A silicon-on-insulator polarization diversity scheme in the mid-infrared.
We propose a silicon-on-insulator (SOI) polarization diversity scheme in the mid-infrared wavelength range. In consideration of absorption loss in silicon dioxide (SiO2), the polarization splitter-rotator (PSR) is designed and optimized with silicon nitride (SiN) upper-cladding and SiO2 lower-cladding. This asymmetry allows the PSR, which consists of mode-conversion tapers and subsequent mode-sorting asymmetric Y-junctions, to be fabricated with a simple one-step etching process. Simulation shows that our PSR has good performance with low mode conversion loss (< 0.25 dB) and low crosstalk (< -18 dB) in a very large wavelength range from 4.0 μm to 4.4 μm. The PSR also exhibits large fabrication tolerance with respect to the size deviations in waveguide width, height and refractive index of the upper-cladding. Additionally, PSR devices based on Y-junctions with SiO2 upper-cladding, and SiN upper- and lower-claddings are designed for potential applications at shorter and longer wavelengths, respectively. These PSR devices could facilitate the development of silicon photonic devices in the mid-infrared.


Design and synthesis of dual BRD4/Src inhibitors for treatment of triple-negative breast cancer.
Triple-negative breast cancer (TNBC) is an extremely aggressive tumor with limited treatment options and effectiveness. Dual-target inhibitors capable of simultaneously suppressing invasion may represent a promising therapeutic approach for TNBC. In this work, we developed a series of dual BRD4/Src inhibitors by connecting JQ1 and dasatinib using various linkers and evaluated their efficacy against TNBC both in vitro and in vivo. Among these compounds, HL403 demonstrated IC50 values of 133 nM for BRD4 inhibition and 4.5 nM for Src inhibition. Most importantly, HL403 not only exhibited potent anti-proliferative capabilities, but also effectively suppressed the invasion of MDA-MB-231 cells in vitro. Finally, the anti-tumor efficacy of HL403 was validated in a mouse MDA-MB-231 xenograft tumor model, achieving a tumor growth inhibition rate (TGI) of 70.7 %, which was superior to the combination of JQ1 and dasatinib (TGI = 54.0 %). Our research provides a promising and feasible new strategy for improving the treatment of TNBC.


Hydrogen-rich saline attenuates radiation-induced male germ cell loss in mice through reducing hydroxyl radicals.
Our recent studies suggest that H2 (hydrogen) has a potential as a novel radioprotector without known toxic side effects. The present study was designed to examine the underlying radioprotective mechanism of H2 and its protective role on irradiated germ cells. Produced by the Fenton reaction and radiolysis of H2O, hydroxyl radicals (•OH) were identified as the free radical species that were reduced by H2. We used a H2 microelectrode to dynamically detect H2 concentration in vivo, and found H2 significantly reduced in situ fluorescence intensity of hydroxyphenyl fluorescein; however, as we treated the mice with H2 after irradiation, the decrease is not significant. We found that pre-treatment of H2 to IR (ionizing radiation) significantly suppressed the reaction of •OH and the cellular macromolecules which caused lipid peroxidation, protein carbonyl and oxidatively damaged DNA. The radioprotective effect of H2 on male germ cells was supported by ameliorated apoptotic findings examined by morphological changes and TUNEL (terminal deoxynucleotidyltransferase-mediated dUTP nick-end labelling) in testicular tissue, and by preserved viability of stem spermatogonia examined for testicular histological parameters, daily sperm production and sperm quality; we used WR-2721 [S-2-(3-aminopropylamino)ethyl phosphorothioic acid] as a reference compound. Our results represent the first in vivo evidence in support of a radioprotective role of H2 by neutralizing •OH in irradiated tissue with no side effects.


Introductory pharmacology for clinical practice.
The average woman will take many medications, both prescription and nonprescription, as well as herbs or other dietary supplements over the course of her lifetime. These chemicals can produce therapeutic benefits but can also cause toxic effects. Pharmacokinetics, pharmacodynamics, pharmacotherapy, adverse drug reactions, and research in pharmacology are discussed as applied to women's health. Clinicians must have a firm understanding of the basic principles of pharmacology so that they can appropriately administer medications, monitor for anticipated effects and adverse reactions, and communicate as needed with women, their families, and other health care team members.


Spatio-temporal processes drive fine-scale genetic structure in an otherwise panmictic seabird population.
When and where animals breed can shape the genetic structure and diversity of animal populations. The importance of drivers of genetic diversity is amplified in island populations that tend to have more delineated gene pools compared to continental populations. Studies of relatedness as a function of the spatial distribution of individuals have demonstrated the importance of spatial organisation for individual fitness with outcomes that are conditional on the overall genetic diversity of the population. However, few studies have investigated the impact of breeding timing on genetic structure. We characterise the fine-scale genetic structure of a geographically-isolated population of seabirds. Microsatellite markers provide evidence for largely transient within-breeding season temporal processes and limited spatial processes, affecting genetic structure in an otherwise panmictic population of sooty terns Onychoprion fuscatus. Earliest breeders had significantly different genetic structure from the latest breeders. Limited evidence was found for localised spatial structure, with a small number of individuals being more related to their nearest neighbours than the rest of the population. Therefore, population genetic structure is shaped by heterogeneities in collective movement in time and to a lesser extent space, that result in low levels of spatio-temporal genetic structure and the maintenance of genetic diversity.


Long-term survival and causes of death in patients with ST-elevation acute coronary syndrome without obstructive coronary artery disease.
We aimed to study survival and causes of death in patients with ST-elevation acute coronary syndrome (STE-ACS) with and without obstructive coronary artery disease (CAD). We included 4793 consecutive patients with STE-ACS triaged for acute coronary angiography at a large cardiac invasive centre (2009-2014). Of these, 88% had obstructive CAD (stenosis ≥50%), 6% had non-obstructive CAD (stenosis 1-49%), and 5% had normal coronary arteries. Patients without obstructive CAD were younger and more often female with fewer cardiovascular risk factors. Median follow-up time was 2.6 years. Compared with patients with obstructive CAD, the short-term hazard of death (≤30 days) was lower in both patients with non-obstructive CAD [hazard ratio (HR) 0.49, 95% confidence interval (CI) 0.27-0.89, P = 0.018] and normal coronary arteries (HR 0.31, 95% CI 0.11-0.83, P = 0.021). In contrast, the long-term hazard of death (>30 days) was similar in patients with non-obstructive CAD (HR 1.15, 95% CI 0.77-1.72, P = 0.487) and higher in patients with normal coronary arteries (HR 2.44, 95% CI 1.58-3.76, P < 0.001), regardless of troponin levels. Causes of death were cardiovascular in 70% of patients with obstructive CAD, 38% with non-obstructive CAD, and 32% with normal coronary arteries. Finally, patients without obstructive CAD had lower survival compared with an age and sex matched general population. STE-ACS patients without obstructive CAD had a long-term risk of death similar to or higher than patients with obstructive CAD. Causes of death were less often cardiovascular. This suggests that STE-ACS patients without obstructive CAD warrant medical attention and close follow-up.


Medical doctors profile in Ethiopia: production, attrition and retention. In memory of 100-years Ethiopian modern medicine & the new Ethiopian millennium.
Although the practice of western medicine in Ethiopia dates back to the time of King Libne Dengel (1520-1535), organized and sustainable modern medical practice started after the battle of Adwa (1896). To review hospitals construction, medical doctors production and attrition, and to suggest alternative medical doctors retention mechanisms in the public sector and production scale up options. In this article, 100 years Ethiopian modern medical history is revised from old and recent medical chronicles. Until December 2006 primary data was collected from 87 public hospitals. Much emphasis is given to medical doctors profile (1906-2006), hospitals profile (1906-2005), medical doctors to population and hospitals ratio (1965-2006), Ethiopian public medical schools 42 years attainment (1964-2006), annual attrition rate (1984-2006), organizational structure of medical faculties & university hospitals, medical doctors remuneration by the Ministry of Health (MOH), Ministry of Education (MOE), NGOs and private health institutions. This article also addresses the way forward from physician training and retention perspectives, multiple alternate mechanisms to increase physicians' motivation to work in government institutions and reveres the loss. Medical doctors production scale up option is also given much emphasis. Most data are presented using line and bar graphs. Literature review showed that the first three hospitals were constructed in 1896 (Russian hospital), 1903 (Harar Ras Mekonnen hospital) and 1906 (Menelik II hospital). In 2005, 139 hospitals (87 public and 52 others) were reported. Remarkable hospital construction was done between 1935 and 1948, and recently between 1995 and 2005; however, in the latter case, private hospitals construction took the lions share. By the time MOH was established (1948), 110 Ethiopian and expatriate medical doctors were working, mainly in the capital, and 46 hospitals constructed. Physician number increment was very slow till 1980 at which time it started to get doubled every five years and reached peak (1658 medical doctors of all type) in 1989 in the public sector. As there was sharp increment in physician number, on the contrary, there was sharp decline in the last 15 years (1990-2006) to nadir 638 doctors in 2006 in the public sector. The last 25 years of Ethiopian modern medical history, in reference to physician number, forms a triangle with the lower and upper base 1980 and 2006, respectively. Since MOH of Ethiopia started registering health professionals with qualifications in 1987, 5743 (76.5% Ethiopian and 23.5% expatriate) medical doctors were registered for the first time. Out of these, 3717 were general practitioners. The three prestigious medical schools (Addis Ababa, Gondar, Jimma) were established in 1964, 1978 and 1984, respectively. Since establishment till 2006, about 3728 medical doctors were graduated with MD degree from the three medical schools. Addis Ababa university medical faculty alone graduated 1890 general practitioners (1964-2006) and 862 clinical specialists (1979-2006). In the 23 years period (1984-2006), the highest and lowest physician to population ratios in the public sector were found to be in 1989 (1:28,000) and 2006 (1:118,000), respectively. In 2006, the physician to population ratio in Amhara, Oromia and SNNPR regional states was computed to be 1:280,000, 1:220,000, and 1:230,000, respectively. The physician deficit analysis in the last 23 years in relation to the WHO standard for developing countries (1:10,000) revealed the lowest record at the national and regional states in the last 12-years. Average physician to hospital ratio in five regional states in December 2006 was 3.6 (Tigray), 4.3 (Amhara), 6.1 (Oromia) and 5.3 (SNNPR). As the December 2006 direct interview with 76 public hospitals outside Addis Ababa showed, there was no specialist in 36 hospitals and no doctor at all in 3 hospitals. Seven public hospitals located in big regional states' town took the lions share of medical doctors. In short, in December 2006, 80.3% of regional hospitals were equipped with 0-2 specialists of one kind, and in 48.7% there were 0-3 General practitioners. Highest medical doctors annual attrition rates (20%-54.3%) were found in 1991-1992, 1998, 2002-2006. In general, in 20 years period (1987-2006), 73.2% of Ethiopian medical doctors left the public sector mainly due to attractive remuneration in overseas countries and local NGOs/private sectors. The number of postgraduate programme in Addis Ababa, Jimma and Gondar medical schools in December 2006 was 22, 12 and 3, respectively. The total number of fully employed academic staff of the medical schools in declining order was Addis Ababa 181, Gondar 118, Jimma 71, Hawassa 63 and Mekele 52: those with second degree and above being 97.2%, 35.6%, 90.1%, 55.6% and 15.4%, respectively. Currently (2006), there are about 416 clinical residents in 3 medical schools. High annual attrition rate, fast population growth, governmental and nongovernmental health institution expansion, low production and increased postgraduate enrollment in the last 3-4 years contributed for extremely low physician-to-population ratio in Ethiopia. Although the Ethiopian government and private sector worked and achieved much on health infrastructure construction and midlevel health professionals training, it does not appear that medical doctors retention mechanisms are sorted out so far. As a result, even despite salary equivalent top up payments in some regions, more than 80% of public hospitals outside Addis Ababa were found ill-equipped with the most important human element--physicians. This implies that the push factors may not invariably correlate with remuneration. It is high time that the government discusses the possible solutions among health professional associations/societies and other health stakeholders, and apply concrete medical doctors retention mechanisms before the public medical schools and hospitals dry off doctors. Among actions to be undertaken from the current Ethiopian perspective: providing land plot for physicians for residential house construction, giving priority to physicians in providing low cost houses, low interest or interest free loan for residential house construction and automobile procurement, allowing duty free automobile procurement, improving the fully employed academic staff taxation system, approving the different remuneration options proposed, adopting the other countries experience of dual employment to academic staff working in teaching hospitals, modifying the academic rank promotion based on year of training, for university hospitals either establishing hospital organizational structure in the Ministry of Education or letting them be under MOH, establishing joint appointment (mutual beneficiary) agreement between medical schools and local hospitals, directing donors and stakeholders to work on the line of reducing internal and external medical doctors brain drain, making independent MOH and higher institutions from Civil Service Agency are proposed as short term solutions. Retention as a strategy & production as a programme, medical doctors production scale up options are proposed as a long term solution to achieve physician to population ratio of 1:15,000. and 1:8,000 by the year 2015 and 2020, respectively.


