Is too much work engagement detrimental? Linear or curvilinear effects on mental health and job performance.
Most studies report a positive relationship of work engagement with health and job performance, but, occasionally, a "dark side of engagement" has also been uncovered. The current study examined two hypotheses: whether work engagement has (1) a U-shaped curvilinear relation with psychological distress and (2) an inverted U-shaped curvilinear relation with job performance (i.e., in-role performance and creative behavior). A two-wave longitudinal Internet survey with a time lag of seven months was conducted among 1,967 Japanese employees. To test our hypotheses, we used a two-wave panel design and examined the lagged and concurrent relations between work engagement and both outcomes. The results confirmed that work engagement had a curvilinear relation with psychological distress concurrently; a favorable effect was found initially, but this disappeared at intermediate levels of work engagement, and, at higher levels, an adverse effect became prominent. In addition, work engagement had a curvilinear relation with in-role performance both concurrently and longitudinally; the higher the levels of work engagement, the stronger the favorable effects on in-role performance. However, contrary to our expectations, work engagement had a linear relation with psychological distress longitudinally and with creative behavior both concurrently and longitudinally. Hence, our results suggest that work engagement plays a different role in health enhancement compared to performance enhancement. Leveling-off and adverse effects of high work engagement were observed for psychological distress in the short and not in a long run. In contrast, no leveling-off effect of high work engagement was observed for job performance. Thus, except for the short-term effect on psychological distress, no dark side of work engagement was observed for psychological distress and job performance.


Multiple intracranial aneurysms and moyamoya disease associated with microcephalic osteodysplastic primordial dwarfism type II: surgical considerations.
Microcephalic osteodysplastic primordial dwarfism type II (MOPD II) is a rare genetic syndrome characterized by extremely small stature and microcephaly, and is associated in 25% of patients with intracranial aneurysms and moyamoya disease. Although aneurysmal subarachnoid hemorrhage and stroke are leading causes of morbidity and death in these patients, MOPD II is rarely examined in the neurosurgical literature. The authors report their experience with 3 patients who presented with MOPD II, which includes a patient with 8 aneurysms (the most aneurysms reported in the literature), and the first report of a patient with both moyamoya disease and multiple aneurysms. The poor natural history of these lesions indicates aggressive microsurgical and/or endovascular therapy. Microsurgery, whether for aneurysm clip placement or extracranial-intracranial bypass, is challenging due to tight surgical corridors and diminutive arteries in these patients, but is technically feasible and strongly indicated when multiple aneurysms must be treated or cerebral revascularization is needed.


Clonal analysis of Staphylococcus epidermidis isolates carrying or lacking biofilm-mediating genes by multilocus sequence typing.
Staphylococcus epidermidis is part of the normal microflora of the human skin but is also a leading cause of device-associated infections in critically ill patients. Commensal and clinical S. epidermidis isolates differ in their ability to form biofilms on medical devices; the synthesis of biofilms is mediated by the icaADBC operon. Currently, the epidemiological relatedness between ica-positive and -negative isolates is not known; neither is it known whether the ica genes can spread to biofilm-negative strains through horizontal gene transfer. In this study, multilocus sequence typing (MLST) was employed for the clonal analysis of 118 S. epidermidis ica-positive and -negative strains. MLST revealed that the majority of ica-positive and -negative strains were closely related and formed a single clonal complex. Within this complex one sequence type (ST27) was identified which contained exclusively ica-positive isolates and represented the majority of clinical strains tested. ST27 and related ica-positive clones carried different SCCmec cassettes (conferring methicillin resistance) and the insertion sequence IS256. The findings suggest that the S. epidermidis infections analyzed in this report are mainly caused by a single clone (ST27) which occurs preferentially in hospitals and differs from clones in the community. It is hypothesized that the successful establishment of ST27 within nosocomial environments has been facilitated by the presence of genes encoding biofilm and resistance traits.


Subgroup analyses in randomised controlled trials: quantifying the risks of false-positives and false-negatives.
Subgroup analyses are common in randomised controlled trials (RCTs). There are many easily accessible guidelines on the selection and analysis of subgroups but the key messages do not seem to be universally accepted and inappropriate analyses continue to appear in the literature. This has potentially serious implications because erroneous identification of differential subgroup effects may lead to inappropriate provision or withholding of treatment. (1) To quantify the extent to which subgroup analyses may be misleading. (2) To compare the relative merits and weaknesses of the two most common approaches to subgroup analysis: separate (subgroup-specific) analyses of treatment effect and formal statistical tests of interaction. (3) To establish what factors affect the performance of the two approaches. (4) To provide estimates of the increase in sample size required to detect differential subgroup effects. (5) To provide recommendations on the analysis and interpretation of subgroup analyses. The performances of subgroup-specific and formal interaction tests were assessed by simulating data with no differential subgroup effects and determining the extent to which the two approaches (incorrectly) identified such an effect, and simulating data with a differential subgroup effect and determining the extent to which the two approaches were able to (correctly) identify it. Initially, data were simulated to represent the 'simplest case' of two equal-sized treatment groups and two equal-sized subgroups. Data were first simulated with no differential subgroup effect and then with a range of types and magnitudes of subgroup effect with the sample size determined by the nominal power (50-95%) for the overall treatment effect. Additional simulations were conducted to explore the individual impact of the sample size, the magnitude of the overall treatment effect, the size and number of treatment groups and subgroups and, in the case of continuous data, the variability of the data. The simulated data covered the types of outcomes most commonly used in RCTs, namely continuous (Gaussian) variables, binary outcomes and survival times. All analyses were carried out using appropriate regression models, and subgroup effects were identified on the basis of statistical significance at the 5% level. While there was some variation for smaller sample sizes, the results for the three types of outcome were very similar for simulations with a total sample size of greater than or equal to 200. With simulated simplest case data with no differential subgroup effects, the formal tests of interaction were significant in 5% of cases as expected, while subgroup-specific tests were less reliable and identified effects in 7-66% of cases depending on whether there was an overall treatment effect. The most common type of subgroup effect identified in this way was where the treatment effect was seen to be significant in one subgroup only. When a simulated differential subgroup effect was included, the results were dependent on the nominal power of the simulated data and the type and magnitude of the subgroup effect. However, the performance of the formal interaction test was generally superior to that of the subgroup-specific analyses, with more differential effects correctly identified. In addition, the subgroup-specific analyses often suggested the wrong type of differential effect. The ability of formal interaction tests to (correctly) identify subgroup effects improved as the size of the interaction increased relative to the overall treatment effect. When the size of the interaction was twice the overall effect or greater, the interaction tests had at least the same power as the overall treatment effect. However, power was considerably reduced for smaller interactions, which are much more likely in practice. The inflation factor required to increase the sample size to enable detection of the interaction with the same power as the overall effect varied with the size of the interaction. For an interaction of the same magnitude as the overall effect, the inflation factor was 4, and this increased dramatically to of greater than or equal to 100 for more subtle interactions of < 20% of the overall effect. Formal interaction tests were generally robust to alterations in the number and size of the treatment and subgroups and, for continuous data, the variance in the treatment groups, with the only exception being a change in the variance in one of the subgroups. In contrast, the performance of the subgroup-specific tests was affected by almost all of these factors with only a change in the number of treatment groups having no impact at all. While it is generally recognised that subgroup analyses can produce spurious results, the extent of the problem is almost certainly under-estimated. This is particularly true when subgroup-specific analyses are used. In addition, the increase in sample size required to identify differential subgroup effects may be substantial and the commonly used 'rule of four' may not always be sufficient, especially when interactions are relatively subtle, as is often the case. CONCLUSIONS--RECOMMENDATIONS FOR SUBGROUP ANALYSES AND THEIR INTERPRETATION: (1) Subgroup analyses should, as far as possible, be restricted to those proposed before data collection. Any subgroups chosen after this time should be clearly identified. (2) Trials should ideally be powered with subgroup analyses in mind. However, for modest interactions, this may not be feasible. (3) Subgroup-specific analyses are particularly unreliable and are affected by many factors. Subgroup analyses should always be based on formal tests of interaction although even these should be interpreted with caution. (4) The results from any subgroup analyses should not be over-interpreted. Unless there is strong supporting evidence, they are best viewed as a hypothesis-generation exercise. In particular, one should be wary of evidence suggesting that treatment is effective in one subgroup only. (5) Any apparent lack of differential effect should be regarded with caution unless the study was specifically powered with interactions in mind. CONCLUSIONS--RECOMMENDATIONS FOR RESEARCH: (1) The implications of considering confidence intervals rather than p-values could be considered. (2) The same approach as in this study could be applied to contexts other than RCTs, such as observational studies and meta-analyses. (3) The scenarios used in this study could be examined more comprehensively using other statistical methods, incorporating clustering effects, considering other types of outcome variable and using other approaches, such as Bootstrapping or Bayesian methods.


Prospective, Multisite, International Comparison of 18F-Fluoromethylcholine PET/CT, Multiparametric MRI, and 68Ga-HBED-CC PSMA-11 PET/CT in Men with High-Risk Features and Biochemical Failure After Radical Prostatectomy: Clinical Performance and Patient Outcomes.
A significant proportion of men with rising prostate-specific antigen (PSA) levels after radical prostatectomy (RP) fail prostate fossa (PF) salvage radiation treatment (SRT). This study was done to assess the ability of 18F-fluoromethylcholine (18F-FCH) PET/CT (hereafter referred to as 18F-FCH), 68Ga-HBED-CC PSMA-11 PET/CT (hereafter referred to as PSMA), and pelvic multiparametric MRI (hereafter referred to as pelvic MRI) to identify men who will best benefit from SRT. Methods: Prospective, multisite imaging studies were carried out in men who had rising PSA levels after RP, high-risk features, and negative/equivocal conventional imaging results and who were being considered for SRT. 18F-FCH (91/91), pelvic MRI (88/91), and PSMA (31/91) (Australia) were all performed within 2 wk. Imaging was interpreted by experienced local/central interpreters who were masked with regard to other imaging results, with consensus being reached for discordant interpretations. Expected management was documented before and after imaging, and data about all treatments and PSA levels were collected for 3 y. The treatment response to SRT was defined as a reduction in PSA levels of >50% without androgen deprivation therapy. Results: The median Gleason score, PSA level at imaging, and PSA doubling time were 8, 0.42 (interquartile range, 0.29-0.93) ng/mL, and 5.0 (interquartile range, 3.3-7.6) months. Recurrent prostate cancer was detected in 28% (25/88) by pelvic MRI, 32% (29/91) by 18F-FCH, and 42% (13/31) by PSMA. This recurrence was found within the PF in 21.5% (19/88), 13% (12/91), and 19% (6/31) and at sites outside the PF (extra-PF) in 8% (7/88), 19% (17/91), and 32% (10/31) by MRI, 18F-FCH, and PSMA, respectively (P < 0.004). A total of 94% (16/17) of extra-PF sites on 18F-FCH were within the pelvic MRI field. Intrapelvic extra-PF disease was detected in 90% (9/10) by PSMA and in 31% (5/16) by MRI. 18F-FCH changed management in 46% (42/91), and MRI changed management in 24% (21/88). PSMA provided additional management changes over 18F-FCH in 23% (7/31). The treatment response to SRT was higher in men with negative results or disease confined to the PF than in men with extra-PF disease (18F-FCH 73% [32/44] versus 33% [3/9] [P < 0.02], pelvic MRI 70% [32/46] versus 50% [2/4] [P was not significant], and PSMA 88% [7/8] versus 14% [1/7] [P < 0.005]). Men with negative imaging results (MRI, 18F-FCH, or PSMA) had high (78%) SRT response rates. Conclusion:18F-FCH and PSMA had high detection rates for extra-PF disease in men with negative/equivocal conventional imaging results and rising PSA levels after RP. These findings affected management and treatment responses, suggesting an important role for PET in triaging men being considered for curative SRT.


