Hypofractionation in non-small cell lung cancer (NSCLC): suggestions from modelling both acute and chronic hypoxia.
Based on experimental estimates for acute and chronic tumour hypoxia, a speculative analysis of the therapeutic ratio dependence on the number of once-daily five-days-per-week fractions (n) for non-small cell lung cancer (NSCLC) radiotherapy is proposed. For this purpose an adapted formulation of the linear-quadratic model has been derived, including the effects of tumour repopulation, inter-tumour alpha-heterogeneity and oxygen enhancement ratio dependence on the dose per fraction. The relation between the curative dose D50, assuring 50% tumour control probability, and n has been computed: for (n, D50) fractionation schemes, the therapeutic ratios have been compared in terms of effective normalized total doses to the lungs (NTD(eff)L), estimated by a few supposed fractions of the normalized total dose to the tumour. Results suggest that D50 is dominated by chronic hypoxia for shortly hypofractionated treatments and by acute hypoxia for multifractionated treatments. Furthermore, the optimum number of fractions depends on the rapidity of the reoxygenation from chronically hypoxic cells, almost independently of the extent of both chronic and acute hypoxia. For NSCLC, both the reduction of n until about 20 fractions in hypofractionated dose-escalation trials, and the extension of extra-cranial stereotactic radiotherapy schedules to include at least 5-10 fractions, seem to be supported by this model.


Spectral-domain optical coherence tomography measurements of choroidal thickness and outer retinal disruption in macular telangiectasia type 2.
To evaluate subfoveal choroidal thickness (CT) and the extent of outer retinal disruption in patients with macular telangiectasia type 2 (MacTel2) compared with healthy eyes. In this prospective, observational, cohort study, 62 patients (62 eyes) with Mac-Tel2 and 130 healthy controls (130 eyes) underwent a complete ophthalmological examination, spectral-domain optical coherence tomography (SD-OCT) imaging, and axial length measurements. Patients in the study group also underwent color fundus photography, fundus autofluorescence, and fluorescein angiography. En face SD-OCT imaging was used to assess abnormalities involving the photoreceptor inner segment/outer segment/ellipsoid zone (IS/OS/EZ). After adjusting for age and axial length, the authors found that eyes with MacTel2 had a mean CT measurement that was greater than control eyes (P = .007). There was a negative correlation between the visual acuity and the area of IS/OS/EZ damage (P = .009), but no statistically significant correlation was seen between CT and the area of IS/OS/EZ damage. Eyes with MacTel2 were found to have thicker CT measurements than control eyes. While the extent of IS/OS/EZ disruption correlated with the loss of visual acuity, this damage did not correlate with CT measurements.


Decreased serum lipocalin-2 levels in human immunodeficiency virus-infected patients: increase during highly active anti-retroviral therapy.
Although neutrophil gelatinase-associated lipocalin (NGAL) may play a pivotal role in the innate immune response, there are currently no data on NGAL levels in human immunodeficiency virus (HIV)-infected patients. In this study we aimed to examine the regulation of NGAL in HIV infection. The regulation of NGAL in HIV infection was examined by different experimental approaches, including studies in peripheral blood and mononuclear cells (MNC) from bone marrow aspirates before and during highly active anti-retroviral therapy (HAART). We found that: before initiating HAART, HIV-infected patients (n = 37) had significantly decreased serum NGAL levels compared with healthy controls (n = 26); (ii) during HAART, there was a gradual and significant increase in NGAL concentrations reaching levels comparable to those in healthy controls after 12 months; (iii) this increase was seen primarily in virological responders to HAART (HIV RNA level <200 copies/ml after 24 months); (iv) phytohaemagglutinin-stimulated NGAL release in MNC cells from bone marrow aspirates was decreased in untreated HIV-infected patients compared with healthy controls, but increased after 26 weeks on HAART; and (v) there was a significant positive correlation between neutrophil counts and NGAL levels at all time-points during HAART. We have shown decreased NGAL levels in HIV-infected patients, potentially reflecting decreased number and function of neutrophils as well as impaired bone marrow myelopoiesis. These abnormalities were reversed by successful HAART. Our findings underscore further the involvement of neutrophils and innate immunity in HIV-related immunodeficiency.


Low frequency noise "pollution" interferes with performance.
To study the possible interference of low frequency noise on performance and annoyance, subjects categorised as having a high- or low sensitivity to noise in general and low frequency noise in particular worked with different performance tasks in a noise environment with predominantly low frequency content or flat frequency content (reference noise), both at a level of 40 dBA. The effects were evaluated in terms of changes in performance and subjective reactions. The results showed that there was a larger improvement of response time over time, during work with a verbal grammatical reasoning task in the reference noise, as compared to the low frequency noise condition. The results further indicated that low frequency noise interfered with a proof-reading task by lowering the number of marks made per line read. The subjects reported a higher degree of annoyance and impaired working capacity when working under conditions of low frequency noise. The effects were more pronounced for subjects rated as high-sensitive to low frequency noise, while partly different results were obtained for subjects rated as high-sensitive to noise in general. The results suggest that the quality of work performance and perceived annoyance may be influenced by a continuous exposure to low frequency noise at commonly occurring noise levels. Subjects categorised as high-sensitive to low frequency noise may be at highest risk.


SPOT-1D-Single: improving the single-sequence-based prediction of protein secondary structure, backbone angles, solvent accessibility and half-sphere exposures using a large training set and ensembled deep learning.
Knowing protein secondary and other one-dimensional structural properties are essential for accurate protein structure and function prediction. As a result, many methods have been developed for predicting these one-dimensional structural properties. However, most methods relied on evolutionary information that may not exist for many proteins due to a lack of sequence homologs. Moreover, it is computationally intensive for obtaining evolutionary information as the library of protein sequences continues to expand exponentially. Here, we developed a new single-sequence method called SPOT-1D-Single based on a large training dataset of 39 120 proteins deposited prior to 2016 and an ensemble of hybrid long-short-term-memory bidirectional neural network and convolutional neural network. We showed that SPOT-1D-Single consistently improves over SPIDER3-Single and ProteinUnet for secondary structure, solvent accessibility, contact number and backbone angles prediction for all seven independent test sets (TEST2018, SPOT-2016, SPOT-2016-HQ, SPOT-2018, SPOT-2018-HQ, CASP12 and CASP13 free-modeling targets). For example, the predicted three-state secondary structure's accuracy ranges from 72.12% to 74.28% by SPOT-1D-Single, compared to 69.1-72.6% by SPIDER3-Single and 70.6-73% by ProteinUnet. SPOT-1D-Single also predicts SS3 and SS8 with 6.24% and 6.98% better accuracy than SPOT-1D on SPOT-2018 proteins with no homologs (Neff = 1), respectively. The new method's improvement over existing techniques is due to a larger training set combined with ensembled learning. Standalone-version of SPOT-1D-Single is available at https://github.com/jas-preet/SPOT-1D-Single. Direct prediction can also be made at https://sparks-lab.org/server/spot-1d-single. The datasets used in this research can also be downloaded from GitHub. Supplementary data are available at Bioinformatics online.


[Academic international cultural exchange: an experience of personal and scientific growth].
This study aimed to describe the student experience to integrate an international academic mobility program during undergraduate nursing. It is reported academic experience in Portugal, Algarve University, made possible by the Federal University of Santa Maria, Rio Grande do Sul, Brazil. The experience of exchange performed within six months, enabled the acquisition of new scientific knowledge and cultural rights, including innovations in healthcare technology, development of research and academic links. As contributions are expected to stimulate and intensify the international mobility especially at undergraduate level, considering its importance to the improvement of academic excellence and the Brazilian higher education through scientific and cultural exchange abroad.


Electronic Media Device Usage and Its Associations With BMI and Obesity in a Rapidly Developing City in South China.
Objective: This study aimed to evaluate the television (TV) viewing and computer and mobile phone usage duration in a community sample of Chinese adults and examine their associations with BMI and obesity. Methods: We conducted a community-based health needs assessment study from February to December 2018 among 2,873 Chinese adults in Nanshan District of Shenzhen, China. We used self-administered questionnaires to collect the data from 24 community health service centers in Nanshan District. The participants individually recorded the time they spent watching TV and using computers and mobile phones. They also answered questions about their sociodemographic and lifestyle factors. We measured their height and weight by using ultrasonic height and weight scales. Then, we calculated and categorized their BMI in accordance with the standards prescribed by the World Health Organization for Asians. Results: Most of the participants were young adults (aged 18-44 years, 74.2%). The overall prevalence of obesity was 15.30%. The prevalence of TV, computer, and mobile phone usage was 75.5, 71.2, and 93.8% in females and 71.1, 75.7, and 94.2% in males, respectively. The youth (18-24 years) and the elderly (65 years or older) had the longest time using mobile phones (3.78 ± 2.51 h) and watching TV programs (2.12 ± 1.38 h), respectively. Longer usage of computers and mobile phones was evident in males (p < 0.05) and participants with a high education level (p < 0.01). The univariate analysis results showed an association between high BMI and obesity and short duration of using computers and mobile phones (all p < 0.05). By contrast, no significant associations were found between the length of TV program viewing and mobile phone usage and BMI (p > 0.05). After we adjusted for potential confounders, we found that computer usage time and the overall usage time of the three electronic devices had an inverse association with BMI (p < 0.05). Conclusions: Mobile phones were the most popular electronic device in Nanshan residents of Shenzhen. Unlike most previous studies, we found an inverse association between screen time and BMI. Nevertheless, scholars should conduct further studies to explore this association. Overall, we strongly encourage the appropriate use of electronic devices.


Dietary strategies to attenuate muscle loss during recovery from injury.
Injuries are an unavoidable aspect of participation in physical activity. Nutrition is important for optimal wound healing and recovery, but little information about nutritional support for injuries exists. Immediately following injury, wound healing begins with an inflammatory response. Excessive anti-inflammatory measures may impair recovery. Many injuries result in limb immobilization. Immobilization results in muscle loss due to increased periods of negative muscle protein balance from decreased basal muscle protein synthesis and resistance to anabolic stimuli, including protein ingestion. Oxidative capacity of muscle is also decreased. Nutrient and energy deficiencies should be avoided. Energy expenditure may be reduced during immobilization, but inflammation, wound healing and the energy cost of ambulation limit the reduction of energy expenditure. There is a theoretical rationale for leucine and omega-3 fatty acid supplementation to help reduce muscle atrophy. During rehabilitation and recovery from immobilization, increased activity, in particular resistance exercise will increase muscle protein synthesis and restore sensitivity to anabolic stimuli. Ample, but not excessive, protein and energy must be consumed to support muscle growth. During rehabilitation and recovery, nutritional needs are very much like those for any athlete desiring muscle growth. The most important consideration is to avoid malnutrition and to apply a risk/benefit approach.