Medical treatment of eosinophilic esophagitis.
Eosinophilic esophagitis (EoE) is a chronic antigen-mediated eosinophilic inflammatory disease isolated to the esophagus. As a clinicopathologic disorder, a diagnosis of EoE requires a constellation of clinical symptoms of esophageal dysfunction and histologic findings (at least 15 eosinophils/high-powered microscope field (eos/hpf)). Current guidelines no longer require the failure of response to proton pump inhibitor medications to establish a diagnosis of EoE, but continue to suggest the exclusion of other etiologies of esophageal eosinophilia. The treatment goals for EoE are improvement in clinical symptoms, resolution of esophageal eosinophilia and other histologic abnormalities, endoscopic improvement, improved quality of life, improved esophageal function, minimized adverse effects of treatment, and prevention of disease progression and subsequent complications. Currently, there is no cure for EoE, making long-term treatment necessary. Standard treatment modalities include dietary modifications, esophageal dilation, and pharmacologic therapy. Effective pharmacologic therapies include corticosteroids, rapidly emerging biological therapies, and proton pump inhibitor medications. To evaluate the efficacy and safety of medical interventions for people with eosinophilic esophagitis. We searched CENTRAL, MEDLINE, Embase, ClinicalTrials.gov, and WHO ICTRP to 3 March 2023. Randomized controlled trials (RCTs) comparing any medical intervention or food elimination diet for the treatment of eosinophilic esophagitis, either alone or in combination, to any other intervention (including placebo). Pairs of review authors independently selected studies and conducted data extraction and risk of bias assessment. We expressed outcomes as a risk ratio (RR) and as the mean or standardized mean difference (MD/SMD) with 95% confidence interval (CI). We assessed the certainty of the evidence using GRADE. Our primary outcomes were: clinical, histological, and endoscopic improvement, and withdrawals due to adverse events. Secondary outcomes were: serious and total adverse events, and quality of life. We included 41 RCTs with 3253 participants. Eleven studies included pediatric patients while the rest recruited both children and adults. Four studies were in patients with inactive disease while the rest were in patients with active disease. We identified 19 intervention comparisons. In this abstract we present the results of the primary outcomes for the two main comparisons: corticosteroids versus placebo and biologics versus placebo, based on the prespecified outcomes defined of the primary studies. Fourteen studies compared corticosteroids to placebo for induction of remission and the risk of bias for these studies was mostly low. Corticosteroids may lead to slightly better clinical improvement (20% higher), measured dichotomously (risk ratio (RR) 1.74, 95% CI 1.08 to 2.80; 6 studies, 583 participants; number needed to treat for an additional beneficial outcome (NNTB) = 4; low certainty), and may lead to slightly better clinical improvement, measured continuously (standard mean difference (SMD) 0.51, 95% CI 0.17 to 0.85; 5 studies, 475 participants; low certainty). Corticosteroids lead to a large histological improvement (63% higher), measured dichotomously (RR 11.94, 95% CI 6.56 to 21.75; 12 studies, 978 participants; NNTB = 3; high certainty), and may lead to histological improvement, measured continuously (SMD 1.42, 95% CI 1.02 to 1.82; 5 studies, 449 participants; low certainty). Corticosteroids may lead to little to no endoscopic improvement, measured dichotomously (RR 2.60, 95% CI 0.82 to 8.19; 5 studies, 596 participants; low certainty), and may lead to endoscopic improvement, measured continuously (SMD 1.33, 95% CI 0.59 to 2.08; 5 studies, 596 participants; low certainty). Corticosteroids may lead to slightly fewer withdrawals due to adverse events (RR 0.64, 95% CI 0.43 to 0.96; 14 studies, 1032 participants; low certainty). Nine studies compared biologics to placebo for induction of remission. Biologics may result in little to no difference in clinical improvement, measured dichotomously (RR 1.14, 95% CI 0.85 to 1.52; 5 studies, 410 participants; low certainty), and may result in better clinical improvement, measured continuously (SMD 0.50, 95% CI 0.22 to 0.78; 7 studies, 387 participants; moderate certainty). Biologics result in better histological improvement (55% higher), measured dichotomously (RR 6.73, 95% CI 2.58 to 17.52; 8 studies, 925 participants; NNTB = 2; moderate certainty). We could not draw conclusions for this outcome when measured continuously (SMD 1.01, 95% CI 0.36 to 1.66; 6 studies, 370 participants; very low certainty). Biologics may result in little to no difference in endoscopic improvement, measured dichotomously (effect not estimable, low certainty). We cannot draw conclusions for this outcome when measured continuously (SMD 2.79, 95% CI 0.36 to 5.22; 1 study, 11 participants; very low certainty). There may be no difference in withdrawals due to adverse events (RR 1.55, 95% CI 0.88 to 2.74; 8 studies, 792 participants; low certainty). Corticosteroids (as compared to placebo) may lead to clinical symptom improvement when reported both as dichotomous and continuous outcomes, from the primary study definitions. Corticosteroids lead to a large increase in histological improvement (dichotomous outcome) and may increase histological improvement (continuous outcome) when compared to placebo. Corticosteroids may or may not increase endoscopic improvement (depending on whether the outcome is measured dichotomously or continuously). Withdrawals due to adverse events (dichotomous outcome) may occur less frequently when corticosteroids are compared to placebo. Biologics (as compared to placebo) may not lead to clinical symptom improvement when reported as a dichotomous outcome and may lead to an increase in clinical symptom improvement (as a continuous outcome), from the primary study definitions. Biologics lead to a large increase in histological improvement when reported as a dichotomous outcome, but this is uncertain when reported as a continuous outcome, as compared to placebo. Biologics may not increase endoscopic improvement (dichotomous outcome), but this is uncertain when measured as a continuous outcome. Withdrawals due to adverse events as a dichotomous outcome may occur as frequently when biologics are compared to placebo.


Hydrogen: Electrolytic Technique for Purifying It and Removing It from a Gas Stream.
Purified hiydrogen is obtained by impressing a potential gradient across a cell containintg two efficient electrodes separated by a barrier electrolyte phase and passing an impure hydrogen stream into the anode compartment. Power requirements are low and can be controlled through thle resistance of the electrolyte. The technique can also be used for pumping and accurate metering.


Perceptions of antenatal iron-folic acid supplements in urban and rural Pakistan: a qualitative study.
In Pakistan, 51% of women are anaemic in pregnancy yet only 44% of women use antenatal iron-folic acid (IFA) supplements. Little information exits on the perception and barriers to the use of IFA supplements during pregnancy in Pakistan. The aim of the study was to understand women and healthcare providers' perceptions, and to investigate the cultural and behavioural factors influencing the use of antenatal IFA supplements in rural and urban settings of Pakistan. We conducted 10 focus group discussions with mothers, 10 in-depth interviews with currently pregnant women, 6 in-depth interviews with Lady Health Workers and 4 in-depth interviews with doctors providing antenatal care services. The study was conducted in two districts of Pakistan--district Swabi and Islamabad for rural and urban samples, respectively. Data was collected between August and November 2012. The majority of women were aware of the perceived benefits of antenatal IFA supplements. However, the rural women had more limited information about the benefits of IFA supplements than the urban women. The facilitating factors for the women's use of supplements were: they had knowledge of benefits; they had trust in the healthcare providers; the supplements were available; they had the financial capacity to buy them; they felt better after taking these supplements; and they received support from family members. The barriers to the women's use of supplements were: they forgot to take them; the non-availability of supplements; their limited financial capacity to buy them; the lack of antenatal care services; family members not allowing use of the supplements; not knowing about the benefits or no education; fear or experience of side effects; considering them as contraceptives; and felt better thus stopped. The coverage of antenatal IFA supplementation can be improved by reducing the barriers related to the use of antenatal IFA supplementation in Pakistan. Interventions focused on providing adequate awareness, good quality counselling, reminder messages, availability of free supplements throughout pregnancy and reducing the side effects should be developed and implemented.