Stability of colloidal dispersions in the presence of protein fibrils.
We studied the stability of monodispersed polystyrene latex dispersions with protein fibrils at different concentrations at pH 2 using microscopy and diffusing wave spectroscopy. At low fibril concentrations, fibrils induced bridging flocculation due to the opposite charges between fibrils and the latex particles. At higher fibril concentration the dispersions were stabilized due to steric and/or electrostatic repulsion. Upon further increasing fibril concentration, we find that the dispersion is destabilized again by depletion interaction. At even higher fibril concentration, the dispersions are stabilized again. These dispersions have a higher stability compared to the dispersions without fibrils. Interestingly, these dispersions contain single particles and small clusters of particles that do not grow beyond a certain size. Although the stabilization mechanism is not clear yet, the results from microscopy and diffusing wave spectroscopy point in the direction of a kinetic barrier that depends on fibril concentration.


From postsynaptic potentials to spikes in the genesis of auditory spatial receptive fields.
Space-specific neurons in the owl's inferior colliculus respond only to a sound coming from a particular direction, which is equivalent to a specific combination of interaural time difference (ITD) and interaural level difference (ILD). Comparison of subthreshold postsynaptic potentials (PSPs) and spike output for the same neurons showed that receptive fields measured in PSPs were much larger than those measured in spikes in both ITD and ILD dimensions. Space-specific neurons fire more spikes for a particular ITD than for its phase equivalents (ITD +/- 1/F, where F is best frequency). This differential response was much less pronounced in PSPs. The two sides of pyramid-shaped ILD curves were more symmetrical in spikes than in PSPs. Furthermore, monaural stimuli that were ineffective in eliciting spikes induced subthreshold PSPs. The main cause of these changes between PSPs and spikes is thresholding. The spiking threshold did not vary with the kind of acoustic stimuli presented. However, the thresholds of sound-induced first spikes were lower than those of later sound-induced and spontaneous spikes. This change in threshold may account for the sharpening of ITD selectivity during the stimulus. Large changes in receptive fields across single neurons are not unique to the owl's space-specific neurons but occur in mammalian visual and somatosensory cortices, suggesting the existence of general principles in the formation of receptive fields in high-order neurons.


The expression pattern of the mafB/kr gene in birds and mice reveals that the kreisler phenotype does not represent a null mutant.
The recessive mouse mutation kreisler affects hindbrain segmentation and inner ear development in homozygous mice. The mouse gene affected by the mutation was found to encode a basic domain leucine-zipper (bZIP)-type transcription factor of the Maf-family named kr (Cordes, S.P. and Barsh, G.S. (1994) Cell 79, 1025-1034). The avian bZIP transcription factor mafB, which shows high homology to kr, has been identified as an interaction partner of c-Ets 1 (Sieweke, M.H., Tekotte, M.H., Frampton, J. and Graf, T. (1996) Cell 85, 49-60). Here we demonstrate by Southern blot analysis that mafB is the avian homologue of kr, and present a detailed pattern of its expression during avian and murine embryonic development. Consistent with the kreisler phenotype, mafB is expressed in avians in the tissues which are affected by the mouse mutation: rhombomeres 5 and 6 (r5 and r6) and the neural crest derived from these rhombomeres. However, our analysis reveals a variety of additional expression sites: mafB/kr expression persists in vestibular and acoustic nuclei and is also observed in differentiating neurons of the spinal cord and brain stem. Restricted expression sites are found in the mesonephros, the perichondrium, and in the hemopoietic system. Since these expression sites are conserved between mouse and chicken we reexamined homozygous kreisler mice for unrevealed phenotypes in the hemopoietic system. However, peritoneal macrophages from homozygous kreisler mice were found to be functionally normal and still expressed mafB/kr. Other adult tissues examined from homozygous kreisler mice had also not lost mafB/kr expression. Our results thus indicate that the kreisler mutation involves a tissue specific gene inactivation and suggest additional roles for mafB/kr in later developmental and differentiation processes that are not revealed by the mutation.


Urinary excretion of lead and mercury after oral administration of meso-2,3-dimercaptosuccinic acid in patients with motor neurone disease.
Amyotrophic lateral sclerosis (ALS) and spinal muscular atrophy (SMA) are progressive neurodegenerative disorders involving motor neurones. The aetiology of the non-familiar forms is still unknown but it has been suggested that long-term exposure to heavy metals such as lead and mercury may play a role in the pathogenesis of these diseases. In 53 patients suffering from ALS (n = 42) and SMA (n = 9) the oral administration of dimercaptosuccinic acid (DMSA, 20 mg/kg) did not result in a greater mobilization of lead and mercury from peripheral depots than in control subjects. Although it cannot be excluded that the amount of lead or mercury excreted after DMSA administration may not be a reflection of the amount accumulated in the motor neurons, this study does not provide support for the hypothesis that heavy metals play a significant role in the occurrence of motor neurone diseases.


Why Should We Have a Periodic Safety and Performance Program for Medical Devices.
Nowadays, more than 10,000 different types of medical devices can be found in hospitals.These devices used in medical centers and hospitals for monitoring and treatment of patients require periodic safety and performance checking in order to have confidence in their functioning and operation. Physicians need better accurate medical measurements in order to better diagnose diseases, monitor patients and deliver treatments, in this way failure to ensure appropriate measurements will certainly have diverse effects. Safety and performance testing of medical devices in the medical sector is a one of the key factor in improving public health. Acquiring results of some investigations indicate a need for new and severe regulations on periodic performance verifications and medical equipment quality control program especially in highrisks instruments. The metrological reliability of four high risk medical devices, Electrosurgical unit, Defibrillator, Syringe pump and Infant incubator in use some hospitals (privates and publics) in one of the province of Iran according to international and national standards was evaluated. Quantitative analysis of Some parameters that impact the safety and performance showed the amount of the obtained results in some equipment are in critical range and have higher values than standard limitations. General electrical safety evaluations for measuring the patient leakage currents and patient auxiliary currents carried out for all of groups,in some cases the amount of leakage currents were over the standard limitations. Acquiring results indicate a need for new and severe regulations on periodic performance verifications and medical equipment quality control program especially in high risk instruments. It is also necessary provide training courses for operating staff in the field of meterology in medicine and what's the critical parameters and how they can get good accuracy results equipment.


Parental separation, loss and psychosis in different ethnic groups: a case-control study.
Numerous studies have reported high rates of psychosis in the Black Caribbean and Black African populations in the UK. However, few studies have investigated the role of specific risk factors in different ethnic groups. We sought to investigate the relationship between long-term separation from, and death of, a parent before the age of 16 and risk of adult psychosis in different ethnic groups. All patients with a first episode of psychosis who made contact with psychiatric services in defined catchment areas in London and Nottingham, UK and a series of community controls were included in the AESOP (Aetiology and Ethnicity in Schizophrenia and Other Psychoses) study. Data relating to clinical and social variables, including parental separation and loss, were collected from patients and controls. Separation from, and death of, a parent before the age of 16 were both strongly associated with a two- to threefold increased risk of psychosis. The strength of these associations were similar for White British and Black Caribbean (but not Black African) subjects. Separation from (but not death of) a parent was more common among Black Caribbean controls than White British controls. Early separation may have a greater impact in the Black Caribbean population, because it is more common, and may contribute to the excess of psychosis in this population.


Post-test comparison of HIV test knowledge and changes in sexual risk behaviour between clients accessing HIV testing online versus in-clinic.
Internet-based HIV testing offers the potential to address privacy-related barriers to testing and increase frequency of testing but may result in missed opportunities related to sexual health education and prevention that typically occur in face-to-face encounters. In this study, we assessed the HIV test knowledge and sexual risk behaviour of clients testing for HIV through GetCheckedOnline, an internet-based sexually transmitted and bloodborne infection testing platform inclusive of HIV testing, in comparison to clients testing through a large sexual health clinic. We concurrently recruited GetCheckedOnline clients and clinic clients from Vancouver, Canada, over the course of a 10-month period during 2015-2016. Participants completed baseline and 3-month questionnaires, anonymous and online. A six-item score was used to estimate knowledge of HIV test concepts typically conveyed during an HIV pretest encounter in a clinic. We used multiple regression to estimate associations between testing modality (online vs clinic based) and two outcomes-HIV test knowledge and change in condom use pre/post-test-with adjustment for relevant background factors. Among 352 participants, online testers demonstrated higher HIV post-test knowledge than clinic-based testers (mean score 4.65/6 vs 4.09/6; p<0.05); this difference was reduced in adjusted analysis (p>0.05). Men who have sex with men, clients with a university degree, those who have lived in Canada >10 years and English speakers had higher HIV post-test knowledge (p<0.05). Eighteen per cent of online testers and 10% of clinic-based testers increased condom use during the 3 months post-test (p>0.05). In this comparative study between online and clinic-based testers, we found no evidence of decreased HIV test knowledge or decreased condom use following HIV testing through GetCheckedOnline. Our findings suggest that with careful design and attention to educational content, online testing services may not lead to missed opportunities for HIV education and counselling.


