Genetic and functional analysis of the bovine uterine microbiota. Part II: Purulent vaginal discharge versus healthy cows

Publication date: Available online 2 March 2017
Source:Journal of Dairy Science
Author(s): M.L.S. Bicalho, S. Lima, C.H. Higgins, V.S. Machado, F.S. Lima, R.C. Bicalho
The aim of this study was to characterize, using metagenomic shotgun DNA sequencing, the intrauterine microbial population and its predicted functional diversity within healthy cows and cows presenting purulent vaginal discharge (PVD). Twenty Holstein dairy cows from a single farm were enrolled in the study at 25 to 35 d postpartum. Purulent vaginal discharge was diagnosed by retrieving and scoring vaginal discharge using the Metricheck device (Simcro, Hamilton, New Zealand). Intrauterine samples for metagenomic analysis were collected by the cytobrush technique from 8 cows diagnosed with PVD and 12 healthy cows. Pair-end sequencing was performed using the Illumina MiSeq platform (Illumina Inc., San Diego, CA). Metagenomic sequences were analyzed using the MG-RAST server (metagenomic rapid annotations using subsystems technology; http://ift.tt/1qOVzJC), and the STAMP software (http://ift.tt/UtpTJL) was used to study statistically significant differential abundance of taxonomic and functional features between the 2 metagenomes. Additionally, the total number of bacterial 16S rDNA copies was estimated by real-time PCR. Taxonomic analysis revealed that Bacteroidetes was the most abundant phylum in the uterine microbiota from cows with PVD, and Fusobacteria was almost completely absent in the healthy uterine microbiota. Moreover, species belonging to the genus Trueperella were present only in the uterine microbiota of PVD cows. The increased abundance of Fusobacteria and the unique presence of Trueperella in the PVD cows highlight the important role of these bacteria in the pathogenesis of PVD. Genes encoding cytolethal distending toxin were exclusive to the microbiota of PVD cows. Similarly, genes associated with lipid A modification were present only in samples from PVD cows; such modification is associated with greater resistance to cationic antimicrobial peptides. Conversely, genes encoding bacteriocins and ribosomally antibacterial peptide were exclusively found in the healthy uterine microbiota and dominated by tolerance to colicin E2. No difference was observed in total bacterial load between the 2 microbiotas. This study provides deep insight into the uterine microbial community in health and disease. The observations that the healthy microbiota is tolerant to colicin E2, whereas the uterine microbiota of PVD cows produces cytolethal distending toxins and modifies its lipopolysaccharides suggest that species-intrinsic factors may be more relevant than bacterial abundance to the development of disease or maintenance of health in the dairy cow postpartum uterus.



http://ift.tt/2miEcT6

Novel approaches to assess the quality of fertility data stored in dairy herd management software

Publication date: Available online 2 March 2017
Source:Journal of Dairy Science
Author(s): K. Hermans, W. Waegeman, G. Opsomer, B. Van Ranst, J. De Koster, M. Van Eetvelde, M. Hostens
Scientific journals and popular press magazines are littered with articles in which the authors use data from dairy herd management software. Almost none of such papers include data cleaning and data quality assessment in their study design despite this being a very critical step during data mining. This paper presents 2 novel data cleaning methods that permit identification of animals with good and bad data quality. The first method is a deterministic or rule-based data cleaning method. Reproduction and mutation or life-changing events such as birth and death were converted to a symbolic (alphabetical letter) representation and split into triplets (3-letter code). The triplets were manually labeled as physiologically correct, suspicious, or impossible. The deterministic data cleaning method was applied to assess the quality of data stored in dairy herd management from 26 farms enrolled in the herd health management program from the Faculty of Veterinary Medicine Ghent University, Belgium. In total, 150,443 triplets were created, 65.4% were labeled as correct, 17.4% as suspicious, and 17.2% as impossible. The second method, a probabilistic method, uses a machine learning algorithm (random forests) to predict the correctness of fertility and mutation events in an early stage of data cleaning. The prediction accuracy of the random forests algorithm was compared with a classical linear statistical method (penalized logistic regression), outperforming the latter substantially, with a superior receiver operating characteristic curve and a higher accuracy (89 vs. 72%). From those results, we conclude that the triplet method can be used to assess the quality of reproduction data stored in dairy herd management software and that a machine learning technique such as random forests is capable of predicting the correctness of fertility data.