An evaluation of interactive projections as digital enrichment for orangutans.
Digital technologies are increasingly being incorporated into the provision of enrichment for captive primates, ranging from the ad-hoc use of iPads to specifically designed hardware installed in the design of new exhibits. In this article, we present a digital enrichment system that utilizes a novel approach with interactive projections, installed in the orangutan exhibit at Melbourne Zoo. While previous research into orangutan enrichment has involved reinforcement using food rewards, this study is the first investigation of digital enrichment for orangutans that does not involve food rewards. Rather, the interactive projections were designed to provide intrinsically rewarding environmental enrichment, instead of a cheaper or more variable way of delivering food. Our observations of orangutans' use of the system supports the approach of using interactive projections, finding that it is more responsive than capacitance or resistive based touch-screen technologies to the variety of ways that orangutans "touch": with whole hands, feet, faces, and with objects. We also present the results from a preliminary study into the effect of this enrichment into orangutan behavior which found presence or use of the device did not affect time spent engaged in the majority of behaviors of the individuals studied.


Surgical Anatomy of the Uterosacral Ligament Colpopexy.
The aim of this study was to describe the relationship of the uterosacral ligament (USL) to the ureter and rectum along a surgeon's target location for suture placement under conditions simulating live surgery. Dissections were performed in 11 unembalmed female cadavers. Steps were taken to identify the USL simulating USL colpopexy. Pins were placed in the midportion of the USL at the level of the IS, and at 1-cm, 2-cm, and 3-cm increments traveling proximally toward the sacrum (Fig. 1). We measured minimum distances from the USL to the ureter and rectum at each target location. In general, the ureters range from 1.3 to 2.0 cm lateral to the USLs along the target length. The rectum ranges from 1.9 to 2.6 cm from the right USL and remains 1.5 cm from the left USL. The mean change in distance between the ureter and USL for every 1 cm advanced toward the sacrum is 0.2 cm (95% confidence interval [CI], 0.19-0.24) on the right and 0.2 cm (95% CI, 0.18-0.27) on the left. The mean change in distance between the rectum and USL for every 1 cm advanced toward the sacrum is 0.2 cm (95% CI, 0.19-0.24) on the right and 0.0 cm (95% CI, 0-0) on the left. For every centimeter traveled along the bilateral USLs from the IS toward the sacrum, the ureter moves 0.2 cm laterally away from the ligament, the rectum moves 0.2 cm medially away from the right USL, but maintains its position from the left USL.


Toxicity of ZnO/TiO2 -conjugated carbon-based nanohybrids on the coastal marine alga Thalassiosira pseudonana.
Increasing consumption of metal-oxide nanoparticles (NPs) and carbon-based nanomaterials has caused significant concern about their potential hazards in aquatic environments. The release of NPs into aquatic environments could result in adsorption of NPs on microorganisms. While metal-oxide NP-conjugated carbon-based nanohybrids (NHs) may exhibit enhanced toxicity toward microorganisms due to their large surface area and the generation of reactive oxygen species (ROS), there is a lack of information regarding the ecotoxicological effects of NHs on marine diatom algae, which are an indicator of marine pollution. Moreover, there is scant information on toxicity mechanisms of NHs on aquatic organisms. In this study, four NHs (ie, ZnO-conjugated graphene oxide [GO], ZnO-conjugated carbon nanotubes [CNTs], TiO2 -conjugated GO, and TiO2 -conjugated CNT) that were synthesized by a hydrothermal method were investigated for their toxicity effects on a Thalassiosira pseudonana marine diatom. The in vitro cellular viability and ROS formation employed at the concentration ranges of 50 and 100 mg/L of NHs over 72 hours revealed that ZnO-GO had the most negative effect on T. pseudonana. The primary mechanism identified was the generation of ROS and GO-induced dispersion that caused electrostatic repulsion, preventing aggregation, and an increase in surface areas of NHs. In contrast to GO-induced dispersion, large aggregates were observed in ZnO/TiO2 -conjugated CNT-based NHs. The scanning electron microscopy images suggest that NHs covered algae cells and interacted with them (shading effects); this reduced light availability for photosynthesis. Detailed in vitro toxicity effects and mechanisms that cause high adverse acute toxicity on T. pseudonana were unveiled; this implied concerns about potential hazards of these mechanisms in aquatic ecosystems.


Translatome analysis of tuberous sclerosis complex 1 patient-derived neural progenitor cells reveals rapamycin-dependent and independent alterations.
Tuberous sclerosis complex (TSC) is an inherited neurocutaneous disorder caused by mutations in the TSC1 or TSC2 genes, with patients often exhibiting neurodevelopmental (ND) manifestations termed TSC-associated neuropsychiatric disorders (TAND) including autism spectrum disorder (ASD) and intellectual disability. Hamartin (TSC1) and tuberin (TSC2) proteins form a complex inhibiting mechanistic target of rapamycin complex 1 (mTORC1) signaling. Loss of TSC1 or TSC2 activates mTORC1 that, among several targets, controls protein synthesis by inhibiting translational repressor eIF4E-binding proteins. Using TSC1 patient-derived neural progenitor cells (NPCs), we recently reported early ND phenotypic changes, including increased cell proliferation and altered neurite outgrowth in TSC1-null NPCs, which were unaffected by the mTORC1 inhibitor rapamycin. Here, we used polysome profiling, which quantifies changes in mRNA abundance and translational efficiencies at a transcriptome-wide level, to compare CRISPR-edited TSC1-null with CRISPR-corrected TSC1-WT NPCs generated from one TSC donor (one clone/genotype). To assess the relevance of identified gene expression alterations, we performed polysome profiling in postmortem brains from ASD donors and age-matched controls. We further compared effects on translation of a subset of transcripts and rescue of early ND phenotypes in NPCs following inhibition of mTORC1 using the allosteric inhibitor rapamycin versus a third-generation bi-steric, mTORC1-selective inhibitor RMC-6272. Polysome profiling of NPCs revealed numerous TSC1-associated alterations in mRNA translation that were largely recapitulated in human ASD brains. Moreover, although rapamycin treatment partially reversed the TSC1-associated alterations in mRNA translation, most genes related to neural activity/synaptic regulation or ASD were rapamycin-insensitive. In contrast, treatment with RMC-6272 inhibited rapamycin-insensitive translation and reversed TSC1-associated early ND phenotypes including proliferation and neurite outgrowth that were unaffected by rapamycin. Our work reveals ample mRNA translation alterations in TSC1 patient-derived NPCs that recapitulate mRNA translation in ASD brain samples. Further, suppression of TSC1-associated but rapamycin-insensitive translation and ND phenotypes by RMC-6272 unveils potential implications for more efficient targeting of mTORC1 as a superior treatment strategy for TAND.


Masked Language Modeling for Resource Constrained Biological Natural Language Processing.
Recent advances in Natural Language Processing (NLP) have produced state of the art results on several sequence to sequence (seq2seq) tasks. Enhancements in embedders and their training methodologies have shown significant improvement on downstream tasks. Word vector models like Word2Vec, FastText & Glove were widely used over one-hot encoded vectors for years until the advent of deep contextualized embedders. Protein sequences consist of 20 naturally occurring amino acids that can be treated as the language of nature. These amino acids in combinations with each other makeup the biological functions. The choice of vector representation and architecture design for a biological task is highly dependent upon the nature of the task. We utilize unlabelled protein sequences to train a Convolution and Gated Recurrent Network (CGRN) embedder using Masked Language Modeling (MLM) technique that shows significant performance boost under resource constraint setting on two downstream tasks i.e., F1-score(Q8) of 73.1% on Secondary Structure Prediction (SSP) & F1-score of 84% on Intrinsically Disordered Region Prediction (IDRP). We also compare different architectures on downstream tasks to show the impact of the nature of biological task on the performance of the model.


The effect of varying footwear configurations on the peroneus longus muscle function following inversion.
The ankle is one of the most commonly injured joints with inversion injury affecting its lateral ligament complex being the commonest of all. Shoes are one of the known risk factors for such an injury. This study seeks to examine the impact of varying shoe configurations on the protective function of the peroneus longus muscle during unanticipated foot inversion. The peak amplitude, latency and post-peak average amplitude of the ipsilateral peroneus longus muscle were recorded by surface electromyography following unanticipated inversion of the feet of 35 subjects in a two-footplate tilting platform from 0° to 20°. The test conditions were barefoot, standard training shoe, shoe with a sole flare, and an above the ankle laced boot. Analysis revealed significant differences in peak muscle contraction between shod and unshod conditions. The standard shoe and the flared sole design showed greater statistically significant differences from the unshod condition, than the boot. The muscle was responding earlier in the shod conditions compared to the barefoot. The post-peak average amplitude with the standard shoe and the flared sole shoe were significantly different from the barefoot condition. Albeit no marked differences could be demonstrated between the tested shoes, the inherent construct of the laced boot probably attempts to protect the ankle-subtalar joint complex evidenced by evoking a less strong peroneus longus muscle's protective response.


Vascular causes of radiculopathy: a literature review.
Vascular etiologies are a significant source of radiculopathy. The entities that are reviewed here include epidural spinal hematoma, subdural spinal hematoma, spinal arteriovenous malformation, vertebral hemangioma, spinal epidural cavernous hemangioma, vertebral artery anomalies (both tortuosity and dissection), aortic aneurysm, hemorrhagic synovial cysts, ligamentum flavum hematoma, and venous varices. The incidence, pathophysiology, typical clinical presentation, appropriate diagnostic workup, and management are summarized. To review and summarize various vascular causes of radiculopathy and how it impacts the management of patients with radicular spinal pain. Literature review. An extensive literature review was done on each of the vascular entities and their presentation as radiculopathy. Vascular etiologies of radiculopathy may be underestimated in the literature. Imaging in addition to the history and physical examination is essential to identifying vascular sources of radiculopathy and establishing correct treatment. In examining patients with radicular pain, extraspinal causes of radiculopathy including vascular entities should be included in the differential. Knowledge of the incidence, pathophysiology, clinical presentation, and management of the possibly underreported various vascular causes of radiculopathy increase the ability of the physician to provide optimal care for the patient. A key aspect to identifying such vascular lesions is appropriate imaging for documenting the source of radiculopathy. Blind injections, done without appropriate imaging for identifying the cause of radiculopathy, and also performed without fluoroscopic guidance, increase not only the risk of misdiagnosis but also the comorbidities associated with spinal injections.