Bypass of DNA lesions generated during anticancer treatment with cisplatin by DNA polymerase eta.
DNA polymerase eta (Pol eta) is a eukaryotic lesion bypass polymerase that helps organisms to survive exposure to ultraviolet (UV) radiation, and tumor cells to gain resistance against cisplatin-based chemotherapy. It allows cells to replicate across cross-link lesions such as 1,2-d(GpG) cisplatin adducts (Pt-GG) and UV-induced cis-syn thymine dimers. We present structural and biochemical analysis of how Pol eta copies Pt-GG-containing DNA. The damaged DNA is bound in an open DNA binding rim. Nucleotidyl transfer requires the DNA to rotate into an active conformation, driven by hydrogen bonding of the templating base to the dNTP. For the 3'dG of the Pt-GG, this step is accomplished by a Watson-Crick base pair to dCTP and is biochemically efficient and accurate. In contrast, bypass of the 5'dG of the Pt-GG is less efficient and promiscuous for dCTP and dATP as a result of the presence of the rigid Pt cross-link. Our analysis reveals the set of structural features that enable Pol eta to replicate across strongly distorting DNA lesions.


Nano-confinement induced chain alignment in ordered P3HT nanostructures defined by nanoimprint lithography.
Control of polymer morphology and chain orientation is of great importance in organic solar cells and field effect transistors (OFETs). Here we report the use of nanoimprint lithography to fabricate large-area, high-density, and ordered nanostructures in conjugated polymer poly(3-hexylthiophene) or P3HT, and also to simultaneously control 3D chain alignment within these P3HT nanostructures. Out-of-plane and in-plane grazing incident X-ray diffraction were used to determine the chain orientation in the imprinted P3HT nanostructures, which shows a strong dependence on their geometry (gratings or pillars). Vertical chain alignment was observed in both nanogratings and nanopillars, indicating strong potential to improve charge transport and optical properties for solar cells in comparison to bulk heterojunction structure. For P3HT nanogratings, pi-pi stacking along the grating direction with an angular distribution of +/-20 degrees was found, which is favorable for OFETs. We propose the chain alignment is induced by the nanoconfinement during nanoimprinting via pi-pi interaction and hydrophobic interaction between polymer chain and mold surfaces.


Regulation of the Epigenome by Vitamin C.
Emerging evidence suggests that ascorbate, the dominant form of vitamin C under physiological pH conditions, influences activity of the genome via regulating epigenomic processes. Ascorbate serves as a cofactor for Ten-eleven translocation (TET) dioxygenases that catalyze the oxidation of 5-methylcytosine (5mC) into 5-hydroxymethylcytosine (5hmC), and further to 5-formylcytosine (5fC) and to 5-carboxylcytosine (5caC), which are ultimately replaced by unmodified cytosine. The Jumonji C (JmjC)-domain-containing histone demethylases also require ascorbate as a cofactor for histone demethylation. Thus, by primarily participating in the demethylation of both DNA and histones, ascorbate appears to be a mediator of the interface between the genome and environment. Furthermore, redox status has a profound impact on the bioavailability of ascorbate in the nucleus. In order to bridge the gap between redox biology and genomics, we suggest an interdisciplinary research field that can be termed redox genomics to study dynamic redox processes in health and diseases. This review examines the evidence and potential molecular mechanism of ascorbate in the demethylation of the genome, and it highlights potential epigenetic roles of ascorbate in various diseases.


Optical imaging of metabolism in HER2 overexpressing breast cancer cells.
The optical redox ratio (fluorescence intensity of NADH divided by that of FAD), was acquired for a panel of breast cancer cell lines to investigate how overexpression of human epidermal growth factor receptor 2 (HER2) affects tumor cell metabolism, and how tumor metabolism may be altered in response to clinically used HER2-targeted therapies. Confocal fluorescence microscopy was used to acquire NADH and FAD auto-fluorescent images. The optical redox ratio was highest in cells overexpressing HER2 and lowest in triple negative breast cancer (TNBC) cells, which lack HER2, progesterone receptor, and estrogen receptor (ER). The redox ratio in ER-positive/HER2-negative cells was higher than what was seen in TNBC cells, but lower than that in HER2 overexpressing cells. Importantly, inhibition of HER2 using trastuzumab significantly reduced the redox ratio in HER2 overexpressing cells. Furthermore, the combinatorial inhibition of HER2 and ER decreased the redox ratio in ER+/HER2+ breast cancer cells to a greater extent than inhibition of either receptor alone. Interestingly, trastuzumab had little impact upon the redox ratio in a cell line selected for acquired resistance to trastuzumab. Taken together, these data indicate that the optical redox ratio measures changes in tumor metabolism that reflect the oncogenic effects of HER2 activity within the cell, as well as the response of the cell to therapeutic inhibition of HER2. Therefore, optical redox imaging holds the promise of measuring response and resistance to receptor-targeted breast cancer therapies in real time, which could potentially impact clinical decisions and improve patient outcome.


Bioassay of 1,5-naphthalenediamine for possible carcinogenicity.
A bioassay of 1,5-naphthalenediamine for possible carcinogenicity was conducted using Fischer 344 rats and B6C3F1 mice. 1,5-Naphthalenediamine was administered in the feed, at either of two concentrations, to groups of 50 male and 50 female animals of each species. The high and low dietary concentrations utilized in the chronic bioassay were, respectively, 0.1 and 0.05 percent for rats and 0.2 and 0.1 percent for mice. The compound was administered in the diet for 103 weeks, followed by up to 4 weeks of observation. Fifty mice of each sex and 25 rats of each sex were placed on test as controls. These animals were observed for up to 110 weeks. There were no significant positive associations between the administered concentrations of 1,5-naphthalenediamine and mortality in either sex of rats or mice. In all groups adequate numbers of animals survived sufficiently long to be at risk from late-developing tumors. Among dosed female rats, a statistically significant increase in endometrial stromal polyps was observed. Several of these tumors underwent malignant transformation to endometrial stromal sarcomas. The incidence of female rats having either adenoma or carcinoma of the clitoral gland was statistically significant. No neoplasms were observed at significantly increased incidences in dosed male rats. Based on lack of clinical signs or weight loss, the male rats may have been able to withstand a higher dose. In mice, dose-related increases in thyroid neoplasms were observed in both sexes. The incidence of thyroid C-cell carcinomas was significant for high dose female mice. The combined incidences of papillary adenomas, follicular-cell adenomas and papillary cystadenomas of the thyroid were significant for mice of both sexes. The incidence of hepatocellular carcinomas and the incidence of alveolar/bronchiolar adenomas were each significant for dosed female mice. Under the conditions of this bioassay, 1,5-naphthalenediamine was carcinogenic in female Fischer 344 rats, causing clitoral and uterine neoplasms. 1,5-Naphthalenediamine was also carcinogenic for B6C3F1 mice, producing thyroid neoplasms in males and neoplasms of the thyroid, liver, and lung in females. Insufficient evidence was provided for the carcinogenicity of the compound in male Fischer 344 rats.


The open reading frame 3a protein of severe acute respiratory syndrome-associated coronavirus promotes membrane rearrangement and cell death.
The genome of the severe acute respiratory syndrome-associated coronavirus (SARS-CoV) contains eight open reading frames (ORFs) that encode novel proteins. These accessory proteins are dispensable for in vitro and in vivo replication and thus may be important for other aspects of virus-host interactions. We investigated the functions of the largest of the accessory proteins, the ORF 3a protein, using a 3a-deficient strain of SARS-CoV. Cell death of Vero cells after infection with SARS-CoV was reduced upon deletion of ORF 3a. Electron microscopy of infected cells revealed a role for ORF 3a in SARS-CoV induced vesicle formation, a prominent feature of cells from SARS patients. In addition, we report that ORF 3a is both necessary and sufficient for SARS-CoV-induced Golgi fragmentation and that the 3a protein accumulates and localizes to vesicles containing markers for late endosomes. Finally, overexpression of ADP-ribosylation factor 1 (Arf1), a small GTPase essential for the maintenance of the Golgi apparatus, restored Golgi morphology during infection. These results establish an important role for ORF 3a in SARS-CoV-induced cell death, Golgi fragmentation, and the accumulation of intracellular vesicles.


Molecular and morphological diversity of Narragansett Bay (RI, USA) Ulva (Ulvales, Chlorophyta) populations.
Macroalgal bloom-forming species occur in coastal systems worldwide. However, due to overlapping morphologies in some taxa, accurate taxonomic assessment and classification of these species can be quite challenging. We investigated the molecular and morphological characteristics of 153 specimens of bloom-forming Ulva located in and around Narragansett Bay, RI, USA. We analyzed sequences of the nuclear internal transcribed spacer 1 region (ITS1) and the chloroplast-encoded rbcL; based on the ITS1 data, we grouped the specimens into nine operational taxonomic units (OTUs). Eight of these OTUs have been previously reported to exist, while one is novel. Of the eight OTUs, all shared sequence identity with previously published sequences or differed by less than 1.5% sequence divergence for two molecular markers. Previously, 10 species names were reported for Ulva in Rhode Island (one blade and nine tube-forming species) based upon morphological classification alone. Of our nine OTUs, three contained blade-forming specimens (U. lactuca, U. compressa, U. rigida), one OTU had a blade with a tubular stipe, and six contained unbranched and/or branched tubular morphologies (one of these six, U. compressa, had both a blade and a tube morphology). While the three blade-forming OTUs in Narragansett Bay can frequently be distinguished by careful observations of morphological characteristics, and spatial/temporal distribution, it is much more difficult to distinguish among the tube-forming specimens based upon morphology or distribution alone. Our data support the molecular species concept for Ulva, and indicate that molecular-based classifications of Ulva species are critical for proper species identification, and subsequent ecological assessment or mitigation of Ulva blooms.


A Paradigm shift in the concept for making dental impressions.
Digital dental impression is a revolutionary technological advancement that so surpasses the accuracy and efficiency of former techniques for obtaining replicas of prepared teeth for the purpose of fabricating restorations that its adoption by dentists is rapidly eclipsing the use of elastomeric impression materials. The ultimate goals of dentists dedicated to quality restorative dentistry are to make their treatment of patients as accurate, stressless, and efficient as possible. By elimination of the everyday problems described above, there is no question that the significant advantages of digital impressions will make intraoral digital scanning standard procedure in most dental offices within the next several years. Furthermore, digital impressions have proven to reduce remakes and returns, as well as increase overall efficiency. The patient also benefits by being provided a far more positive experience. Finally, through the use of digital impression making, it has been determined that laboratory products become more consistent and require less chair time at insertion.


