[Expression of nephrin, podocin and a-actinine-4 in renal tissue of patients with proteinuria].
Slit diaphragm and/or podocyte's cytoskeleton alterations are related to proteinuria and nephrotic syndrome. In our population, focal and segmental glomerulosclerosis causing nephrotic syndrome is the more frequent biopsy demonstrated glomerulopathy. Our aim was search for alterations in some slit diaphragm-associated proteins in patients with nephrotic range proteinuria. Renal tissue from 40 patients with nephrotic range proteinuria, 10 patients with non-nephrotic proteinuria, 3 with isolated hematuria, and 10 samples of normal renal tissue (deceased donors) were studied, by indirect immunofluorescence, for expression of nephrin, podocin, and alpha-actinin-4. Expression of these proteins was lineal, homogeneous, in the glomerular capillary walls in normal renal tissue and in patients with isolated hematuria. In nephrotic proteinuria this normal appearance was altered and immunostaining showed a fine granular appearance. In 18 cases (45%) of patients with nephrotic proteinuria and 3 cases (30%) of patients with non-nephrotic proteinuria there was loss of at least one of these proteins (p = 0.49). These alterations were found in the diverse glomerulopathies more frequently causing nephrotic syndrome. In nephrotic range proteinuria redistribution or loss of expression of slit diaphragm-associated proteins is very frequent. In many of our cases this fact could be more a consequence than a cause of proteinuria. These alterations can be also evidenced in patients with non-nephrotic proteinuria.


Prevention of Cholesterol Gallstone Formation by Lactobacillus acidophilus ATCC 43121 and Lactobacillus fermentum MF27 in Lithogenic Diet-Induced Mice.
The objective of this study was to evaluate the effects of Lactobacillus acidophilus ATCC 43121 and L. fermentum MF27 on biochemical indices in the serum, cholesterol metabolism in the liver and mucin expression in the gallbladder in lithogenic diet (LD)-induced C57BL/6J mice to determine the preventive effects of lactobacilli on gallstone formation. By the end of 4 wk of the experimental period, mice fed on a LD with high-fat and high-cholesterol exhibited higher levels of total and low-density lipoprotein cholesterol in the serum compared to mice fed on control diet or LD with L. acidophilus ATCC 43121 (LD+P1; p<0.05). Cholesterol-lowering effects observed in the LD+P1 and LD with L. fermentum MF27 (LD+P2) groups were associated with reduced expression of 3-hydroxy-3-methylglutaryl coenzyme A reductase in the liver compared to the LD group (p<0.05). Furthermore, expression of the gel-forming mucin, including MUC5AB and MUC5B, was suppressed in the LD+P1 and LD+P2 groups compared to the LD group (p<0.05). Therefore, steady intake of both L. acidophilus ATCC 43121 and L. fermentum MF27 may have the ability to prevent the formation of cholesterol gallstones in LD-induced C57BL/6J mice.


The role of c-MYB in benzene-initiated toxicity.
Chronic exposure to benzene has been correlated with increased oxidative stress and leukemia. Oncogene activation, including c-Myb activation, is one of the earliest steps leading to the formation of leukemic cells, however the molecular mechanisms involved in these events are poorly understood. Given that oxidative stress can alter the activity and fate of cell signaling pathways we hypothesize that the bioactivation of benzene leads to the formation of reactive oxygen species (ROS), which if not detoxified can alter the c-Myb signaling pathway. Using chicken erythroblast HD3 cells we have shown that exposure to the benzene metabolites catechol, benzoquinone, and hydroquinone leads to increased c-Myb activity, increased phosphorylation of c-Myb and increased production of ROS supporting our hypothesis. Activation of the aryl hydrocarbon receptor (AhR) by environmental contaminants has also been associated with carcinogenesis and mice lacking this receptor are resistant to benzene-initiated hematotoxicity. Using wild type and AhR deficient cells we are investigating the role of this receptor in benzene-initiated alterations in the c-Myb signaling pathway. We have found that both wild type and AhR deficient cells are sensitive to catechol and hydroquinone-initiated increases in c-Myb activity while both cell types are resistant to benzene-initiated alterations leaving the role of the AhR still undetermined. Interestingly, protein expression of c-Myb is increased after catechol exposure in AhR deficient cells while decreased in wild-type cells. Further studies on the role of the AhR in benzene-initiated alterations on the c-Myb signaling pathway are on going.


Inhibition of neural crest migration in Xenopus using antisense slug RNA.
Based primarily on studies in the chick, it has been assumed that the zinc finger transcription factor Slug is required for neural crest migration. In the mouse, however, Slug is not expressed in the premigratory neural crest, which forms normally in Slug -/- animals. To study the role of Slug in Xenopus laevis, we used the injection of XSlug antisense RNA and tissue transplantation. Injection of Slug antisense RNA did not suppress the early expression of the related gene XSnail, but led to reduced expression of both XSlug and XSnail in later stage embryos, whereas the expression of another neural crest marker, XTwist, was not affected. Down-regulation of XSlug and XSnail was associated with the inhibition of neural crest cell migration and the reduction or loss of many neural crest derivatives. In particular, the formation of rostral cartilages was often highly aberrant, whereas the posterior cartilages were less frequently affected. The effects of Slug antisense RNA on neural crest migration and cartilage formation were rescued by the injection of either XSlug or XSnail mRNA. These studies indicate that XSlug is required for neural crest migration, that XSlug and XSnail may be functionally redundant, and that both genes are required to maintain each other's expression in the neural crest development of xenopus laevis.


Genetic parameters estimated with multitrait and linear spline-random regression models using Gelbvieh early growth data.
Estimates of direct and maternal genetic parameters in beef cattle were obtained with a random regression model with a linear spline function (SFM) and were compared with those obtained by a multitrait model (MTM). Weight data of 18,900 Gelbvieh calves were used, of which 100, 75, and 17% had birth (BWT), weaning (WWT), and yearling (YWT) weights, respectively. The MTM analysis was conducted with a three-trait maternal animal model. The MTM included an overall linear partial fixed regression on age at recording for WWT and YWT, and direct-maternal genetic and maternal permanent environmental effects. The SFM included the same effects as MTM, plus a direct permanent environmental effect and heterogeneous residual variance. Three knots, or breakpoints, were set to 1, 205, and 365 d. (Co)variance components in both models were estimated with a Bayesian implementation via Gibbs sampling using flat priors. Because BWT had no variability of age at recording, there was good agreement between corresponding components of variance estimated from both models. For WWT and YWT, with the exception of the sum of direct permanent environmental and residual variances, there was a general tendency for SFM estimates of variances to be lower than MTM estimates. Direct and maternal heritability estimates with SFM tended to be lower than those estimated with MTM. For example, the direct heritability for YWT was 0.59 with MTM, and 0.48 with SFM. Estimated genetic correlations for direct and maternal effects with SFM were less negative than those with MTM. For example, the direct-maternal correlation for WWT was -0.43 with MTM and -0.33 with SFM. Estimates with SFM may be superior to MTM due to better modeling of age in both fixed and random effects.


Na,K-ATPase inhibitor from guinea pig brain is not ouabain-like.
An acetone HCl extract of guinea pig brain reported to have ouabain-like activity was prepared. The extract was column-chromatographed on Sephadex G-10 and fractions were tested for ion content and "apparent ouabain-like" activity, i.e., ability to displace ouabain from its binding site and to inhibit Na,K-ATPase under conditions that optimize one or the other assay. "Apparent ouabain-like" activity eluted with the salt peak. When the ability of these fractions to inhibit ouabain binding was correlated with their potassium concentrations, the displacement could be entirely accounted for by the potassium content. The Na,K-ATPase inhibiting activity of these salt peak fractions was demonstrable in a Tris assay medium but not in a histidine (metal chelator)assay medium. Thorough desalting of fractions with "apparent ouabain-like" activity abolished this activity. Additional Na,K-ATPase inhibitory activity appeared in fractions distinct from the salt peak. However, these fractions did not display ouabain displacing ability. It is concluded that the "apparent ouabain-like" activity of the guinea pig brain extract is not truly ouabain-like and can be explained by interfering ions.


A novel, low-cost, high performance dissolved methane sensor for aqueous environments.
A new method for in-situ detection and measurement of dissolved methane in aqueous media/environments with a limit of detection of 0.2 nM (3 sigma, and t90 approxiamtely 110s) and range (1-300 nM) is presented. The detection method is based on refractive index (RI) modulation of a modified PolyDiMethylSiloxane (PDMS) layer incorporating molecules of cryptophane-A [1] which have a selective and reversible affinity for methane [2]. The refractive index is accurately determined using surface plasmon resonance (SPR) [3]. A prototype sensor has been repeatedly tested, using a dissolved gas calibration system under a range of temperature and salinity regimes. Laboratory-based results show that the technique is specific, sensitive, and reversible. The method is suitable for miniaturization and incorporation into in situ sensor technology.


