 positivity (P = .024). This association remained significant after adjustment for UTUC-BCa. TLS were also associated with a greater number of infiltrating PD-1-positive lymphocytes (P = .013). This study is one of the first comparative studies of the TME in UTUC and UTUC-BCa. PD-L1 is expressed in a subset of UTUC and is associated with TLS. The presence of TLS is an inherent characteristic of UTUC and not secondary to the presence of BCa.


Safety evaluation of synthetic beta-carotene.
The safety of beta-carotene was reassessed by evaluating the relevant literature on the beneficial and adverse effects of beta-carotene on cancer and, in particular, by evaluating the results of toxicity studies. Beta-carotene appeared neither genotoxic nor reprotoxic or teratogenic, and no signs of organ toxicity have been found in subacute, subchronic, or chronic oral toxicity studies in experimental animals receiving doses of up to 1000 mg/day beta-carotene per kg body weight via the diet. Synthetic beta-carotene did not exert any carcinogenic effect in Sprague-Dawley rats or in CD1 mice. An enhanced risk of lung cancer was found in two human intervention studies. Although dose and (timing of) exposure, smoking status, and imbalance of antioxidant defense have been recognized as potential factors accounting for the outcome of these studies, a conclusive explanation has not yet been found. It is concluded that exposure to beta-carotene resulting in mean plasma concentrations of no more than 2.2 micromol/l (1.2 microg/ml) is safe to the general population. By contrast, in heavy smokers higher plasma concentrations may be associated with a higher lung cancer risk.


Aspergillus endocarditis in chronic granulomatous disease.
We report the first case, to our knowledge, of Aspergillus endocarditis in chronic granulomatous disease in a patient who also had an atrial septal defect. A diagnosis was made on culture of the organism from the mass despite extensive prior investigation. The presence of distinctive skin lesions as a diagnostic clue of fungaemia is highlighted. Possible advances in diagnosis by detection of fungal cell wall components and in prophylaxis by use of itraconazole are referred to. We conclude that fungal endocarditis should be considered in this condition, especially in the presence of a structural heart defect.


An IL-1 receptor antagonist blocks a morphine-induced attenuation of locomotor recovery after spinal cord injury.
Morphine is one of the most commonly prescribed medications for the treatment of chronic pain after a spinal cord injury (SCI). Despite widespread use, however, little is known about the secondary consequences of morphine use after SCI. Unfortunately, our previous studies show that administration of a single dose of morphine, in the acute phase of a moderate spinal contusion injury, significantly attenuates locomotor function, reduces weight gain, and produces symptoms of paradoxical pain (Hook et al., 2009). The current study focused on the cellular mechanisms that mediate these effects. Based on data from other models, we hypothesized that pro-inflammatory cytokines might play a role in the morphine-induced attenuation of function. Experiment 1 confirmed that systemic morphine (20 mg/kg) administered one day after a contusion injury significantly increased expression levels of spinal IL-1β 24 h later. Experiment 2 extended these findings, demonstrating that a single dose of morphine (90 μg, i.t.) applied directly onto the spinal cord increased expression levels of spinal IL-1β at both 30 min and 24 h after administration. Experiment 3 showed that administration of an interleukin-1 receptor antagonist (IL-1ra, i.t.) prior to intrathecal morphine (90 μg), blocked the adverse effects of morphine on locomotor recovery. Further, pre-treatment with 3 μg IL-1ra prevented the increased expression of at-level neuropathic pain symptoms that was observed 28 days later in the group treated with morphine-alone. However, the IL-1ra also had adverse effects that were independent of morphine. Treatment with the IL-1ra alone undermined recovery of locomotor function, potentiated weight loss and significantly increased tissue loss at the injury site. Overall, these data suggest that morphine disrupts a critical balance in concentrations of pro-inflammatory cytokines in the spinal cord, and this undermines recovery of function.