Variability in standard care for cytomegalovirus prevention and detection in pediatric lung transplantation: survey of eight pediatric lung transplant programs.
Cytomegalovirus (CMV) infection after pediatric lung transplantation is a significant risk factor for morbidity and mortality in the first year after transplantation. Multiple strategies have been reported for CMV prevention among adult lung transplant programs. In contrast, little information has been reported regarding protocols for prevention and detection of CMV from pediatric programs. We conducted a survey to better understand the range of practice patterns for CMV prevention and detection at pediatric lung transplant centers. A self-administered questionnaire was distributed to 11 pediatric lung transplant centers identified through the International Pediatric Lung Transplant Collaborative in September 2002. A member of the lung transplant team from each institution was asked to provide the methods of CMV prevention and surveillance. Eight of 11 centers surveyed responded to the questionnaire accounting for 45.6% (26 of 57) and 100% (three of three) of the pediatric lung transplants performed in the US and UK in 2001, respectively. All centers used prophylactic therapy against CMV with either ganciclovir or valganciclovir with duration ranging from 3.5 wk to indefinitely. Most centers (six of eight) prescribed a prophylactic regimen based on donor and recipient CMV serostatus. Half (four of eight) of the centers report using CMV hyperimmune globulin in addition to an antiviral agent. Method for CMV detection varied widely, including use of conventional viral culture (n = 1), antigenemia (n = 7), and polymerase chain reaction (n = 2). A wide range of strategies is used to prevent and detect CMV in pediatric lung transplant recipients with little empiric evidence demonstrating the optimal approach. A retrospective analysis among these centers is being conducted to evaluate the efficacy of these approaches.


Application of the Hough transform for the automatic determination of soot aggregate morphology.
We report a new method for automated identification and measurement of primary particles within soot aggregates as well as the sizes of the aggregates and discuss its application to high-resolution transmission electron microscope (TEM) images of the aggregates. The image processing algorithm is based on an optimized Hough transform, applied to the external border of the aggregate. This achieves a significant data reduction by decomposing the particle border into fragments, which are assumed to be spheres in the present application, consistent with the known morphology of soot aggregates. Unlike traditional techniques, which are ultimately reliant on manual (human) measurement of a small sample of primary particles from a subset of aggregates, this method gives a direct measurement of the sizes of the aggregates and the size distributions of the primary particles of which they are composed. The current version of the algorithm allows processing of high-resolution TEM images by a conventional laptop computer at a rate of 1-2 ms per aggregate. The results were validated by comparison with manual image processing, and excellent agreement was found.


The influence of paraquat on the in vitro oxygen consumption of rabbit lung.
The effects of paraquat on the aerobic metabolism of rabbit lung slices were investigated. The oxygen consumption of lung slices was examined at different oxygen tensions and in the presence or absence of glucose as substrate in a Krebs-Ringer phosphate medium. In an air phase, the oxygen consumption of control lung tissue with glucose in the medium was practically the same as the oxygen consumption without glucose over a 3-hour period. In a 100% oxygen phase, glucose induced a marked increase in oxygen uptake, which persisted for about 2 hours. Thereafter, a decrease occurred in oxygen consumption which was notably faster than that of lung tissue without added glucose. With 10 mM glucose in the medium, paraquat (10(-5)M and 10(-3)M) immediately stimulated the oxygen consumption of lung slices. Although the initial increase in aerobic metabolism seemed to be glucose-dependent, the secondary inhibitory effect of paraquat appeared to be of the same magnitude in the presence or absence of glucose. Both the initial stimulation as well as the secondary inhibition were much more pronounced in a 100% oxygen atmosphere than in an air phase. These results indicate that the rabbit lung is sensitive to paraquat toxicity in vitro, and confirm that oxygen and paraquat supplement each other's toxicity in the lung.


Management strategies and medication use for treating paediatric patients with concussions.
To assess management strategies for paediatric patients suffering from concussions. A 17-item questionnaire was distributed to 1305 section members of the American Academy of Pediatrics Sections on Adolescent Health, Sports Medicine and Fitness, Community Pediatrics and School Health. The use of medications, neuropsychological testing, neuroimaging and published guidelines in concussion management was queried. Two hundred and twenty respondents (17%) completed the questionnaire, of which 64% had been an attending for greater than 10 years. A majority of respondents (92%) managed patients with concussions, with 26% treating more than 24 patients per year. Most paediatricians (84%) reported using a published guideline. The majority of respondents (89%) manage the symptoms of concussed patients with medications, most commonly acetaminophen (62%) or nonsteroidal anti-inflammatory medications (54%). The use of prescriptions medications such as tricyclic antidepressants (23%), amantadine (10%) and methylphenidate (8%) was also commonly reported. Paediatricians treating >16 patients per year with concussion were more likely to prescribe tricyclic antidepressants, stimulants and agents used for sleep disturbance. Paediatricians nationwide routinely use medications when managing patients with concussions. The pharmacological agents used differ according to number of patients treated per year. In addition, most paediatricians use published guidelines in concussion management.


Outpatient surgery centers draw cases away from hospitals, impact resident training volume.
Across the United States, ambulatory surgery centers (ASCs) are increasing in both number and surgical volume. This trend has been the focus of debate regarding reimbursement and patient safety, as well as surgical productivity and efficiency. However, the impact on surgical resident training caused by this shift toward outpatient surgery in nonhospital settings has not been studied. We reviewed data reported by our hospital and by local surgery centers as well as the case logs of the surgical residents at our institution to determine whether a negative effect on resident case volume has occurred. We conducted a retrospective review of our PGY-1 through PGY-3 level surgical residents' case logs for 3 consecutive academic years, from July 2004 through June 2007. We evaluated a group of common outpatient procedures that are now also being performed in stand-alone surgical centers in our area, such as breast biopsies, incision and drainage, hernia repair, colonoscopy, and esophagogastroduodenoscopy (EGD). The data were tallied by academic year and compared over time. In addition, we analyzed data reported to state agencies by our hospital and local surgery centers over the last 6 calendar years for any trends in case volume. By evaluating 2 different independent data sets for the same endpoint, we could evaluate our hypothesis twice. When evaluating state-reported data for the defined cases, a significant decrease was observed in the total number of cases performed at Easton Hospital, Easton, Pennsylvania, each year between 2003 and 2006 (p < 0.0001). When reported cases by procedure category for 2003 versus 2005 only (because of incomplete data from ASCs in 2004 and 2006), a significant decrease was observed as well for certain specific procedures as follows: colonoscopy (p < 0.0001), inguinal/femoral hernia (p = 0.04), excision of skin lesion (p = 0.0022), and incision/drainage (p < 0.0001). When comparing resident reported data, significant decreases were observed in the number of hemorrhoidectomies, breast biopsies, skin grafts, carpal tunnel releases, and excision of skin lesions performed by residents during each academic year from July 2004 to June 2007. Our residents historically have gained all of their outpatient surgery experience from procedures performed at our home institution. With the recent surge of stand-alone surgical centers, many outpatient procedures are being performed outside of the hospital in centers where our residents do not rotate. Although current residents in our program are performing enough cases to fulfill the ACGME required minimums, the number of cases is significantly decreased because of cases performed by stand-alone surgical centers.


Dipole soliton-vortices.
On universal symmetry grounds, we analyze the existence of a new type of discrete-symmetry vortex solitons that can be considered as coherent states of dipole solitons carrying a nonzero topological charge. Remarkably, they can be also interpreted as excited angular Bloch states. The stability of new soliton states is elucidated numerically.


A 'Split-Gene' Transketolase From the Hyper-Thermophilic Bacterium Carboxydothermus hydrogenoformans: Structure and Biochemical Characterization.
A novel transketolase has been reconstituted from two separate polypeptide chains encoded by a 'split-gene' identified in the genome of the hyperthermophilic bacterium, Carboxydothermus hydrogenoformans. The reconstituted active α2β2 tetrameric enzyme has been biochemically characterized and its activity has been determined using a range of aldehydes including glycolaldehyde, phenylacetaldehyde and cyclohexanecarboxaldehyde as the ketol acceptor and hydroxypyruvate as the donor. This reaction proceeds to near 100% completion due to the release of the product carbon dioxide and can be used for the synthesis of a range of sugars of interest to the pharmaceutical industry. This novel reconstituted transketolase is thermally stable with no loss of activity after incubation for 1 h at 70°C and is stable after 1 h incubation with 50% of the organic solvents methanol, ethanol, isopropanol, DMSO, acetonitrile and acetone. The X-ray structure of the holo reconstituted α2β2 tetrameric transketolase has been determined to 1.4 Å resolution. In addition, the structure of an inactive tetrameric β4 protein has been determined to 1.9 Å resolution. The structure of the active reconstituted α2β2 enzyme has been compared to the structures of related enzymes; the E1 component of the pyruvate dehydrogenase complex and D-xylulose-5-phosphate synthase, in an attempt to rationalize differences in structure and substrate specificity between these enzymes. This is the first example of a reconstituted 'split-gene' transketolase to be biochemically and structurally characterized allowing its potential for industrial biocatalysis to be evaluated.


Uniform large-area x-ray imaging at 9 keV using a backlit pinhole.
The development and application of point backlighting at high x-ray energies is an essential step in diagnosing radiation-driven experiments. The point-backlighting technique provides uniform backlighter irradiance over a large field of view. This technique circumvents the large laser energy required for area backlighters at energies of 9 keV and above. We present the results of a Zn 9 keV point-backlighter source using the technique of pinhole aperturing to define the source size and hence the resolution. Details of the design and application of this technique to an undriven gold-walled hohlraum are described.