Estimating age-specific vaccine effectiveness using data from a large measles outbreak in Berlin, Germany, 2014/15: evidence for waning immunity.
BackgroundMeasles elimination is based on 95% coverage with two doses of a measles-containing vaccine (MCV2), high vaccine effectiveness (VE) and life-long vaccine-induced immunity. Longitudinal analysis of antibody titres suggests existence of waning immunity, but the relevance at the population-level is unknown.AimWe sought to assess presence of waning immunity by estimating MCV2 VE in different age groups (2-5, 6-15, 16-23, 24-30 and 31-42 years) in Berlin.MethodsWe conducted a systematic literature review on vaccination coverage and applied the screening-method using data from a large measles outbreak (2014/15) in Berlin. Uncertainty in input variables was incorporated by Monte Carlo simulation. In a scenario analysis, we estimated the proportion vaccinated with MCV2 in those 31-42 years using VE of the youngest age group, where natural immunity was deemed negligible.ResultsOf 773 measles cases (median age: 20 years), 40 had received MCV2. Average vaccine coverage per age group varied (32%-88%). Estimated median VE was > 99% (95% credible interval (CrI): 98.6-100) in the three youngest age groups, but lower (90.9%, 95% CrI: 74.1-97.6) in the oldest age group. In the scenario analysis, the estimated proportion vaccinated was 98.8% (95% CrI: 96.5-99.8).ConclusionVE for MCV2 was generally high, but lower in those aged 31-42 years old. The estimated proportion with MCV2 should have led to sufficient herd immunity in those aged 31-42 years old. Thus, lower VE cannot be fully explained by natural immunity, suggesting presence of waning immunity.


The W(sh), W(57), and Ph Kit expression mutations define tissue-specific control elements located between -23 and -154 kb upstream of Kit.
The Kit and PDGFRa receptor tyrosine kinases are encoded in close proximity at the murine white spotting (W) and patch (Ph) loci. Whereas W mutations affect hematopoiesis, melanogenesis, and gametogenesis, the Ph mutation affects melanogenesis and causes early lethality in homozygotes. The W(sh), W(57), and Ph mutations diminish Kit expression in certain cell types such as mast cells and enhance it in others. The W(sh), W(57), and Ph mutations arose from deletions and inversions affecting sequences in between the Kit and PDGFRa genes. We have determined the precise location of the breakpoint of the W(sh) inversion and the endpoints of the W(57) deletion upstream of the Kit transcription start site and examined the effect of these mutations on Kit expression in mast cells and hematopoietic stem cells and lineage progenitors. Our results indicate that positive elements controlling Kit expression in mast cells mapping in between -23 and -154 kb from the transcription start site can be dissociated from negative elements controlling Kit misexpression during embryonic development in the vicinity of the PDGFRa gene. In addition, we have identified two clusters of hypersensitive sites in mast cells at -23 -28 kb and -147 -154 kb from the Kit gene transcription start site. Analysis of these hypersensitive sites in mutant mast cells indicates a role for HS4-6 in Kit expression in mast cells. These findings provide a molecular basis for the phenotype of these Kit expression mutations and they provide insight into the complex mechanisms governing the regulation of Kit expression.


Performance of an in situ passive sampling system for metals in stormwater.
A passive sampler has been developed and is demonstrated in situ for urban runoff. The passive sampler is compared to conventional composite (time-dependent and flow-weighted) bottle sampling during and between storm events. The sampling was carried out at established stormwater stations; before and after a stormwater detention pond. In situ deployment of the passive sampler provides the metal concentrations, corresponding to the electrochemically available fraction of total metal, for time-dependent samples collected in parallel. The sampler provides improved accuracy compared to bottle sampling because contamination during sample transport and handling is minimised. Laboratory handling is reduced by direct analysis of the accumulated metals on the receiving membrane by laser ablation inductively coupled plasma mass spectrometry. Passive sampling also solves the problem of metal speciation change during transport to the laboratory, which is a potential problem for bottle samples. The low cost and convenience of the passive sampler and subsequent analysis should allow significantly more extensive spatial and temporal monitoring of metals in the aquatic environment than has previously been possible.


Preparation of N-sulfonyl- and N-carbonyl-11-azaartemisinins with greatly enhanced thermal stabilities: in vitro antimalarial activities.
As the clinically used artemisinins do not withstand the thermal stress testing required to evaluate shelf life for storage in tropical countries where malaria is prevalent, there is a need to develop thermally more robust artemisinin derivatives. Herein we describe the attachment of electron-withdrawing arene- and alkanesulfonyl and -carbonyl groups to the nitrogen atom of the readily accessible Ziffer 11-azaartemisinin to provide the corresponding N-sulfonyl- and -carbonylazaartemisinins. Two acylurea analogues were also prepared by treatment of the 11-azaartemisinin with arylisocyanates. Several of the N-sulfonylazaartemisinins have melting points above 200 degrees C and possess substantially greater thermal stabilities than the artemisinins in current clinical use, with the antimalarial activities of several of the arylsulfonyl derivatives being similar to that of artesunate against the drug-sensitive 3D7 clone of the NF54 isolate and the multidrug-resistant K1 strain of P. falciparum. The compounds possess relatively low cytotoxicities. The carbonyl derivatives are less crystalline than the N-sulfonyl derivatives, but are generally more active as antimalarials. The N-nitroarylcarbonyl and arylurea derivatives possess sub-ng ml(-1) activities. Although several of the azaartemisinins possess log P values below 3.5, the compounds have poor aqueous solubility (<1 mg L(-1) at pH 7). The greatly enhanced thermal stability of our artemisinins suggests that strategic incorporation of electron-withdrawing polar groups into both new artemisinin derivatives and totally synthetic trioxanes or trioxolanes may assist in the generation of practical new antimalarial drugs which will be stable to storage conditions in the field, while retaining favorable physicochemical properties.


Methodological challenges in union commitment studies.
Methodological problems in studies of union commitment were identified and illustrated with data from 4,641 members and 479 stewards in 297 local teachers' unions. Using a 20-item union commitment scale, results confirmed the existence of 3 substantive factors and 1 method factor at the individual level of analysis: loyalty to the union, responsibility to the union, willingness to work for the union, and a factor of negatively worded items. Tests of measurement invariance showed that the scale captured commitment for rank-and-file members but not for union stewards. The authors also found partial measurement invariance between long-time and newer members and full measurement invariance between men and women. Finally, the authors found that violation of the statistical assumption of independence reduced model fit when individual commitment scores were analyzed without attention to the hierarchical nature of the data.


The need for revised standards for skinfold thickness in infancy.
Skinfold standards provide a useful indication of subcutaneous fat. To evaluate skinfold thickness of 252 Cambridge infants over the first 2 years of age, SD scores relative to the Tanner standards were calculated, taking account of skewness in the standards. Cambridge SD scores were low, varying according to age from -1.2 to -1.8 for triceps and -0.6 to -1.2 for subscapular skinfolds. The Tanner skinfold standards were last revised 30 years ago, at a time of high prevalence of infantile obesity, and the present and other studies indicate that infants are now thinner. There is a need for new skinfold standards to reflect this change. Since the Cambridge infants contributed to the recent British height and weight references, it is suggested that their skinfold measurements could also serve as reference points.


The pattern of histogenesis and growth of tooth plates in larval stages of extant lungfish.
Comparison of new data obtained in this study on Protopterus aethiopicus with that published on Protopterus aethiopicus and Neoceratodus forsteri has confirmed the suggestion that the pattern of histogenesis of tooth plates in the early larval stages is very similar in the two genera. These similarities are more apparent both when a common terminology is adopted, based on a topogenic classification, and when the fundamental assumption is made that a single morphogenetic system operates for all odontodes. The model to explain the structure of all vertebrate dentitions with separate teeth in single or multiple tooth rows has been found to apply to dipnoan dentitions with fused teeth in a composite tooth plate. In this model, the epithelial invagination surrounding the margins of the tooth plate represents the dental lamina and, where this is in contact with mesenchymal cells, cell clusters (protogerms) arise. From these protogerms new odontodes (teeth) may develop if factors to inhibit differentiation are not present. Sites for initiation of odontodes become restricted to the labial margins of existing ridges on the tooth plate. Experimental studies on mammalian tooth germs are discussed and a model proposed for control of odontogenesis and histogenesis in dipnoan dentitions. Patterns of growth of hypermineralised petrodentine have been analysed and shown to depend initially on the arrangement of odontodes, and subsequently upon the ability of special cells in the pulp to generate new and wider layers of petrodentine. The initial pattern of petrodentine depends upon the position of odontodes in the forming ridges of the tooth plate. Subsequent patterns of petrodentine depend upon the extent of replacement growth beneath the tritural surface. Specialised cells, petroblasts, secrete the petrodentine within a shell of dentine. These cells differentiate from cells of the dental papilla after odontoblasts have begun to form dentine. They are regarded as a unique type of cell for dipnoans.


When may road fatalities start to decrease?
The comparative analysis of macroscopic trends in road safety has been a popular research topic. The objective of this research is to propose a simple and, at the same time, reliable multiple regime model framework for international road safety comparisons, allowing for the identification of slope changes of personal risk curves and respective breakpoints. The trends of road traffic fatalities in several EU countries have been examined through the temporal evolution of elementary socioeconomic indicators, namely motorized vehicle fleet and population, at the country level. Piece-wise linear regression models have been fitted, using a methodology that allows the simultaneous estimation of all slopes and breakpoints. The number and location of breakpoints, as well as the slope of the connecting trends, vary among countries, thus indicating different road safety evolution patterns. Macroscopic analysis of road accident trends may be proved beneficial for the identification of best examples and the implementation of appropriate programmes and measures, which will lead to important benefits for the society and the economy through the reduction of road fatalities and injuries. Best performing countries and the related programmes and measures adopted may concern several safety improvements at the processes of the road, the vehicle and the insurance industries. Lessons from the analysis of the past road safety patterns of developed countries provide some insight into the underlying process that relates motorization levels with personal risk and can prove to be beneficial for predicting the road safety evolution of developing countries that may have not yet reached the same breakpoints. Furthermore, the presented framework may serve as a basis to build more elaborate models, including more reliable exposure indicators (such as vehicle-km driven).