Fluorescence thermometry application of photoluminescence image from Cr-doped YAG.
The temperature dependence of a photoluminescence (PL) image from a Cr-doped Y3Al5O12 (YAG:Cr) crystal was evaluated for a fluorescence thermometry application. The brightness of a red PL image of YAG:Cr was found to be increased by increasing the temperature. Temperature variations of the peak intensity and peak area of red PL related to the PL peaks at λ = 680, 690, 708, and 725 nm were evaluated to reveal the cause of the increase in brightness of the PL image. While intensities of the PL peaks at λ = 690, 690, and 708 nm decreased with temperature due to thermal quenching, the peak intensity at λ = 725 nm and the peak area increased with temperature up to 250 °C. The latter factor greatly affects the temperature variation of the brightness of PL images. The increase in the brightness of the PL image with temperature is evidently linked with the thermal excitation of electrons at the 2E level responsible for the increase in the PL peak area. Cr concentration suitable for the temperature sensor is suggested based on the temperature variation of the PL peak area.


Principles and framework for eHealth strategy development.
Significant investment in eHealth solutions is being made in nearly every country of the world. How do we know that these investments and the foregone opportunity costs are the correct ones? Absent, poor, or vague eHealth strategy is a significant barrier to effective investment in, and implementation of, sustainable eHealth solutions and establishment of an eHealth favorable policy environment. Strategy is the driving force, the first essential ingredient, that can place countries in charge of their own eHealth destiny and inform them of the policy necessary to achieve it. In the last 2 years, there has been renewed interest in eHealth strategy from the World Health Organization (WHO), International Telecommunications Union (ITU), Pan American Health Organization (PAHO), the African Union, and the Commonwealth; yet overall, the literature lacks clear guidance to inform countries why and how to develop their own complementary but locally specific eHealth strategy. To address this gap, this paper further develops an eHealth Strategy Development Framework, basing it upon a conceptual framework and relevant theories of strategy and complex system analysis available from the literature. We present here the rationale, theories, and final eHealth strategy development framework by which a systematic and methodical approach can be applied by institutions, subnational regions, and countries to create holistic, needs- and evidence-based, and defensible eHealth strategy and to ensure wise investment in eHealth.


Structural and Functional Changes and Possible Molecular Mechanisms in Aged Skin.
Skin aging is a complex process influenced by intrinsic and extrinsic factors. Together, these factors affect the structure and function of the epidermis and dermis. Histologically, aging skin typically shows epidermal atrophy due to decreased cell numbers. The dermis of aged skin shows decreased numbers of mast cells and fibroblasts. Fibroblast senescence contributes to skin aging by secreting a senescence-associated secretory phenotype, which decreases proliferation by impairing the release of essential growth factors and enhancing degradation of the extracellular matrix through activation of matrix metalloproteinases (MMPs). Several molecular mechanisms affect skin aging including telomere shortening, oxidative stress and MMP, cytokines, autophagic control, microRNAs, and the microbiome. Accumulating evidence on the molecular mechanisms of skin aging has provided clinicians with a wide range of therapeutic targets for treating aging skin.