A Highly Characterized Yeast Toolkit for Modular, Multipart Assembly.
Saccharomyces cerevisiae is an increasingly attractive host for synthetic biology because of its long history in industrial fermentations. However, until recently, most synthetic biology systems have focused on bacteria. While there is a wealth of resources and literature about the biology of yeast, it can be daunting to navigate and extract the tools needed for engineering applications. Here we present a versatile engineering platform for yeast, which contains both a rapid, modular assembly method and a basic set of characterized parts. This platform provides a framework in which to create new designs, as well as data on promoters, terminators, degradation tags, and copy number to inform those designs. Additionally, we describe genome-editing tools for making modifications directly to the yeast chromosomes, which we find preferable to plasmids due to reduced variability in expression. With this toolkit, we strive to simplify the process of engineering yeast by standardizing the physical manipulations and suggesting best practices that together will enable more straightforward translation of materials and data from one group to another. Additionally, by relieving researchers of the burden of technical details, they can focus on higher-level aspects of experimental design.


Pulmonary-vein isolation for atrial fibrillation in patients with heart failure.
Pulmonary-vein isolation is increasingly being used to treat atrial fibrillation in patients with heart failure. In this prospective, multicenter clinical trial, we randomly assigned patients with symptomatic, drug-resistant atrial fibrillation, an ejection fraction of 40% or less, and New York Heart Association class II or III heart failure to undergo either pulmonary-vein isolation or atrioventricular-node ablation with biventricular pacing. All patients completed the Minnesota Living with Heart Failure questionnaire (scores range from 0 to 105, with a higher score indicating a worse quality of life) and underwent echocardiography and a 6-minute walk test (the composite primary end point). Over a 6-month period, patients were monitored for both symptomatic and asymptomatic episodes of atrial fibrillation. In all, 41 patients underwent pulmonary-vein isolation, and 40 underwent atrioventricular-node ablation with biventricular pacing; none were lost to follow-up at 6 months. The composite primary end point favored the group that underwent pulmonary-vein isolation, with an improved questionnaire score at 6 months (60, vs. 82 in the group that underwent atrioventricular-node ablation with biventricular pacing; P<0.001), a longer 6-minute-walk distance (340 m vs. 297 m, P<0.001), and a higher ejection fraction (35% vs. 28%, P<0.001). In the group that underwent pulmonary-vein isolation, 88% of patients receiving antiarrhythmic drugs and 71% of those not receiving such drugs were free of atrial fibrillation at 6 months. In the group that underwent pulmonary-vein isolation, pulmonary-vein stenosis developed in two patients, pericardial effusion in one, and pulmonary edema in another; in the group that underwent atrioventricular-node ablation with biventricular pacing, lead dislodgment was found in one patient and pneumothorax in another. Pulmonary-vein isolation was superior to atrioventricular-node ablation with biventricular pacing in patients with heart failure who had drug-refractory atrial fibrillation. (ClinicalTrials.gov number, NCT00599976.)


Environmental determinants of polycystic ovary syndrome.
In this review, we summarize existing research on a variety of environmental factors potentially involved in the etiology, prevalence, and modulation of polycystic ovary syndrome (PCOS), and we suggest avenues for future research. The main environmental factors we consider include environmental toxins, diet and nutrition, socioeconomic status, and geography. There is some evidence that environmental toxins play a role in disrupting reproductive health, but there is limited research as to how these toxins may affect the development of PCOS. Although research has also shown that PCOS symptoms are reduced with certain dietary supplements and with weight loss among obese women, additional research is needed to compare various approaches to weight loss, as well as nutritional factors that may play a role in preventing or mitigating the development of PCOS. Limited studies indicate some association of low socioeconomic status with certain PCOS phenotypes, and future research should consider socioeconomic conditions during childhood or adolescence that may be more relevant to the developmental onset of PCOS. Finally, the limited scope of comparable international studies on PCOS needs to be addressed, because global patterns of PCOS are potentially valuable indicators of cultural, environmental, and genetic factors that may contribute to excess risk in certain regions of the world.


Inferior parietal rtms affects performance in an addition task.
Neuropsychological and neuroimaging studies strongly suggest that the inferior parietal cortex is important for calculation. However, the evidence from neuroimaging experiments for a left hemispheric dominance in calculation is not as clear as one would expect from the studies of patients. Often a concomitant activation of the homologous inferior parietal region of the right hemisphere is reported in the same tasks. The objective of this study was to replicate basic findings of acalculic patients and to investigate discrepancies between data from patients and results from neuroimaging studies in an addition task. Repetitive transcranial magnetic stimulation (rTMS) was applied over inferior parietal areas and the adjacent intraparietal sulcus (IPS) while subjects solved double-digit addition tasks. From studies of acalculic patients it was hypothesised that left hemispheric rTMS stimulation should result in longer reaction times (RTs) in the addition task. On addition trials without TMS subjects showed the classical problem size effect with longer RTs the larger the sum of the two operands. Magnetic stimulation over left inferior parietal areas disrupted performance significantly. The effect was specific to the left hemisphere stimulation. There was no increase in RTs for rTMS stimulation over the right hemisphere.


TEM-like optical mode of a coaxial nanowaveguide.
We study propagation of electromagnetic waves in a nanocoaxial waveguide for frequencies around and below the surface plasmon frequency. We show, that for frequencies sufficiently lower than the surface plasmon frequency, the waveguide supports a plasmon polariton mode that resembles, and indeed reduces to the conventional TEM mode of the conventional coaxial transmission line, known in the radiotechnology.


Abnormal growth factor and cytokine expression in Dupuytren's contracture.
To analyse patterns of gene expression for peptide regulatory factors in patients with Dupuytren's contracture. Tissue samples (palmer fascia) from 12 patients with Dupuytren's contracture and 12 controls were studied using the reverse transcription/polymerase chain reaction (RT/PCR) technique. Tissue from patients with Dupuytren's contracture expressed a higher percentage of peptide regulatory factors than that of controls: interleukin-1 alpha (83% v 16%; p < 0.01); interleukin-1 beta (66% v 8%; p < 0.01); transforming growth factor beta (75% v 25%; p < 0.02); and basic fibroblast growth factor (66% v 25%; p < 0.05). Platelet derived growth factors alpha and beta were also expressed more commonly (66% v 33% and 25% v 16%, respectively), but these differences were not significant. The increased prevalence of expression for the above mRNAs in Dupuytren's tissue is relevant as interleukin-1, basic fibroblast growth factor, and transforming growth factor beta stimulate the growth of fibroblasts and transforming growth factor beta also enhances production of collagen and other extracellular matrix proteins. Excessive local release of these peptide regulatory factors may have an important role in the pathogenesis of Dupuytren's contracture.


Deep TMS in a resistant major depressive disorder: a brief report.
Repetitive transcranial magnetic stimulation (rTMS) has proven effective. Recently, a greater intracranial penetration coil has been developed. We tested the efficacy of the coil in the treatment of resistant major depression. Our sample included seven patients suffering from major depression who were treated using Brainsway's H1-coil connected to a Magstim rapid 2 stimulator. Deep TMS treatment was given to each patient in five sessions per week over a period of 4 weeks. Patients were treated with 120% intensity of the motor threshold and a frequency of 20 HZ with a total of 1,680 pulses per session. Five patients completed 20 sessions: one attained remission (Hamilton Depression Rating Scale (HDRS)=9); three patients reached a reduction of more than 50% in their pre-treatment HDRS; and one patient achieved a partial response (i.e., the HDRS score dropped from 21 to 12). Average HDRS score dropped to 12.6 and average Hamilton Anxiety Rating Scale score dropped to 9.Two patients dropped out: one due to insomnia and the second due to a lack of response. Compared to the pooled response and remission rates when treating major depression with rTMS, deep TMS as used in this study is at least similarly effective. Still, a severe limitation of this study is its small sample size, which makes the comparison of the two methods in terms of their effectiveness or side effects impossible. Greater numbers of subjects should be studied to achieve this aim. An H1 deep TMS coil could be used as an alternative treatment for major depressive disorder.


Dorsotonals/homothorax, the Drosophila homologue of meis1, interacts with extradenticle in patterning of the embryonic PNS.
The homeotic genes of the bithorax complex are required, among other things, for establishing the patterns of sensory organs in the embryonic peripheral nervous system (PNS). However, the molecular mechanisms by which these genes affect pattern formation in the PNS are not understood and other genes that function in this pathway are not characterized. Here we report the phenotypic and molecular analysis of one such gene, homothorax (hth; also named dorsotonals). Mutations in the hth gene seem to alter the identity of the abdominal chordotonal neurons, which depend on Abd-A for their normal development. However, these mutations do not alter the expression of the abd-A gene, suggesting that hth may be involved in modulating abd-A activity. We have generated multiple mutations in the hth locus and cloned the hth gene. hth encodes a homeodomain-containing protein that is most similar to the murine proto-oncogene meis1. The hth gene is expressed throughout embryonic development in a spatially restricted pattern, which is modulated in abdominal segments by abd-A and Ubx. The spatial distribution of the HTH protein during embryonic development is very similar to the distribution of the Extradenticle (EXD) protein, a known modulator of homeotic gene activity. Here we show that the PNS phenotype of exd mutant embryos is virtually indistinguishable from that of hth mutant embryos and does not simply follow the homeotic transformations observed in the epidermis. We also show that the HTH protein is present in extremely low levels in embryos lacking exd activity as compared to wild-type embryos. In contrast, the EXD protein is present in fairly normal levels in hth mutant embryos, but fails to accumulate in nuclei and remains cytoplasmic. Ectopic expression of hth can drive ectopic nuclear localization of EXD. Based on our observations we propose that the genetic interactions between hth and exd serve as a novel mechanism for regulating homeotic protein activity in embryonic PNS development.


One-Pot Preparation of HCPT@IRMOF-3 Nanoparticles for pH-Responsive Anticancer Drug Delivery.
Metal-organic frameworks (MOFs) are considered to be promising materials for drug delivery. In this work, a Zinc-based MOF nanocomposite IRMOF-3 was introduced as a drug carrier for 10-hydroxycamptothecine (HCPT). Without an extra drug-loading process, a nanoscale drug delivery material HCPT@IRMOF-3 was prepared via one-pot synthesis. The composition and structure of the material were investigated, and the drug release character was measured. Compared with preparing IRMOF-3 first and loading the drug, the one-pot-prepared HCPT@IRMOF-3 exhibited a higher drug-loading capacity. The material presented pH-responsive release. The HCPT release rate at pH 5.0 was significantly higher than that at pH 7.4. The cytotoxicity experiments showed that IRMOF-3 was non-toxic, and HCPT@IRMOF-3 exhibited notable cytotoxicity to Hela and SH-SY5Y cells. One-pot synthesis is a simple and rapid method for the preparation of an MOF drug delivery system, and IRMOF-3 can be potentially used in pH-responsive drug delivery systems.