Clinical characteristics, outcomes, and risk factors for mortality in patients with cancer and COVID-19 in Hubei, China: a multicentre, retrospective, cohort study.
Patients with cancer are a high-risk population in the COVID-19 pandemic. We aimed to describe clinical characteristics and outcomes of patients with cancer and COVID-19, and examined risk factors for mortality in this population. We did a retrospective, multicentre, cohort study of 205 patients with laboratory-confirmed severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection and with a pathological diagnosis of a malignant tumour in nine hospitals within Hubei, China, from Jan 13 to March 18, 2020. All patients were either discharged from hospitals or had died by April 20, 2020. Clinical characteristics, laboratory data, and cancer histories were compared between survivors and non-survivors by use of χ2 test. Risk factors for mortality were identified by univariable and multivariable logistic regression models. Between Jan 13 and Mar 18, 2020, 205 patients with cancer and laboratory-confirmed SARS-CoV-2 infection were enrolled (median age 63 years [IQR 56-70; range 14-96]; 109 [53%] women). 183 (89%) had solid tumours and 22 (11%) had haematological malignancies. The median duration of follow-up was 68 days (IQR 59-78). The most common solid tumour types were breast (40 [20%] patients), colorectal (28 [14%]), and lung cancer (24 [12%]). 54 (30%) of 182 patients received antitumour therapies within 4 weeks before symptom onset. 30 (15%) of 205 patients were transferred to an intensive care unit and 40 (20%) died during hospital admission. Patients with haematological malignancies had poorer prognoses than did those with solid tumours: nine (41%) of 22 patients with haematological malignancies died versus 31 (17%) of 183 patients with solid tumours (hazard ratio for death 3·28 [95% CI 1·56-6·91]; log rank p=0·0009). Multivariable regression analysis showed that receiving chemotherapy within 4 weeks before symptom onset (odds ratio [OR] 3·51 [95% CI 1·16-10·59]; p=0·026) and male sex (OR 3·86 [95% CI 1·57-9·50]; p=0·0033) were risk factors for death during admission to hospital. Patients with cancer and COVID-19 who were admitted to hospital had a high case-fatality rate. Unfavourable prognostic factors, including receiving chemotherapy within 4 weeks before symptom onset and male sex, might help clinicians to identify patients at high risk of fatal outcomes. National Natural Science Foundation of China.


Performance of currently available risk models in a cohort of mechanically supported high-risk percutaneous coronary intervention--From the PROTECT II randomized trial.
Procedural risk scores facilitate clinical decision making by using individual patient characteristics to estimate the risk of adverse events. The performance of PCI-based risk scores is not well-described among patients undergoing hemodynamically supported high risk PCI. A total of 427 patients with unprotected left main disease, last remaining vessel or three-vessel disease with severely reduced left ventricular function underwent supported high-risk PCI with an intra-aortic balloon pump (IABP, N = 211) or a left ventricular assist device (Impella 2.5, N = 216) as part of the PROTECT II trial. We examined the performance of the additive Euroscore, logistic Euroscore, STS mortality score, STS morbidity and mortality score, Mayo Clinic risk score and New York state PCI risk score on the endpoint of 90-day mortality in this unique high-risk population. Mean age was 67.2 ± 10.9 years; 65.8% of patients were in NYHA class III/IV, and mean LVEF was 24%. All-cause 90-day mortality was 10.4%. The scores were generally correlated (p < 0.0001 for all comparisons), with R(2) values ranging from 0.28 (STS morbidity/mortality and Mayo Clinic) to 0.68 (logistic Euroscore and STS mortality). However, receiver-operator curves for 90-day all-cause mortality for all risk scores demonstrated poor discriminatory performance with c-statistics of 0.542-0.616. Calibration of the risk scores was not poor, but varied according to the specific score examined. The discriminatory capacity of currently available risk models is suboptimal when applied to a cohort of mechanically supported complex high-risk PCI. A risk score designed specifically for this population could help to further refine risk assessment.


In vivo quantification of cell coupling in plants with different phloem-loading strategies.
Uptake of photoassimilates into the leaf phloem is the key step in carbon partitioning and phloem transport. Symplasmic and apoplasmic loading strategies have been defined in different plant taxa based on the abundance of plasmodesmata between mesophyll and phloem. For apoplasmic loading to occur, an absence of plasmodesmata is a sufficient but not a necessary criterion, as passage of molecules through plasmodesmata might well be blocked or restricted. Here, we present a noninvasive, whole-plant approach to test symplasmic coupling and quantify the intercellular flux of small molecules using photoactivation microscopy. Quantification of coupling between all cells along the prephloem pathways of the apoplasmic loader Vicia faba and Nicotiana tabacum showed, to our knowledge for the first time in vivo, that small solutes like sucrose can diffuse through plasmodesmata up to the phloem sieve element companion cell complex (SECCC). As expected, the SECCC was found to be symplasmically isolated for small solutes. In contrast, the prephloem pathway of the symplasmic loader Cucurbita maxima was found to be well coupled with the SECCC. Phloem loading in gymnosperms is not well understood, due to a profoundly different leaf anatomy and a scarcity of molecular data compared with angiosperms. A cell-coupling analysis for Pinus sylvestris showed high symplasmic coupling along the entire prephloem pathway, comprising at least seven cell border interfaces between mesophyll and sieve elements. Cell coupling together with measurements of leaf sap osmolality indicate a passive symplasmic loading type. Similarities and differences of this loading type with that of angiosperm trees are discussed.


Distribution of polymorphisms rs12979860, rs8099917 and rs12980275 IL28B in patients with chronic hepatitis C.
The prognosis concerning the treatment of patients with chronic hepatitis C (CHC) is closely related to the genotype of the virus as well as to the factors dependent on the patient. It was proved that polymorphisms of the gene encoding interleukin 28B (IL28B) are associated with sustained viral response, which in the case of profitable variants of IL28B polymorphisms may reach up to 87% of the patients. The aim of the study is to determine the prevalence of alleles and distribution of IL28B polymorphisms genotypes in the examined group of patients with CHC in Wielkopolska Province. A total of 710 people with diagnosed hepatitis C virus were examined in order to determine the distribution of polymorphisms of gene IL28B rs12979860, rs8099917 and rs12980275. The polymorphisms were evaluated by sequencing of PCR products. The most often noted profitable variant was genotype TT for polymorphism rs8099917 present in 43.5% of the patients, next was AA rs12980275 in 22.5%. The rarest was the profitable variant CC of the polymorphism rs12979860 present in 17.5% of the patients. An occurrence of at least 2 IL28B polymorphisms in the preferred variants (homozygote CC, TT, AA) was found in 239 out of 710 (34%) patients, among which 117 patients had favorable genotypes for all 3 examined polymorphisms. The SNP distribution of gene IL28 with fixed prognostic value in the population of patients with chronic hepatitis C is different from the general population, and shows the need to evaluate polymorphisms prior to treatment.


Cutaneous alternariosis treated with miconazole.
A 40-year-old man had had a chronic, slowly spreading inflammatory process in the skin over the right knee of ten years' duration. Culture of a biopsy specimen grew Alternaria sp. Antimycotic susceptibility testing with flucytosine, amphotericin B, and miconazole nitrate was performed. The Alternaria was highly sensitive to the last two drugs. Miconazole nitrate, 1.2 g daily intravenously for 31 days, did not induce a clinical response. The lesions were then infiltrated with 10 mg of miconazole nitrate in 10 mL of polyethoxylated castor oil (Cremophor EL) twice weekly, with clearing of the lesions during a several-month period.


Smooth muscle α-actin is a direct target of PLZF: effects on the cytoskeleton and on susceptibility to oncogenic transformation.
Changes in cell morphology and rearrangements of the actin cytoskeleton are common features accompanying cell transformation induced by various oncogenes. In this study, we show that promyelocytic leukemia zinc finger protein (PLZF) binds to the promoter of smooth muscle α-actin, reducing mRNA and protein levels encoded by this gene and resulting in a reorganization of the actin cytoskeleton. In cultures of chicken embryo fibroblasts (CEF), this effect on α-actin expression is correlated with a change in cellular phenotype from spindle shaped to polygonal and flattened. This morphological change is dependent on Ras function. The polygonal, flattened CEF show a high degree of resistance to the transforming activity of several oncoproteins. Our results support the conclusion that reorganization of the actin cytoskeleton plays an important role in tumor suppression by PLZF.


Time, symptom burden, androgen deprivation, and self-assessed quality of life after radical prostatectomy or watchful waiting: the Randomized Scandinavian Prostate Cancer Group Study Number 4 (SPCG-4) clinical trial.
Quality-of-life outcomes are important in the choice of treatment strategy for men with localized prostate cancer. To evaluate how follow-up time, number of physical symptoms, and presence of androgen deprivation affected quality of life among men randomized to radical prostatectomy or watchful waiting. The study group was composed of all 376 living men included in the Swedish part of the Scandinavian Prostate Cancer Group Study Number 4 (SPCG-4) between January 1, 1989, and February 29, 1996. Quality-of-life data were collected after a mean follow-up time of 4.1 yr. All patients were randomly assigned to radical prostatectomy or watchful waiting. Forty-five men were androgen deprived. Data of specific symptoms, symptom-induced stress, sense of well-being, and self-assessed quality of life were obtained by means of a questionnaire. Psychological symptoms were assessed using seven-point visual digital scales. In analyses stratified on the basis of the numbers of physical symptoms, anxiety and depressed mood were less common, and sense of well-being and self-assessed quality of life were better throughout in the radical prostatectomy group than in the watchful waiting group. As the number of physical symptoms increased, all psychological variables became worse and more prominent in the watchful waiting group. After a follow-up time of 6-8 yr, a significant decrease in quality of life (p=0.03) was seen in the watchful waiting group. Twenty-four percent of androgen-deprived patients assigned to watchful waiting reported high self-assessed quality of life compared with 60% in the radical prostatectomy group. Eighty-eight percent of patients had clinically detected tumors. Androgen deprivation negatively affected self-assessed quality of life in men assigned to watchful waiting. The number of physical symptoms was associated with the level of quality of life. Quality of life was lower with longer follow-up time in both groups and was statistically significant in the watchful waiting group (p=0.03).