Two new bifunctional protein modification reagents and their application to the study of parvalbumin.
2-[(Trifluoroacetoxy)mercuri]-4-fluorophenol (MFP) and 4-(acetoxymercuri)phenyl azide (MPA) have been prepared and characterized. The sulfhydryl-specific reagents MFP and MPA have been used to study the structure of parvalbumin. The 19F resonance of the derivative of parvalbumin with MFP was studied at 94.6 and 235.2 MHz. At 94.6 MHz, the 19F signal line width is 25 Hz and T1 = 0.28 s, while at 235.2 MHz, the line width is 35 Hz and T1 = 0.39 s. T1 was not affected by substitution of D2O for H2O as a solvent. Removal of calcium from the parvalbumin derivative resulted in the upfield shift of the 19F signal and a decrease in T1 at 94.6 MHz. The pKa for phenolic titration of the MFP derivative was 10.75 compared with 9.5 for the free reagent. These results are interpreted in terms of lodgment of the aryl group in a cleft in the surface of the parvalbumin. The derivative with MPA was photolyzed and subjected to amino acid analysis. Comparison of the analysis of parvalbumin, parvalbumin-MPA, and the photolyzed product indicated destruction of aspartic acid during the photolysis. Asp-22 is a reasonable candidate for the residue attacked, based on comparison with the published crystal structure of parvalbumin.


miR-145-5p Targets Sp1 in Non-Small Cell Lung Cancer Cells and Links to BMI1 Induced Pemetrexed Resistance and Epithelial-Mesenchymal Transition.
Pemetrexed is a folic acid inhibitor used as a second-line chemotherapeutic agent for the treatment of locally advanced or metastatic non-small cell lung cancer (NSCLC), which accounts for 85% of lung cancers. However, prolonged treatment with pemetrexed may cause cancer cells to develop resistance. In this study, we found increased expressions of BMI1 (B Lymphoma Mo-MLV insertion region 1 homolog) and Sp1 and a decreased expression of miR-145-5p was found in pemetrexed-resistant A400 cells than in A549 cells. Direct Sp1 targeting activity of miR-145-5p was demonstrated by a luciferase based Sp1 3'-UTR reporter. Changed expression of miR-145-5p in A400 or A549 cells by transfection of miR-145-5p mimic or inhibitor affected the sensitivity of the cells to pemetrexed. On the other hand, the overexpression of Sp1 in A549 cells caused the decreased sensitivity to pemetrexed, induced cell migratory capability, and epithelial-mesenchymal transition (EMT) related transcription factors such as Snail Family Transcriptional Repressor 1 and Zinc Finger E-Box Binding Homeobox 1. In addition, the overexpression of BMI1 in the A549 cells resulted in an increase in Sp1 and a decrease in miR-145-5p accompanied by the elevations of cell proliferation and EMT transcription factors, which could be reduced by the overexpression of miR-145-5p or by treatment with the Sp1 inhibitor of mithramycin A. In conclusion, the results of this study suggest that the downregulation of miR-145-5p by BMI1 overexpression could lead to the enhanced expression of Sp1 to induce the EMT process in pemetrexed-resistant NSCLC cells. These results suggest that increasing miR-145-5p expression by delivering RNA drugs may serve as a sensitizing agent for pemetrexed-resistant NSCLC patients.


Biological response to intravenously administered endotoxin in patients with advanced cancer.
The purpose of the study was to evaluate the toxicity and biological activity of highly purified lipopolysaccharide (LPS) administered intravenously to cancer patients in order to establish an optimum dosage scheme. An initial subtoxic dose was increased in weekly increments in accordance with individual regimens that maintained patient reaction at a safe and acceptable level. Purified LPS from Salmonella abortus equi was administered to 11 patients with advanced solid tumors on a weekly schedule with intraindividually escalating dosage as determined by patient response. Biological response was monitored by complete blood count, C-reactive protein, and cytokine measurements at different time points after LPS injection. Tumor necrosis factor-alpha (TNF) and interleukin-1 beta serum levels were measured by enzyme-linked immunosorbent assay and interleukin-6 (IL-6) by bioassay. Dose-limiting toxicities including chills and fever (WHO grade III) were reached at 1.0 ng/kg of body weight (maximal tolerated dose-1, MTD-1). Pretreatment with ibuprofen (1,600 mg) abrogated these side effects, allowing further escalation of LPS doses up to 10 ng/kg of body weight. At dose levels greater than 8.0 ng/kg of body weight (MTD-2), the aforementioned side effects occurred again and, additionally, hepatic toxicity (WHO grade III) was observed. Hematological changes included neutropenia followed by a pronounced neutrophilia contributed to by up to 30% bands, marked monocytopenia for 3 h, and retarded lymphopenia. By 24 h, all hematological parameters returned to pretreatment values. TNF serum levels increased from 10 pg/ml before treatment to 7,000 pg/ml as a function of dosage. Maximum serum levels were reached at 60 to 90 min after LPS injection. Similarly, IL-6 serum concentrations increased from less than 4 to 2,500 U/ml; peak levels were obtained 30 min after TNF peak values. Prior administration of ibuprofen had no effect on the above-mentioned hematological changes nor on cytokine release. LPS can be administered intravenously in weekly intervals at escalating doses from 0.15-10.0 ng/kg of body weight, when patients are protected by pretreatment with ibuprofen at dose levels above 1.0 ng/kg of body weight. Cytokine release as measured by TNF and IL-6 increased in a dose-dependent manner although the constitutional symptoms are completely attenuated.


Carryover of aflatoxin from feed to milk in dairy cows with low or high somatic cell counts.
Aflatoxin M1 (AFM1) residues in milk are regulated in many parts of the world and can cost dairy farmers significantly due to lost milk sales. Additionally, due to the carcinogenicity of this compound contaminated milk can be a major public health concern. Thirty-four lactating dairy cows were utilised to investigate the relationship between somatic cell counts (SCC), milk yield and conversion of dietary aflatoxin B1 (AFB1) into milk AFM1 (carryover (CO)). The AFM1 in milk increased as soon as the first milking after animal ingestion with a pattern of increment up to the observed plateau (between 7th and 12th days of AFB1 ingestion). There was a significant (P < 0.01) effect of the milk yield whereas no effect could be attributed to the SCC levels or to the milk yield × SCC interaction. Similarly, the main effect of milk yield was also observed (P < 0.01) on the total amount of AFM1 excreted during the ingestion period. Although the plasma concentration of gamma-glutamyl transferase was significantly affected by aflatoxin administration, levels of this liver enzyme were within the normal range for lactating dairy cows. The current data suggest that milk yield is the major factor affecting the total excretion of AFM1 and that SCC as an indicator of mammary gland permeability was not related to an increase in AFM1 CO.


The role of vasoactive agents in the resuscitation of microvascular perfusion and tissue oxygenation in critically ill patients.
The clinical use of vasoactive drugs is not only intended to improve systemic hemodynamic variables, but ultimately to attenuate derangements in organ perfusion and oxygenation during shock. This review aims (1) to discuss basic physiology with respect to manipulating vascular tone and its effect on the microcirculation, and (2) to provide an overview of available clinical data on the relation between vasoactive drugs and organ perfusion, with specific attention paid to recent developments that have enabled direct in vivo observation of the microcirculation and concepts that have originated from it. A MedLine search was conducted for clinical articles in the English language over the last 15 years pertainig to shock, sepsis, organ failure, or critically ill patients in combination with vasoactive drugs and specific variables of organ perfusion/oxygenation (e.g., tonometry, indocyanine clearance, laser Doppler, and sidestream dark field imaging). Eighty original papers evaluating the specific relationship between organ perfusion/oxygenation and the use of vasoactive drugs were identified and are discussed in light of physiological theory of vasomotor tone. Solid clinical data in support of the idea that increasing blood pressure in shock improves microcirculatory perfusion/oxygenation seem to be lacking, and such a concept might not be in line with physiological theory of microcirculation as a low-pressure vascular compartment. In septic shock no beneficial effect on microcirculatory perfusion above a mean arterial pressure of 65 mmHg has been reported, but a wide range in inter-individual effect seems to exist. Whether improvement of microcirculatory perfusion is associated with better patient outcome remains to be elucidated.


The association of perceived discrimination with low back pain.
A handful of recent studies have documented perceived discrimination as a correlate of poor physical and mental health status among ethnic and racial minority groups. To date, however, despite a proliferation of research on ethnic disparities in the severity and impact of a number of persistent pain conditions, there have been no reports on associations between perceived discrimination and pain-related symptoms. Using data from a national survey (the National Survey of Midlife Development in the United States; MIDUS), we explore the relationships between perceived discriminatory events and the report of back pain among African-American and white men and women. As expected, African-American participants reported substantially greater perceptions of discrimination than white participants. Moreover, in models that included a variety of physical and mental health variables, episodes of major lifetime discriminatory events were the strongest predictors of back pain report in African-Americans, and perceived day-to-day discrimination was the strongest predictor of back pain report specifically in African-American women. Among white participants, perceptions of discrimination were minimally related or unrelated to back pain. To our knowledge, these are the first data documenting an association between perceived discrimination and report of back pain; the fact that perceptions of discrimination were stronger predictors than physical health variables highlights the potential salience and adverse impact of perceived discrimination in ethnic and racial minority groups.


Efficiency and finite size effects in enhanced transmission through subwavelength apertures.
We investigate transmission efficiency and finite size effects for the subwavelength hole arrays. Experiments and simulations show how the finite size effects depend strongly on the hole diameter. The transmission efficiency reaches an asymptotic upper value when the array is larger than the surface plasmon propagation length on the corrugated surface. By comparing the transmission of arrays with that of the corresponding single holes, the relative enhancement is found to increase as the hole diameter decreases. In the conditions of the experiments the enhancement is one to two orders of magnitude but there is no fundamental upper limit to this value.


Effects of endotoxaemia and carbohydrate overload on glucose and insulin dynamics and the development of laminitis in horses.
Insulin resistance (IR) is a risk factor for pasture-associated laminitis in equids and alimentary carbohydrate overload may trigger laminitis. Whether glucose metabolism responses to carbohydrate overload are more pronounced in insulin-resistant horses requires further study. Horses pretreated with endotoxin to alter insulin sensitivity differ significantly in their glucose and insulin responses to carbohydrate overload. Horses (n=24) were divided into 3 groups. A lipopolysaccharide (LPS; n=8) group that received endotoxin as an 8 h 7.5 ng/kg bwt/h i.v. continuous rate infusion, an oligofructose (OF; n=8) group that received an infusion of saline followed by 5 g/kg bwt OF via nasogastric intubation, and a LPS/OF (n=8) group that received LPS followed 16 h later by OF. Glucose and insulin dynamics were evaluated at -24 h and 48 h using the frequently sampled i.v. glucose tolerance test and minimal model analysis. Physical examinations and haematology were performed and the severity of laminitis assessed. Horses receiving LPS developed leucopenia and both LPS and OF induced clinical signs consistent with systemic inflammation. Insulin sensitivity significantly decreased (P < 0.001) over time, but responses did not differ significantly among groups. Time (P < 0.001) and treatment x time (P = 0.038) effects were detected for the acute insulin response to glucose, with mean values significantly increasing in LPS and LPS/OF groups, but not the OF group. Five horses in the LPS/OF group developed clinical laminitis compared with 0 and 2 horses in the LPS and OF groups, respectively. Endotoxaemia and carbohydrate overload reduce insulin sensitivity in horses. Endotoxin pretreatment does not affect the alterations in glucose metabolism induced by carbohydrate overload. Insulin sensitivity decreases after carbohydrate overload in horses, which may be relevant to the development of pasture-associated laminitis.