Molecular mechanisms of thrombin-induced endothelial cell permeability.
Confluent endothelium serves as a selective barrier between the vascular space of blood vessels and underlying tissues. Compromised barrier function of the endothelium in response to inflammation mediators, such as thrombin, is accompanied by reversible cell rounding and interendothelial gap formation. Endothelial barrier integrity substantially depends on the cytoskeleton, which ensures actin stress fiber formation and via actomyosin-driven contraction regulates cell shape and adhesion. Recent studies have shown the sequence of events that mediate signal transduction in endothelial cells. Binding of thrombin with its receptor initiates activation of heterotrimeric G-proteins, which, in turn, entails a decrease in cAMP level in the cell, increase in intracellular Ca2+ and diacylglycerol concentration, and activation of the small G-protein Rho. Phosphorylation of myosin light chains as a result of activation of myosin light chain kinase and inactivation of myosin phosphatases stimulates stress fiber formation and triggers actomyosin contraction. In addition, thrombin-induced rearrangement in the endothelial cytoskeleton is regulated by Ca2+/calmodulin-dependent protein kinase, protein kinase C, and tyrosine protein kinases. This review focuses on presently known biochemical mechanisms of cell response to thrombin and their role in endothelial barrier dysfunction.


Ceramide content is higher in type I compared to type II fibers in obesity and type 2 diabetes mellitus.
This study investigated fiber-type-specific muscle ceramide content in obese subjects and type 2 diabetes patients. Two substudies, one which compared type 2 diabetes patients to both lean- and obese BMI-matched subjects and the other study which compared lean body-matched post-obese, obese, and control subjects, were performed. A fasting blood sample was obtained and plasma insulin and glucose determined. A muscle biopsy was obtained from deltoideus and vastus lateralis, and fiber-type ceramide content was determined by fluorescence immunohistochemistry. Insulin sensitivity estimated by Quicki index was higher in lean compared to type 2 diabetes patients and obese controls. Also in control and post-obese subjects, a higher insulin sensitivity was observed compared to obese subjects. Ceramide content was consistently higher in type I than in type II muscle fibers and higher in deltoideus than vastus lateralis across all groups. No significant differences between groups were observed in ceramide content in either of the two substudies. In human skeletal muscle, ceramide content was higher in type I than in type II fibers in patients with type 2 diabetes and in obese subjects, but overall ceramide muscle fiber content was not different compared to controls.


LDL-cholesterol concentrations: a genome-wide association study.
LDL cholesterol has a causal role in the development of cardiovascular disease. Improved understanding of the biological mechanisms that underlie the metabolism and regulation of LDL cholesterol might help to identify novel therapeutic targets. We therefore did a genome-wide association study of LDL-cholesterol concentrations. We used genome-wide association data from up to 11,685 participants with measures of circulating LDL-cholesterol concentrations across five studies, including data for 293 461 autosomal single nucleotide polymorphisms (SNPs) with a minor allele frequency of 5% or more that passed our quality control criteria. We also used data from a second genome-wide array in up to 4337 participants from three of these five studies, with data for 290,140 SNPs. We did replication studies in two independent populations consisting of up to 4979 participants. Statistical approaches, including meta-analysis and linkage disequilibrium plots, were used to refine association signals; we analysed pooled data from all seven populations to determine the effect of each SNP on variations in circulating LDL-cholesterol concentrations. In our initial scan, we found two SNPs (rs599839 [p=1.7x10(-15)] and rs4970834 [p=3.0x10(-11)]) that showed genome-wide statistical association with LDL cholesterol at chromosomal locus 1p13.3. The second genome screen found a third statistically associated SNP at the same locus (rs646776 [p=4.3x10(-9)]). Meta-analysis of data from all studies showed an association of SNPs rs599839 (combined p=1.2x10(-33)) and rs646776 (p=4.8x10(-20)) with LDL-cholesterol concentrations. SNPs rs599839 and rs646776 both explained around 1% of the variation in circulating LDL-cholesterol concentrations and were associated with about 15% of an SD change in LDL cholesterol per allele, assuming an SD of 1 mmol/L. We found evidence for a novel locus for LDL cholesterol on chromosome 1p13.3. These results potentially provide insight into the biological mechanisms that underlie the regulation of LDL cholesterol and might help in the discovery of novel therapeutic targets for cardiovascular disease.


In vitro activity of seven azole compounds against some clinical isolates of non-dermatophytic filamentous fungi and some dermatophytes.
The in vitro activity of seven azole compounds viz clotrimozole, isoconazole, bifanazole, fluconazole oxyconazole, Bay n 7133 and Bay L 9139 was investigated against 47 clinical isolates of pathogenic non-dermatophytic filamentous fungi and dermatophytic fungi. The isolates included Hendersonula toruloidea-26, Scytalidium hyalinium-5, Scytalidium japonicum-1, Trichophyton rubrum-5, Trichophyton tonsurans-3, Trichophyton mentagrophytes var. mentagrophytes-4, Epidemophyton floccosum-2, Microporum gypseum-2 isolates. The drugs were significantly more active against the dermatophytes (MIC range 0.025-1.56 micrograms/ml) than non-dermatophytes (MIC range 0.39-6.25 micrograms/ml). Isoconazole showed more activity than the rest of the azole compound tested. Clotrimazole, fluconazole, oxyconazole, bifonazole were comparable in their inhibitory activity against both dermatophytes and non-dermatophytes. The azole derivatives, Bay n 7133 and Bay L 9139 showed higher MIC range i.e. gave a range of 0.39-1.56 micrograms/ml for dermatophytes and 1.56-6.25 micrograms/ml for non-dermatophytic filamentous fungi. The minimal fungicidal concentration (MFC) of all the drugs tested were mostly within 2-8 times their MIC values.


Role of transplantation in the management of hepatic malignancy.
The acceptance of liver transplantation in the management of hepatic malignancy declined after early poor outcomes. Despite recent developments, including stricter selection criteria and improved adjuvant therapies, the role of liver transplantation in the management of cancer remains controversial. This review explores the evidence for the current role of liver transplantation in the management of hepatic malignancy in the context of recent advances in surgical resection and non-surgical treatments. A literature search was conducted using the Cochrane Library and Ovid MEDLINE and EMBASE, using terms for hepatic malignancy and interventions that included liver transplantation, percutaneous interventions, chemotherapy and surgical resection. In patients with primary hepatocellular carcinoma, improved selection has led to outcomes equivalent to those from surgical resection and comparable to those in patients transplanted for non-malignant indications. Recent studies suggest that selection criteria may be refined further. Surgical resection or percutaneous therapies may reduce the risk of progression while waiting for a transplant. Recent improvements have occurred in neoadjuvant therapies for cholangiocarcinoma. Nevertheless, a number of questions regarding the role of liver transplantation for hepatic malignancy remain.


Identical-dual-bandpass sampled fiber Bragg grating and its application to ultranarrow filters.
We have theoretically proposed and experimentally demonstrated a new kind of ultranarrow identical-dual-bandpass sampled fiber Bragg gratings (SFBGs) with a pi phase shift technique. The spacing of two bandpasses of the proposed grating can be flexibly adjusted by changing the sampled period, and any desired spacing can be achieved in principle. An experimental example shows that the transmission peaks of two narrow transmission-band are near 1549.1 and 1550.1 nm. Based on the proposed SFBG, an ultranarrow identical-dual-channel filter is designed. Two channels of the proposed filter have an equal bandwidth, an even strength, and the same group delay. The bandwidth of each channel of our filter is as small as 1 pm and up to 10(-3) pm (corresponding to approximately 0.1 MHz), which is less than the bandwidth of the conventional SFBG filters by a factor of 10(2)-10(4). The proposed grating and filter can find potential applications with slow light and dual-wavelength single-longitudinal-mode fiber lasers.


A putative consensus sequence for the nucleotide-binding site of annexin A6.
Reaction-induced infrared difference spectroscopy (RIDS) has been used to investigate the nature of interactions of human annexin A6 (ANXA6) with nucleotides. RIDS results for ANXA6, obtained after the photorelease of GTP-gamma-S, ATP, or P(i) from the respective caged compounds, were identical, suggesting that the interactions between the nucleotide and ANXA6 were dominated by the phosphate groups. Phosphate-induced structural changes in ANXA6 were small and affected only seven or eight amino acid residues. The GTP fluorescent analogue, 2'(3')-O-(2,4,6-trinitrophenyl)guanosine 5'-triphosphate (TNP-GTP), quenched tryptophan fluorescence of ANXA6 when bound to the protein. A binding stoichiometry of 1 mol of nucleotide/mol ANXA6 was established with a K(D) value of 2.8 microM for TNP-GTP. The bands observed on RIDS of ANXA6 halves (e.g., N-terminal half, ANXA6a, and C-terminal half, ANXA6b) were similar to those of the whole molecule. However, their amplitudes were smaller by a factor of 2 compared to those of whole ANXA6. TNP-GTP bound to both fragments of ANXA6 with a stoichiometry of 0.5 mol/mol. However, the binding affinities of ANXA6a and ANXA6b differed from that of ANXA6. Simulated molecular modeling revealed a nucleotide-binding site which was distributed in two distinct domains. Residues K296, Y297, K598, and K644 of ANXA6 were less than 3 A from the bound phosphate groups of either GTP or ATP. The presence of two identical sequences in ANXA6 with the F-X-X-K-Y-D/E-K-S-L motif, located in the middle of ANXA6, at residues 293-301 (within ANXA6a) and at 641-649 (within ANXA6b), suggested that the F-X-X-K-Y-D/E-K-S-L motif was the putative sequence in ANXA6 for nucleotide binding.