Control processes underlying elbow flexion movements may be independent of kinematic and electromyographic patterns: experimental study and modelling.
Using a non-linear dynamic model based on the lambda version of the equilibrium-point hypothesis, we investigated the shape and duration of the control patterns underlying discrete elbow movements. The model incorporates neural control variables, time-, position- and velocity-dependent intrinsic muscle and reflex properties. Two control variables (R and C) specify a positional frame of reference for activation of flexor and extensor motoneurons. The variable R (reciprocal command) specifies the referent joint angle (R) at which the transition of net flexor to extensor active torque or vice versa can be observed during changes in the actual joint angle elicited by an external force. The variable C (coactivation command) surrounds the transition angle by an angular range in which flexor and extensor muscles may be simultaneously active (if C > 0) or silent (if C < or = 0). An additional, time-dimensional control variable (mu command) influences the dependency of the threshold of the stretch reflex on movement velocity. This control variable is responsible for the reflex damping. Changes in the R command result in shifts in the equilibrium state of the system, a dynamical process leading to electromyographic modifications and movement production. Commands C and mu provide movement stability and effective energy dissipation preventing oscillations at the end of movement. A comparison of empirical and model data was carried out. A monotonic ramp-shaped pattern of the R command can account for the empirical kinematic and electromyographic patterns of the fastest elbow flexion movements made with or without additional inertia, as well as of self-paced movements. The rate of the shifts used in simulation was different for the three types of movements but independent of movement distance (20-80 degrees). This implies that, for a given type of movement, the distance is encoded by the duration of shift in the equilibrium state. The model also reproduces the kinematic and electromyographic patterns of the fastest uncorrected movements opposed in random trials by a high load (80-90% of the maximal) generated by position feedback to a torque motor. The following perturbation effects were simulated: a substantial decrease in the arm displacement (from 60-70 degrees to 5-15 degrees) and movement duration (to about 100 ms) so that these movements ended near the peak velocity of those which were not perturbed; a prolongation of the first agonist electromyographic burst as long as the load was applied; the suppression of the antagonist burst during the dynamic and static phases of movements: the reappearance of the antagonist burst in response to unloading accompanied by a short-latency suppression of agonist activity. These kinematic and electromyographic features of both perturbed and non-perturbed movements were reproduced by using the same control patterns which elicited a monotonic shift in the equilibrium state of the system ending before the peak velocity of non-perturbed movements. Our results suggest that the neural control processes underlying the fastest unopposed changes in the arm position are completed long before the end of the movement and phasic electromyographic activity. Neither the timing nor the amplitude of electromyographic bursts are planned but rather they represent the long-lasting dynamic response of central, reflex and mechanical components of the system to a monotonic, short-duration shift in the system's equilibrium state.


Fixation strategy influences the ability to focus attention on two spatially separate objects.
The ability to devote attention simultaneously to multiple visual objects plays an important role in domains ranging from everyday activities to the workplace. Yet, no studies have systematically explored the fixation strategies that optimize attention to two spatially distinct objects. Assuming the two objects require attention nearly simultaneously, subjects either could fixate one object or they could fixate between the objects. Studies measuring the breadth of attention have focused almost exclusively on the former strategy, by having subjects simultaneously perform one attention-demanding task at fixation and another in the periphery. We compared performance when one object was at fixation and the other was in the periphery to a condition in which both objects were in the periphery and subjects fixated between them. Performance was better with two peripheral stimuli than with one central and one peripheral stimulus, meaning that a strategy of fixating between stimuli permitted greater attention breadth. Consistent with the idea that both measures tap attention breadth, sport experts consistently outperformed novices with both fixation strategies. Our findings suggest a way to improve performance when observers must pay attention to multiple objects across spatial regions. We discuss possible explanations for this performance advantage.


The effect of a selective estrogen receptor modulator on the progression of spontaneous autoimmune disease in MRL lpr/lpr mice.
The MRL lpr/lpr mouse strain is an animal model for the autoimmune disorder systemic lupus erythematosus (SLE). Pathologic changes in the mice include a severe proliferative glomerulonephritis, lymph node and spleen enlargement, increase in autoantibody titers, and shortened life spans. In the present investigation, female MRL lpr/lpr mice have been dosed po daily for 7 months with the selective estrogen receptor modulator (SERM) LY139478 (4 mg/kg) or 17alpha-ethinylestradiol (EE2, 1 mg/kg) and compared to vehicle control animals. The LY139478 group had an increase in survival (73% survival at 7 months, P = 0.02) but the EE2-treated animals did not (53% survival at 7 months, P = 0.4) when compared to the control group (32% survival at 7 months). Although there were no reductions in autoantibody levels as determined by anti-DNA antibody ELISA, histological analysis of kidney tissue indicated that both LY139478 and EE2 mitigated the progression of glomerular nephritis which was evident in the controls. In contrast, there were no significant differences in lymph node size although the LY139478 and EE2 groups retained a well-defined sinusoidal region. Finally, flow cytometric analysis documented that thymuses from animals treated for 7 months with LY139478 but not with EE2 contained predominantly CD4+/CD+ T cells consistent with a normal thymic phenotype observed in non-MRL lpr/lpr mouse strains. These studies demonstrate that SERMs may be potentially useful for the treatment of autoimmune disorders.


Distinct, constitutively active MAPK phosphatases function in Xenopus oocytes: implications for p42 MAPK regulation In vivo.
Xenopus oocyte maturation requires the phosphorylation and activation of p42 mitogen-activated protein kinase (MAPK). Likewise, the dephosphorylation and inactivation of p42 MAPK are critical for the progression of fertilized eggs out of meiosis and through the first mitotic cell cycle. Whereas the kinase responsible for p42 MAPK activation is well characterized, little is known concerning the phosphatases that inactivate p42 MAPK. We designed a microinjection-based assay to examine the mechanism of p42 MAPK dephosphorylation in intact oocytes. We found that p42 MAPK inactivation is mediated by at least two distinct phosphatases, an unidentified tyrosine phosphatase and a protein phosphatase 2A-like threonine phosphatase. The rates of tyrosine and threonine dephosphorylation were high and remained constant throughout meiosis, indicating that the dramatic changes in p42 MAPK activity seen during meiosis are primarily attributable to changes in MAPK kinase activity. The overall control of p42 MAPK dephosphorylation was shared among four partially rate-determining dephosphorylation reactions, with the initial tyrosine dephosphorylation of p42 MAPK being the most critical of the four. Our findings provide biochemical and kinetic insight into the physiological mechanism of p42 MAPK inactivation.


The effect of automatic gain control structure and release time on cochlear implant speech intelligibility.
Nucleus cochlear implant systems incorporate a fast-acting front-end automatic gain control (AGC), sometimes called a compression limiter. The objective of the present study was to determine the effect of replacing the front-end compression limiter with a newly proposed envelope profile limiter. A secondary objective was to investigate the effect of AGC speed on cochlear implant speech intelligibility. The envelope profile limiter was located after the filter bank and reduced the gain when the largest of the filter bank envelopes exceeded the compression threshold. The compression threshold was set equal to the saturation level of the loudness growth function (i.e. the envelope level that mapped to the maximum comfortable current level), ensuring that no envelope clipping occurred. To preserve the spectral profile, the same gain was applied to all channels. Experiment 1 compared sentence recognition with the front-end limiter and with the envelope profile limiter, each with two release times (75 and 625 ms). Six implant recipients were tested in quiet and in four-talker babble noise, at a high presentation level of 89 dB SPL. Overall, release time had a larger effect than the AGC type. With both AGC types, speech intelligibility was lower for the 75 ms release time than for the 625 ms release time. With the shorter release time, the envelope profile limiter provided higher group mean scores than the front-end limiter in quiet, but there was no significant difference in noise. Experiment 2 measured sentence recognition in noise as a function of presentation level, from 55 to 89 dB SPL. The envelope profile limiter with 625 ms release time yielded better scores than the front-end limiter with 75 ms release time. A take-home study showed no clear pattern of preferences. It is concluded that the envelope profile limiter is a feasible alternative to a front-end compression limiter.


Hypoenergetic diet-induced reductions in myofibrillar protein synthesis are restored with resistance training and balanced daily protein ingestion in older men.
Strategies to enhance weight loss with a high fat-to-lean ratio in overweight/obese older adults are important since lean loss could exacerbate sarcopenia. We examined how dietary protein distribution affected muscle protein synthesis during energy balance (EB), energy restriction (ER), and energy restriction plus resistance training (ER + RT). A 4-wk ER diet was provided to overweight/obese older men (66 ± 4 yr, 31 ± 5 kg/m(2)) who were randomized to either a balanced (BAL: 25% daily protein/meal × 4) or skewed (SKEW: 7:17:72:4% daily protein/meal; n = 10/group) pattern. Myofibrillar and sarcoplasmic protein fractional synthetic rates (FSR) were measured during a 13-h primed continuous infusion of l-[ring-(13)C6]phenylalanine with BAL and SKEW pattern of protein intake in EB, after 2 wk ER, and after 2 wk ER + RT. Fed-state myofibrillar FSR was lower in ER than EB in both groups (P < 0.001), but was greater in BAL than SKEW (P = 0.014). In ER + RT, fed-state myofibrillar FSR increased above ER in both groups and in BAL was not different from EB (P = 0.903). In SKEW myofibrillar FSR remained lower than EB (P = 0.002) and lower than BAL (P = 0.006). Fed-state sarcoplasmic protein FSR was reduced similarly in ER and ER + RT compared with EB (P < 0.01) in both groups. During ER in overweight/obese older men a BAL consumption of protein stimulated the synthesis of muscle contractile proteins more effectively than traditional, SKEW distribution. Combining RT with a BAL protein distribution "rescued" the lower rates of myofibrillar protein synthesis during moderate ER.


Does physical activity and BMI mediate the association between DCD and internalizing problems in early childhood? A partial test of the Environmental Stress Hypothesis.
Young children with Developmental Coordination Disorder (DCD) are more likely to experience internalizing problems, such as depression and anxiety, than typically developing (TD) children. Currently, the underlying mechanisms resulting in increased internalizing problems in DCD remains unknown; however, a previous study based on the Environmental Stress Hypothesis (ESH) indicated that physical inactivity and obesity may mediate the relationship between DCD and internalizing problems. The purpose of this study is to investigate the relationships among DCD, internalizing problems, physical activity, and BMI, and the role of sex in these relationships in preschool children, a population for which we currently have very limited data. Young children between the ages of 4 and 5 years enrolled in the Coordination and Activity Tracking in CHildren (CATCH) study comprised the sample (n = 589). Of these, 288 (193 boys, 67.0%) were classified as at risk for DCD (rDCD), based on scoring at or below 16th percentile on the Movement Assessment Battery for Children - Second Edition. Physical activity was measured using accelerometers and height and weight were measured by trained research assistants, while parents completed the Child Behavior Checklist to rate internalizing problems. The mediating effects of physical activity and BMI were tested on the relationship between rDCD and internalizing problems. Children with rDCD reported more internalizing problems than TD children. While there was a direct effect of rDCD on internalizing problems, neither physical activity nor BMI were found to mediate this relationship. The findings from this study support co-occurring internalizing problems in preschool children with DCD, and extend these findings to demonstrate that this relationship is not explained by physical activity or BMI in early childhood. Further research should be directed toward other psychosocial factors identified in the ESH to better understand the underlying mechanisms between DCD and co-occurring internalizing problems.