Both prostaglandin E2 and nitric oxide sequentially mediate the tumor necrosis factor alpha-induced inhibition of surfactant synthesis by human type II pneumocytes.
Tumor necrosis factor alpha (TNF-alpha)-induced inhibition of surfactant synthesis seems to participate in the pathogenesis of the adult respiratory distress syndrome. To examine the ability of human type II pneumocytes to produce nitric oxide (NO) in the presence of TNF-alpha and, in addition, to explore the role of this radical in the transduction of the cytokine signal. Multiple organ donors were the source of lung tissue specimens. Type II pneumocytes were isolated by enzymatic digestion, adherence separation of macrophages, and gradient purification. After 24-hour preculture, cells were cultured for 24 hours in the presence or absence of TNF-alpha (100 ng/mL), sodium nitroprusside (100 mumol/L), N omega-nitro-L-arginine methyl ester (NAME) (1 mmol/L), methylene blue (10 mumol/L),8-bromo-3',5'-cyclic guanosine monophosphate (8-Br-cGMP) (1 mmol/L), prostaglandin E2 (PGE2) (0.1 mumol/L), indomethacin (30 mumol/L), and combinations. The NO release to the medium and cGMP and PGE2 contents of the cells were measured. The incorporation of 14C-labeled glucose (D-[U-14C]glucose) into phosphatidylcholine and phosphatidylglycerol was selectively inhibited either by 8-Br-cGMP or in the presence of TNF-alpha, PGE2, or nitroprusside, all of which caused an increase in the intracellular levels of cGMP. The inhibitory effect of TNF-alpha was partially reverted by indomethacin, NAME, N-monomethyl arginine, or methylene blue. The inhibitory effect of PGE2 was partially reverted by NAME, while that of nitroprusside was reverted by methylene blue, but not by indomethacin. Tumor necrosis factor alpha induced an increase in PGE2 (4.31 +/- 0.27 vs 1.65 +/- 0.17-pg/microgram protein, n = 10, P < .01) and cGMP (0.238 +/- 0.012 vs 0.109 +/- 0.014-pmol/microgram protein, n = 10, P < .01) cell content and in the NO release to the medium (3.10 +/- 0.14 vs 1.19 +/- 0.11-nmol/microgram protein, n = 10, P < .01). The basal NO release to the medium was also increased in the presence of PGE2. The NAME, which blocked NO generation and cGMP increase, did not affect PGE2 production in response to TNF-alpha. However, indomethacin, which blocked PGE2 production, also blunted NO generation and cGMP increase. The NO generation, secondary to PGE2 production, seems responsible for the TNF-alpha-induced inhibition of phosphatidylcholine synthesis by human type II pneumocytes. Nitric oxide seems to exert this effect through activation of guanylyl cyclase.


The Traditional/Acoustic Music Project: a study of vocal demands and vocal health.
The Traditional/Acoustic Music Project seeks to identify the musical and performance characteristics of traditional/acoustic musicians and determine the vocal demands they face with the goals of (1) providing information and outreach to this important group of singers and (2) providing information to physicians, speech-language pathologists, and singing teachers who will enable them to provide appropriate services. Descriptive cross-sectional study. Data have been collected through administration of a 53-item questionnaire. The questionnaire was administered to artists performing at local venues in Knoxville, Tennessee and also to musicians attending the 2008 Folk Alliance Festival in Memphis, Tennessee. Approximately 41% of the respondents have had no vocal training, whereas approximately 34% of the respondents have had some form of formal vocal training (private lessons or group instruction). About 41% of the participants had experienced a tired voice, whereas about 30% of the participants had experienced either a loss of the top range of the voice or a total loss of voice at least once in their careers. Approximately 31% of the respondents had no health insurance. Approximately 69% of the respondents reported that they get their information about healthy singing practices solely from fellow musicians or that they do not get any information at all. Traditional/acoustic musicians are a poorly studied population at risk for the development of voice disorders. Continued research is necessary with the goal of a large sample that can be analyzed for associations, identification of subpopulations, and formulation of specific hypotheses that lend themselves to experimental research. Appropriate models of information and service delivery tailored for the singer-instrumentalist are needed.