Immunomodulatory effects in a phase II study of lenalidomide combined with cetuximab in refractory KRAS-mutant metastatic colorectal cancer patients.
This study assessed the immunomodulatory effects in previously treated KRAS-mutant metastatic colorectal cancer patients participating in a phase II multicenter, open-label clinical trial receiving lenalidomide alone or lenalidomide plus cetuximab. The main findings show the T cell immunostimulatory properties of lenalidomide as the drug induced a decrease in the percentage CD45RA(+) naïve T cells 3-fold while increasing the percentage HLA-DR(+) activated T helper cells and percentage total CD45RO(+) CD8(+) memory T cytotoxic cells, 2.6- and 2.1-fold respectively (p<0.0001). In addition, lenalidomide decreased the percentage of circulating CD19(+) B cells 2.6-fold (p<0.0001). Lenalidomide increased a modest, yet significant, 1.4-fold change in the percentage of circulating natural killer cells. Our findings indicate that lenalidomide significantly activates T cells, suggestive of an immunotherapeutic role for this drug in settings of maintenance therapy and tumor immunity. Furthermore, reported for the first time is the effect of lenalidomide in combination with cetuximab on T cell function, including increases in circulating naïve and central memory T cells. In summary, lenalidomide and cetuximab have significant effects on circulating immune cells in patients with colorectal carcinoma. ClinicalTrials.gov NCT01032291.


Group II introns designed to insert into therapeutically relevant DNA target sites in human cells.
Mobile group II intron RNAs insert directly into DNA target sites and are then reverse-transcribed into genomic DNA by the associated intron-encoded protein. Target site recognition involves modifiable base-pairing interactions between the intron RNA and a >14-nucleotide region of the DNA target site, as well as fixed interactions between the protein and flanking regions. Here, we developed a highly efficient Escherichia coli genetic assay to determine detailed target site recognition rules for the Lactococcus lactis group II intron Ll.LtrB and to select introns that insert into desired target sites. Using human immunodeficiency virus-type 1 (HIV-1) proviral DNA and the human CCR5 gene as examples, we show that group II introns can be retargeted to insert efficiently into virtually any target DNA and that the retargeted introns retain activity in human cells. This work provides the practical basis for potential applications of targeted group II introns in genetic engineering, functional genomics, and gene therapy.


[The effect of finoptin on platelet aggregation, blood coagulability and fibrinolysis in patients with ischemic heart disease during physical exercise].
Exercise platelet aggregation, blood coagulability, and fibrinolysis were examined in patients prior myocardial infarction 2-3 months following the disease. Seventy patients of whom 35 had received placebo and 35 finoptin prior to exercise were studied. The platelet aggregation, blood coagulability, and fibrinolysis were explored before and after bicycle ergometric test. Following exercise, the patients on placebo showed higher platelet aggregation, blood coagulability and lower fibrinolysis, whereas those on finoption exhibited a lower increase in platelet aggregation and blood coagulability and activated fibrinolysis.


Beyond the Condon limit: Condensed phase optical spectra from atomistic simulations.
While dark transitions made bright by molecular motions determine the optoelectronic properties of many materials, simulating such non-Condon effects in condensed phase spectroscopy remains a fundamental challenge. We derive a Gaussian theory to predict and analyze condensed phase optical spectra beyond the Condon limit. Our theory introduces novel quantities that encode how nuclear motions modulate the energy gap and transition dipole of electronic transitions in the form of spectral densities. By formulating the theory through a statistical framework of thermal averages and fluctuations, we circumvent the limitations of widely used microscopically harmonic theories, allowing us to tackle systems with generally anharmonic atomistic interactions and non-Condon fluctuations of arbitrary strength. We show how to calculate these spectral densities using first-principles simulations, capturing realistic molecular interactions and incorporating finite-temperature, disorder, and dynamical effects. Our theory accurately predicts the spectra of systems known to exhibit strong non-Condon effects (phenolate in various solvents) and reveals distinct mechanisms for electronic peak splitting: timescale separation of modes that tune non-Condon effects and spectral interference from correlated energy gap and transition dipole fluctuations. We further introduce analysis tools to identify how intramolecular vibrations, solute-solvent interactions, and environmental polarization effects impact dark transitions. Moreover, we prove an upper bound on the strength of cross correlated energy gap and transition dipole fluctuations, thereby elucidating a simple condition that a system must follow for our theory to accurately predict its spectrum.


Plant-based oral tolerance to hemophilia therapy employs a complex immune regulatory response including LAP+CD4+ T cells.
Coagulation factor replacement therapy for the X-linked bleeding disorder hemophilia is severely complicated by antibody ("inhibitor") formation. We previously found that oral delivery to hemophilic mice of cholera toxin B subunit-coagulation factor fusion proteins expressed in chloroplasts of transgenic plants suppressed inhibitor formation directed against factors VIII and IX and anaphylaxis against factor IX (FIX). This observation and the relatively high concentration of antigen in the chloroplasts prompted us to evaluate the underlying tolerance mechanisms. The combination of oral delivery of bioencapsulated FIX and intravenous replacement therapy induced a complex, interleukin-10 (IL-10)-dependent, antigen-specific systemic immune suppression of pathogenic antibody formation (immunoglobulin [Ig] 1/inhibitors, IgE) in hemophilia B mice. Tolerance induction was also successful in preimmune mice but required prolonged oral delivery once replacement therapy was resumed. Orally delivered antigen, initially targeted to epithelial cells, was taken up by dendritic cells throughout the small intestine and additionally by F4/80(+) cells in the duodenum. Consistent with the immunomodulatory responses, frequencies of tolerogenic CD103(+) and plasmacytoid dendritic cells were increased. Ultimately, latency-associated peptide expressing CD4(+) regulatory T cells (CD4(+)CD25(-)LAP(+) cells with upregulated IL-10 and transforming growth factor-β (TGF-β) expression) as well as conventional CD4(+)CD25(+) regulatory T cells systemically suppressed anti-FIX responses.


Abscess formation in Gartner's duct cysts associated with ipsilateral renal agenesis. A report of two cases.
The occurrence of both urinary and genital abnormalities in the same female patient is well known. Two cases of abscess formation in a Gartner's duct cyst associated with ipsilateral renal agenesis are presented. Marsupialization is recommended as a satisfactory method of treatment.


Are lutein and zeaxanthin conditionally essential nutrients for eye health?
The carotenoids lutein and zeaxanthin are found in the macula in high concentrations and may play a role in the pathogenesis of age-related macular degeneration (ARMD). Lutein and zeaxanthin may protect the macula and photoreceptor outer segments throughout the retina from oxidative stress and play a role in an antioxidant cascade that safely disarms the energy of reactive oxygen species. Although lutein and zeaxanthin are not essential nutrients, studies are beginning to suggest that they fit the criteria for conditionally essential nutrients. Low plasma lutein and zeaxanthin concentrations or dietary intake are associated with low macular pigment density and increased risk of ARMD. Dietary deprivation of lutein and zeaxanthin in primates causes pathological changes in the macula. Should controlled clinical trials show lutein and/or zeaxanthin supplementation protects against the development or progression of ARMD and other eye diseases, then lutein and zeaxanthin could be considered as conditionally essential nutrients for humans.


Emerging therapies for the treatment of sepsis.
Sepsis affects patients of all ages with multiple comorbidities and underlying diagnoses, and is the result of infection by many potential pathogens infecting various organs or sites. Many molecules have been clinically tested in recent years for their potential immunomodulatory effects, but have been shown to have no beneficial effects on outcomes in heterogeneous populations of patients with sepsis. There are, therefore, no specific antisepsis therapies and mortality and morbidity rates remain high despite improved overall management of these patients. This review covers promising agents currently used in clinical trials. There are several candidates currently undergoing early and later phase of clinical testing, including thrombomodulin, alkaline phosphatase, interferon-beta, and selepressin. Other approaches including immunoglobulins, extracorporeal therapies, and pharmaconutrients will also be discussed. Despite multiple trials of potential therapies for sepsis, no strategies have yet been persistently shown to have beneficial effects on outcomes. The main reason for the disappointing results is that patient populations in these studies have been too heterogeneous. Selecting patients on the basis of general symptoms is not enough. Rather patients should be selected according to the likely action of the drug in question. To achieve this, improved biomarkers of sepsis and of the immune response are needed and the activities of the individual agents need to be carefully characterized. New candidates are being developed and the results of ongoing and recent clinical trials of immunomodulatory therapies are eagerly awaited as new therapies for sepsis are urgently needed.


Mcl-1 protects eosinophils from apoptosis and exacerbates allergic airway inflammation.
Eosinophils are key effector cells in allergic diseases. Here we investigated Mcl-1 (an anti-apoptotic protein) in experimental allergic airway inflammation using transgenic overexpressing human Mcl-1 mice (hMcl-1) and reducing Mcl-1 by a cyclin-dependent kinase inhibitor. Overexpression of Mcl-1 exacerbated allergic airway inflammation, with increased bronchoalveolar lavage fluid cellularity, eosinophil numbers and total protein, and an increase in airway mucus production. Eosinophil apoptosis was suppressed by Mcl-1 overexpression, with this resistance to apoptosis attenuated by cyclin-dependent kinase inhibition which also rescued Mcl-1-exacerbated allergic airway inflammation. We propose that targeting Mcl-1 may be beneficial in treatment of allergic airway disease.


Telomere shortening in breast cancer correlates with the pathological features of tumor progression.
Telomeres are involved in the maintenance of genomic stability. Telomere alteration has been observed in most human cancer types, and is known to be a feature of malignancy. The aim of the present study was to evaluate whether the telomere length of breast cancer cells correlates with TNM stage and several pathological features. We investigated a total of 44 breast cancers, including 17 scirrhous, 15 papillotubular and 12 solid-tubular carcinomas. Telomere lengths were determined by tissue quantitative fluorescence in situ hybridization (Q-FISH), and compared according to the TNM stage, histological tumor size, lymph node metastases, vascular invasion and immunohistochemical status (ER, PR, HER2 status and Ki67 labeling index). In all histological types, telomeres of cancer cells were significantly shorter than those of normal epithelial cells. Mean telomere length was significantly less in patients with TNM stage III, and in those with large tumors, lymph node metastases and vascular invasion. Our results suggest that the telomere length of cancer cells is strongly correlated with the degree of cancer progression.