Multifunctional ion transport properties of human SLC4A11: comparison of the SLC4A11-B and SLC4A11-C variants.
Congenital hereditary endothelial dystrophy (CHED), Harboyan syndrome (CHED with progressive sensorineural deafness), and potentially a subset of individuals with late-onset Fuchs' endothelial corneal dystrophy are caused by mutations in the SLC4A11 gene that results in corneal endothelial cell abnormalities. Originally classified as a borate transporter, the function of SLC4A11 as a transport protein remains poorly understood. Elucidating the transport function(s) of SLC4A11 is needed to better understand how its loss results in the aforementioned posterior corneal dystrophic disease processes. Quantitative PCR experiments demonstrated that, of the three known human NH2-terminal variants, SLC4A11-C is the major transcript expressed in human corneal endothelium. We studied the expression pattern of the three variants in mammalian HEK-293 cells and demonstrated that the SLC4A11-B and SLC4A11-C variants are plasma membrane proteins, whereas SLC4A11-A is localized intracellularly. SLC4A11-B and SLC4A11-C were shown to be multifunctional ion transporters capable of transporting H+ equivalents in both a Na+-independent and Na+-coupled mode. In both transport modes, SLC4A11-C H+ flux was significantly greater than SLC4A11-B. In the presence of ammonia, SLC4A11-B and SLC4A11-C generated inward currents that were comparable in magnitude. Chimera SLC4A11-C-NH2-terminus-SLC4A11-B experiments demonstrated that the SLC4A11-C NH2-terminus functions as an autoactivating domain, enhancing Na+-independent and Na+-coupled H+ flux without significantly affecting the electrogenic NH3-H(n)+ cotransport mode. All three modes of transport were significantly impaired in the presence of the CHED causing p.R109H (SLC4A11-C numbering) mutation. These complex ion transport properties need to be addressed in the context of corneal endothelial disease processes caused by mutations in SLC4A11.


Early embryo loss is associated with local production of nitric oxide by decidual mononuclear cells.
In early embryo loss, the fetus may be considered to be an allograft and, therefore, may be rejected by maternal immunocytes. However, the cytotoxic mechanisms involved are still poorly understood. We have previously shown the involvement of natural killer (NK) cells and mononuclear cells expressing Mac-1 (CD11b) and F4/80 in resorbing compared to nonresorbing embryos. In this study, the role of nitric oxide (NO) in the mechanism of early embryo loss was studied. Pregnant CBA/J females mated with DBA/2 males (20-30% early embryo loss) and CD1 females mated with CD1 males (5-10% early embryo loss) were studied on days 8, 10, and 12 of gestation. Cells from the implantation sites of individual embryos were tested for the production of nitrite and nitrate with or without in vitro challenge with lipopolysaccharide (LPS) to determine whether decidual macrophages were primed in situ. On day 12 of gestation, when resorption was clearly visible, resorbing embryos showed more than a fivefold increase in both basal- and LPS-induced nitrite and nitrate production compared to nonresorbing embryos in both mouse strains tested, indicating that the decidual mononuclear cells were primed. Furthermore, more than 20% of CBA/J embryos showed a significant nitrate release on days 8 and 10 of gestation before any signs of embryo cytopathology. This percentage corresponded to the spontaneous resorption rate seen in CBA/J female X DBA/2 male matings. Similarly, 4% of the embryos from pregnant CD1 mice on days 8 and 12 of gestation produced a significant amount of nitrate, which again correlated with the low incidence of resorption observed in these mice. Using immunohistochemistry, the presence of inducible nitric oxide synthase (iNOS) was detected at implantation sites. Furthermore, decidual cells positive for both iNOS and the macrophage marker Mac-1 were demonstrated in implantation sites by double immunostaining. This strongly suggests that decidual macrophages could be the cellular source of NO production. Aminoguanidine, a selective inhibitor of the iNOS, inhibited the in vitro production of nitric oxide by cells isolated from individual implantation sites, and more strikingly, significantly reduced early embryo losses in CBA/J females mated by DBA/2 males when given orally or parenterally to the gravid females starting on day 6 of gestation. In addition, aminoguanidine-treated pregnant mice showed a significant increase in average litter size when the pregnancies were allowed to proceed to term.(ABSTRACT TRUNCATED AT 400 WORDS)


Values and preferences in oral anticoagulation in patients with atrial fibrillation, physicians' and patients' perspectives: protocol for a two-phase study.
Oral anticoagulation prevents strokes in patients with atrial fibrillation but, for reasons that remain unclear, less than 40% of all patients with atrial fibrillation receive warfarin. The literature postulates that patient and clinician preferences may explain this low utilization. The proposed research seeks to answer the following questions: i) When assessed systematically, do patients' and clinicians' preferences explain the utilization of warfarin to prevent strokes associated with atrial fibrillation? ii) To what extent do patients' and clinicians' treatment preferences differ? iii) What factors explain any differences that exist in treatment preferences between patients and clinicians? To answer these questions we will conduct a two-phase study of patient and clinician preferences for health states and treatments. In the first phase of this study we will conduct structured interviews to determine their treatment preferences for warfarin vs. aspirin to prevent strokes associated with atrial fibrillation using the probability trade-off technique. In the same interview, we will conduct preference-elicitation exercises using the feeling thermometer to identify the utilities that patients place on taking medication (warfarin and aspirin), and on having a mild stroke, a severe stroke, and a major bleed. In the second phase of the study we will convene focus groups of clinicians and patients to explore their answers to the exercises in the first phase. This is a study of patient and clinician preferences for health states and treatments. Because of its clinical importance and our previous work in this area, we will conduct our study in the clinical context of the decision to use antithrombotic agents to reduce the risk of stroke in patients with non-valvular chronic atrial fibrillation.


Three-dimensional angioarchitecture of spinal dural arteriovenous fistulas, with special reference to the intradural retrograde venous drainage system.
There have been significant advances in understanding the angioarchitecture of spinal dural arteriovenous fistulas (AVFs). However, the major intradural retrograde venous drainage system has not been investigated in detail, including the most proximal sites of intradural radiculomedullary veins as they connect to the dura mater, which are the final targets of interruption in both microsurgical and endovascular treatments. Between April 1984 and March 2011, 27 patients with 28 AVFs were treated for spinal dural AVFs at the authors' university hospital. The authors assessed vertebral levels of feeding arteries and dural AVFs by using conventional digital subtraction angiography. They also assessed 3D locations of the most proximal sites of intradural radiculomedullary veins and the 3D positional relationship between the major intradural retrograde venous drainage system and intradural neural structures, including the spinal cord, spinal nerves, and the artery of Adamkiewicz, by using operative video recordings plus 3D rotational angiography and/or 3D computer graphics. In addition, they statistically assessed the clinical results of 27 cases. Of these lesions, 23 were treated with open microsurgery and the rest were treated with endovascular methods. Feeding arteries consisted of T2-10 intercostal arteries with 19 lesions, T-12 subcostal arteries with 3 lesions, and L1-3 lumbar arteries with 6 lesions. The 3D locations of the targets of interruption (the most proximal sites of intradural radiculomedullary veins as they connect to the dura mater) were identified at the dorsolateral portion of the dura mater adjacent to dorsal roots in all 19 thoracic lesions, whereas they were identified at the ventrolateral portion of the dura mater adjacent to ventral roots in 7 (78%) of 9 cases of conus medullaris/lumbar lesions (p < 0.001). The major intradural retrograde venous drainage system was located dorsal to the spinal cord in all 19 thoracic lesions, whereas it was located ventral to the spinal cord in 4 (44%) of 9 cases of conus/lumbar lesions (p = 0.006). In 3 (11%) of 27 cases, AVFs had a common origin of the artery of Adamkiewicz. In 2 lumbar lesions, the artery of Adamkiewicz ascended very close to the vein because of its ventral location. Although all lesions were successfully obliterated without major complications and both gait and micturition status significantly improved (p = 0.005 and p = 0.015, respectively), conus/lumbar lesions needed careful differential diagnosis from ventral intradural perimedullary AVFs, because the ventral location of these lesions contradicted the Spetzler classification system. The angioarchitecture of spinal dural AVFs in the thoracic region is strikingly different from that in conus/lumbar regions with regard to the intradural retrograde venous drainage system. One should keep in mind that spinal dural AVFs are not always dorsal types, especially in conus/lumbar regions.


Self-healing of tightly focused scalar and vector Bessel-Gauss beams at the focal plane.
The property of self-healing at the focal plane for both scalar and vector Bessel-Gauss (BG) beams is investigated in the tight focusing condition. For the BG beam, which is partially obstructed at the pupil plane, the spatial intensity distribution at the focal plane is well recovered. Furthermore, recovery of not only intensity but also polarization distribution is observed for an obstructed vector BG beam. This self-healing effect for both the intensity and polarization components is recognized even when the half of the beam is obstructed by a semicircular obstacle. The effect of the size of the obstacle on recovery of polarization and intensity distribution is studied. The role of the beam size at the pupil plane is also discussed.


Hematopoietic stem cell development and regulatory signaling in zebrafish.
Hematopoietic stem cells (HSCs) are a population of multipotent cells that can self-renew and differentiate into all blood lineages. HSC development must be tightly controlled from cell fate determination to self-maintenance during adulthood. This involves a panel of important developmental signaling pathways and other factors which act synergistically within the HSC population and/or in the HSC niche. Genetically conserved processes of HSC development plus many other developmental advantages make the zebrafish an ideal model organism to elucidate the regulatory mechanisms underlying HSC programming. This review summarizes recent progress on zebrafish HSCs with particular focus on how developmental signaling controls hemogenic endothelium-derived HSC development. We also describe the interaction of different signaling pathways during these processes. The hematopoietic stem cell system is a paradigm for stem cell studies. Use of the zebrafish model to study signaling regulation of HSCs in vivo has resulted in a great deal of information concerning HSC biology in vertebrates. These new findings facilitate a better understanding of molecular mechanisms of HSC programming, and will provide possible new strategies for the treatment of HSC-related hematological diseases, such as leukemia. This article is part of a Special Issue entitled Biochemistry of Stem Cells.


The Italian version of rheumatoid arthritis pain scale (IT-RAPS): psychometric properties on community and clinical samples.
This paper describes the validation process of the Italian version of the Rheumatoid Arthritis Pain Scale (ITRAPS), describing its translation and adaptation to Italian culture. The cultural adaptation and validation were based on data from a sample of people affected by rheumatoid arthritis (RA). The process required a forward and backward translation of the original language, reviewed by an expert panel. The adapted version of the RAPS was then tested on a community and clinical sample, in order to test its psychometric properties. The IT-RAPS was submitted to 122 people affected by RA. The data was analyzed using Cronbach's coefficient alpha and the Pearson product-moment correlation coefficients. The IT-RAPS showed an internal consistency reliability coefficient of 0.96. This is the first study reporting the validation and cross-cultural adaptation of the RAPS in Italian. The study's findings provided support for the IT-RAPS as a reliable and valid measurement of multidimensional pain in RA patients.