Customized Deep Eutectic Solvents as Green Extractants for Ultrasonic-Assisted Enhanced Extraction of Phenolic Antioxidants from Dogbane Leaf-Tea.
This study evaluates the application of eco-friendly deep eutectic solvents (DESs) in the extraction of phenolic antioxidants from dogbane leaf-tea (DLT). The results showed DESs with lower viscosity allowed an efficient extraction of significantly higher contents of total phenolics or flavonoids. An innovative and high-efficient solvent, choline chloride-levulinic acid (ChCl-LevA), was screened and used in ultrasonic-assisted extraction (UAE) of phenolic compounds from DLT. According to full factorial design experimental results, total phenolic content (TPC), total flavonoid content (TFC), antioxidant activity, and anti-α-glucosidase activity (α-GIA) of the DLT extracts were simultaneously optimized by response surface methodology. Sonication temperature and water content in ChCl-LevA were found to be the major factors affecting the TPC, TFC, antioxidant activity, and α-GIA of DLT extracts. Under the optimum parameters (water content in ChCl-LevA was 45%, sonication temperature was 50 °C, and extraction time was 30 min), the measured results for all the responses were obtained as follows: TPC-91.38 ± 7.20 mg GAE/g DW, TFC-84.12 ± 3.47 mg RE/g DW, ABTS+-492 ± 7.33 mmol TE/g DW, FRAP-6235 ± 121 μmol Fe(II)/g DW and α-GIA-230 ± 7.59 mmol AE/g DW, which were consistent with the predicted values. In addition, strongly significant positive correlations were observed between TPC/TFC and bio-activities of the DLT extracts. HPLC results indicated high contents of (-)-epigallocatechin (4272 ± 84.86 μg/g DW), catechin (5268 ± 24.53 μg/g DW), isoquercitrin (3500 ± 86.07 μg/g DW), kaempferol 3-O-rutinoside (3717 ± 97.71 μg/g DW), and protocatechuic acid (644 ± 1.65 μg/g DW) were observed in the DLT extracts. In contrast to other extraction methods, ChCl-LevA-based UAE yielded higher TPC, TFC, individual phenolic contents, stronger antioxidant activity, and α-GIA. Scanning electron microscope (SEM) analysis further confirmed that ChCl-LevA-based UAE enhanced the disruption of cell wall structure, thereby making more phenolic antioxidants released from DLT. In short, ChCl-LevA-based UAE was confirmed to be an innovative and high-efficient method for extraction of phenolic antioxidants from DLT. Dogbane leaves can be considered as a good tea source rich in natural antioxidants.


Equipment modification is associated with fewer stress fractures in female Israel border police recruits.
The study was planned to assess whether combat equipment weight reduction would lead to a reduction in the incidence of stress fractures in female border police infantry recruits taking a 4-month course of basic combat training. 213 female border police recruits, 18-19 years of age, undergoing 16 weeks of basic combat training with lighter rifle and lighter closely fitted combat vest, (total 9.4 kg) were followed prospectively for stress fracture (SFx) incidence, compared to a historical control group of 1,210 recruits who trained with traditional equipment (12.5 kg). Equipment modification was associated with a significant reduction in SFx from 18.3% in the control group to 8.0% in the intervention group (p < 0.0001). This study implies that equipment weight reduction may achieve a significant effect in SFx reduction, Approximating fighting gear to body center of gravity may enhance this effect.


Hyperthermostable surface layer protein tetrabrachion from the archaebacterium Staphylothermus marinus: evidence for the presence of a right-handed coiled coil derived from the primary structure.
The scaffold of the surface layer covering the hyperthermophilic archaebacterium Staphylothermus marinus is formed by an extended filiform glycoprotein complex, tetrabrachion, which is anchored in the cell membrane at one end of a 70 nm stalk and branches at the other end into four arms of 24 nm length. The arms from a canopy-like meshwork by end-to-end contacts, enclosing a "quasi-periplasmic space". The primary structure of the complex, obtained by an approach based entirely on the polymerase chain reaction, shows that the light and the heavy chains are encoded in this order in a single gene and are generated by internal proteolytic cleavage. One light chain associates with the N-terminal part of a heavy chain to form one of the four arms of the complex, comprising about 1000 residues. Following a glycine-rich linker of about ten residues, the C-terminal 500 residues of the four heavy chains converge to form a four-stranded parallel coiled coil, which ends in a transmembrane segment. The sequence of the coiled coil is exceptional in that the heptad repeat of hydrophobic residues typical for left-handed coiled coils shifts to an undecad repeat after an internal proline residue, indicating that the C-terminal part of the sequence forms a right-handed coiled coil. Such a periodicity has not been detected in coiled coils to date. The almost flawless pattern of aliphatic residues, mainly leucine and isoleucine, throughout the hydrophobic core of the stalk provide one explanation for its exceptional stability.