Loss of myocardial ischemic postconditioning in adenosine A1 and bradykinin B2 receptors gene knockout mice.
Ischemic postconditioning (PostC) is a recently described cardioprotective modality against reperfusion injury, through series of brief reflow interruptions applied at the very onset of reperfusion. It is proposed that PostC can activate a complex cellular signaling cascade, in which cell membrane receptors could serve as the upstream triggers of PostC. However, the exact subtypes of such receptors remain controversial or uninvestigated. To this context, the purpose of present study was to determine the definitive role of adenosine A(1) and bradykinin B(1) and B(2) receptors in PostC. The hearts isolated from adult male C57BL/6J wild-type mice or the mice lacking adenosine A(1), or bradykinin B(1) or B(2) receptors subjected to zero-flow global ischemia and reperfusion in a Langendorff model. PostC, consisting of 6 cycles of 10 seconds of reperfusion and 10 seconds of ischemia, demonstrated significantly reduced myocardial infarct size (22.8+/-3.1%, mean+/-SEM) as compared with the non-PostC wild-type controls (35.1+/-2.8%, P<0.05). The infarct-limiting protection of PostC was absent in adenosine A(1) receptor knockout mice (34.9+/-2.7%) or bradykinin B(2) receptor knockout mice (33.3+/-1.7%) and was partially attenuated in bradykinin B(1) receptor-deficient mice (25.6+/-2.9%; P>0.05). On the other hand, PostC did not significantly alter postischemic cardiac contractile function and coronary flow. With the use of three distinctive strains of gene knockout mice, the current study has provided the first conclusive evidence showing PostC-induced infarct-limiting cardioprotection could be triggered by activation of multiple types of cell membrane receptors, which include adenosine A(1) and bradykinin B(2) receptors.


Utilizing a PTPN22 gene signature to predict response to targeted therapies in rheumatoid arthritis.
Despite the development of several targeted therapies for rheumatoid arthritis (RA), there is still no reliable drug-specific predictor to assist rheumatologists in selecting the most effective targeted therapy for each patient. Recently, a gene signature caused by impaired induction of PTPN22 in anti-CD3 stimulated peripheral blood mononuclear cells (PBMC) was observed in healthy at-risk individuals. However, the downstream target genes of PTPN22 and the molecular mechanisms regulating its expression are still poorly understood. Here we report that the PTPN22 gene signature is also present in PBMC from patients with active RA and can be reversed after effective treatment. The expression of PTPN22 correlates with that of more than 1000 genes in Th cells of anti-CD3 stimulated PBMC of healthy donors and is inhibited by TNFα or CD28 signals, but not IL-6, through distinct mechanisms. In addition, the impaired induction of PTPN22 in PBMC of patients with active RA can be normalized in vitro by several targeted therapies. More importantly, the in vitro normalization of PTPN22 expression correlates with clinical response to the targeted therapies in a longitudinal RA cohort. Thus, in vitro normalization of PTPN22 expression by targeted therapies can potentially be used to predict clinical response.


Does breathing disturb coordination in butterfly?
This study quantified the effects of breathing compared to non-breathing and "race pace" on arm to leg coordination in the butterfly stroke. Twelve elite male swimmers swam at four paces: 400 m, 200 m, 100 m and 50 m. The arm and leg stroke phases were identified by video analysis to calculate the total time gap (TTG), which is the sum of T1 (hands' entry in the water/high point of first kick), T2 (beginning of the hands' backward movement/low point of first kick), T3 (hands' arrival in a vertical plane to the shoulders/high point of second kick) and T4 (hands' release from the water/low point of second kick). Two strokes with breathing were compared to two strokes with breath-holding. The TTG was greater with breathing (23.3% VS. 19%), showing less propulsive continuity between arm and leg actions (p<0.05). This was due to the shorter downward leg kick and longer arm catch and upward leg kick that led to longer glide time. Conversely, breathing leads to greater coupling between the hand exit and the end of leg propulsion, which was due to a shorter arm push phase to facilitate the head exit to breathe.


Access point analysis on the state of health care services in South African prisons: a qualitative exploration of correctional health care workers' and inmates' perspectives in Kwazulu-Natal and Mpumalanga.
This paper explores inmates and prison health care workers perceptions of the state of health care services in four correctional facilities in South Africa. Structural and organisational issues are explored in terms of how they impact the delivery and provision of health care to inmates within correctional facilities. Additionally, the study forms an access point analysis of prisons as a health care setting as part of the development and testing of a STI/HIV health education intervention for soon to be released inmates. Focus group discussions (6-8 participants per group) were conducted with male inmates in four facilities in KwaZulu-Natal and Mpumalanga provinces. Individual face-to-face interviews were conducted with eight health personnel to get a view from both providers and end users of health care in correctional settings. Data were analysed thematically. We found strong evidence of prison being a strategic point to increase access to health services for offenders. Curative services within prisons were well established and running despite the presence of certain challenges varying across institutions. Prevention programmes emerged as an area that requires stronger emphasis to facilitate imparting skills and promoting safer practices for inmates upon release. Peer-led education programmes emerged as a key aspect of preparation for release and community reintegration amongst inmates.


Autonomous Visualization of Damage in Polymers by Metal-Free Polymerizations of Microencapsulated Activated Alkynes.
The development of autonomous materials with desired performance and built-in visualizable sensing units is of great academic and industrial significance. Although a wide range of damage indication methods have been reported, the "turn-on" sensing mechanism by damaging events based on microcapsule systems, especially those relying on chemical reactions to elicit a chromogenic response, are still very limited. Herein, a facile and metal-free polymerization route with an interesting reaction-induced coloration effect is demonstrated. Under the catalysis of 1,4-diazabicyclo[2.2.2]octane (DABCO), the polymerizations of difunctional or trifunctional activated alkynes proceed very quickly at 0 °C in air. A series of polymers composed of stereoregular enyne structure (major unit) and divinyl ether structure (minor unit) are obtained. Both the catalyst and monomers are colorless while the polymerized products are deep-colored. This process can be applied for the damage visualization of polymers using the microencapsulation technique. Microcapsules containing the reactive alkyne monomer are prepared and mixed in a DABCO-dispersed polymer film. Both the external and internal damage regions of this composite film can be readily visualized once the reaction is initiated from the ruptured microcapsules. Moreover, the newly formed polymer automatically seals the cracks with an additional protection function.


Autoantibodies to aberrantly glycosylated MUC1 in early stage breast cancer are associated with a better prognosis.
Detection of serum biomarkers for early diagnosis of breast cancer remains an important goal. Changes in the structure of O-linked glycans occur in all breast cancers resulting in the expression of glycoproteins that are antigenically distinct. Indeed, the serum assay widely used for monitoring disease progression in breast cancer (CA15.3), detects a glycoprotein (MUC1), but elevated levels of the antigen cannot be detected in early stage patients. However, since the immune system acts to amplify the antigenic signal, antibodies can be detected in sera long before the antigen. We have exploited the change in O-glycosylation to measure autoantibody responses to cancer-associated glycoforms of MUC1 in sera from early stage breast cancer patients. We used a microarray platform of 60mer MUC1 glycopeptides, to confirm the presence of autoantibodies to cancer associated glycoforms of MUC1 in a proportion of early breast cancer patients (54/198). Five positive sera were selected for detailed definition of the reactive epitopes using on chip glycosylation technology and a panel of glycopeptides based on a single MUC1 tandem repeat carrying specific glycans at specific sites. Based on these results, larger amounts of an extended repertoire of defined MUC1 glycopeptides were synthesised, printed on microarrays, and screened with sera from a large cohort of breast cancer patients (n = 395), patients with benign breast disease (n = 108) and healthy controls (n = 99). All sera were collected in the 1970s and 1980s and complete clinical follow-up of breast cancer patients is available. The presence and level of autoantibodies was significantly higher in the sera from cancer patients compared with the controls, and a highly significant correlation with age was observed. High levels of a subset of autoantibodies to the core3MUC1 (GlcNAcβ1-3GalNAc-MUC1) and STnMUC1 (NeuAcα2,6GalNAc-MUC1) glycoforms were significantly associated with reduced incidence and increased time to metastasis. Autoantibodies to specific cancer associated glycoforms of MUC1 are found more frequently and at higher levels in early stage breast cancer patients than in women with benign breast disease or healthy women. Association of strong antibody response with reduced rate and delay in metastases suggests that autoantibodies can affect disease progression.


Pelvic skeletal metastasis of hepatocellular carcinoma with sarcomatous change: a case report.
Sarcomatoid hepatocellular carcinoma (HCC) is a very rare histologic variant of HCC. The characteristic of skeletal metastatic sarcomatoid hepatocellular carcinoma has never been reported. We reported a patient with sarcomatoid hepatocellular carcinoma pelvic metastasis who presented with huge pelvic metastasis that had relatively small osteolytic lesion centrally located accompanied by huge bipeduncular invasive expansile lesions into surrounding soft tissue. The lesion showed almost non-isotope uptake in 99mTc-methylene diphosphonate bone scintigraphy study. He underwent radiotherapy and tumor excision but the tumor rapidly recurred. In addition, serum alpha-fetoprotein level was never elevated beyond normal limit (< 20 ng/mL) through the whole course of treatment. We considered sarcomatoid hepatocellular carcinoma bone metastasis a highly aggressive lesion with unusual metastatic pattern. Surgical treatment with adequate safe margin in such a huge tumor with hypervascularity and extensive invasion in the pelvis was difficult; and radiotherapy maybe refractory regarding the sarcomatous nature. Therefore, debulking operation with local symptoms control may provide a better quality of life. And the clinical course suggests sarcomatoid hepatocellular carcinoma is derived from the transition of an ordinary hepatocellular carcinoma.


Hair testing of GHB: an everlasting issue in forensic toxicology.
In this paper, the authors present a critical review of different studies regarding hair testing of endogenous γ-hydroxybutyrate (GHB), concentrations in chronic users, and values measured after a single GHB exposure in drug facilitated sexual assault (DFSA) cases together with the role of a recently identified GHB metabolite, GHB-glucuronide. The following databases (up to March 2017) PubMed, Scopus and Web of Science were used, searching the following key words: γ-hydroxybutyrate, GHB, GHB glucuronide, hair. The main key words "GHB" and "γ-hydroxybutyrate" were searched singularly and then associated individually to each of the other keywords. Of the 2304 sources found, only 20 were considered appropriate for the purpose of this paper. Summing up all the studies investigating endogenous GHB concentration in hair, a very broad concentration range from 0 to 12 ng/mg was found. In order to detect a single GHB dose in hair it is necessary to commonly wait 1 month for collecting hair and a segmental analysis of 3 or 5 mm fragments and the calculation of a ratio between the targeted segment and the others represent a reliable method to detect a single GHB intake considering that the ratios presently proposed vary from 3 and 10. The only two studies so far performed, investigating GHB-Glucuronide in hair, show that the latter does not seem to provide any diagnostic information regarding GHB exposure. A practical operative protocol is proposed to be applied in all suspected cases of GHB-facilitated sexual assault (GHB-FSA).