Myeloid cell-specific disruption of Period1 and Period2 exacerbates diet-induced inflammation and insulin resistance.
The circadian clockworks gate macrophage inflammatory responses. Given the association between clock dysregulation and metabolic disorders, we conducted experiments to determine the extent to which over-nutrition modulates macrophage clock function and whether macrophage circadian dysregulation is a key factor linking over-nutrition to macrophage proinflammatory activation, adipose tissue inflammation, and systemic insulin resistance. Our results demonstrate that 1) macrophages from high fat diet-fed mice are marked by dysregulation of the molecular clockworks in conjunction with increased proinflammatory activation, 2) global disruption of the clock genes Period1 (Per1) and Per2 recapitulates this amplified macrophage proinflammatory activation, 3) adoptive transfer of Per1/2-disrupted bone marrow cells into wild-type mice potentiates high fat diet-induced adipose and liver tissue inflammation and systemic insulin resistance, and 4) Per1/2-disrupted macrophages similarly exacerbate inflammatory responses and decrease insulin sensitivity in co-cultured adipocytes in vitro. Furthermore, PPARγ levels are decreased in Per1/2-disrupted macrophages and PPARγ2 overexpression ameliorates Per1/2 disruption-associated macrophage proinflammatory activation, suggesting that this transcription factor may link the molecular clockworks to signaling pathways regulating macrophage polarization. Thus, macrophage circadian clock dysregulation is a key process in the physiological cascade by which diet-induced obesity triggers macrophage proinflammatory activation, adipose tissue inflammation, and insulin resistance.


Production of hepatitis B virus particles in Hep G2 cells transfected with cloned hepatitis B virus DNA.
The hepatoblastoma cell line Hep G2 was transfected with a plasmid carrying the gene that confers resistance to G418 and four 5'-3' tandem copies of the hepatitis B virus (HBV) genome positioned such that two dimers of the genomic DNA are 3'-3' with respect to one another. Cells of one clone that grew in the presence of G418 produce high levels of hepatitis B e antigen and of hepatitis B surface antigen. HBV DNA is carried by these cells as chromosomally integrated sequences and episomally as relaxed circular, covalently closed, and incomplete copies of the HBV genome. Viral DNA was detected also in conditioned growth medium at the buoyant densities characteristic for infectious Dane and immature core particles. Finally, HBV-specific components morphologically identical to the 22-nm spherical and filamentous hepatitis B surface antigen particles as well as 42-nm Dane particles were visualized by immunoelectron microscopic analysis. Therefore, we have demonstrated that the Hep G2 cell line can support the assembly and secretion not only of several of the replicative intermediates of HBV DNA but also of Dane-like particles. This in vitro system can now be used to study the life cycle of HBV and the reaction of immunocompetent cells with cells carrying HBV.


A safe technique for the delivery of "Mini' Gianturco stainless steel coils.
A modified introducer guide wire for the delivery of mini-coils is described. Mini-coils can now be used through standard tapered catheters with an inside diameter large enough to let pass of 0.97-mm (0.038-in) guide wire.


Seroprevalence of SARS-CoV-2 in urban settings in three sub-Saharan African countries (SeroCoV): a study protocol for a household-based cross-sectional prevalence study using two-stage cluster sampling.
The current COVID-19 pandemic has impacted the entire world with increasing morbidity and mortality and has resulted in serious economic and social consequences. Assessing the burden of COVID-19 is essential for developing efficient pandemic preparedness and response strategies and for determining the impact of implemented control measures. Population-based seroprevalence surveys are critical to estimate infection rates, monitor the progression of the epidemic and to allow for the identification of persons exposed to the infection who may either have been asymptomatic or were never tested. This is especially important for countries where effective testing and tracking systems could not be established and where non-severe cases or under-reported deaths might have blurred the true burden of COVID-19. Most seroprevalence surveys performed in sub-Saharan Africa have targeted specific high risk or more easily accessible populations such as healthcare workers or blood donors, and household-based estimates are rarely available. Here, we present the study protocol for a SARS-CoV-2 seroprevalence estimation in the general population of Burkina Faso, Ghana and Madagascar in 2021. The SeroCoV study is a household-based cross-sectional prevalence investigation in persons aged 10 years and older living in urban areas in six cities using a two-stage geographical cluster sampling method stratified by age and sex. The presence of anti-SARS-CoV-2 IgG antibodies will be determined using a sensitive and specific SARS-CoV-2 IgG ELISA. In addition, questionnaires will cover sociodemographic information, episodes of diseases and history of testing and treatment for COVID-like symptoms, travel history and safety measures. We will estimate the seroprevalence of SARS-CoV-2, taking into account test performance and adjusting for the age and sex of the respective populations. Ethical approval was received for all participating countries. Results will be disseminated through reports and presentations at the country level as well as peer-reviewed publications and international scientific conferences presentations.


Decreased levels of heat shock proteins in gut epithelial cells after exposure to plant lectins.
The enterocytes of the intestinal epithelium are regularly exposed to potentially harmful substances of dietary origin, such as lectins. Expression of heat shock proteins (HSPs) by this epithelium may be part of a protective mechanism developed by intestinal epithelial cells to deal with noxious components in the intestinal lumen. To investigate if the lectins PHA, a lectin from kidney beans (Phaseolus vulgaris) and WGA, a lectin from wheat germ (Triticum aestivum) could modify the heat shock response in gut epithelial cells and to establish the extent of this effect. Jejunal tissue sections from PHA and WGA fed rats were screened for expression of HSP70, HSP72, and HSP90 using monoclonal antibodies. Differentiated Caco-2 cells, the in vitro counterpart of villus enterocytes, were exposed to 100 microg/ml of PHA-E(4) or WGA for 48 hours and investigated for changes in DNA and protein synthesis by double labelling with [2-(14)C]thymidine and L-[methyl-(3)H]methionine. The relative concentrations of HSP60, HSP70, HSP72, and HSP90 and binding protein (BiP) in these cells exposed to lectins were analysed by polyacrylamide gel electrophoresis and immunoblotting. To establish if lectin exposed differentiated Caco-2 cells were still capable of producing a heat shock response, these cells received a heat shock (40 degrees C, 41 degrees C, and 42 degrees C) for one hour and were allowed to recover for six hours at 37 degrees C. During heat shock and recovery periods, lectin exposure was continued. Constitutive levels of HSPs were measured in the intestinal cells of lactalbumin fed (control) rats, as may be expected from the function of this tissue. However, in PHA and WGA fed rats a marked decline in the binding of antibodies against several HSPs to the intestinal epithelium was found. These results were confirmed by in vitro experiments using differentiated Caco-2 cells exposed to PHA-E(4) and WGA. However, after exposure to lectins, these cells were still capable of heat induced heat shock protein synthesis, and total protein synthesis was not impaired indicating specific inhibition of HSP synthesis in non-stressed cells. We conclude that PHA and WGA decrease levels of stress proteins in rat gut and enterocyte-like Caco-2 cells, leaving these cells less well protected against the potentially harmful content of the gut lumen.


Increased expression of complement decay-accelerating factor during activation of human neutrophils.
Decay-accelerating factor (DAF) is a membrane protein that protects blood cells from damage by autologous complement. Using monoclonal antibodies in both direct-binding studies and flow cytometry, we found that resting neutrophils (polymorphonuclear leukocytes [PMN]) expressed 10(4) DAF molecules on their surface, and that surface DAF expression more than doubled when the cells were activated. Upregulation of surface DAF occurred within minutes, paralleled the upregulation of complement receptor types 1 and 3 (CR1 and CR3), and was not dependent on new protein synthesis. It was unaffected by EDTA but was inhibited by 10 microM trifluoperazine, suggesting involvement of intracellular Ca2+ and calmodulin or protein kinase C. Upon activation, the affected PMN lacking surface DAF from patients with paroxysmal nocturnal hemoglobulinuria failed to increase DAF expression. In contrast, these cells increased CR1 and CR3 expression normally, suggesting that DAF deficiency in affected cells involves abnormal synthesis or packaging of DAF for intracellular storage. Translocation of DAF to the cell surface induced by chemoattractants may be important in allowing PMN to survive and function at inflammatory sites where there is rapid complement turnover.


Distribution of polyphenolic and sugar compounds in different buckwheat plant parts.
The aim of this study was to provide information on the phenolic and sugar profiles of different parts of the buckwheat plant, which can define that buckwheat is a functional food, with a high nutritional value and very useful for human health. Therefore, the extracts of buckwheat leaf, stem, and flower, as well as buckwheat grain were analysed for the content of polyphenol and antioxidant tests. The identification of a notable number of phenolic compounds and quantification of sugars in different parts of buckwheat indicates that buckwheat is a highly valuable plant. A total of 60 phenolic compounds were identified (18 cinnamic acid derivatives, 14 flavonols, 13 flavan-3-ols (including proanthocyanidins), 10 hydroxybenzoic acid derivatives, and 5 flavones) using ultra-high-performance liquid chromatography (UHPLC), coupled with a hybrid mass spectrometer which combines the Linear Trap Quadrupole (LTQ) and OrbiTrap mass analyzer. The highest number of phenolic compounds was found in the analysed buckwheat flower sample, and then in the leaf, followed by the grain and the stem. In addition, the sugar profile of buckwheat leaf, stem, flower and grain, as well as the buckwheat pollen and the nectar was analysed. Hence, 16 sugars and 5 sugar alcohols were detected by the high-performance anion exchange chromatography (HPAEC) with a pulsed amperometric detector (PAD). Sucrose was found in a significant amount with the highest content in buckwheat leaf. Trisaccharides had similar accumulation in the sample extracts, while disaccharides dominated in buckwheat leaf, followed by nectar and pollen. The sugar alcohols showed the highest content in buckwheat grain, where erythritol was predominant. The obtained results show that buckwheat is very rich in phenolic compounds and sugars. In addition to grain, the other parts of the buckwheat plant can be used as a very good source of different classes of phenolic compounds. This study provides useful information on the distribution of phytochemicals in different parts of the buckwheat plant, which contribute to the maintaining of the status of buckwheat as a functional food.