Management of proximal biceps tendon pathology.
The long head of the biceps tendon is widely recognized as an important pain generator, especially in anterior shoulder pain and dysfunction with athletes and working individuals. The purpose of this review is to provide a current understanding of the long head of the biceps tendon anatomy and its surrounding structures, function, and relevant clinical information such as evaluation, treatment options, and complications in hopes of helping orthopaedic surgeons counsel their patients. An understanding of the long head of the biceps tendon anatomy and its surrounding structures is helpful to determine normal function as well as pathologic injuries that stem proximally. The biceps-labral complex has been identified and broken down into different regions that can further enhance a physician's knowledge of common anterior shoulder pain etiologies. Although various physical examination maneuvers exist meant to localize the anterior shoulder pain, the lack of specificity requires orthopaedic surgeons to rely on patient history, advanced imaging, and diagnostic injections in order to determine the patient's next steps. Nonsurgical treatment options such as anti-inflammatory medications, physical therapy, and ultrasound-guided corticosteroid injections should be utilized before entertaining surgical treatment options. If surgery is needed, the three options include biceps tenotomy, biceps tenodesis, or superior labrum anterior to posterior repair. Specifically for biceps tenodesis, recent studies have analyzed open vs arthroscopic techniques, the ideal location of tenodesis with intra-articular, suprapectoral, subpectoral, extra-articular top of groove, and extra-articular bottom of groove approaches, and the best method of fixation using interference screws, suture anchors, or cortical buttons. Orthopaedic surgeons should be aware of the complications of each procedure and respond accordingly for each patient. Once treated, patients often have good to excellent clinical outcomes and low rates of complications.


Quasi-Linear Viscoelastic Characterization of Soft Tissue-Mimicking Materials.
We present a novel method based on the quasi-linear viscoelastic (QLV) theory to describe the time-dependent behavior of soft materials. Unlike previous methods for deriving QLV parameters, we characterize the elastic and viscous behavior of materials separately by using two different sets of experiments. To model the nonlinear elastic behavior, we fit the elastic stress response with a one-term Ogden model. Then, we model the relaxation behavior with a Prony series to compare the stress relaxation of the material at different timescales. This new method allows us to characterize materials with narrow confidence intervals (high accuracy), independently from the loading conditions. We validate our model using samples made of phantom materials that mimic normal and cancerous prostate tissues in terms of Young's modulus. Our model is shown to distinguish materials with similar elastic (viscous) properties but different viscous (elastic) properties. Drawing a precise distinction between the phantoms, this method could be useful for prostate cancer (PCa) diagnosis; but significant clinical studies will be needed in the future.


Ciclesonide: a closer look at its systemic and oropharyngeal safety profile.
Inhaled corticosteroids (ICS) are recommended first-line therapy for patients with asthma of all severities. Prolonged exposure to high-dose ICS can cause systemic and oropharyngeal adverse events. Minimizing ICS-related adverse events by selecting an ICS with an improved safety profile may increase patients' adherence to their asthma treatment. Ciclesonide, a novel ICS currently under development, is a parent compound that is converted in the lungs by endogenous esterases to its active metabolite, desisobutyryl-ciclesonide. Reported data suggest that ciclesonide is well tolerated, with no observed effect on hypothalamic-pituitary-adrenal (HPA)-axis function and a low incidence of oropharyngeal adverse events (comparable with placebo). These safety benefits, observed in children and adults with asthma, may be due to ciclesonide's favorable pharmacokinetic/pharmacodynamic properties. The lack of HPA-axis function suppression may be due to the low oral bioavailability, high serum protein binding and rapid apparent systemic clearance reported with desisobutyryl-ciclesonide. The low incidence of oropharyngeal adverse events may be attributed to the low oral deposition of ciclesonide in the oropharynx and its limited conversion to desisobutyryl-ciclesonide. The favorable safety profile of ciclesonide suggests a conferred benefit to asthma patients treated with this novel ICS.