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Association of single nucleotide polymorphisms in candidate genes previously related to genetic variation in fertility with phenotypic measurements of reproductive function in Holstein cows

Publication date: Available online 2 March 2017
Source:Journal of Dairy Science
Author(s): M. Sofia Ortega, Anna C. Denicol, John B. Cole, Daniel J. Null, Jeremy F. Taylor, Robert D. Schnabel, Peter J. Hansen
Many genetic markers related to health or production traits are not evaluated in populations independent of the discovery population or related to phenotype. Here we evaluated 68 single nucleotide polymorphisms (SNP) in candidate genes previously associated with genetic merit for fertility and production traits for association with phenotypic measurements of fertility in a population of Holstein cows that was selected based on predicted transmitting ability (PTA) for daughter pregnancy rate (DPR; high, ≥1, n = 989; low, ≤ −1.0, n = 1,285). Cows with a high PTA for DPR had higher pregnancy rate at first service, fewer services per conception, and fewer days open than cows with a low PTA for DPR. Of the 68 SNP, 11 were associated with pregnancy rate at first service, 16 with services per conception, and 19 with days open. Single nucleotide polymorphisms in 12 genes (BDH2, BSP3, CAST, CD2, CD14, FUT1, FYB, GCNT3, HSD17B7, IBSP, OCLN, and PCCB) had significant associations with 2 fertility traits, and SNP in 4 genes (CSPP1, FCER1G, PMM2, and TBC1D24) had significant associations with each of the 3 traits. Results from this experiment were compared with results from 2 earlier studies in which the SNP were associated with genetic estimates of fertility. One study involved the same animals as used here, and the other study was of an independent population of bulls. A total of 13 SNP associated with 1 or more phenotypic estimates of fertility were directionally associated with genetic estimates of fertility in the same cow population. Moreover, 14 SNP associated with reproductive phenotype were directionally associated with genetic estimates of fertility in the bull population. Nine SNP (located in BCAS, BSP3, CAST, FUT1, HSD17B7, OCLN, PCCB, PMM2, and TBC1D24) had a directional association with fertility in all 3 studies. Examination of the function of the genes with SNP associated with reproduction in more than one study indicates the importance of steroid hormones and immune function as determinants of reproductive function. All but 1 of the 68 evaluated SNP were variable in 11 breeds besides Holstein, indicating the potential effects of these SNP on reproductive function across breeds of cattle.



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Representing interconversions among volatile fatty acids in the Molly cow model