Fucosylation of N-glycans regulates the secretion of hepatic glycoproteins into bile ducts.
Fucosylated alpha-fetoprotein (AFP) is a highly specific tumor marker for hepatocellular carcinoma (HCC). However, the molecular mechanism by which serum level of fucosylated AFP increases in patients with HCC remains largely unknown. Here, we report that the fucosylation of glycoproteins could be a possible signal for secretion into bile ducts in the liver. We compared oligosaccharide structures on glycoproteins in human bile with those in serum by several types of lectin blot analyses. Enhanced binding of biliary glycoproteins to lectins that recognize a fucose residue was observed over a wide range of molecular weights compared with serum glycoproteins. A structural analysis of oligosaccharides by two-dimensional mapping high performance liquid chromatography and matrix-assisted laser desorption ionization time-of flight mass spectrometry confirmed the increases in the fucosylation of biliary glycoproteins. Purification followed by structural analysis on alpha1-antitrypsin, alpha1-acid glycoprotein and haptoglobin, which are synthesized in the liver, showed higher fucosylation in bile than in serum. To find direct evidence for fucosylation and sorting signal into bile ducts, we used alpha1-6 fucosyltransferase (Fut8)-deficient mice because fucosylation of glycoproteins produced in mouse liver was mainly an alpha1-6 linkage. Interestingly, the levels of alpha1-antitrypsin and alpha1-acid glycoprotein were quite low in bile of Fut8-deficient mice as compared with wild-type mice. An immunohistochemical study showed dramatic changes in the localization of these glycoproteins in the liver of Fut8-deficient mice. Taken together, these results suggest that fucosylation is a possible signal for the secretion of glycoproteins into bile ducts in the liver. A disruption in this system might involve an increase in fucosylated AFP in the serum of patients with HCC.


How to measure maturation: a comparison of probabilistic methods used to test for genotypic variation and plasticity in the decision to mature.
Maturation is a developmental trait that plays a key role in shaping organisms' life-history. However, progress in understanding how maturation phenotypes evolve has been held back by confusion over how best to model maturation decisions and a lack of studies comparing genotypic variation in maturation. Here, we fitted probabilistic maturation reaction norms (PMRNs) to data collected from five clones of Daphnia magna and five of Daphnia pulex collected from within and between different populations. We directly compared the utility of modeling approaches that assume maturation to be a process with an instantaneous rate with those that do not by fitting maturation rate and logistic regression models, and emphasize similarities and differences between them. Our results demonstrate that in Daphnia, PMRNs using a logistic regression approach were simpler to use and provided a better fit to the data. The decision to mature was plastic across a range of growth trajectories and dependent upon both body size and age. However, the age effect was stronger in D. magna than D. pulex and varied considerably between clones. Our results support the idea that maturation thresholds can evolve but also suggest that the notion of a threshold based on a single fixed state is an oversimplification that underestimates the adaptability of these important traits.


Prenatal cocaine exposure related to cortisol stress reactivity in 11-year-old children.
Determine the association between prenatal cocaine exposure and postnatal environmental adversity on salivary cortisol stress reactivity in school-aged children. Subjects included 743 11-year-old children (n = 320 cocaine-exposed; 423 comparison) followed since birth in a longitudinal prospective multisite study. Saliva samples were collected to measure cortisol at baseline and after a standardized procedure to induce psychological stress. Children were divided into those who showed an increase in cortisol from baseline to post stress and those who showed a decrease or blunted cortisol response. Covariates measured included site, birthweight, maternal pre and postnatal use of alcohol, tobacco or marijuana, social class, changes in caretakers, maternal depression and psychological symptoms, domestic and community violence, child abuse, and quality of the home. With adjustment for confounding variables, cortisol reactivity to stress was more likely to be blunted in children with prenatal cocaine exposure. Children exposed to cocaine and who experienced domestic violence showed the strongest effects. The combination of prenatal cocaine exposure and an adverse postnatal environment could downregulate the hypothalamic-pituitary-adrenal axis resulting in the blunted cortisol response to stress possibly increasing risk for later psychopathology and adult disease.


Breathing room: Toward next-generation tracheal engineering.
The creation of an engineered trachea with robust phenotype and sufficient mechanical properties for clinical application remains a challenge. In their work, Tang et al.1 propose a stacked approach of alternating cartilaginous and fibrous rings to form a tracheal segment, which integrated and retain patency in rabbits for 8 weeks.


The Mitochondrial Peptidase Pitrilysin Degrades Islet Amyloid Polypeptide in Beta-Cells.
Amyloid formation and mitochondrial dysfunction are characteristics of type 2 diabetes. The major peptide constituent of the amyloid deposits in type 2 diabetes is islet amyloid polypeptide (IAPP). In this study, we found that pitrilysin, a zinc metallopeptidase of the inverzincin family, degrades monomeric, but not oligomeric, islet amyloid polypeptide in vitro. In insulinoma cells when pitrilysin expression was decreased to 5% of normal levels, there was a 60% increase in islet amyloid polypeptide-induced apoptosis. In contrast, overexpression of pitrilysin protects insulinoma cells from human islet amyloid polypeptide-induced apoptosis. Since pitrilysin is a mitochondrial protein, we used immunofluorescence staining of pancreases from human IAPP transgenic mice and Western blot analysis of IAPP in isolated mitochondria from insulinoma cells to provide evidence for a putative intramitochondrial pool of IAPP. These results suggest that pitrilysin regulates islet amyloid polypeptide in beta cells and suggest the presence of an intramitochondrial pool of islet amyloid polypeptide involved in beta-cell apoptosis.


[The activity of components of system of matrix metalloproteinase and tissue inhibitors of metalloproteinase on different stages of chronic cardiac failure].
The article discusses the results of detection of activity of matrix metalloproteinase-9, tissue inhibitors of metalloproteinases-4, endotoxin, tumor necrosis factor-alpha, C-reactive protein, sE-selectine in blood of patients with chronic cardiac failure depending on severity of disease. It is demonstrated that progressing of chronic cardiac failure is associated with the increase of level of endotoxemia, activation of systemic inflammation and misbalance in the system of matrix metalloproteinase and tissue inhibitors of metalloproteinase.


The spinal component to skin blood flow abnormalities in reflex sympathetic dystrophy.
To determine whether the mechanisms of reflex sympathetic dystrophy, a neuropathic pain syndrome characterized by skin blood flow abnormalities associated with sympathetic vasoconstrictor and antidromic vasodilator mechanisms, are solely of peripheral origin or have an additional spinal component and act exclusively through neural or also involve humoral pathways. The 54 patients with unilateral reflex sympathetic dystrophy were divided into the following three stages according to their perception of skin temperature in the clinically affected hand: stage I, stationary warmth sensation; stage II, intermittent warmth and cold sensation; and stage III, stationary cold sensation. Investigation of basal skin blood flow and vasoconstrictive response to dependency of skin microvessels in the clinically unaffected hand and the clinically affected hand of patients with reflex sympathetic dystrophy and the left hand of 16 control subjects. Microcirculation was investigated at the predominantly neurally controlled thermoregulatory level (Doppler laser flowmetry) and at the predominantly humorally controlled nutritive level (capillary microscopy). In the clinically unaffected hand, at the thermoregulatory level of the microcirculation: (1) basal skin blood flow was increased at stage I compared with the control subjects, whereas no differences could be observed at this stage compared with the clinically affected hand; (2) the vasoconstrictive response to dependency (defined as skin blood flow at heart level divided by skin blood flow in the dependent position) was attenuated at stage I compared with the control subjects, whereas no differences could be observed at this stage compared with the clinically affected hand; and (3) basal skin blood flow and the vasoconstrictive response to dependency did not differ from the control subjects at stages II and III. In the clinically unaffected hand, at the nutritive level, no differences could be observed at any stage of the syndrome compared with the control subjects. This study indicates that there is a spinal component to microcirculatory abnormalities at stage I of the reflex sympathetic dystrophy syndrome that most likely acts through neural (antidromic vasodilator) mechanisms and that may be initiated by traumatic excitation of a peripheral nerve on the clinically affected side.


Lateral sensitivity modulation explains the flanker effect in contrast discrimination.
We used a dual-masking paradigm to study how contrast discrimination can be influenced by the presence of adjacent stimuli. The task of the observer was to detect a target superimposed on a pedestal in the presence of flankers. The flankers (i) reduce the target threshold at zero pedestal contrast, (ii) shift the target threshold versus pedestal contrast (TvC) function horizontally to the left on a log-log plot at high pedestal contrasts, and (iii) reduce the size of pedestal facilitation at low pedestal contrasts. The horizontal shift at high pedestal contrasts suggests that the flanker effect is a multiplicative factor that cannot be explained by previous models of contrast discrimination. We extend the divisive inhibition model of contrast discrimination by implementing the flanker effect as a lateral multiplicative sensitivity modulation. This extended model provides a good account of the data.


Reactivation of genito-urinary tuberculosis: a retrospective review of 838 cases.
Relapse occurred in 53 cases out of 838 consecutive cases of genito-urinary tuberculosis, followed for periods of 11 months to 27 years. Initial and recurrent disease is classified, and the treatment given is described. Length of inactivity is estimated, and correlated with the type of therapy given. Sensitivity of the organisms to chemotherapy is stated. Conclusions suggest that a life-long follow-up, at yearly intervals, of patients with genito-urinary tuberculosis is justified, as recurrences are diagnosed early, destructive changes are than unlikely, further surgery may be avoided, and a repeat course of chemotherapy is usually curative.


Preschoolers' perception of other people's looking: photographs and drawings.
Children aged 3-4 years were tested for their ability to decide which of two photographs or drawings of a face depicted the act of fixating on a target object; in each control photograph or drawing the same face and object were present without fixation. Performance was above chance on both stimulus types, but low enough to call into question conclusions from previous research. The same children were also tested on their ability to discriminate between photographs/drawings depicting two faces fixating the same object (joint visual attention) and the same two faces fixating different objects. While discrimination of joint visual attention depicted in drawings was as good as discrimination of fixation in the single-face tasks, the ability to reliably choose between a photograph of two people attending to a common object and a control photograph was significantly poorer. The results suggest that, while young infants and children may be highly sensitive to face-on gaze, even well into the fourth year of life children are unable consistently to interpret (1) direction of non-self-directed gaze in static faces and (2) joint visual attention by others.