A critique of the use of the Kolmogorov-Smirnov (KS) statistic for the analysis of BOLD fMRI data.
The use of the Kolmogorov-Smirnov (KS) statistic for testing hypotheses regarding activation in blood oxygenation level-dependent functional MRI data is critiqued on both theoretical and empirical grounds. Theoretically, it is argued that the KS test is formally unable to support inferences of interest to most neuro-imaging studies and has reduced sensitivity compared with parametric alternatives. Empirically, false-positive rates yielded by the KS test in human data collected under the null hypothesis were significantly in excess of tabular values. These excessive false-positive rates could be explained by the presence of temporal autocorrelation. We also present evidence that the distribution of blood oxygenation level-dependent functional MRI data is only slightly nonnormal, questioning the initial impetus for the use of the KS test in this context. Finally, it is noted that parametric alternatives exist that do provide adequate control of the false-positive rate and can support inferences of interest.


Multiphasic analysis of the temporal development of the distal gut microbiota in patients following ileal pouch anal anastomosis.
The indigenous gut microbiota are thought to play a crucial role in the development and maintenance of the abnormal inflammatory responses that are the hallmark of inflammatory bowel disease. Direct tests of the role of the gut microbiome in these disorders are typically limited by the fact that sampling of the microbiota generally occurs once disease has become manifest. This limitation could potentially be circumvented by studying patients who undergo total proctocolectomy with ileal pouch anal anastomosis (IPAA) for the definitive treatment of ulcerative colitis. A subset of patients who undergo IPAA develops an inflammatory condition known as pouchitis, which is thought to mirror the pathogenesis of ulcerative colitis. Following the development of the microbiome of the pouch would allow characterization of the microbial community that predates the development of overt disease. We monitored the development of the pouch microbiota in four patients who underwent IPAA. Mucosal and luminal samples were obtained prior to takedown of the diverting ileostomy and compared to samples obtained 2, 4 and 8 weeks after intestinal continuity had been restored. Through the combined analysis of 16S rRNA-encoding gene amplicons, targeted 16S amplification and microbial cultivation, we observed major changes in structure and function of the pouch microbiota following ileostomy. There is a relative increase in anaerobic microorganisms with the capacity for fermentation of complex carbohydrates, which corresponds to the physical stasis of intestinal contents in the ileal pouch. Compared to the microbiome structure encountered in the colonic mucosa of healthy individuals, the pouch microbial community in three of the four individuals was quite distinct. In the fourth patient, a community that was much like that seen in a healthy colon was established, and this patient also had the most benign clinical course of the four patients, without the development of pouchitis 2 years after IPAA. The microbiota that inhabit the ileal-anal pouch of patients who undergo IPAA for treatment of ulcerative colitis demonstrate significant structural and functional changes related to the restoration of fecal flow. Our preliminary results suggest once the pouch has assumed the physiologic role previously played by the intact colon, the precise structure and function of the pouch microbiome, relative to a normal colonic microbiota, will determine if there is establishment of a stable, healthy mucosal environment or the reinitiation of the pathogenic cascade that results in intestinal inflammation.


Design of a system for monitoring antimicrobial resistance in pathogenic, zoonotic and indicator bacteria from food animals.
DANMAP is a Danish programme for integrated monitoring of and research on antimicrobial resistance in bacteria from food animals, food and humans. The paper describes how bacteria from broilers, pigs, and cattle are collected, as well as the procedures for data handling and presentation of results. The bacteria from animals include certain pathogens, selected so that they are representative for submissions to Danish diagnostic laboratories, as well as zoonotic bacteria (Campylobacter, Salmonella and Yersinia) and indicator bacteria (E. coli, E. faecium and E. faecalis), from samples collected at abattoirs. The latter samples are selected so that they are representative of the respective animal populations. Therefore, the apparent prevalence of antimicrobial resistance in the populations may be calculated. The isolates are identified to species level and the results of susceptibility testing are stored as continuous variables. All isolates are maintained in a strain collection so that they are available for subsequent research projects. The data handling facilities makes it possible to present results as percent resistant isolates or as the apparent prevalence of resistance in the population, or alternatively as graphical distributions of mm inhibition zones or MIC values. Computer routines have been established that make it possible to detect specific phenotypic expressions of resistance that may be of particular interest.


Gastric outlet obstruction caused by a giant gastroduodenal artery aneurysm: a case report.
Gastric outlet (GO) obstruction in an adult is usually caused by intrinsic gastric or duodenal lesions or pancreatic tumours. This study describes a case of a 77-year-old man who developed GO obstruction due to extrinsic compression from a large gastroduodenal artery aneurysm under rupture. This cause of GO obstruction has never previously been reported in the literature.


[Characteristics of the expression of t12 mutation in mouse embryos with structural aberrations of chromosome 17].
Developmental profiles of mouse embryos with deletions, duplications of nullisomy for the proximal part AB of chromosome 17, including genes of the T-t complex, were studied in mice with marker translocations Rb (16.17)7Bnr or T(16;17)43H and heterozygous for lethal t12 mutation. The embryos t12t12 and t12t12-(Dp17CDE; Dl16) were shown to be eliminated at the morula stage; embryos t12+, t12+ + or t12t12+ survive during preimplantation and early postimplantation stages: t12 embryos (hemizygous for all genes of the 17AB region, including all t-alleles) have quite normal cleavage, blastulation and implantation, but die soon thereafter. The embryos with nullisomy 17AB combined with deletion 17CDE survive up to the morula stage. These data are in line with previously proposed hypothetical mechanism for mutual activation of homologous chromosomes and their segments during initial stages of embryogenesis in mice. The system of marker chromosomes Rb7Bnr and T43H in combination with various alleles of the T complex might be recommended as a useful tool in analysis of primary developmental effects of different t-alleles in mice.


[Availability of essential drugs in two regions of Minas Gerais, Brazil].
To investigate the availability of essential drugs in municipalities with a human development index < 0.699. We surveyed 69 institutions, including municipal pharmacies, public clinics, private and philanthropic health units, and commercial pharmacies, in 19 municipalities of the state of Minas Gerais, Brazil. The municipalities were chosen according to the following selection criteria: (1) a human development index (HDI) < 0.699 (the HDI for the entire state of Minas Gerais in 1991) in the microregion where the municipality was located; (2) the municipality had to be the seat of government for the microregion where it was located; (3) there had to be at least two eligible institutions (belonging to the public, private, or philanthropic sectors) in full functioning in the municipality during the survey period. Health professionals who were directly responsible for stock control and drug dispensation at the institutions surveyed were interviewed. Institutional documents and records were also reviewed. A list of 21 tracer essential drugs, which were selected among the drugs most widely employed in the State of Minas Gerais' Basic Pharmacy Program, was used to measure availability. The availability of each tracer drug was calculated at the time of the site visit and for the 12-month period immediately before the survey. In addition, the availability of tracer drugs was calculated for each type of institution surveyed. The availability of essential drugs in municipal pharmacies was 52.0%; in public health clinics, 46.9%; and in philanthropic and private health units, 41.0% and 38.1%, respectively. In commercial pharmacies, the availability of essential drugs reached 81.2%. The availability of essential drugs in public facilities is low and varies widely, with the result that persons who need such drugs the most are often those who lack access to them. Private pharmacies are the main source of essential drugs. The results of this study point to the need to seek increased awareness and implementation of the concept of essential drugs throughout the country.


Autistic Cognition: Charting Routes to Anxiety.
Autism Spectrum Conditions are typified by a divergence in cognitive style from that of the non-autistic population. Cognitive differences in autism may underlie significant strengths, but also increase vulnerability to psychopathology such as anxiety, which is a major problem for many autistic people. Many autistic people also do not respond to typical psychotherapeutic interventions, suggesting that autism-specific models and interventions are needed. We advance a theoretical model explaining how three constructs, attenuated predictions, intolerance of uncertainty, and 'black and white thinking', may interact to lead to anxiety in autism. We hope to start a dialogue surrounding how we can best address specific autistic cognitive differences that may lead to distress by developing appropriate models, measurements, and psychotherapeutic interventions.


Biomechanical comparison of expandable cages for vertebral body replacement in the cervical spine.
Recently, expandable cages for vertebral body replacement in the cervical spine have been developed. The purpose of this study was to compare the biomechanical properties of expandable cages with those of a tricortical iliac crest graft and a nonexpandable cage. Forty human cervical spines (C3-5) were tested in flexion, extension, axial rotation, and lateral bending. First all motion segments were evaluated intact. After corpectomy of C4 the spines were divided into five groups of eight and the following stabilization techniques were used: 1) autologous iliac crest bone graft; 2) mesh titanium cage; 3) anterior distraction device; 4) Synex-C titanium; and 5) Synex-C PEEK. Additionally, anterior plating and anterior plating plus posterior screw/rod fixation were applied. Stiffness, range of motion, and neutral and elastic zones were determined. In comparison with the intact motion segment all implants significantly increased stiffness in flexion and bending, but decreased stiffness in extension. There were no biomechanical differences between the nonexpandable and expandable cages. Furthermore, there were no biomechanical differences between the tricortical iliac crest graft and the cages, except for Synex-C in rotation. Additional anterior plating significantly increased biomechanical stiffness in all test modes; particularly in rotation mode, combined anterior-posterior stabilization increased stiffness by up to 102% compared with anterior plating alone. In comparison to a tricortical iliac crest bone graft and a nonexpandable cage, expandable cages have no biomechanical advantages. Due to the low extension and rotational stiffness, none of the implants can be recommended as a stand-alone device. Additional anterior plating increased biomechanical stability adequately. Therefore, additional posterior stabilization should only be considered in cases of severe rotational instability of the cervical spine.