Deep bite: a case report with chewing pattern and electromyographic activity before and after therapy with function generating bite.
The purpose of this case report is the concurrent evaluation of the masticatory pattern and the electromyographic activity, recorded during mastication, before and after therapy of deep bite malocclusion. An 11-year-old boy, affected by deep bite (overbite = 5 mm) was treated by the use of a functional appliance (Function Generating Bite for Deep bite correction = FGB-D). Mandibular movements during mastication of a soft and a hard bolus were recorded both before and 10 months after correction of the malocclusion. Electromyographic activity (EMG) of the masseters and anterior temporalis muscles were recorded at the same time. Chewing cycles and EMG activity were recorded with the K7 I kinesiograph (Myotronics Inc., Seattle, WA-USA). Before therapy a higher EMG activity was recorded for both masseters and anterior temporalis muscles in comparison with the results after therapy. The results showed a great decrease of the EMG activity of masseter and anterior temporalis muscles. Moreover, the height and width of the chewing cycles in the frontal plane increased after therapy. The functional improvement showed after therapy with FGB-D showed that the functional appliance is able to correct the dental malocclusion and the masticatory function. The orthodontic treatment should consider not only the repositioning of teeth within the dental arches but also the effects on function, especially when the malocclusion involves the muscular and skeletal structures.


A presynaptic role for the cytomatrix protein GIT in synaptic vesicle recycling.
Neurotransmission involves the exo-endocytic cycling of synaptic vesicles (SVs) within nerve terminals. Exocytosis is facilitated by a cytomatrix assembled at the active zone (AZ). The precise spatial and functional relationship between exocytic fusion of SVs at AZ membranes and endocytic SV retrieval is unknown. Here, we identify the scaffold G protein coupled receptor kinase 2 interacting (GIT) protein as a component of the AZ-associated cytomatrix and as a regulator of SV endocytosis. GIT1 and its D. melanogaster ortholog, dGIT, are shown to directly associate with the endocytic adaptor stonin 2/stoned B. In Drosophila dgit mutants, stoned B and synaptotagmin levels are reduced and stoned B is partially mislocalized. Moreover, dgit mutants show morphological and functional defects in SV recycling. These data establish a presynaptic role for GIT in SV recycling and suggest a connection between the AZ cytomatrix and the endocytic machinery.


Sleep and Circadian Disturbances in Children With Neurodevelopmental Disorders.
Sleep problems are highly prevalent in those with neurodevelopmental disorders (NDDs). We propose this is secondary to multiple factors that directly and indirectly negatively impact sleep and circadian processes in those with NDDs, which in turn, further perturbs development, resulting in a "developmental and sleep/circadian-related encephalopathy." In this review, we discuss select NDDs with known or suspected sleep and circadian phenotypes. We also highlight important considerations when evaluating and treating sleep and circadian disorders in these populations.


Protein attributes contribute to halo-stability, bioinformatics approach.
Halophile proteins can tolerate high salt concentrations. Understanding halophilicity features is the first step toward engineering halostable crops. To this end, we examined protein features contributing to the halo-toleration of halophilic organisms. We compared more than 850 features for halophilic and non-halophilic proteins with various screening, clustering, decision tree, and generalized rule induction models to search for patterns that code for halo-toleration. Up to 251 protein attributes selected by various attribute weighting algorithms as important features contribute to halo-stability; from them 14 attributes selected by 90% of models and the count of hydrogen gained the highest value (1.0) in 70% of attribute weighting models, showing the importance of this attribute in feature selection modeling. The other attributes mostly were the frequencies of di-peptides. No changes were found in the numbers of groups when K-Means and TwoStep clustering modeling were performed on datasets with or without feature selection filtering. Although the depths of induced trees were not high, the accuracies of trees were higher than 94% and the frequency of hydrophobic residues pointed as the most important feature to build trees. The performance evaluation of decision tree models had the same values and the best correctness percentage recorded with the Exhaustive CHAID and CHAID models. We did not find any significant difference in the percent of correctness, performance evaluation, and mean correctness of various decision tree models with or without feature selection. For the first time, we analyzed the performance of different screening, clustering, and decision tree algorithms for discriminating halophilic and non-halophilic proteins and the results showed that amino acid composition can be used to discriminate between halo-tolerant and halo-sensitive proteins.


A divide and conquer approach for imbalanced multi-class classification and its application to medical decision making.
Many real world data contains more than two categories and the number of instances in each category differs greatly. Such as in medical diagnostic data, there may be several types of cancer and each with tens instances, but contains even more normal instances. Similarly, there may be very few abnormal samples in pharmaceutical test but which may cause great harm. Classification of such type of data is often summarized as imbalanced multi-class classification. Most existing researches study multi-class classification and imbalanced data classification separately, few study in a combination way, in particular for medical diagnosis data classification. In the context of medical diagnosis and pharmaceutical test, in this paper, we propose a divide and conquer approach to partition multi-class data and a self-adaptive data resample method for imbalanced data. The proposed methods are tested on 23 UCI datasets in medical, pharmaceutical and other fields. Experiment results show that the proposed methods outperform other compared methods, in particular on those medical and pharmaceutical dataset.


[Recanalization of lower-limb deep veins as an index of efficacy of treatment for acute venous thrombosis].
The authors analysed the results of examination and treatment of a total of 102 patients presenting with iliofemoral venous thrombosis. During treatment, ultrasonographic duplex scanning was used to determine the localization of the proximal margin of thrombotic masses, the time of appearing of the first signs of recanalization, its degree at various levels of the deep venous system, as well as alteration in velocity of the venous blood flow in the deep veins of the lower limbs. The dynamics of clinical symptoms was assessed by the visual analogue scale. Clinical and instrumental examination was performed on day 10, and then 1, 3, 6 and 12 months after the beginning of treatment. The patients were subdivided into three groups. Group One comprised 38 patients receiving therapy with low-molecular-weight heparin (enoxaprin) followed by switching to indirect anticoagulants (warfarin) combined with venotonics (original highly-purified diosmin 600 mg once daily). Group Two was composed of 33 patients receiving rivaroxaban at a dose of 15 mg twice daily for 3 weeks, followed by 20 mg once daily. Group Tree patients (n=31) were also given rivaroxaban according to the above-described standard regimen but in combination with venotonics (original highly-purified diosmin 600 mg once daily). The obtained findings showed that prescribing rivaroxaban to patients from the first day of the disease made it possible to considerably improve and accelerate the processes of restoration of patency of deep veins of lower extremities as compared with the patients taking vitamin K antagonists (warfarin). In patients receiving rivaroxaban, there were no cases of residual thrombotic occlusions of the major veins, and recanalization in three fourths of patients was assessed as good and in the remaining third as moderate. In the warfarin group, occlusion in the iliac veins was noted to persist persisted in 13% of patients, with good recanalization observed only in half of the patients. Addition of venotonics (original highly-purified diosmin) to anticoagulants from the first day demonstrated safety of this therapeutic regimen (with no cases of clinically significant haemorrhagic complications revealed) and its high efficacy as compared with monotherapy with rivaroxaban. A combination of diosmin with rivaroxaban turned out more efficient than a combination of diosmin with warfarin.


Fish oil suppresses cell growth and metastatic potential by regulating PTEN and NF-κB signaling in colorectal cancer.
Homeostasis in eukaryotic tissues is tightly regulated by an intricate balance of the prosurvival and antisurvival signals. The tumor suppressor PTEN (phosphatase and tensin homolog deleted on chromosome 10), a dual-specificity phosphatase, plays a functional role in cell cycle arrest and apoptosis. NF-κB and its downstream regulators (such as VEGF) play a central role in prevention of apoptosis, promotion of inflammation and tumor growth. Therefore, we thought to estimate the expression of PTEN, Poly-ADP-ribose polymerase (PARP), NF-κBp50, NF-κBp65 and VEGF to evaluate the effect of supplementation of fish oil on apoptotic and inflammatory signaling in colon carcinoma. Male wistar rats in Group I received purified diet while Group II and III received modified diet supplemented with FO∶CO(1∶1)&FO∶CO(2.5∶1) respectively. These were further subdivided into controls receiving ethylenediamine-tetra acetic-acid and treated groups received dimethylhydrazine-dihydrochloride (DMH)/week for 4 weeks. Animals sacrificed 48 hours after last injection constituted initiation phase and that sacrificed after 16 weeks constituted post-initiation phase. We have analysed expression of PTEN, NF-κBp50, NF-κBp65 by flowcytometer and nuclear localization of NF-κB by immunofluorescence. PARP and VEGF were assessed by immunohistochemistry. In the initiation phase, animals receiving DMH have shown increased % of apoptotic cells, PTEN, PARP, NF-κBp50, NF-κBp65 and VEGF however in post-initiation phase no significant alteration in apoptosis with decreased PTEN and increased PARP, NF-κBp50, NF-κBp65 and VEGF were observed as compared to control animals. On treatment with both ratios of fish oil in both the phases, augmentation in % of apoptotic cells, decreased PTEN, PARP, NF-κBp50, NF-κBp65 and VEGF were documented with respect to DMH treated animals with effect being more exerted with higher ration in post-initiation phase. Hence, fish oil activates apoptosis, diminishes DNA damage and inhibits inflammatory signalling in a dose and time dependent manner so as to inhibit progression of colon cancer.


A review of the history, development and application of auditory weighting functions in humans and marine mammals.
This document reviews the history, development, and use of auditory weighting functions for noise impact assessment in humans and marine mammals. Advances from the modern era of electroacoustics, psychophysical studies of loudness, and other related hearing studies are reviewed with respect to the development and application of human auditory weighting functions, particularly A-weighting. The use of auditory weighting functions to assess the effects of environmental noise on humans-such as hearing damage-risk criteria-are presented, as well as lower-level effects such as annoyance and masking. The article also reviews marine mammal auditory weighting functions, the development of which has been fundamentally directed by the objective of predicting and preventing noise-induced hearing loss. Compared to the development of human auditory weighting functions, the development of marine mammal auditory weighting functions have faced additional challenges, including a large number of species that must be considered, a lack of audiometric information on most species, and small sample sizes for nearly all species for which auditory data are available. The review concludes with research recommendations to address data gaps and assumptions underlying marine mammal auditory weighting function design and application.