A randomized clinical trial indicates that levamisole increases the time to relapse in children with steroid-sensitive idiopathic nephrotic syndrome.
Levamisole has been considered the least toxic and least expensive steroid-sparing drug for preventing relapses of steroid-sensitive idiopathic nephrotic syndrome (SSINS). However, evidence for this is limited as previous randomized clinical trials were found to have methodological limitations. Therefore, we conducted an international multicenter, placebo-controlled, double-blind, randomized clinical trial to reassess its usefulness in prevention of relapses in children with SSINS. The efficacy and safety of one year of levamisole treatment in children with SSINS and frequent relapses were evaluated. The primary analysis cohort consisted of 99 patients from 6 countries. Between 100 days and 12 months after the start of study medication, the time to relapse (primary endpoint) was significantly increased in the levamisole compared to the placebo group (hazard ratio 0.22 [95% confidence interval 0.11-0.43]). Significantly, after 12 months of treatment, six percent of placebo patients versus 26 percent of levamisole patients were still in remission. During this period, the most frequent serious adverse event (four of 50 patients) possibly related to levamisole was asymptomatic moderate neutropenia, which was reversible spontaneously or after treatment discontinuation. Thus, in children with SSINS and frequent relapses, levamisole prolonged the time to relapse and also prevented recurrence during one year of treatment compared to prednisone alone. However, regular blood controls are necessary for safety issues.


Complex formation between protein C inhibitor and prostate-specific antigen in vitro and in human semen.
Protein C inhibitor (PCI), a serine-proteinase inhibitor first purified from human blood plasma, occurs at high concentrations (3-4 microM) in seminal fluid in both a high-molecular-mass and low-molecular-mass form. Immunochemical data have previously suggested that PCI in seminal plasma forms complexes with the most abundant serine proteinase in semen, prostate-specific antigen (PSA). To provide a structural characterization of the PCI target, immunodetected as PSA, a procedure was developed to isolate low-molecular-mass and high-molecular-mass-forms of PCI from seminal fluid. The high-molecular-mass form of PCI, recognized by monoclonal antibodies against PSA, was dissociated by alkaline treatment into the low-molecular-mass form of PCI and a 33-kDa protein identified as PSA by 25 conclusive steps of N-terminal sequence analysis. We developed a sensitive immunofluorometric assay (IFMA) to measure PCI-PSA complexes in body fluids and investigated the rate at which purified PSA may form complexes with purified PCI. Formation of complexes detected by this IFMA and the appearance of SDS-stable approximately 90-kDa complexes paralleled loss of PSA activity recorded with chromogenic substrates. The rate of complex formation was slow compared to that reported for PCI and activated protein C, but was enhanced up to sixfold in the presence of heparin. Less than 10% of the initial PSA activity remained after 3 h incubation with a sevenfold molar excess of PCI and in the presence of heparin. In freshly collected ejaculates, the rate of PCI-PSA complex formation measured by IFMA was similar to that observed between the purified proteins, and paralleled the appearance of SDS-stable complexes by immunoblotting. During gel dissolution in freshly collected ejaculates, approximately 40% of immunodetected PCI becomes complexed to PSA. Although PCI is a slow inhibitor of PSA, complexes between PCI and PSA are detected at levels that correspond to an inactivation of up to 5% of the PSA activity in the ejaculate.


Systematic Review on the Therapeutic Options for COVID-19: Clinical Evidence of Drug Efficacy and Implications.
A novel coronavirus-2 (SARS-CoV-2) was first identified in Wuhan, China, and quickly spread globally. Several treatments have been proposed, many of which have proven ineffective. Consequently, there is a need to review the published evidence of drug clinical trials to guide future prescribing. A systematic review of published clinical trials and retrospective observational studies was carried out. The search was made using PubMed, Embase, MEDLINE, and China National Knowledge Infrastructure (CNKI) databases. Articles published between January 2020 and October 2020 and written in the English language were retrieved and included in the study. Researches that used traditional medicine, in-vitro and in-vivo animal studies, as well as reviews were excluded. Seventy-three relevant articles that fulfilled the inclusion criteria were finally selected and reviewed. Hydroxychloroquine, chloroquine, and azithromycin produced no clinical evidence of efficacy in randomized controlled clinical trials (RCT). However, retrospective observational studies reported the efficacy of remdesivir and lopinavir/ritonavir in reducing viral load, although there have been concerns with lopinavir/ritonavir and, more recently, remdesivir. Recently, tocilizumab, dexamethasone, and methylprednisolone significantly relieved lung inflammation and decreased mortality in patients with severe COVID-19. In addition, convalescent plasma was effective in boosting strong immunity among patients with mild COVID-19. There is currently no single worldwide approved therapeutic option for patients with COVID-19 despite the initial hype with medicines, including hydroxychloroquine. Nonetheless, dexamethasone has shown promise in symptomatic treatment and convalescent plasma in boosting immunity. New treatments are currently being researched, and the findings will be reported accordingly to provide evidence-based guidance for prescribers and policymakers.


Hepatoprotective efficacy of Spirulina platensis against lead-induced oxidative stress and genotoxicity in catfish; Clarias gariepinus.
Lead (Pb) is a toxic environmental pollutant that induces a broad range of biochemical and physiological hazards in living organisms. We investigated the possible hepatoprotective effects of Spirulina platensis (SP) in counteracting the Pb-induced oxidative damage. Ninety-six adult African catfish were allocated into four equal groups. The 1st group (control) fed basal diet while the 2nd group (Pb-treated) fed on basal diet and exposed to 1mg Pb(NO3)2/L. The 3rd and 4th groups fed SP-supplemented basal diets at levels of 0.25% and 0.5%, respectively and exposed to Pb. Serum samples were used to analyze hepatic function biomarkers, electrolytes, and oxidant and antioxidant status. Lipid peroxidation and DNA fragmentation were determined in the liver tissues. Pb exposure induced hepatic dysfunction, electrolytes (Na+, K+, Ca+2, and Cl-) imbalance, as well a significant decrease in GSH content, and LDH, AChE, SOD, CAT and GST enzymes activity. SP supplementation reverted these biochemical and genetic alterations close to control levels. This amelioration was higher with 0.5% SP and at the 4th week of exposure, showing concentration- and time-dependency. Thus, the current study suggests that SP could protect the catfish liver against lead-induced injury by scavenging ROS, sustaining the antioxidant status and diminishing DNA oxidative damage. The dietary inclusion of SP can be used as a promising protective agent to counteract oxidative stress-mediated diseases and toxicities.


Intimate partner violence and mental health effects: a population-based study among married women in Karachi, Pakistan.
Intimate partner violence (IPV) is recognized all over the world for its association with mental health problems in women. In Pakistan, such violence occurs commonly, but detailed information on mental health effects is scarce. The purpose of this study is to focused on married couples in urban Karachi to investigate mental health effects associated with physical, sexual and psychological violence perpetrated by husbands towards wives. Disclosure rates and health care-seeking behaviour were also investigated. This cross-sectional study involved 759 women between the ages of 25 and 60 years, selected using a multi-stage random sampling technique. The women were interviewed by trained community midwives using a structured questionnaire. In the total population of women, mental symptoms were prevalent. Women subjected to any form of violence reported, however, considerably poorer mental health than unexposed women. A statistically significant difference for almost all of the studied health parameters persisted even after controlling for socio-demographic factors. The strongest associations were found for suicidal thoughts and physical violence (OR 4.41; 3.18-6.12), sexual abuse (OR 4.39; 3.17-6.07) and psychological abuse (OR 5.17; 3.28-8.15). The interviews revealed that only 27% of the women subjected to violence had disclosed this to anyone, in most cases to their parents. The findings in this study highlight that the violence women have to face contributes to the development of multiple forms of psychological stress and serious mental health problems. Women's restrictive life circumstances seriously hamper women's empowerment. Reliable health surveillance system and health care services are needed to serve abused women. Policy initiatives focused on IPV and gender inequality in Pakistan should be initiated.


Generating conformation-specific synthetic antibodies to trap proteins in selected functional states.
A set of phage display sorting strategies and validation methodologies are presented that are capable of producing high performance synthetic antibodies (sABs) with customized properties. Exquisite control of antigen and conditions during the phage display selection process can yield sABs that: (1) recognize conformational states, (2) target specific regions of the surface of a protein, (3) induce conformational changes, and (4) capture and stabilize multi-protein complexes. These unique capabilities open myriad opportunities to study complex macromolecular processes inaccessible to traditional affinity reagent technology. We present detailed protocols for de novo isolation of binders, as well as examples of downstream biophysical characterization. The methods described are generalizable and can be adapted to other in vitro direct evolution approaches based on yeast or mRNA display.


The aurora kinase A inhibitor MLN8237 enhances cisplatin-induced cell death in esophageal adenocarcinoma cells.
Esophageal adenocarcinomas are poorly responsive to chemotherapeutics. This study aimed to determine the levels of Aurora kinase A (AURKA) and the therapeutic potential of MLN8237, an investigational AURKA inhibitor, alone and in combination with cisplatin. Using quantitative real-time PCR, we detected frequent AURKA gene amplification (15 of 34, 44%) and mRNA overexpression (37 of 44, 84%) in esophageal adenocarcinomas (P < 0.01). Immunohistochemical analysis showed overexpression of AURKA in more than two-thirds of esophageal adenocarcinoma tissue samples (92 of 132, 70%; P < 0.001). Using FLO-1, OE19, and OE33 esophageal adenocarinoma cell lines, with constitutive AURKA overexpression and mutant p53, we observed inhibition of colony formation with a single treatment of 0.5 μmol/L MLN8237 (P < 0.05). This effect was further enhanced in combination with 2.5 μmol/L cisplatin (P < 0.001). Twenty-four hours after treatment with the MLN8237 or MLN8237 and cisplatin, cell-cycle analyses showed a sharp increase in the percentage of polyploid cells (P < 0.001). This was followed by an increase in the percentage of cells in the sub-G(1) phase at 72 hours, concordant with the occurrence of cell death (P < 0.001). Western blot analysis showed higher induction of TAp73β, PUMA, NOXA, cleaved caspase-3, and cleaved PARP with the combined treatment, as compared with a single-agent treatment. Using xenograft models, we showed an enhanced antitumor role for the MLN8237 and cisplatin combination, as compared with single-agent treatments (P < 0.001). In conclusion, this study shows frequent overexpression of AURKA and suggests that MLN8237 could be an effective antitumor agent, which can be combined with cisplatin for a better therapeutic outcome in esophageal adenocarcinomas.