A Markovian decision model for beef cattle replacement that considers spring and fall calving.
In certain production environments, beef cows are mated during a breeding season that starts in early summer. Cows found not to be pregnant at the end of the breeding season could either be culled or retained and remated the following summer. Alternatively, nonpregnant cows could be mated in the winter. This option would result in having both a spring and a fall calving herd. The purpose of our study was to determine the optimal replacement policy (maximizing long-run average net returns) for a specific production environment by determining for each age of cow, reproductive status (nonpregnant or pregnant), and season of pregnancy checking (spring or fall) whether the cow should be retained to the next breeding season (summer or winter) or be replaced by a pregnant heifer. The problem was formulated as a Markovian decision process and the optimal policy was found by linear programming. The optimal policy was one in which nonpregnant cows were always culled and replaced by heifers in the summer breeding herd, resulting in spring calving only.


Prevalence of dental erosion and the implications for oral health.
The literature on dental erosion is made up of a number of areas, some of which are reviewed here: there are anecdotal case reports, linking a small number of clinical cases with a possible etiological factor. The second area of evidence concentrates on case control studies in which there is a strong association with dental erosion, for example, patients with bulimia. The final piece of epidemiological evidence is beginning to emerge as prevalence studies, but the information from prevalence data worldwide is scanty. It is difficult to compare prevalence studies because of the different indices used in the various studies and also because of the different teeth assessed in the sample. Standardization of indices used would overcome some of these differences as would the reporting of results in a comparable way. However, identifying the true prevalence of erosion per se is fraught with difficulty because there may be more than one etiological factor operating and attrition and or abrasion may complicate the picture, especially in older populations. Case reports frequently associate evidence of erosion with excessive use of particular drinks or foodstuffs so they must be viewed with caution. There is an increasing awareness, amongst the dental profession, of the potential for this particular form of tooth wear to occur. It is important that the dental team is vigilant and instigates preventive measures before tooth tissue loss becomes clinically significant. The importance of erosion in dental health promotion should not be overlooked.


Direct evidence for singlet-singlet energy transfer in Escherichia coli DNA photolyase.
The active form of native Escherichia coli DNA photolyase contains 1,5-dihydro-FAD (FADH2) plus 5,10-methenyltetrahydropteroylpolyglutamate [5,10-CH(+)-H4Pte(Glu)n]. Enzyme containing FADH2 and/or 5,10-methyltetrahydrofolate (5,10-CH(+)-H4folate) can be prepared in reconstitution experiments. Fluorescence quantum yield measurements at various wavelengths with native or reconstituted enzyme provide a simple method for detecting singlet-singlet energy transfer from pterin to FADH2, a key step in the proposed catalytic mechanism. The data satisfy the following criteria: (1) Wavelength-independent quantum yield values are observed for 5,10-CH(+)-H4folate in the absence (0.434) or presence (3.57 X 10(-2)) of FADH2, for 5,10-CH(+)-H4Pte(Glu)n in the presence of FADH2 (5.58 X 10(-2)) and for FADH2 in the absence of pterin (5.34 X 10(-3)); (2) The observed decrease in pterin fluorescence quantum yield in the presence of FADH2 can be used to estimate the efficiency of pterin fluorescence quenching (EQ = 0.918 or 0.871 with 5,10-CH(+)-H4folate or 5,10-CH(+)-H4Pte(Glu)n, respectively); (3) The fluorescence quantum yield of FADH2 is increased in the presence of pterin and varies depending on the excitation wavelength, in agreement with the predicted effect of energy transfer on acceptor fluorescence quantum yield [phi acceptor (+ donor)/phi acceptor (alone) = 1 + EET(epsilon donor/epsilon acceptor), where EET is the efficiency of the energy transfer process]. With 5,10-CH(+)-H4Pte(Glu)n in native enzyme the value obtained for EET (0.92) is similar to EQ, whereas with 5,10-CH(+)-H4folate in reconstituted enzyme the value obtained for EET (0.46) is 2-fold smaller than EQ.(ABSTRACT TRUNCATED AT 250 WORDS)