Sulfakinin Signalling Influences Fatty Acid Levels and Composition in Tenebrio Molitor Beetle.
Sulfakinins are arthropod neuropeptides that are structurally and functionally similar to vertebrate gastrin-cholecystokinin. Sulfakinins with sulfated tyrosine (sSK) or nonsulfated tyrosine (nSK) in the C-terminated heptapeptide XY(SO3H)GHMRFamide display different biological functions, including myotropic activity, inhibition of food intake, stimulation of digestive enzymes and regulation of carbohydrate and lipid content. To reveal the mechanisms by which sulfakinin signalling modulates lipid homeostasis, we analysed the changes in the level and composition of fatty acids and organic compounds in the fat body and haemolymph of Tenebrio molitor larvae after nSK and sSK treatment. Fatty acids in fat body and haemolymph of insects were analysed using Gas Chromatography - Mass Spectrometry (GC-MS). The direction of the changes observed for major fatty acids, 18:1 and 18:2, and the less abundant fatty acids, 16:0, 18:0, 16:1 and 14:0, was the same for unsaturated (UFAs) and saturated (SFAs) fatty acids, and elevated after nSK application in both analysed tissues. However, the action of sSK in fat body tissue evoked distinct effects and induced either significant decreases in individual fatty acids or UFAs and SFAs. Administration of nSK and sSK significantly increased the level of total organic compounds in the haemolymph, contrary to the effect of sSK in fat body, where the level of total organic compounds decreased, although changes differ between individual chemicals. Sulfakinins are engaged in the precise modulation of fatty acid levels and composition, but their action depends on the presence of sulfate group on the tyrosyl residue of the peptide what determines the different roles of these peptides in insect physiology.


Two novel proteins in the mitochondrial outer membrane mediate beta-barrel protein assembly.
Mitochondrial outer and inner membranes contain translocators that achieve protein translocation across and/or insertion into the membranes. Recent evidence has shown that mitochondrial beta-barrel protein assembly in the outer membrane requires specific translocator proteins in addition to the components of the general translocator complex in the outer membrane, the TOM40 complex. Here we report two novel mitochondrial outer membrane proteins in yeast, Tom13 and Tom38/Sam35, that mediate assembly of mitochondrial beta-barrel proteins, Tom40, and/or porin in the outer membrane. Depletion of Tom13 or Tom38/Sam35 affects assembly pathways of the beta-barrel proteins differently, suggesting that they mediate different steps of the complex assembly processes of beta-barrel proteins in the outer membrane.


Hypercalcemia and parathyroid hormone-related peptide expression in a dog with thyroid carcinoma and histiocytic sarcoma.
A 9.5-year-old, male castrated Walker Hound was presented for evaluation of progressive weakness, anorexia, and weight loss. Imaging revealed multiple abdominal and thoracic masses and ascites; fine-needle aspirates of mesenteric and splenic masses confirmed malignancy, most likely histiocytic sarcoma. Laboratory analyses revealed increased ionized calcium and parathyroid hormone-related peptide (PTH-rP) concentrations, and concurrent low-normal parathyroid hormone concentration, consistent with humoral hypercalcemia of malignancy. Necropsy was performed after euthanasia. The dog had disseminated histiocytic sarcoma, including sarcomatosis, as well as bilateral thyroid carcinoma. PTH-rP immunostaining was positive in the thyroid carcinoma but negative in the histiocytic neoplasm. These results suggest that thyroid carcinoma-associated hypercalcemia can be caused by tumor secretion of PTH-rP.


Rotation driven translational diffusion of polyatomic ions in water: A novel mechanism for breakdown of Stokes-Einstein relation.
While most of the existing theoretical and simulation studies have focused on simple, spherical, halide and alkali ions, many chemically, biologically, and industrially relevant electrolytes involve complex non-spherical polyatomic ions like nitrate, chlorate, and sulfate to name only a few. Interestingly, some polyatomic ions in spite of being larger in size show anomalously high diffusivity and therefore cause a breakdown of the venerable Stokes-Einstein (S-E) relation between the size and diffusivity. Here we report a detailed analysis of the dynamics of anions in aqueous potassium nitrate (KNO3) and aqueous potassium acetate (CH3COOK) solutions. The two ions, nitrate (NO3-) and acetate (CH3CO2-), with their similar size show a large difference in diffusivity values. We present evidence that the translational motion of these polyatomic ions is coupled to the rotational motion of the ion. We show that unlike the acetate ion, nitrate ion with a symmetric charge distribution among all periphery oxygen atoms shows a faster rotational motion with large amplitude rotational jumps which enhances its translational motion due to translational-rotational coupling. By creating a family of modified-charge model systems, we have analysed the rotational motion of asymmetric polyatomic ions and the contribution of it to the translational motion. These model systems help clarifying and establishing the relative contribution of rotational motion in enhancing the diffusivity of the nitrate ion over the value predicted by the S-E relation and also over the other polyatomic ions having asymmetric charge distribution like the acetate ion. In the latter case, reduced rotational motion results in lower diffusivity values than those with symmetric charge distribution. We propose translational-rotational coupling as a general mechanism of the breakdown of the S-E relation in the case of polyatomic ions.


[The effect of benzodiazepine derivatives on the formation of brain edema-swelling in the dynamics of the posttraumatic period].
In experiments on rats the influence of diazepam, cinazepam, and phenazepam (0.5 mg/kg, intraperitoneally) on the development of brain edema in the dynamics of acute craniocerebral injury was studied. The diazepam and phenazepam prevented the development of brain edema in the post-traumatic period. The antiedematous effect of benzodiazepines was connected with their influence on metabolism of the brain.


Fatty acid binding protein type 7 deficiency preserves auditory function in noise-exposed mice.
Fatty acid-binding protein 7 (FABP7) is vital for uptake and trafficking of fatty acids in the nervous system. To investigate the involvement of FABP7 in noise-induced hearing loss (NIHL) pathogenesis, we used Fabp7 knockout (KO) mice generated via CRISPR/Cas9 in the C57BL/6 background. Initial auditory brainstem response (ABR) measurements were conducted at 9 weeks, followed by noise exposure at 10 weeks. Subsequent ABRs were performed 24 h later, with final measurements at 12 weeks. Inner ears were harvested 24 h after noise exposure for RNA sequencing and metabolic analyses. We found no significant differences in initial ABR measurements, but Fabp7 KO mice showed significantly lower thresholds in the final ABR measurements. Hair cell survival was also enhanced in Fabp7 KO mice. RNA sequencing revealed that genes associated with the electron transport chain were upregulated or less impaired in Fabp7 KO mice. Metabolomic analysis revealed various alterations, including decreased glutamate and aspartate in Fabp7 KO mice. In conclusion, FABP7 deficiency mitigates cochlear damage following noise exposure. This protective effect was supported by the changes in gene expression of the electron transport chain, and in several metabolites, including excitotoxic neurotransmitters. Our study highlights the potential therapeutic significance of targeting FABP7 in NIHL.


Single-Leg Squat Performance and Its Relationship to Extensor Mechanism Strength After Anterior Cruciate Ligament Reconstruction.
Performance in strength and functional testing is important when considering return to sport after anterior cruciate ligament (ACL) reconstruction. Both knee extensor strength and the single-leg squat (SLS) have been used in this context. To evaluate the relationship between knee extensor strength and SLS performance after primary ACL reconstruction. Cohort study (Diagnosis); Level of evidence, 3. A prospective cohort of 100 patients was assessed 6 and 12 months after primary ACL reconstruction with a hamstring tendon autograft. Knee extensor peak torque was measured. Three sequential SLSs were performed, and the maximum flexion angle identified from frame-by-frame video analysis was used as the measure of squat performance. A limb symmetry index (LSI) was calculated and satisfactory performance defined as ≥90%. Extensor mechanism strength deficits were seen in 75% of patients at 6 months and 57% at 12 months postoperatively. Mean extensor mechanism strength showed a large improvement between 6 and 12 months (123.6 vs 147.8 N·m, respectively; P < .001; Cohen d = 1.10), and while there was also a statistically significant improvement in the mean maximum flexion angle, the change was small (66.1° vs 68.1°, respectively; P = .011; Cohen d = 0.26). There was a weak positive correlation between knee extensor strength and the SLS maximum flexion angle at 6 months (r = 0.342; P < .001) and 12 months (r = 0.245; P = .014). An SLS LSI <90% was 80% specific and 35% sensitive for extensor mechanism weakness at 6 months and 79% specific and 18% sensitive at 12 months. Extensor mechanism strength deficits are common after ACL reconstruction but reduce between 6 and 12 months. The SLS maximum flexion angle has a weak linear relationship to knee extensor strength. SLS performance has high specificity but low sensitivity in identifying extensor mechanism strength deficits. The SLS maximum flexion angle is therefore a suboptimal surrogate test to identify extensor mechanism strength deficits as diagnosed by isokinetic dynamometric testing. However, unsatisfactory SLS performance indicates a very high chance of underlying extensor mechanism weakness.


Feasibility and Preliminary Responses to a Screening and Brief Intervention Program for Maternal Mental Disorders Within the Context of Primary Care.
There is little evidence of the feasibility and acceptability of integrating screening, brief intervention and referral to treatment services that address depression and alcohol, tobacco and other drug (ATOD) use into antenatal care in South Africa. Data were extracted from program records on the number of eligible women screened and number meeting criteria for depression and self-reported ATOD use. 70 women completed a questionnaire examining their preliminary responses and five MOU personnel were interviewed to identify potential barriers to implementation. Of the 3407 eligible women, 1468 (43 %) women were screened for depression or ATOD use, of whom 302 (21.4 %) screened at risk for depression, 388 (26.4 %) disclosed smoking tobacco, and 29 (2 %) disclosed alcohol or other drugs (AOD). Seventy participants completed the three month follow-up interview. Depression scores decreased significantly following the intervention (t (69) = 8.51, p < 0.001) as did self-reported tobacco use (t (73) = 3.45, p < 0.001), however self-reported AOD use remained unchanged.