Language Exposure Relates to Structural Neural Connectivity in Childhood.
Neuroscience research has elucidated broad relationships between socioeconomic status (SES) and young children's brain structure, but there is little mechanistic knowledge about specific environmental factors that are associated with specific variation in brain structure. One environmental factor, early language exposure, predicts children's linguistic and cognitive skills and later academic achievement, but how language exposure relates to neuroanatomy is unknown. By measuring the real-world language exposure of young children (ages 4-6 years, 27 male/13 female), we confirmed the preregistered hypothesis that greater adult-child conversational experience, independent of SES and the sheer amount of adult speech, is related to stronger, more coherent white matter connectivity in the left arcuate and superior longitudinal fasciculi on average, and specifically near their anterior termination at Broca's area in left inferior frontal cortex. Fractional anisotropy of significant tract subregions mediated the relationship between conversational turns and children's language skills and indicated a neuroanatomical mechanism underlying the SES "language gap." Post hoc whole-brain analyses revealed that language exposure was not related to any other white matter tracts, indicating the specificity of this relationship. Results suggest that the development of dorsal language tracts is environmentally influenced, specifically by early, dialogic interaction. Furthermore, these findings raise the possibility that early intervention programs aiming to ameliorate disadvantages in development due to family SES may focus on increasing children's conversational exposure to capitalize on the early neural plasticity underlying cognitive development.SIGNIFICANCE STATEMENT Over the last decade, cognitive neuroscience has highlighted the detrimental impact of disadvantaged backgrounds on young children's brain structure. However, to intervene effectively, we must know which proximal aspects of the environmental aspects are most strongly related to neural development. The present study finds that young children's real-world language exposure, and specifically the amount of adult-child conversation, correlates with the strength of connectivity in the left hemisphere white matter pathway connecting two canonical language regions, independent of socioeconomic status and the sheer volume of adult speech. These findings suggest that early intervention programs aiming to close the achievement gap may focus on increasing children's conversational exposure to capitalize on the early neural plasticity underlying cognitive development.


An evaluation of relugolix/estradiol/norethindrone acetate for the treatment of heavy menstrual bleeding associated with uterine fibroids in premenopausal women.
Uterine Fibroids (UFs) are the most predominant benign tumor in women who are coming of reproductive age, and causes intense economic load priced in billions of US dollars. Historically, surgery has been the main definitive treatment, albeit less attractive nowadays, especially for women with future fertility plans. Therefore, studies to explore the pharmacological treatment options are increasing especially as those that are currently available are limited for short-term use only. This drug evaluation features the clinical results from previous and ongoing studies of relugolix, in combination with the add back therapy of estradiol (E2) and norethindrone acetate (NETA), as a novel, orally administered, nonpeptide antagonist of gonadotropin-releasing hormone (GnRH) for the management of heavy menstrual bleeding (HMB) in premenopausal women with UFs. The combination of relugolix/E2/NETA is an encouraging, well-tolerated and noninvasive pharmacological option for UFs patients. Relugolix induced a concentration-dependent decrease in HMB. However, it should be used with hormonal add-back therapy (E2+ NETA) to avoid induced hypoestrogenic side effects, importantly bone mineral density loss. Moreover, symptoms will likely resume shortly after the termination of the relugolix combination administration.


[Dilemmas in the treatment of psychiatric symptoms in patients with Parkinson's disease].
Psychiatric symptoms are common in Parkinson's disease (PD) and may complicate treatment. To review the prevalence and treatment options of psychiatric symptoms in PD patients and discuss the dilemmas that may arise. Literature review. Psychiatric complaints, including depression, anxiety, apathy, impulse control disorders, hallucinations, delusions, sleep disturbances, and cognitive symptoms, frequently occur in PD patients. These symptoms have a great influence on the general functioning and quality of life of the patient. When treating these symptoms, adjusting neurological treatment and starting or adjusting psychotherapeutic or psychopharmacological treatment may be necessary. Even if individual symptoms can often be treated adequately, unwanted side effects in other symptom domains have to be taken into consideration. Adequate treatment of neuropsychiatric symptoms in PD patients is complex, and requires close multidisciplinary collaboration, especially in more advanced disease stages.


Killing the killer: predation between protists and predatory bacteria.
Predation by microbes is one of the main drivers of bacterial mortality in the environment. In most ecosystems multiple micropredators compete at least partially for the same bacterial resource. Predatory interactions between these micropredators might lead to shifts within microbial communities. Integrating these interactions is therefore crucial for the understanding of ecosystem functioning. In this study, we investigated the predation between two groups of micropredators, i.e. phagotrophic protists and Bdellovibrio and like organisms (BALOs). BALOs are obligate predators of Gram-negative bacteria. We hypothesised that protists can prey upon BALOs despite the small size and high swimming speed of the latter, which makes them potentially hard to capture. Predation experiments including three protists, i.e. one filter feeder and two interception feeder, showed that BALOs are a relevant prey for these protists. The growth rate on BALOs differed for the respective protists. The filter feeding ciliate was growing equally well on the BALOs and on Escherichia coli, whereas the two flagellate species grew less well on the BALOs compared to E. coli. However, BALOs might not be a favourable food source in resource-rich environments as they are not enabling all protists to grow as much as on bacteria of bigger volume.


Genome-Wide Identification and Transcriptional Analysis of Arabidopsis DUF506 Gene Family.
The Domain of unknown function 506 (DUF506) family, which belongs to the PD-(D/E)XK nuclease superfamily, has not been functionally characterized. In this study, 266 DUF506 domain-containing genes were identified from algae, mosses, and land plants showing their wide occurrence in photosynthetic organisms. Bioinformatics analysis identified 211 high-confidence DUF506 genes across 17 representative land plant species. Phylogenetic modeling classified three groups of plant DUF506 genes that suggested functional preservation among the groups based on conserved gene structure and motifs. Gene duplication and Ka/Ks evolutionary rates revealed that DUF506 genes are under purifying positive selection pressure. Subcellular protein localization analysis revealed that DUF506 proteins were present in different organelles. Transcript analyses showed that 13 of the Arabidopsis DUF506 genes are ubiquitously expressed in various tissues and respond to different abiotic stresses and ABA treatment. Protein-protein interaction network analysis using the STRING-DB, AtPIN (Arabidopsis thaliana Protein Interaction Network), and AI-1 (Arabidopsis Interactome-1) tools indicated that AtDUF506s potentially interact with iron-deficiency response proteins, salt-inducible transcription factors, or calcium sensors (calmodulins), implying that DUF506 genes have distinct biological functions including responses to environmental stimuli, nutrient-deficiencies, and participate in Ca(2+) signaling. Current results provide insightful information regarding the molecular features of the DUF506 family in plants, to support further functional characterizations.


Rinderpest virus isolates of different virulence vary in their capacity to infect bovine monocytes and macrophages.
Three isolates of rinderpest virus (RPV) with different in vivo virulence were able to infect and productively replicate in bovine monocytic cells. They differed in their kinetics of replication and the morphological changes induced in infected cultures. The highly virulent RPV-Saudi infected > 80% of cells within 6 days p.i. (m.o.i. = 0.1 TCID50 per cell). Under identical conditions, > 50% of cells were infected by the 'mild' (causes minimal mortality in vivo) isolate RPV-Egypt, whereas only 25% were infected by the avirulent RPV-RBOK. Infection by all three viruses produced infectious progeny, induced the formation of syncytia and stellate cells with long processes, and down-regulated MHC class II expression; there was no apparent effect on MHC class I nor LFA-1. RPV-Saudi was the most efficient at generating progeny virus and producing syncytia. While RPV-RBOK was the least efficient at inducing syncytia, RPV-Egypt was the least efficient for progeny virus production. In contrast, RPV-Egypt was particularly efficient at inducing stellate cell formation and down-regulating MHC class II expression. These results indicate a relationship between in vivo virulence and the characteristics of replication and induced morphological changes in monocytes/macrophages. The down-regulation of MHC class II expression would offer a means by which the virus could evade immune recognition. This would be particularly useful for the more cell-associated, but less efficient at maturing, RPV-Egypt.


Mild senile dementia of Alzheimer type: research diagnostic criteria, recruitment, and description of a study population.
Clinical investigations of senile dementia of the Alzheimer type require establishment of explicit clinical diagnostic criteria before histological confirmation is possible. Criteria for selection of mildly impaired subjects with senile dementia of Alzheimer type, free of other major disease, are proposed. Problems of recruitment of this select population for a longitudinal study are discussed. A study population with matched healthy control subjects has been enrolled and described. Short term follow-up has provided preliminary support for the diagnostic criteria.


[Invasive aspergillosis in immunocompromised patients].
To analyse results of treatment of invasive aspergillesis in immunocompromised patients for 2000-2002. The study was made of patients who, when treated with antibiotics, exhibited foci in the lungs typical for invasive aspergillesis. Aspergillas were detected in the sputum, bronchoalveolar lavage, bronchial wash-ups, aspergilla antigen (galactomannan) was detected in the blood. Invasive aspergillesis was diagnosed in 25 patients. 13 (52%) patients were treated with adjuvant glucocorticoids. 19 (76%) patients had neutropenia. All the patients had fever. Foci in the lungs were in 24 patients. Aspergillas were detected in 15 patients, a positive antigen galactomannan in 7 patients. A. Fumigatus, A flavus, A. Niger occurred in 67, 26.5 and 6.5% patients, respectively. All the patients received amphotericin B (median of the treatment reached 38 days, total dose 880-3500 mg). In 5 patients amphotericin B was replaced for liposomal amphotericin B because of high creatinine. 7 patients continued with itraconasol in a dose 400-600 mg/day. The foci were removed in 3 patients. The cure was achieved in 12 patients, 13 patients, 13 patients died (cause of death--respiratory insufficiency). Lethality in invasive aspergillesis in immunocompromised patients remains high--52%. Cultural detection of mycelial fungi was, as a rule, delayed. Early diagnosis of the disease requires monitoring of the aspergilla antigen in the blood and computer tomography of the chest especially in fever persisting in the treatment of wide-spectrum antibiotics.