Age-dependent biphasic changes in ischemic sensitivity in the striatum of Huntington's disease R6/2 transgenic mice.
We used the oxygen/glucose deprivation (OGD) model of ischemia in corticostriatal brain slices to test the hypothesis that metabolic deficiencies in R6/2 transgenic Huntington's disease (HD) mice will impair their recovery from an ischemic challenge. Corticostriatal extracellular field excitatory postsynaptic potentials (fEPSPs) were evoked in transgenic and wild-type (WT) mice in three age groups: 3-4 wk, before the overt behavioral phenotype develops; 5-9 wk, as overt behavioral symptoms begin; and 10-15 wk when symptoms were most severe. OGD for 8 min completely and reversibly inhibited fEPSPs. Although responses of 3-4 wk WTs showed a tolerance to ischemia and recovered rapidly, ischemic sensitivity developed progressively; at 5-9 and 10-15 wk, responses recovered more slowly from OGD. In contrast, although 3-4 wk R6/2 transgenic fEPSPs showed significantly more ischemic sensitivity than their WT counterparts, the R6/2 fEPSPs maintained a relative tolerance to ischemia at 5-9 and 10-15 wk. As a result, a "crossover" point occurred, roughly coinciding with the development of the overt behavioral phenotype (5-9 wk), after which time R6/2 fEPSPs were significantly more resistant to ischemia than WT responses. The increased ischemic sensitivity in 3-4 wk R6/2 responses was not due to excessive glutamate release during OGD as it persisted in the presence of the glutamate receptor antagonist kynurenic acid (1 mM). Although the mechanism for development of ischemic resistance in R6/2 transgenics remains unknown, it correlates with metabolic and biochemical changes described in this model and in HD patients.


Rescue of coagulation factor VII function by the U1+5A snRNA.
Our previous studies with genomic minigenes have demonstrated that an engineered small nuclear RNA-U1 (U1+5a) partially rescued coagulation factor VII (FVII) mRNA processing impaired by the 9726+5G>A mutation. Here, to evaluate the U1+5a effects on FVII function, we devised a full-length FVII splicing-competent construct (pSCFVII-wt). This construct drove in COS-1 cells the synthesis of properly processed FVII transcripts and of secreted functional FVII (23 +/- 4 ng/mL), which were virtually undetectable upon introduction of the 9726+5G>A mutation (pSCFVII-9726+5a). Cotransfection of pSCFVII-9726+5a with pU1+5a resulted in a partial rescue of FVII splicing and protein biosynthesis. The level increase in medium was dose dependent and, with a molar excess (1.5x) of pU1+5a, reached 9.5% plus or minus 3.2% (5.0 +/- 2.8 ng/mL) of FVII-wt coagulant activity. These data provide the first insights into the U1-snRNA-mediated rescue of donor splice sites at protein level, thus further highlighting its therapeutic implications in bleeding disorders, which would benefit even from tiny increase of functional levels.


Evaluation of the Factors Impacting the Accuracy of 13C NMR Chemical Shift Predictions using Density Functional Theory-The Advantage of Long-Range Corrected Functionals.
The various factors influencing the accuracy of 13C NMR calculations using density functional theory (DFT), including the basis set, exchange-correlation (XC) functional, and isotropic shielding calculation method, are evaluated. A wide selection of XC functionals (over 70) were considered, and it was found that long-range corrected functionals offer a significant improvement over the other classes of functionals. Based on a thorough study, it is recommended that for calculating NMR chemical shifts (δ) one should use the CSGT method, the COSMO solvation model, and the LC-TPSSTPSS exchange-correlation functional in conjunction with the cc-pVTZ basis set. A selection of problems in natural product identification are considered in light of the newly recommended level of theory.