Atypical swallowing: a review.
Atypical swallowing is a myofunctional problem consisting of an altered tongue position during the act of swallowing. High incidence in population, multifactorial etiology and the recurring connection with the presence of malocclusions made it a topic of strong interest and discussion in science. The purpose of this review is to illustrate the current orientation on the topic of atypical swallowing, trying in particular to answer two questions: 1) what kind of connection is there between atypical swallowing and malocclusion; 2) what kind of therapy should be used to solve it. This review was conducted on the Medline database [www.ncbi.nim.nih.gov/pubmed] searching for the keywords "atypical swallowing" and "tongue thrust". We examined all the documents from the year 1990 onwards, excluding the ones about syndromic cases of the central motor system. The causal relation between the two problems seems to be biunique: some authors affirm that this oral habit starts as a compensation mechanism for a preexisting malocclusion (especially in case of open-bite); other texts show that it has a tendency to exacerbate cases of malocclusion; it is also proven that a non-physiological tongue thrust can negatively influence the progress of an ongoing orthodontic therapy. Thereby, the best therapeutic approach seems to be a multidisciplinary one: beside orthodontics, which is necessary to correct the malocclusion, it is essential to set up a myofunctional rehabilitation procedure to correct the oral habit, therefore granting long time permanent results. There is also proof of a substantial difference between the results obtained from early (deciduous or primary mixed dentition) or later treatments. The biunique causal relation between atypical swallowing and malocclusion suggests a multidisciplinary therapeutic approach, orthodontic and myofunctional, to temporarily solve both problems. An early diagnosis and a prompt intervention have a significantly positive influence on the therapy outcome.


[Mathematical modeling to assess the effects of organic waste dumping on sanitary conditions of environmental waters].
This article provides an overview of emerging trends in environmental approaches to wastewater management in the developing world. Organic matter present in wastewater is one of the main sources of water pollution. In large amounts it can increase the number of microorganisms and thus the excessive consumption of dissolved oxygen in the metabolic processes of its utilization and stabilization, leading to the disappearance and extinction of aerobic organisms, favoring the appearance of other life forms in anaerobic conditions, often producing toxic residues. The main focus of this article is to analyze the presence of organic matter in urban sewage using a water quality model whose main parameters are dissolved oxygen and biochemical oxygen demand. Wastewater management methodologies can contribute to public health, sanitation, and improved conservation of water resources.


Effect of nitrogen-containing compounds on polychlorinated dibenzo-p-dioxin/dibenzofuran formation through de novo synthesis.
An experimental study was conducted to clarify the suppression effect of nitrogen-containing compounds, that is, ammonia and urea, on the formation of polychlorinated dibenzo-p-dioxins (PCDDs) and polychlorinated dibenzofurans (PCDFs) through the de novo synthesis reaction. In the experiment, graphite and copper chloride contained in a mixture were used as sources of carbon and chlorine, respectively. The granulated sample mixture was charged as a packed-bed in the glass tube and heated at 300 degrees C in the flow of Ar-O2 gas mixture. In some cases, urea was added as aqueous solution to the sample, while ammonia was added to the gas flowed through the sample bed. The amount of PCDD/Fs formed decreases significantly by the addition of both ammonia and urea. Particularly, the addition of urea reduces the amount of PCDD/Fs discharged in the outlet gas by approximately 90%. The oxidation rate of carbon in the early stage of the experiment, that is, the heating period, is promoted bythe addition of nitrogen-containing compounds. However, soon afterthe temperature reaches 300 degrees C, the formation rate becomes lower than that of the case without the addition of nitrogen-containing compounds. On the other hand, organic compounds containing amino (-NH2) or cyanide (-CN) groups and those containing nitrogen within the carbon ring frame were detected in the outlet gas in the case of urea addition. Typically observed aromatic compounds are chlorobenzonitriles, chlorobenzeneamines, and chloropyridines. This suggests a possibility that hydrogen and/or chlorine combined with PCDD/Fs are also substituted by such nitrogen-containing groups, and this decreases the formation rate of their frame of carbon rings. This phenomenon was also consistent with the fact that a significant reduction was observed in the amount of PCDD/Fs released to the outlet gas when urea was added.


Critical Roles of Calpastatin in Ischemia/Reperfusion Injury in Aged Livers.
Ischemia/reperfusion (I/R) injury unavoidably occurs during hepatic resection and transplantation. Aged livers poorly tolerate I/R during surgical treatment. Although livers have a powerful endogenous inhibitor of calpains, calpastatin (CAST), I/R activates calpains, leading to impaired autophagy, mitochondrial dysfunction, and hepatocyte death. It is unknown how I/R in aged livers affects CAST. Human and mouse liver biopsies at different ages were collected during in vivo I/R. Hepatocytes were isolated from 3-month- (young) and 26-month-old (aged) mice, and challenged with short in vitro simulated I/R. Cell death, protein expression, autophagy, and mitochondrial permeability transition (MPT) between the two age groups were compared. Adenoviral vector was used to overexpress CAST. Significant cell death was observed only in reperfused aged hepatocytes. Before the commencement of ischemia, CAST expression in aged human and mouse livers and mouse hepatocytes was markedly greater than that in young counterparts. However, reperfusion substantially decreased CAST in aged human and mouse livers. In hepatocytes, reperfusion rapidly depleted aged cells of CAST, cleaved autophagy-related protein 5 (ATG5), and induced defective autophagy and MPT onset, all of which were blocked by CAST overexpression. Furthermore, mitochondrial morphology was shifted toward an elongated shape with CAST overexpression. In conclusion, CAST in aged livers is intrinsically short-lived and lost after short I/R. CAST depletion contributes to age-dependent liver injury after I/R.


Nonalcoholic Fatty Liver Disease and Associated Risk Factors in a Community-Based Sample of Mexican-Origin Adults.
The incidence of nonalcoholic fatty liver disease (NAFLD) is highest among Mexican-origin (MO) adults. Few studies have estimated the prevalence of NAFLD in this subpopulation, particularly by sex and age. We assessed the prevalence of NAFLD in a community sample of MO adults residing in a border region of southern Arizona and determined risk factors associated with NAFLD. A total of 307 MO adults (n = 194 women; n = 113 men) with overweight or obesity completed an in-person study visit, including vibration-controlled transient elastography (FibroScan) for the assessment of NAFLD status. A continuous attenuation parameter score of ≥288 dB/m (≥5% hepatic steatosis) indicated NAFLD status. Multivariable logistic regression was used to estimate odds ratios (ORs) and 95% confidence intervals (CIs) for NAFLD. We identified 155 participants (50%) with NAFLD, including 52% of women and 48% of men; there were no sex differences in steatosis (men, 287.8 dB/m; women, 288.4 dB/m). Sex, age, patatin-like phospholipase domain containing 3 (PNPLA3) risk allele carrier status, comorbidities, and cultural and behavioral variables were not associated with NAFLD status. There was some evidence for effect modification of body mass index (BMI) by sex (Pinteraction = 0.08). The estimated OR for an increase in BMI of 5 kg/m2 was 3.36 (95% CI, 1.90, 5.91) for men and 1.92 (95% CI, 1.40, 2.64) for women. In post hoc analyses treating steatosis as a continuous variable in a linear regression, significant effect modification was found for BMI by sex (Pinteraction = 0.03), age (P = 0.05), and PNPLA3 risk allele carrier status (P = 0.02). Conclusion: Lifestyle interventions to reduce body weight, with consideration of age and genetic risk status, are needed to stem the higher rates of NAFLD observed for MO populations.


Prevalence and genotypic characterization of Salmonella spp. from chicken meats marketed in the province of Skikda, Algeria.
Here, we aim to determine the prevalence of Salmonella contamination of poultry meat from butcheries of the province of Skikda and to investigate antibiotic resistance. Salmonella spp. isolates were screened from 70 samples, including chicken breasts (n = 40 samples) and chicken thighs (n = 30 samples) collected from 14 butcheries. All suspected Salmonella colonies from selective media were confirmed by MALDI-TOF MS and serotyped. The susceptibility profile to 16 antibiotics was studied. According to the antibiotic susceptibility results, resistance genes were investigated by standard PCR targeting various genes such as blaSHV, blaTEM, aac3, aac6-Ibcr, aad, qnrA and qnrB. Of the 14 butcheries studied, samples from eight butcheries were contaminated with Salmonella (57.14%). 19 Salmonella strains were isolated, including five serotypes with a predominance of Kentucky serotype (n = 9), Enteridis (n = 3), followed by Heidelberg (n = 3), Virchow (n = 3), and Manhattan (n= 1). All isolates were resistant to Rifampicin (100%; n = 19), and to other antibiotics such as Ciprofloxacin (47.36%), Amoxicillin-clavulanic acid (47.36%; n = 9), Amoxicillin, (47.36%; n = 9), Ticarcillin-clavulanic acid (47.36%; n = 9), and Gentamycin (47.36%; n = 9). All isolates showing multidrug resistance (47.36%; n = 9) were positive by PCR to the blaTEM-1 β-lactamase gene, from which 8 strains carried the aminoglycoside resistance aad7 gene. However, none was positive for the tested blaSHV, Aac3, Aac6-Ibcr, qnrA, qnrB, ArmA and ArmB genes. Our findings show a worrying rate of Salmonella contamination of poultry meats.


On being the right size: the impact of population size and stochastic effects on the evolution of drug resistance in hospitals and the community.
The evolution of drug resistant bacteria is a severe public health problem, both in hospitals and in the community. Currently, some countries aim at concentrating highly specialized services in large hospitals in order to improve patient outcomes. Emergent resistant strains often originate in health care facilities, but it is unknown to what extent hospital size affects resistance evolution and the resulting spillover of hospital-associated pathogens to the community. We used two published datasets from the US and Ireland to investigate the effects of hospital size and controlled for several confounders such as antimicrobial usage, sampling frequency, mortality, disinfection and length of stay. The proportion of patients acquiring both sensitive and resistant infections in a hospital strongly correlated with hospital size. Moreover, we observe the same pattern for both the percentage of resistant infections and the increase of hospital-acquired infections over time. One interpretation of this pattern is that chance effects in small hospitals impede the spread of drug-resistance. To investigate to what extent the size distribution of hospitals can directly affect the prevalence of antibiotic resistance, we use a stochastic epidemiological model describing the spread of drug resistance in a hospital setting as well as the interaction between one or several hospitals and the community. We show that the level of drug resistance typically increases with population size: In small hospitals chance effects cause large fluctuations in pathogen population size or even extinctions, both of which impede the acquisition and spread of drug resistance. Finally, we show that indirect transmission via environmental reservoirs can reduce the effect of hospital size because the slow turnover in the environment can prevent extinction of resistant strains. This implies that reducing environmental transmission is especially important in small hospitals, because such a reduction not only reduces overall transmission but might also facilitate the extinction of resistant strains. Overall, our study shows that the distribution of hospital sizes is a crucial factor for the spread of drug resistance.