BRAFK601E Mutation in a Follicular Thyroid Adenoma: A Case Report.
BRAF mutations represent the most common genetic alteration in papillary thyroid carcinoma (PTC). The p.V600E mutation is specific for the classic and tall-cell variants of PTC and has been associated with a more aggressive biologic behavior. On the other hand, the p.K601E mutation is peculiar to the follicular variant of PTC, and seems to be a favorable prognostic indicator. A 12-year-old boy presented with a 10-mm left-sided thyroid nodule. Fine-needle aspiration cytology reported the lesion as suspicious for a follicular neoplasm (Bethesda category IV). The patient underwent lobectomy, and histopathology revealed a follicular adenoma with normal surrounding tissue. The cytological smear was found to be positive for BRAF p.K601E mutation, and this was later confirmed on the corresponding paraffin block. This case was independently revised by 4 expert pathologists, all of whom confirmed the benign nature of the thyroid lesion. This article describes the presence of a BRAF mutation in a benign thyroid lesion. To the authors' knowledge, this is the fourth case of follicular adenoma carrying BRAFK601E reported in literature to date. BRAFK601E mutation can occur in benign thyroid lesions. This finding, in the context of the current literature and the recently proposed reclassification of the noninvasive encapsulated follicular variant of papillary thyroid carcinoma into a benign lesion, confirms the importance of preoperative BRAF p.K601E testing in offering patients a tailored treatment plan and avoiding overtreatment.


The clinico-epidemiological, diagnostic and therapeutic aspects of methanol poisoning: A five-year retrospective study, northern Iran.
Methanol poisoning (MP) is a serious health issue that has become more prevalent in recent years and has resulted in numerous deaths. Early detection and timely treatment are critical for preventing fatalities and reducing the incidence of neurological complications. This study was designed as a retrospective investigation with the purpose of analyzing the clinico-epidemiological, diagnostic, and therapeutic aspects of patients who were admitted to two training hospitals in northern Iran due to MP. The selection of samples for this study was based on a pre-defined checklist. Following the completion of the treatment period in the hospital, the patients were categorized into three groups based on their clinical outcome. All relevant variables for each group were recorded and reported separately, using the SciPy library in the Python programming language. The majority of the patients (88.12%) were male (P=0.012), the average age was 41.46, and mostly (82.18%) lived in urban regions (P=0.025). The primary clinical complaint reported was visual disorders, accounting for 75.25% of the cases, followed by nervous, gastrointestinal, respiratory, and chest pain accordingly. The average hospitalization length for the patients was 5.065 days. Out of the 101 patients, 65 (64.36%) were discharged without any complications, 17 (16.83%) were discharged with complications, and unfortunately, 19 (18.81%) were died. A decreased level of consciousness and severe metabolic acidosis are commonly associated with unfavorable outcomes in MP. The use of systemic corticosteroids as a treatment method has a significant association with reducing mortality rates.


Neural computation approach for developing a 3D shape reconstruction model.
The shape from shading problem refers to the well-known fact that most real images usually contain specular components and are affected by unknown reflectivity. In this paper, these limitations are addressed and a new neural-based 3D shape reconstruction model is proposed. The idea behind this approach is to optimize a proper reflectance model by learning the parameters of the proposed neural reflectance model. In order to do this, new neural-based reflectance models are presented. The feedforward neural network (FNN) model is able to generalize the diffuse term, while the RBF model is able to generalize the specular term. A hybrid structure of FNN-based and RBF-based models is also presented because most real surfaces are usually neither Lambertian models nor ideally specular models. Experimental results, including synthetic and real images, are presented to demonstrate the performance of our approach given different specular effects, unknown illuminate conditions, and different noise environments.


The Role of the Hindfoot in Total Knee Arthroplasty Alignment.
Limb alignment is a critically important factor to consider in the management of the patient with knee arthritis. Abnormal alignment is associated with the accelerated progression of osteoarthritis and, if not addressed at the time of surgery, may contribute to early failure of knee replacement implants. The contribution of the hindfoot to overall limb alignment has received limited attention in the context of deformity correction in total knee arthroplasty (TKA). In this review, we present evidence supporting the inclusion of the hindfoot in the consideration of overall limb alignment for TKA and propose a management algorithm.


Physical activity and predisposition for hip fractures: a review.
Studies on the association between physical activity and hip fractures are reviewed. All the studies, which comprise four follow-up studies, one nested case-control study and 17 case-control studies, suggest a protective effect of physical activity with regard to hip fractures. The association is strong and consistent with physical activity in leisure, weaker with respect to physical activity at work. The association is present for physical activity from childhood to adult age, and it is consistent in study populations from the USA, Australia, Asia and Northern and Southern Europe, in spite of very different hip fractures incidences in these populations. The magnitude of the association is difficult to assess because of varying criteria for exposure, but to be among the physically active seems to reduce the risk of later hip fracture by up to 50%. It seems that even daily chores, such as climbing stairs and walking, protect against hip fracture.


Profilin 1 associates with stress granules and ALS-linked mutations alter stress granule dynamics.
Mutations in the PFN1 gene encoding profilin 1 are a rare cause of familial amyotrophic lateral sclerosis (ALS). Profilin 1 is a well studied actin-binding protein but how PFN1 mutations cause ALS is unknown. The budding yeast, Saccharomyces cerevisiae, has one PFN1 ortholog. We expressed the ALS-linked profilin 1 mutant proteins in yeast, demonstrating a loss of protein stability and failure to restore growth to profilin mutant cells, without exhibiting gain-of-function toxicity. This model provides for simple and rapid screening of novel ALS-linked PFN1 variants. To gain insight into potential novel roles for profilin 1, we performed an unbiased, genome-wide synthetic lethal screen with yeast cells lacking profilin (pfy1Δ). Unexpectedly, deletion of several stress granule and processing body genes, including pbp1Δ, were found to be synthetic lethal with pfy1Δ. Mutations in ATXN2, the human ortholog of PBP1, are a known ALS genetic risk factor and ataxin 2 is a stress granule component in mammalian cells. Given this genetic interaction and recent evidence linking stress granule dynamics to ALS pathogenesis, we hypothesized that profilin 1 might also associate with stress granules. Here we report that profilin 1 and related protein profilin 2 are novel stress granule-associated proteins in mouse primary cortical neurons and in human cell lines and that ALS-linked mutations in profilin 1 alter stress granule dynamics, providing further evidence for the potential role of stress granules in ALS pathogenesis.


Nursing care of people with dementia in an orthopaedic acute care setting: An integrative literature review.
This integrative literature review is to collect what is known about the care of people with dementia when they require a hospital admission for an orthopaedic surgical procedure and to contribute to developing an evidence-base to support nursing practice when caring for people with dementia in an orthopaedic setting. People with a dementia diagnosis are increasingly common in acute orthopaedic care settings and the admission exposes people with dementia to risks during their hospital stay. In addition, nurses find people with dementia challenging to care for due to the complexity of dual conditions. Little is known specifically about the care requirements for people with dementia in orthopaedic settings. Integrative literature review. An integrative literature review and qualitative deductive content analysis using McCormack and McCance's theoretical nursing framework (Person-Centred Nursing Framework) of nine studies were undertaken. The process of the review was guided by PRISMA checklist. The care environment and resistance either in passive form, or through physical intervention, is common in orthopaedic nursing. Planning and delivering care for physical, cognitive and emotional needs is identified as being difficult, resulting in a lack of inclusion for patients, partly due to communication challenges. Finding ways to implement tailored care plans within standard ward routines proves difficult, and the consequence is a less than optimal care experience with adverse effects on patients characterised by an increase in dementia symptoms. Care for people with dementia in an orthopaedic setting is complex. It needs to be further studied so that more evidence and supporting literature can contribute to improved care for this group of patients. This study describes the complexity of providing fundamental care for people with dual conditions of dementia and orthopaedic injury and suggests opportunities for improvement.


Early pandemic influenza (2009 H1N1) in Ho Chi Minh City, Vietnam: a clinical virological and epidemiological analysis.
To date, little is known about the initial spread and response to the 2009 pandemic of novel influenza A ("2009 H1N1") in tropical countries. Here, we analyse the early progression of the epidemic from 26 May 2009 until the establishment of community transmission in the second half of July 2009 in Ho Chi Minh City (HCMC), Vietnam. In addition, we present detailed systematic viral clearance data on 292 isolated and treated patients and the first three cases of selection of resistant virus during treatment in Vietnam. Data sources included all available health reports from the Ministry of Health and relevant health authorities as well as clinical and laboratory data from the first confirmed cases isolated at the Hospital for Tropical Diseases in HCMC. Extensive reverse transcription (RT)-PCR diagnostics on serial samples, viral culture, neuraminidase-inhibition testing, and sequencing were performed on a subset of 2009 H1N1 confirmed cases. Virological (PCR status, shedding) and epidemiological (incidence, isolation, discharge) data were combined to reconstruct the initial outbreak and the establishment of community transmission. From 27 April to 24 July 2009, approximately 760,000 passengers who entered HCMC on international flights were screened at the airport by a body temperature scan and symptom questionnaire. Approximately 0.15% of incoming passengers were intercepted, 200 of whom tested positive for 2009 H1N1 by RT-PCR. An additional 121 out of 169 nontravelers tested positive after self-reporting or contact tracing. These 321 patients spent 79% of their PCR-positive days in isolation; 60% of PCR-positive days were spent treated and in isolation. Influenza-like illness was noted in 61% of patients and no patients experienced pneumonia or severe outcomes. Viral clearance times were similar among patient groups with differing time intervals from illness onset to treatment, with estimated median clearance times between 2.6 and 2.8 d post-treatment for illness-to-treatment intervals of 1-4 d, and 2.0 d (95% confidence interval 1.5-2.5) when treatment was started on the first day of illness. The patients described here represent a cross-section of infected individuals that were identified by temperature screening and symptom questionnaires at the airport, as well as mildly symptomatic to moderately ill patients who self-reported to hospitals. Data are observational and, although they are suggestive, it is not possible to be certain whether the containment efforts delayed community transmission in Vietnam. Viral clearance data assessed by RT-PCR showed a rapid therapeutic response to oseltamivir.