Activation of corticotropin-releasing factor receptors in the rostral ventrolateral medulla is required for glucose-induced sympathoexcitation.
Energy expenditure is determined by metabolic rate and diet-induced thermogenesis. Normally, energy expenditure increases due to neural mechanisms that sense plasma levels of ingested nutrients/hormones and reflexively increase sympathetic nerve activity (SNA). Here, we investigated neural mechanisms of glucose-driven sympathetic activation by determining contributions of neuronal activity in the hypothalamic paraventricular nucleus (PVN) and activation of corticotropin-releasing factor (CRF) receptors in the rostral ventrolateral medulla (RVLM). Glucose was infused intravenously (150 mg/kg, 10 min) in male rats to raise plasma glucose concentration to a physiological postprandial level. In conscious rats, glucose infusion activated CRF-containing PVN neurons and TH-containing RVLM neurons, as indexed by c-Fos immunofluorescence. In α-chloralose/urethane-anesthetized rats, glucose infusion increased lumbar and splanchnic SNA, which was nearly prevented by prior RVLM injection of the CRF receptor antagonist astressin (10 pmol/50 nl). This cannot be attributed to a nonspecific effect, as sciatic afferent stimulation increased SNA and ABP equivalently in astressin- and aCSF-injected rats. Glucose-stimulated sympathoexcitation was largely reversed during inhibition of PVN neuronal activity with the GABA-A receptor agonist muscimol (100 pmol/50 nl). The effects of astressin to prevent glucose-stimulated sympathetic activation appear to be specific to interruption of PVN drive to RVLM because RVLM injection of astressin prior to glucose infusion effectively prevented SNA from rising and prevented any fall of SNA in response to acute PVN inhibition with muscimol. These findings suggest that activation of SNA, and thus energy expenditure, by glucose is initiated by activation of CRF receptors in RVLM by descending inputs from PVN.


General seizures revealing macro-adenomas secreting prolactin or prolactin and growth hormone in men.
Epilepsy is a heterogeneous condition with numerous etiologies. Pituitary tumors are rarely responsible for generalized convulsions except when they are very large. Apart from anecdotic cases, only one study concerning epilepsy frequency in male macroprolactinomas is available in Medline. Our aim was to analyze epilepsy frequency and conditions under which seizures appear and disappear in men harboring macroprolactinomas or somatolactotroph adenomas. We retrospectively analyzed 90 men with macro-adenomas (>1 cm) secreting prolactin (PRL) (n = 82) or both PRL and growth hormone (n = 8) to look for generalized seizures. We took into account familial and personal medical history, clinical examination, routine and hormonal analyzes, and radiological assessment based on cerebral magnetic resonance imaging. Between 1992 and 2012, we collected eight cases (8.9%): Seven were hospitalized for recent generalized seizures; one had epilepsy after conventional radiotherapy given in 1992 because of neurosurgery failure and resistance to bromocriptine. Their median age was 33.75 years (22-58), median PRL was 9,198 ng/ml and median tumor height was 74 mm (41-110). The temporal lobe was invaded in six cases. After tumor reduction, epilepsy disappeared and never relapsed after a follow-up varying between 1 and 20 years. Epilepsy, which is a life-threatening condition, can be the first presentation in men with prolactinomas or somatolactotroph adenomas, especially those involving the supra sellar area, and the brain. Convulsions can also appear after radiotherapy. That one should be avoided, if possible, before tumor reduction by surgery or medical treatment.


Autonomic Small-Fiber Pathology in Patients With Fibromyalgia.
In this clinical and skin biopsy study, we aimed to investigate whether fibromyalgia-associated small-fiber pathology (SFP), consisting of an intraepidermal nerve fiber loss, implies damage of dermal autonomic nerve fibers and how this damage is associated with autonomic symptoms that patients with fibromyalgia syndrome experience. Using skin biopsy, we investigated intraepidermal nerve fiber density, piloerector muscle, and sweat gland nerve fiber density (SGNFD) in 138 participants, that is, 58 patients with fibromyalgia syndrome, 48 healthy subjects, and 32 patients with small-fiber neuropathy. In patients with fibromyalgia-associated SFP, we also investigated how the different skin biopsy variables correlated with autonomic symptoms, as assessed with the Composite Autonomic Symptom Score 31 questionnaire. We found that in patients with fibromyalgia-associated SFP, the piloerector muscle and SGNFD were lower than that in healthy subjects. However, the autonomic small-fiber damage had no correlation with autonomic symptoms severity. In patients with SFP, the intraepidermal, piloerector muscle, and SGNFD were higher than that in patients with small-fiber neuropathy. Our clinical and skin biopsy study shows that patients with fibromyalgia have a reduction of dermal autonomic small fibers paralleling the intraepidermal nerve fiber loss, thus indicating that SFP also implies autonomic small nerve fiber damage. However, the autonomic small-fiber damage we found had no correlation with the severity of autonomic symptoms, and thus its clinical impact is still undetermined. PERSPECTIVE: In patients with fibromyalgia, SFP also affects autonomic fibers. These novel data provide additional insights into the pathophysiology of fibromyalgia syndrome, highlighting the complex role of small-fiber damage in the clinical picture of fibromyalgia.


New protocols for the synthesis of 5-amino-7-(4-phenyl)-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidine-6-carboxylate esters using an efficient additive.
This work introduces a new additive named 4,4'-trimethylenedipiperidine for the practical and ecofriendly preparation of ethyl 5-amino-7-(4-phenyl)-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidine-6-carboxylate derivatives. This chemical is commercially available and easy to handle. It also possesses a low melting point and a broad liquid range temperature, high thermal stability, and good solubility in water. Based on green chemistry principles, the reaction was performed in a) a mixture of green solvents i.e. water and ethanol (1:1 v/v) at reflux temperature, and b) the additive was liquefied at 65 °C and the reaction was conducted in the liquid state of the additive. High yields of the desired triazolo-pyrimidines were obtained under both aforementioned conditions. Our results demonstrated that this additive, containing 2 Lewis base sites and able to act as an acceptor-donor hydrogen bonding group, is a novel and efficient alternative to piperidine, owing to its unique properties such as its reduced toxicity, nonflammable nature, nonvolatile state, broad liquid range temperature, high thermal stability, and ability to be safely handled. Furthermore, this additive could be completely recovered and exhibited high recyclability without any change in its chemical structure and no significant reduction in its activity. The current methodology has several advantages: (a) it avoids the use of hazardous materials, as well as toxic, volatile, and flammable solvents, (b) it does not entail tedious processes, harsh conditions, and the multistep preparation of catalysts, (c) it uses a metal-free and noncorrosive catalyst, and (d) reduces the generation of hazardous waste and simple work-up processes. The most important result of this study is that 4,4'-trimethylenedipiperidine can be a promising alternative for toxic, volatile, and flammable base reagents in organic synthesis owing to its unique properties.


Drainage, rupture, and lifetime of deionized water films: effect of dissolved gases?
Gas bubbles coalesce in deionized (DI) water because the water (foam) films between the bubbles are not stable. The so-called hydrophobic attraction has been suggested as the cause of the film instability and the bubble coalescence. In this work, microinterferometry experiments show that foam films of ultrapure DI water can last up to 10 s and the contact time between the two gas bubble surfaces at close proximity (approximately 1 microm separation distance) significantly influences the film drainage, rupture, and lifetime. Specifically, when the two bubbles were first brought into contact, the films instantly ruptured at 0.5 microm thickness. However, the film drainage rate and rupture thickness sharply decreased and the film lifetime steeply increased with increasing contact time up to 10 min, but then they leveled off. The constant thickness of film rupture was around 35 nm. Possible contamination was vigorously investigated and ruled out. It is argued that migration of gases inherently dissolved in water might cause the transient behavior of the water films at the short contact time. The film drainage rate and instability at the long contact time were analyzed employing Eriksson et al.'s phenomenological theory of long-range hydrophobic attraction (Eriksson, J. C.; Ljunggren, S.; Claesson, P. M., J. Chem. Soc., Faraday Trans. 2 1989, 85, 163-176) and the hypothesis of water molecular structure modified by dissolved gases, and the extended Stefan-Reynolds theory by incorporating the mobility of the air-DI-water interfaces.


[Cardiac transplantation with bicaval anastomosis and prophylactic graft tricuspid annuloplasty].
This study aims to evaluate the effects of prophylactic heart donor tricuspid annuloplasty in patients after heart transplantation with bicaval anastomosis. From 2002 to 2005, 20 patients undergoing heart transplantation with bicaval anastomosis and with a survival rate over 6 months were deliberately selected. Patients were divided into two groups: Group I - 10 patients who underwent prophylactic heart donor tricuspid annuloplasty by the De Vega technique; and Group II - 10 patients did not undergo annuloplasty. In both groups, presurgical clinical characteristics were the same. The tricuspid regurgitation degree was evaluated by transthoracic Doppler echocardiography and it was qualified from 0 to 3 (0=absent, 1=mild, 2=moderated, 3=severe). Myocardial performance was evaluated by ventricular ejection fraction and invasive hemodynamic study performed during routine endomyocardial biopsies. Mean clinical follow-up was 14.6+/-4.3 (6 and 16) months. There was only one death in group II. It was not related to annuloplasty. Mean degree of tricuspid regurgitation in Group I was 0.4+/-0.6 and in Group II was 1.6+/-0.8 (p < 0.05). There was a statistically significant difference between both groups in right atrium pressure, which was higher in Group II. In view of the limitations of the study, the prophylactic tricuspid annuloplasty in heart donor reduced the degree of valvar regurgitation in the medium term after heart transplantation with bicaval anastomosis, in spite of not interfering with the allograft hemodynamic performance in the period under consideration.


Catalyst-free assembly of a polyfunctionalized 1,2,4-triazole-fused N-heterocycle, 6-acylated pyrrolo[1,2-a][1,2,4]triazolo[5,1-c]pyrazine.
A catalyst-free synthetic route to a new 1,2,4-triazole-pyrrolo[1,2-a]pyrazine hybrid system, 6-acylpyrrolo[1,2-a][1,2,4]triazolo[5,1-c]pyrazine, was realized upon sequential exposure of pyrrole-2-carbonitrile-derived substrates to DMF-DMA and acyl hydrazide through which acylated pyrazine and 1,2,4-triazole rings were consecutively formed with the formation of multiple bonds (one C-C bond and three C-N bonds). This annulative functionalization approach towards N-fused polycycles enabled us to install various substituents at specific positions on the core skeleton.