The role of nanotechnology-based approaches for clinical infectious diseases and public health.
Given the high incidence of infection and the growing resistance of bacterial and viral infections to the traditional antiseptic, the need for novel antiseptics is critical. Therefore, novel approaches are urgently required to reduce the activity of bacterial and viral infections. Nanotechnology is increasingly being exploited for medical purposes and is of significant interest in eliminating or limiting the activity of various pathogens. Due to the increased surface-to-volume ratio of a given mass of particles, the antimicrobial properties of some naturally occurring antibacterial materials, such as zinc and silver, increase as particle size decreases into the nanometer regime. However, the physical structure of a nanoparticle and the way it interacts with and penetrates the bacteria also appear to provide unique bactericidal mechanisms. To measure the efficacy of nanoparticles (diameter 100 nm) as antimicrobial agents, it is necessary to comprehend the range of approaches for evaluating the viability of bacteria; each of them has its advantages and disadvantages. The nanotechnology-based disinfectants and sensors for SARS-CoV-2 provide a roadmap for creating more effective sensors and disinfectants for detecting and preventing coronaviruses and other infections. Moreover, there is an increasing role of nanotechnology-based approaches in various infections, including wound healing and related infection, nosocomial infections, and various bacterial infections. To meet the demand for patient care, nanotechnology-based disinfectants need to be further advanced with optimum approaches. Herein, we review the current burden of infectious diseases with a focus on SARS-CoV-2 and bacterial infection that significantly burdens developed healthcare systems and small healthcare communities. We then highlight how nanotechnology could aid in improving existing treatment modalities and diagnosis of those infectious agents. Finally, we conclude the current development and future perspective of nanotechnology for combating infectious diseases. The overall goal is to update healthcare providers on the existing role and future of nanotechnology in tackling those common infectious diseases.


Characteristics to consider when selecting a positive control material for an in vitro assay
The use of in vitro assays to inform decision-making requires robust and reproducible results across studies, laboratories, and time. Experiments using positive control materials are an integral component of an assay procedure to demonstrate the extent to which the measurement system is performing as expected. This paper reviews ten characteristics that should be considered when selecting a positive control material for an in vitro assay: 1) the biological mechanism of action, 2) ease of preparation, 3) chemical purity, 4) verifiable physical properties, 5) stability, 6) ability to generate responses spanning the dynamic range of the assay, 7) technical or biological interference, 8) commercial availability, 9) user toxicity, and 10) disposability. Examples and a case study of the monocyte activation test are provided to demonstrate the application of these characteristics for identification and selection of potential positive control materials. Because specific positive control materials are often written into testing standards for in vitro assays, selection of the positive control material based on these characteristics can aid in ensuring the long-term relevance and usability of these standards.


Cardioembolic stroke in atrial fibrillation-rationale for preventive closure of the left atrial appendage.
Atrial fibrillation is the most common cardiac arrhythmias, and a major cause of morbidity and mortality due to cardioembolic stroke. The left atrial appendage is the major site of thrombus formation in non-valvular atrial fibrillation. Loss of atrial systole in atrial fibrillation and increased relative risk of associated stroke point strongly toward a role for stasis of blood in left atrial thrombosis, although thrombus formation is multifactorial, and much more than blood flow irregularities are implicated. Oral anticoagulation with vitamin-K-antagonists is currently the most effective prophylaxis for stroke in atrial fibrillation. Unfortunately, this treatment is often contraindicated, particularly in the elderly, in whom risk of stroke is high. Moreover, given the risk of major bleeding, there is reason to be skeptical of the net benefit when warfarin is used in those patients. This work reviews the pathophysiology of cardioembolic stroke and critically spotlights the current status of preventive anticoagulation therapy. Various techniques to exclude the left atrial appendage from circulation were discussed as a considerable alternative for stroke prophylaxis.