Production of Long Chain Fatty Alcohols Found in Bumblebee Pheromones by Yarrowia lipolytica.
Fatty alcohols (FA-OH) are aliphatic unbranched primary alcohols with a chain of four or more carbon atoms. Besides potential industrial applications, fatty alcohols have important biological functions as well. In nature, fatty alcohols are produced as a part of a mixture of pheromones in several insect species, such as moths, termites, bees, wasps, etc. In addition, FA-OHs have a potential for agricultural applications, for example, they may be used as a suitable substitute for commercial insecticides. The insecticides have several drawbacks associated with their preparation, and they exert a negative impact on the environment. Currently, pheromone components are prepared mainly through the catalytic hydrogenation of plant oils and petrochemicals, which is an unsustainable, ecologically unfriendly, and highly expensive process. The biotechnological production of the pheromone components using engineered microbial strains and through the expression of the enzymes participating in the biosynthesis of these components is a promising approach that ensures ecological sustenance as well. The present study was aimed at evaluating the production of FA-OHs in the oleaginous yeast, Yarrowia lipolytica, with different lengths of fatty-acyl chains by expressing the fatty acyl-CoA reductase (FAR) BlapFAR4 from B. lapidarius, producing C16:0-OH, C16:1Δ9-OH, and lower quantities of both C14:0-OH and C18:1Δ9-OH, and BlucFAR1 from B. lucorum, producing FA-OHs with a chain length of 18-26 carbon atoms, in this yeast. Among the different novel Y. lipolytica strains used in the present study, the best results were obtained with JMY7086, which carried several lipid metabolism modifications and expressed the BlucFAR1 gene under the control of a strong constitutive promoter 8UAS-pTEF. JMY7086 produced only saturated fatty alcohols with chain lengths from 18 to 24 carbon atoms. The highest titer and accumulation achieved were 166.6 mg/L and 15.6 mg/g DCW of fatty alcohols, respectively. Unlike JMY7086, the BlapFAR4-expressing strain JMY7090 produced only 16 carbon atom-long FA-OHs with a titer of 14.6 mg/L.


Searching for convergent evolution in manganese superoxidase dismutase using hydrophobic cluster analysis.
There are numerous examples of convergent evolution in nature. Major ecological adaptations such as flight, loss of limbs in vertebrates, pesticide resistance, adaptation to a parasitic way of life, etc., have all evolved more than once, as seen by their analogous functions in separate taxa. But what about protein evolution? Does the environment have a strong enough influence on intracellular processes that enzymes and other functional proteins play, to evolve similar functional roles separately in different organisms? Manganese Superoxide Dismutase (MnSOD) is a manganesedependant metallo-enzyme which plays a crucial role in protecting cells from anti-oxidative stress by eliminating reactive (superoxide) oxygen species. It is a ubiquitous housekeeping enzyme found in nearly all organisms. In this study we compare phylogenies based on MnSOD protein sequences to those based on scores from Hydrophobic Cluster Analysis (HCA). We calculated HCA similarity values for each pair of taxa to obtain a pair-wise distance matrix. A UPGMA tree based on the HCA distance matrix and a common tree based on the primary protein sequence for MnSOD was constructed. Differences between these two trees within animals, enterobacteriaceae, planctomycetes and cyanobacteria are presented and cited as possible examples of convergence. We note that several residue changes result in changes in hydrophobicity at positions which apparently are under the effect of positive selection.


Molecular switches as photocontrollable "smart" receptors.
This critical review focuses on the development of photochromic compounds as sensors for cations, anions, and biologically important molecules. The review commences with a brief description of photochromism and the strategies to exploit photochromic molecular switches' properties for sensing application. This is followed by a summary of photoswitchable receptors emerged to date and classified according to the photochromic structure they are based on. These include azobenzenes, fulgides, dithienylethenes, dihydroindolizines, chromenes and spiropyrans.


Preparing to work with big data in proteomics - a report on the HUPO-PSI Spring Workshop: April 15-17, 2013, Liverpool, UK.
The Human Proteome Organisation Proteomics Standards Initiative (HUPO-PSI) was established in 2002 with the aim of defining community standards for data representation in proteomics and facilitating data comparison, exchange and verification. The 2013 annual spring workshop was hosted by the University of Liverpool, UK and concentrated on updating and refining the existing standards in the light of new methodologies and technologies. To control the inflation of file sizes, strategies for file compression, particularly for mzML files, were explored. Best practices for encoding information such as protein grouping and PTM localisation were refined and documented. Additional example files for the mzQuantML format were designed to provide support for selected reaction monitoring techniques. Enhancements to the PSI Common Query Interface (PSICQUIC) and PSI-MI XML were discussed. Finally, the group engaged in discussion on how the existing work of the HUPO-PSI can be leveraged by the Metabolomics Standards Initiative to improve the capture of metabolite data.


Phylogenetic analysis of the rompB genes of Rickettsia felis and Rickettsia prowazekii European-human and North American flying-squirrel strains.
The rickettsial outer membrane protein B (rompB) gene encodes the major surface antigens of Rickettsia species. We undertook sequencing and molecular analysis of the rompB gene of Rickettsia felis and a comparison with its homologs in spotted fever group (SFG) and typhus group (TG) rickettsiae, including the complete sequences of two North American flying squirrel strains and two European human strains of Rickettsia prowazekii. We sequenced 5,226 base pairs (bp) of the R. felis rompB, encoding a protein of 1,654 amino acids. We also sequenced 5,015 bp of rompB of the flying squirrel strains, encoding a protein of 1,643 amino acids. Analysis of the R. felis rompB gene sequence showed 10-13% divergence from SFG rickettsiae and 18% divergence from the TG rickettsiae. The rompB of all sequenced strains of R. prowazekii showed an overall similarity of 99.7-99.9%.


Lung epithelial cells and extracellular matrix components induce expression of Pneumocystis carinii STE20, a gene complementing the mating and pseudohyphal growth defects of STE20 mutant yeast.
Pneumocystis carinii causes severe pneumonia in immunocompromised hosts. The binding of P. carinii to alveolar epithelial cells and extracellular matrix constituents such as fibronectin and vitronectin is a central feature of infection, which initiates proliferation of the organism. Herein, we demonstrate that P. carinii binding to lung cells specifically alters the gene expression of the organism, regulating fungal growth. Subtractive hybridization was performed to isolate P. carinii genes expressed following binding to mammalian extracellular matrix constituents. P. carinii STE20 (PCSTE20), a gene participating in mating and pseudohyphal growth of other fungi, was identified following adherence to the extracellular matrix constituents fibronectin, vitronectin, collagen, and lung epithelial cells. The expression of PCSTE20 and a related P. carinii mitogen-activated protein kinase (MAPK) kinase gene, also implicated in signaling of mating, were both specifically upregulated by binding to matrix protein. The expression of general cyclin-dependent kinases and other MAPKs not involved in mating pathways were not altered by organism binding. PCSTE20 expression was also strongly enhanced following organism attachment to A549 lung epithelial cells. When expressed in a Saccharomyces cerevisiae ste20Delta mutant, PCSTE20 suppressed defects in both mating and pseudohyphal growth. These findings are consistent with the observed proliferation and filopodial extension of Pneumocystis organisms adherent to the epithelium in the lungs of immunocompromised hosts. PCSTE20 expression appears to represent a significant component in the regulation of the life cycle of this intractable opportunistic pathogen.


Comparison of human metapneumovirus, respiratory syncytial virus and influenza A virus lower respiratory tract infections in hospitalized young children.
We compared the clinical and demographic features of children with lower respiratory tract infection (LRI) caused by human metapneumovirus (HMPV), respiratory syncytial virus (RSV) and influenza A virus and sought to determine whether coinfection by HMPV and other respiratory viruses leads to increased disease severity. Nasal wash specimens were prospectively obtained from 516 children hospitalized for LRI during a 1-year period and tested for the presence of HMPV by reverse transcription-polymerase chain reaction and for RSV and influenza A by direct immunofluorescence. HMPV was detected in 68 (13%) patients and was the third most common viral pathogen; 16 of 68 HMPV-positive children (24%) had coinfection with other respiratory viruses (HMPVco).HMPV patients were older than RSV patients (17.6 +/- 16.8 months versus 10.5 +/- 11.8 months, P = 0.02). HMPV was associated with wheezing and hypoxemia at a rate similar to that of RSV and higher than that of influenza A. Atelectasis was more common among HMPV (40%) than among RSV and influenza patients (13%, P < 0.05 for each). HMPV infection was more often associated with a diagnosis of pneumonia than RSV and influenza A and was more often associated with a diagnosis of asthma and less often associated with a diagnosis of bronchiolitis than RSV infection (P < 0.05 for each), even when corrected for age. Children with HMPVco had a higher rate of gastrointestinal symptoms but did not show a more severe respiratory picture. The clinical pattern of HMPV more closely resembles that of RSV than that of influenza A LRI, yet the differences in age, radiographic findings and clinical diagnosis suggest that HMPV pathogenesis may differ from that of RSV.


A bio-functions integration microcosm: Self-immobilized biochar-pellets combined with two strains of bacteria to remove atrazine in water and mechanisms.
A self-immobilization method for microorganisms was developed based on fungal pellets. Generally, pellets have some problems such as cell leakage, cell loading limitation and low mechanical strength. Therefore, biochar was applied to overcome these disadvantages. Atrazine degradable microorganism Arthrobacter sp. ZXY-2 was immobilized by Aspergillus niger Y3 pellets. After adding biochar with optimal dosage (0.006 g biochar for 0.3 g pellets with ZXY-2), the self-immobilized biomixture (SIB) removed 50 mg /L atrazine rapidly within 1 h, which was 61% higher compared to pellets without biochar. The kinetic adsorption results showed that the biosorption of biochar by pellets followed a pseudo-second-order kinetic model. The ATZ removal ability and reusability of SIB were significantly increased by biochar. The results showed that the addition of biochar could enhance the connection between ZXY-2 and pellets based carrier, and the favorable biodegradation pH of ZXY-2 changed to 6 and 10. Several analyses such as ζ-potential measurements, FTIR, XPS, SEM-EDS, and elemental analyses were performed to evaluate the mechanism of action of SIB. To enhance the ATZ degradation by single strain, Agrobacterium, sp WL-1 was isolated and added. The metabolic pathways and their function complementation were studied. Furthermore, a biomass integration model for wastewater treatment was proposed herein.