The evolution of the terrestrial vertebrates: environmental and physiological considerations.
Physiological evidence has long been used to suggest that the gnathostomous vertebrates (those possessing jaws) were primitively fresh water. The same was also the case for the Osteichthyes (bony fish) and the Tetrapoda (Amphibia, Reptilia, Aves, Mammalia). However, the geological evidence favours a marine origin for the vertebrates as a whole, and, for the gnathostomes and the osteichthyans in particular. Some of the earliest amphibian remains may be associated with tidally influenced sediments. Furthermore, during the early part of the Devonian, fresh water chemistry may well have been different from that of today, lessening the divide between marine and non-marine environments. Urea formation via the ornithine cycle, and urea retention in the body fluids, are useful adaptations for terrestrial life. They prevent excessive water loss associated with the elimination of nitrogenous waste. These abilities may have been primitive for the gnathostomes, and were developed in the marine environment to reduce osmotic dehydration. In the aqueous medium, gaseous exchange is effected by the gills. These organs are, on the whole, useless in air. For vertebrates, air-breathing is effected by an inflatable sac, with moist linings, and an internal location. Some form of air-breathing sac was primitive for the osteichthyans, and may have been primitive for the gnathostomes. Again, this adaptation for terrestrial life developed in response to conditions experienced in the marine, aquatic environment. A new model of tetrapod evolution is proposed in the light of the basic marine origin and character of the ancestors of the tetrapods.


Rapid screening method for intact glucosinolates in Chinese medicinal herbs by using liquid chromatography coupled with electrospray ionization ion trap mass spectrometry in negative ion mode.
An optimized method using liquid chromatography coupled with electrospray ionization ion trap mass spectrometry (LC/ESI-ITMS) in negative ion mode has been developed for screening different structural classes of intact glucosinolates in six Chinese medicinal herbs. The glucosinolates were extracted with hot methanol/water (70:30 v/v) and separation of the individual glucosinolates was achieved using a reversed-phase C18 column with an aqueous ammonium acetate/methanol gradient. Identification of the intact glucosinolates was based on the detection of compounds with a constant neutral loss of 242 Da corresponding to the combined loss of anhydroglucose (162 Da) and sulfur trioxide (80 Da) in collision-induced dissociation. The structures of the identified glucosinolates were confirmed with the use of group-specific product ions at m/z 195, 241, 259, 275 in their corresponding MS/MS product ion spectra. Differentiation of intact glucosinolates was achieved through their respective retention times and molecular masses as well as the characteristic product ions. The limits of detection were at the low nanogram level per injection, based on constant neutral loss scans. Significant variation in the compositions of intact glucosinolates was identified in the cruciferous herbs. This method was applied in the differentiation and quality control of two pairs of easily confused herbs.


[Effects of transgenic crops on soil microorganisms: a review].
The worldwide cultivation of transgenic crops not only provides tremendous economic benefits, but also induces the concern about the potential risks of transgenic crops on soil ecosystem in which microorganisms are involved. The potential effects of transgenic crops on soil microorganisms include the direct effects of the transgenic proteins on non-target soil microorganisms, and the indirect effects of the unintentional changes in the chemical compositions of root exudates induced by the introduction of the exogenous transgenic proteins. Most of the studies on transgenic crops suggested that transgenic crops could affect the quantity and structure of soil microbial populations. However, the perceivable effects on the soil microorganisms are inconsistent, with some in significant and others in non-significant, or some with persistent and others with non-persistent. This paper summarized the effects of different transgenic crops on soil microorganisms, and discussed the factors affecting the assessment reliability, including the species of transgenic crops and the experimental technologies and principles. Some issues needed to be paid special attention to in the future studies were put forward.


Decalcification of schistosome eggs during staining of tissue sections: a potential source of diagnostic error.
Miracidial nuclei within calcified schistosome eggs are frequently mummified. Calcified eggs are often decalcified prior to or during the routine processing and staining of tissues, and mummified miracidial nuclei then stain clearly and are often arranged in the patterns seen in live eggs. These miracidia may be incorrectly assumed to be alive or to have died recently and thus be taken as evidence of an active infection requiring chemotherapy. The present observations demonstrate the decalcification of mineralized eggs by hematoxylin preparations used for routine staining of tissue sections.


Identification of receptor-tyrosine-kinase-signaling target genes reveals receptor-specific activities and pathway branchpoints during Drosophila development.
Receptor tyrosine kinases (RTKs) are an important family of signaling molecules with the unusual property that they are able to transduce their signals using the same downstream pathways. This has led to an unresolved debate as to whether individual receptors are interchangeable, or if each receptor can mediate specific downstream responses. To address this question, we have conducted a screen to identify target genes whose expression is differentially modulated by RTKs and their downstream pathway components. Using whole-mount in situ hybridization in Drosophila embryos exposed to constitutively active RTK pathway signaling, along with quantitative RT-PCR, we found that a significant fraction of target genes respond differentially in a spatial and/or quantitative manner. This includes differential responses to EGF receptor vs. fibroblast growth factor receptor signaling as well as to more downstream components such as Ras1 and pointed. We show that not only genes but also individual alternative transcripts can respond differently to signaling, and we present evidence that the differential responses can be mediated at the transcriptional level. Our results demonstrate that different RTKs can elicit distinct transcriptional responses, and the target genes obtained from our screen provide a valuable resource for further exploration of the mechanisms underlying this signaling specificity.


Retinoids differentially regulate the proliferation of colon cancer cell lines.
In this study, the proliferative effects of retinoids were examined in the MC-26 and LoVo colon adenocarcinoma cell lines. The proliferation of the LoVo cell line was not altered in the presence of the retinoids all trans-retinoic acid (atRA) and 9-cis-retinoic acid (9-cis-RA). Both retinoids, however, stimulated the growth, as measured by cell proliferation, of MC-26 cells. atRA and 9-cis-RA were equipotent in increasing MC-26 cell proliferation, suggesting that the growth stimulation is mediated by one or more retinoic acid receptors (RARs). To determine the RAR which might be responsible for this growth stimulatory effect, we characterized the RAR subtypes which were present in both cell lines. mRNA for the RAR alpha, RAR beta, and RAR gamma were detected in the MC-26 cell. Of the RARs present in MC-26 cells, the RAR alpha does not mediate the growth stimulatory effects of retinoids, for a selective RAR alpha antagonist was unable to prevent the retinoid-induced increase in MC-26 cell growth. RAR alpha, RAR beta, and RAR gamma mRNA are also expressed in the LoVo cell line; the lack of growth-stimulation by retinoids in LoVo cells, therefore, does not seem to be due to the absence of RARs. The results obtained in these experiments demonstrate that the growth response elicited by retinoids can vary between colon cancer cells and that the differences in response may not be solely determined by the RAR subtypes which are expressed in a colon cancer cell line.


Diet-microbiota interactions and their implications for healthy living.
It is well established that diet influences the health of an individual and that a diet rich in plant-based foods has many advantages in relation to the health and well-being of an individual. What has been unclear until recently is the large contribution of the gut microbiota to this effect. As well as providing basic nutritional requirements, the long-term diet of an animal modifies its gut microbiota. In adults, diets that have a high proportion of fruit and vegetables and a low consumption of meat are associated with a highly diverse microbiota and are defined by a greater abundance of Prevotella compared to Bacteroides, while the reverse is associated with a diet that contains a low proportion of plant-based foods. Furthermore, it is becoming increasingly clear that the effect of the microbial ecology of the gut goes beyond the local gut immune system and is implicated in immune-related disorders, such as IBS, diabetes and inflamm-ageing. In this review, we investigate the evidence that a balanced diet leads to a balanced, diverse microbiota with significant consequences for healthy ageing by focusing on conditions of interest.


Control of hydroxyproline catabolism in Sinorhizobium meliloti.
Hydroxyproline (Hyp) in decaying organic matter is a rich source of carbon and nitrogen for microorganisms. A bacterial pathway for Hyp catabolism is known; however, genes and function relationships are not established. In the pathway, trans-4-hydroxy-L-proline (4-L-Hyp) is epimerized to cis-4-hydroxy-D-proline (4-D-Hyp), and then, in three enzymatic reactions, the D-isomer is converted via Δ-pyrroline-4-hydroxy-2-carboxylate (HPC) and α-ketoglutarate semialdehyde (KGSA) to α-ketoglutarate (KG). Here a transcriptional analysis of cells growing on 4-L-Hyp, and the regulation and functions of genes from a Hyp catabolism locus of the legume endosymbiont Sinorhizobium meliloti are reported. Fourteen hydroxyproline catabolism genes (hyp), in five transcripts hypR, hypD, hypH, hypST and hypMNPQO(RE)XYZ, were negatively regulated by hypR. hypRE was shown to encode 4-hydroxyproline 2-epimerase and a hypRE mutant grew with 4-D-Hyp but not 4-L-Hyp. hypO, hypD and hypH are predicted to encode 4-D-Hyp oxidase, HPC deaminase and α-KGSA dehydrogenase respectively. The functions for hypS, hypT, hypX, hypY and hypZ remain to be determined. The data suggest 4-Hyp is converted to the tricarboxylic acid cycle intermediate α-ketoglutarate via the pathway established biochemically for Pseudomonas. This report describes the first molecular characterization of a Hyp catabolism locus.


Differentiation and characterization of rat adipose tissue mesenchymal stem cells into endothelial-like cells.
In this study, mesenchymal stem cells were isolated from rat adipose tissue (AD-MSCs) to characterize and differentiate them into endothelial-like cells. AD-MSCs were isolated by mechanical and enzymatic treatments, and their identity was verified by colony-forming units (CFU) test and by differentiation into cells of mesodermal lineages. The endothelial differentiation was induced by plating another aliquot of cells in EGM-2 medium, enriched with specific endothelial growth factors. Five subcultures were performed. The expression of stemness genes (OCT4, SOX2 and NANOG) was investigated. The presence of CD90 and the absence of the CD45 were evaluated by flow cytometry. The endothelial-like cells were characterized by the evaluation of morphological changes and gene expression analysis for endothelial markers (CD31, CD144, CD146). Characterization of AD-MSCs showed their ability to form clones, to differentiate in vitro and the OCT-4, SOX-2, NANOG genes expression. Immunophenotypic characterization showed the CD90 presence and the CD45 absence. The endothelial-like cells showed morphological changes, the expression of CD31, CD144, CD146 genes and the presence of CD31 membrane receptor. Matrigel assay showed their ability to form network and vessels-like structures. This study lays the foundations for future evaluation of the potential AD-MSCs pro-angiogenic and therapeutic role.