Mesenchymal stem cells from CD34(-) human umbilical cord blood.
Umbilical cord blood (UCB) is well known to be a rich source of stem cells especially for haematopoietic stem cells (HSCs). Recently, mesenchymal stem cells (MSCs) have also been shown to exist in cord blood. Although MSCs have been described by a subset of surface antigens after expansion, little is known about the cell surface phenotype of undifferentiated MSCs. The aim of this study therefore was to clarify whether undifferentiated MSCs are resident among CD34(-) UCB cells. CD34(+) cells were separated from UCB mononuclear cells (MNCs) by magnetic sorting and the CD34(-) cell fractions were cultured in Dulbecco's modified Eagle's medium (DMEM) with 10% foetal calf serum (FCS) and basic-fibroblast growth factor. Isolated CD34(+) cells were also cultured in the same medium. Adherent fibroblast-like cells at passage 3-4 were analyzed by fluorescence-activated cell sorting (FACS) for MSC marker expression , and standard adipogenic, osteogenic and chondrogenic assays were used to investigate their differentiation potentials. After 4-5 weeks in culture, the cells from the CD34(-) fraction became confluent with flat and fibroblast-like morphology. These cells were positively stained for the mesenchymal cell markers CD29, CD73 and CD105. In adipogenic differentiation, the cells showed oil red O positive and expressed FABP4, adipsin and proliferation-activated receptor gamma-2 (PPARgamma2 genes) associated with adipogenesis. In osteogenic differentiation, calcium accumulation and osteocalcin were detected. The cells grown in chondrogenic conditions were positively stained for human aggrecan and expressed collagen type II and Sox-9 genes. In contrast, cells from the CD34(+) fraction failed to generate any cells with MSC morphology under the same culture conditions. Our results showed that UCB contained MSCs which are only resident in the CD34(-) fraction. The MSCs could be induced to differentiate into at least three lineage cell types, adipocytes, osteoblasts and chondrocytes.


Maternal endocrine activation during pregnancy alters neurobehavioral state in primate infants.
The purpose of this study was to investigate whether maternal endocrine activation during pregnancy would affect the neurobehavioral state of primate offspring in a manner similar to that observed in human infants from pregnancies involving maternal substance abuse or maternal stress. Twenty-two rhesus monkey (Macaca mulatta) infants were derived from females administered either adrenocorticotrophic hormone (ACTH), which increased the mother's endocrine activity, or saline solutions for 14 consecutive days during mid-pregnancy. On days 15 and 30 postpartum, infants underwent brief separations from their mothers and were videotaped for later evaluation of neurobehavioral state. Infants from mothers administered ACTH spent significantly more time in a drowsy state than did controls (p < .04), and the increased drowsiness tended to be most pronounced during the postseparation period, when acute stress was highest. In contrast, controls remained in a more active alert state (p < .03), presumably searching for their mother, a species-typical adaptive response to maternal separation. Female infants spent more time in distressed state than did males on day 15, and the proportion of time in distressed state decreased in all infants after administration of .2 ml of 24% sucrose solution. The results demonstrate that neurobehavioral state alterations are found in infants from mothers with increased endocrine activity during pregnancy. Neurobehavioral state disorganization can have an adverse impact on the human infant's concurrent and subsequent occupational performance. These findings establish the usefulness of the nonhuman primate model for advancing knowledge on early contributions to the development of human infant occupational behavior.


The whereabouts of 2D gels in quantitative proteomics.
Two-dimensional gel electrophoresis has been instrumental in the development of proteomics. Although it is no longer the exclusive scheme used for proteomics, its unique features make it a still highly valuable tool, especially when multiple quantitative comparisons of samples must be made, and even for large samples series. However, quantitative proteomics using 2D gels is critically dependent on the performances of the protein detection methods used after the electrophoretic separations. This chapter therefore examines critically the various detection methods (radioactivity, dyes, fluorescence, and silver) as well as the data analysis issues that must be taken into account when quantitative comparative analysis of 2D gels is performed.


Nanostructures in superhydrophobic Ti6Al4V hierarchical surfaces control wetting state transitions.
This paper mainly reports the wetting state of liquid droplets on a Ti6Al4V micro-nanoscale hierarchical structured hydrophobic surface. In this work, the detailed action mechanism of the secondary nanostructure in the hierarchical structure on the wetting-state transition (from the Wenzel state to the Cassie state) was revealed and discussed. The variation of micro-morphology of the sample surface was observed using a field emission scanning electron microscope (FE-SEM). Furthermore, the apparent contact angle and sliding angle of the droplets on the surfaces were measured via a contact angle measurement instrument. The theoretical and experimental results indicated that the one-dimensional nanowire structure, which was planted on the microstructure surface by the hydrothermal method, effectively changed the wetting state of liquid droplets on the surface from the Wenzel state to the Cassie state owing to its good size synergies with microscale structure. This process not only increased the apparent contact angle of liquid droplets on the solid surface (to 161°), but also decreased the sliding angle significantly (to 3°) and contact angle hysteresis (to ∼2°), demonstrating the robust non-wetting property.


The subunits of the stimulatory regulatory component of adenylate cyclase. Resolution, activity, and properties of the 35,000-dalton (beta) subunit.
The stimulatory guanine nucleotide-binding regulatory component (G/F) of adenylate cyclase is activated by exposure to guanine nucleotide analogs or to Al3+ + F-. Activated G/F can reconstitute adenylate cyclase activity when mixed with the catalytic moiety of the enzyme system in the absence of an effective free concentration of stimulatory ligand. Activation is explained by dissociation of the alpha (45,000-Da) and beta (35,000-Da) subunits of G/F. The beta subunit of G/F facilitates reversal of the activated state of the regulatory protein. This phenomenon, which has been exploited as an assay for the resolved beta subunit, has the following properties. 1) Addition of the resolved beta subunit to fluoride-activated G/F increases the initial rate of deactivation from a t 1/2 of 10 min to less than 0.5 min. 2) The enhancement of the rate of deactivation is a saturable process with a K 1/2 value of 60 ng/ml (approximately 2 nM). 3) G/F does not display beta subunit activity unless the alpha subunit has been inactivated or the subunits have been resolved. beta Subunit activity is measurable in detergent extracts of rabbit liver membranes or plasma membranes from S49 cell clones. The activity in such extracts is similar to that found with purified G/F, in that incubation at 30 degrees C in the presence of Mg2+ is required for its expression. However, cyc-, UNC, and H21a (S49 cell mutants with deficient or altered G/F activity) have amounts of beta subunit activity similar to that found in wild type S49 cells. Furthermore, the amount of beta subunit activity exceeds by 5- to 10-fold the amount expected based on the quantity of G/F in wild type extracts. All of the beta subunit activity in detergent extracts of liver membranes can be purified as a 35,000-Da polypeptide that is indistinguishable from the beta subunit of G/F. The beta subunit activity in extracts of cyc- membranes is expressed after incubation with guanine nucleotide analogs, implying association of the beta subunit with a GTP-binding protein. By analysis of the chromatographic behavior of G/F and the recently identified 41,000/35,000-Da heterodimeric substrate for the islet-activating protein from Bordetella pertussis, we have identified the 41,000-Da subunit of the substrate for islet-activating protein as the GTP-binding protein with which the majority of the beta subunit activity associates. These data have direct bearing on the mechanisms of hormonal activation and inhibition of adenylate cyclase.


Two highly conserved cysteine residues in HPV16 L2 form an intramolecular disulfide bond and are critical for infectivity in human keratinocytes.
Minor capsid protein L2 performs an indispensable but uncharacterized role in human papillomavirus infections. A neutralizing B cell epitope has recently been mapped to the N-terminus of HPV16 L2, residues 17-36, and exposure of this region of L2 has been implicated in translocation of incoming virions from the endo/lysosomal compartment to the cellular cytoplasm. Here we examine the redox state of Cys22 and Cys28 two highly conserved cysteines located within this epitope. We also investigate the infectivity of virions containing L2 single and double cysteine point mutants. Denaturing/non-reducing gel analysis and thiol labeling experiments of wild type and cysteine mutant HPV16 virion particles strongly support the existence of a buried intramolecular C22-C28 disulfide bond. The disulfide was confirmed by tandem mass spectrometry of L2 protein from non-reduced virions. Single C22S and C28S and the double C22/28S mutants were non-infectious but had no apparent defects in cell binding, endocytosis, or trafficking to lysosomes by 8 h post infection. During infection with L2 mutant particles, there was a marked decrease in L2 levels compared to wild type L2-containing virions, suggesting a failure of mutant L2/genome complexes to exit the endo/lysosomal compartment. L2 residues C22 and C28 are bound as an intramolecular disulfide bond in HPV16 virions and are necessary for infectivity. Previous work has suggested that the furin-dependent exposure of the 17-36 epitope and subsequent interaction of this region with an unknown receptor is necessary for egress from the endo/lysosomal compartment and infection. Identification of the C22-C28 disulfide suggests that reduction of this disufide bond may be necessary for exposure of 17-36 and HPV16 infection.


Endotoxin biomarkers, hepatic fat fraction, liver volume and liver stiffness among adolescents at high-risk for non-alcoholic fatty liver disease: The HEROES study.
Non-alcoholic fatty liver disease (NAFLD) is on the rise among youth. Identifying biomarkers of NAFLD progression/risk can aid in prevention efforts. This pilot study investigated associations of two endotoxin biomarkers-lipopolysaccharide-binding protein (LBP) and anti-endotoxin core immunoglobulin G (EndoCab)-with markers of NAFLD among 99 Latino/Latina adolescents (11-19 years) with obesity. We used linear regression to examine associations of each endotoxin biomarker (per 1-SD) with hepatic fat fraction (HFF), liver volume, and liver stiffness. We found positive associations of LBP with HFF and liver volume. Each 1-SD increment in LBP corresponded with 2.35% (95% CI: 0.46%, 4.23%) higher HFF and 0.14 (0.06, 0.23) L greater liver volume after adjusting for age, sex, and maternal education. Accounting for abdominal adiposity and Tanner stage did not change results. Excluding 72 participants with NAFLD attenuated associations of LBP with HFF but associations with liver volume persisted (0.11 [0.01, 0.21] L). EndoCab was not associated with any liver outcomes. Neither endotoxin biomarker predicted liver stiffness. While additional research is warranted, our results support LBP as a biomarker of NAFLD risk/progression in high-risk youth.