The vitamin D status of Canadians relative to the 2011 Dietary Reference Intakes: an examination in children and adults with and without supplement use.
The 2011 Dietary Reference Intakes (DRIs) for vitamin D use 25-hydroxyvitamin D [25(OH)D] concentrations to define vitamin D deficiency (<30 nmol/L), the Estimated Average Requirement (40 nmol/L), and the Recommended Dietary Allowance (RDA; 50 nmol/L). The Canadian population has not yet been assessed according to these recommendations. We determined the prevalence of meeting DRI recommendations and the role of vitamin D supplement use among Canadians aged 6-79 y. Plasma 25(OH)D from a representative sample of Canadians in the Canadian Health Measures Survey-Cycle 1 (n = 5306) were used. Supplement use was assessed by household interview. Concentrations of 25(OH)D were compared in supplement users and nonusers by season and race. Overall, 5.4%, 12.7%, and 25.7% of the participants had 25(OH)D concentrations below the 30-, 40-, and 50-nmol/L cutoffs, respectively. In white Canadians, plasma 25(OH)D concentrations ranged from an undetectable percentage with concentrations <30 nmol/L in summer to 24.5% with concentrations <50 nmol/L in winter; the corresponding values ranged from 12.5% to 53.1% in nonwhite Canadians. Supplement users had significantly higher 25(OH)D concentrations than did nonusers, and no seasonal differences were found. In nonsupplement users, the prevalence of 25(OH)D concentrations <50 nmol/L in winter was 37.2% overall and was 60.7% in nonwhites. One-quarter of Canadians did not meet the RDA, but the use of vitamin D supplements contributed to a better 25(OH)D status. Nonwhite Canadians had the highest risk of not achieving DRI recommendations. More than one-third of Canadians not using supplements did not meet the RDA in winter. This suggests that current food choices alone are insufficient to maintain 25(OH)D concentrations of 50 nmol/L in many Canadians, especially in winter.


Effect of pre- and postpartum supplementation with lipid-encapsulated conjugated linoleic acid on reproductive performance and the growth hormone-insulin-like growth factor-I axis in multiparous high-producing dairy cows.
The objective of the study was to evaluate the effect of prepartum and postpartum (PP) supplementation with 2 isomers of conjugated linoleic acid (CLA) on reproductive parameters and some related metabolic factors in dairy cows. High-producing, multiparous Holstein Friesian cows (n = 60) were allotted to 3 treatment groups: the CLA1 group (n = 20) was supplemented with 70 g of lipid-encapsulated CLA providing 7 g each of cis-9,trans-11 and trans-10,cis-12 CLA from d 21 (d 21) before expected calving until d 7 after artificial insemination (AI), that is, until 77 to 91 d PP; the CLA2 group (n = 20) was supplemented with the same amount of CLA beginning at calving until d 7 after AI; and the control group (n = 20) received an isocaloric, isonitrogenous, and isolipidic diet. Blood samples were taken weekly to measure glucose, insulin, insulin-like growth factor-I (IGF-I), and leptin. Liver biopsy was performed in 10 cows per group for growth hormone receptor 1A and IGF-I mRNA analyses. At d 49 to 63 PP, ovulation was synchronized with the Pre-Synch protocol followed by fixed-time AI. Milk progesterone was monitored from calving until d 35 post-AI. Cows returning to estrus following AI were inseminated. Supplementation with CLA before calving improved the recovery of plasma leptin levels in the early PP period (from the day of calving until wk 3 PP; treatment effect). Later PP (wk 5), plasma IGF-I, and leptin remained significantly higher in both CLA1 and CLA2 groups compared with control, although hepatocellular IGF-I mRNA was not different among groups. Plasma IGF-I levels remained higher in both CLA-treated groups on the day of AI. Growth hormone receptor 1A mRNA levels in hepatic tissue decreased in all groups, reaching a nadir in the first week PP. Days to first PP ovulation did not differ between groups; however, both supplemented groups conceived earlier compared with control (d 97 ± 19, d 97 ± 23, and d 113 ± 30 for CLA1, CLA2, and control, respectively). Plasma progesterone concentration was higher in both supplemented groups on d 2 to 5 following the synchronized ovulation than in controls. We concluded that CLA supplementation around calving alters PP metabolic signals as reflected by higher plasma leptin and IGF-I levels. Conjugated linoleic acid stimulated early luteal function and reduced the PP interval to conception.


Reversal of diurnal cortisol rhythm and suppression of plasma testosterone in obstetric residents on call.
The study was undertaken to quantify the psychologic and physiologic responses to the stresses of an obstetrics/gynecology residency program. Six male residents were studied on four occasions: one day during the first 2 weeks of their residency, one day immediately following a vacation period, one day after a night on call in obstetrics, and one day while in the gynecology clinic. Stress was evaluated by validated psychologic instruments and by levels of plasma testosterone, luteinizing hormone (LH), cortisol, and prolactin in morning and afternoon blood samples. Self-reported stress was significantly elevated during the first 2 weeks of the residency after a night on call. Anxiety scores were significantly elevated after a night on call as were depression subscores for some residents. Plasma testosterone was highly significantly suppressed after the obstetrics night on call and during the first 2 weeks of the residency in comparison with the vacation period. Luteinizing hormone levels were also significantly lower after the obstetrics on-call experience. Plasma cortisol levels after a night on call were suppressed in the morning and normal or elevated in the afternoon. In comparison, the gynecology rotation was associated with normal levels of testosterone and lower levels of cortisol. In this small sample of residents, we observed an inverse relationship between self-reported stress levels and the concentrations of plasma testosterone and LH. The high levels of stress and anxiety expressed after a night on call also disrupted the normal pattern of plasma cortisol levels.


A randomized, double-blind, placebo-controlled trial of eszopiclone for the treatment of insomnia in patients with chronic low back pain.
Insomnia, which is very common in patients with chronic low back pain (LBP), has long been viewed as a pain symptom that did not merit specific treatment. Recent data suggest that adding insomnia therapy to pain-targeted treatment should improve outcome; however, this has not been empirically tested in LBP or in any pain condition treated with a standardized pain medication regimen. We sought to test the hypothesis that adding insomnia therapy to pain-targeted treatment might improve sleep and pain in LBP. Double-blind, placebo-controlled, parallel-group, 1-mo trial. Duke University Medical Center Outpatient Sleep Clinic. Fifty-two adult volunteers with LBP of at least 3 mo duration who met diagnostic criteria for insomnia (mean age: 42.5 y; 63% females). Subjects were randomized to eszopiclone (ESZ) 3 mg plus naproxen 500 mg BID or matching placebo plus naproxen 500 mg twice a day. ESZ SIGNIFICANTLY IMPROVED TOTAL SLEEP TIME (MEAN INCREASE: ESZ, 95 min; placebo, 9 min) (primary outcome) and nearly all sleep measures as well as visual analog scale pain (mean decrease: ESZ, 17 mm; placebo, 2 mm) (primary pain outcome), and depression (mean Hamilton Depression Rating Scale improvement ESZ, 3.8; placebo, 0.4) compared with placebo. Changes in pain ratings were significantly correlated with changes in sleep. The addition of insomnia-specific therapy to a standardized naproxen pain regimen significantly improves sleep, pain, and depression in patients with chronic low back pain (LBP). The findings indicate the importance of administering both sleep and pain-directed therapies to patients with LBP in clinical practice and provide strong evidence that improving sleep disturbance may improve pain. clinicaltrials.gov identifier: NCT00365976.


STIM1 Is Required for Remodeling of the Endoplasmic Reticulum and Microtubule Cytoskeleton in Steering Growth Cones.
The spatial and temporal regulation of calcium signaling in neuronal growth cones is essential for axon guidance. In growth cones, the endoplasmic reticulum (ER) is a significant source of calcium signals. However, it is not clear whether the ER is remodeled during motile events to localize calcium signals in steering growth cones. The expression of the ER-calcium sensor, stromal interacting molecule 1 (STIM1) is necessary for growth cone steering toward the calcium-dependent guidance cue BDNF, with STIM1 functioning to sustain calcium signals through store-operated calcium entry. However, STIM1 is also required for growth cone steering away from semaphorin-3a, a guidance cue that does not activate ER-calcium release, suggesting multiple functions of STIM1 within growth cones (Mitchell et al., 2012). STIM1 also interacts with microtubule plus-end binding proteins EB1/EB3 (Grigoriev et al., 2008). Here, we show that STIM1 associates with EB1/EB3 in growth cones and that STIM1 expression is critical for microtubule recruitment and subsequent ER remodeling to the motile side of steering growth cones. Furthermore, we extend our data in vivo, demonstrating that zSTIM1 is required for axon guidance in actively navigating zebrafish motor neurons, regulating calcium signaling and filopodial formation. These data demonstrate that, in response to multiple guidance cues, STIM1 couples microtubule organization and ER-derived calcium signals, thereby providing a mechanism where STIM1-mediated ER remodeling, particularly in filopodia, regulates spatiotemporal calcium signals during axon guidance.SIGNIFICANCE STATEMENT Defects in both axon guidance and endoplasmic reticulum (ER) function are implicated in a range of developmental disorders. During neuronal circuit development, the spatial localization of calcium signals controls the growth cone cytoskeleton to direct motility. We demonstrate a novel role for stromal interacting molecule 1 (STIM1) in regulating microtubule and subsequent ER remodeling in navigating growth cones. We show that STIM1, an activator of store-operated calcium entry, regulates the dynamics of microtubule-binding proteins EB1/EB3, coupling ER to microtubules, within filopodia, thereby steering growth cones. The STIM1-microtubule-ER interaction provides a new model for spatial localization of calcium signals in navigating growth cones in the nascent nervous system.


[hrGH treatment of glucocorticoid-induced short stature in children].
The treatment of systemic diseases with glucocorticoids is often associated with decreased height velocity (HV), and can result in shorter final height. Interactions between adrenal and GH-IGF axis have been described and can occur at hypothalamic-pituitary level or at the regulation of IGF system, including the IGF1R signaling. The clinical state of these patients may be considered as an absolute and/or functional IGF-1 deficiency. Interventions aiming to restore the normal function of GH-IGF axis might reduce the glucocorticoids-induced growth suppression in these children. It has been shown that recombinant human GH (hrGH) induces an increase in HV and a decrease in protein loss in patients with juvenile idiopathic arthritis treated with glucocorticoids. Significant increment in HV was also described after hrGH treatment in children under glucocorticoid therapy due to inflammatory bowel disease or renal transplantation. There is a positive correlation between HV and the dose of hrGH. The results support that the IGF-1 deficiency in these children may be counteract by hrGH therapy. The effect of hrGH is observed only during the treatment period and depends on the replacement strategy, nutritional status and disease control.