Publication date: Available online 2 March 2017
Source:Journal of Dairy Science
Author(s): S. Ghimire, R.A. Kohn, P. Gregorini, R.R. White, M.D. Hanigan
The Molly cow model uses fixed stoichiometric coefficients for predicting volatile fatty acid (VFA) production from the fermented individual dietary nutrient fractions of forage and concentrate. We previously showed that predictions of VFA production had large errors and hypothesized that it was due to a lack of representation of carbon exchange among VFA. The objectives of the present study were to add VFA interconversion equations based on thermodynamics to the Molly cow model and evaluate the effect of these additions on model accuracy and precision of VFA predictions. Previously described thermodynamic equations were introduced to represent interconversions among VFA. The model was further modified to predict de novo acetate, propionate, and butyrate production coefficients based on forage-to-concentrate ratios rather than discrete, fixed sets of coefficients for forage-based, concentrate-based, and mixed diets. Both the original model and the modified one were reparameterized and evaluated against a common data set containing 8 studies reporting pH, VFA concentration, and VFA production rates using isotope dilution techniques and 62 studies reporting VFA concentrations and pH. Evaluations after parameter estimation revealed that predictions of VFA production rates were not improved, with root mean squared prediction errors (RMSPE) of 77, 60, and 51% for acetate, propionate, and butyrate, respectively, for the revised model versus 75, 63, and 55, respectively, for the original model. The RMSPE for predictions of VFA concentrations were reduced from 28, 46, and 40% to 22, 31, and 26% for acetate, propionate, and butyrate, respectively, simply by rederiving the VFA coefficients, but minimal further improvement was achieved with the addition of thermodynamically driven interconversion equations (RMSPE of 21, 32, and 27% for acetate, propionate, and butyrate, respectively). Thus, the results indicate that thermodynamically driven interchanges among VFA, as represented in this study, may not be a primary determinant for the accuracy of predictions of net production rates. Including the effect of pH on VFA absorption reduced the mean bias of propionate production and slope bias of acetate production, but not the overall RMSPE. The larger prediction errors for VFA production as compared with concentrations suggest the data quality may not be high, or that our representation of VFA production and absorption as well as ruminal digestion is inadequate. Additional data are required to discriminate among these hypotheses.



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Citizens' views on the practices of zero-grazing and cow-calf separation in the dairy industry: Does providing information increase acceptability?

Publication date: Available online 2 March 2017
Source:Journal of Dairy Science
Author(s): Maria J. Hötzel, Clarissa S. Cardoso, Angélica Roslindo, Marina A.G. von Keyserlingk
The primary aim of this study was to assess the influence of provision of information on lay citizens' opinions regarding 2 common management practices, zero-grazing and cow-calf separation. To aid in the interpretation of the findings, our secondary aim was to explore the awareness and opinions of Brazilian citizens about these practices. We surveyed a convenience sample of Brazilian citizens (192 men and 208 women), recruited in a public place, with the majority stating that they were largely unfamiliar with animal production and lived in urban environments. Participants were presented short scenarios with information on the primary production factors and welfare concerns for and against zero-grazing (n = 200) or cow-calf separation (n = 200). Participants were then asked to state their position (reject, indifferent, or support), and to provide the reason(s) justifying their position. Immediately following, participants were provided a short statement describing either zero-grazing or cow-calf separation, depending on what question they responded to in the first part. Two closed questions (Q) followed each of these statements: (Q1) "Are you aware of this practice?" with choices yes, somewhat, or no, and (Q2) "What is your position regarding this practice?" with choices reject, indifferent, or support. Only 31 and 33% of the respondents were aware of zero-grazing and cow-calf separation, respectively. Previous awareness of existence of practice did not influence levels of support. Provision of information resulted in more people rejecting the practices of zero-grazing and cow-calf separation. Participants' main justifications to reject zero-grazing and cow-calf separation focused on perceived negative effects of practices on farm animal welfare and product quality, and loss of naturalness. Survey participants, Brazilians living in urban environments, with little or no association with dairy production, were generally unaware that many cows do not have access to pasture and that cows are separated from their calf at birth. Independent of provision of additional information, most participants did not support these practices. Provision of brief explanatory information played a minor role in influencing people's views, but failed to result in general acceptance.



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Evaluation of the National Research Council (2001) dairy model and derivation of new prediction equations. 1. Digestibility of fiber, fat, protein, and nonfiber carbohydrate

Publication date: Available online 2 March 2017
Source:Journal of Dairy Science
Author(s): R.R. White, Y. Roman-Garcia, J.L. Firkins, M.J. VandeHaar, L.E. Armentano, W.P. Weiss, T. McGill, R. Garnett, M.D. Hanigan
Evaluation of ration balancing systems such as the National Research Council (NRC) Nutrient Requirements series is important for improving predictions of animal nutrient requirements and advancing feeding strategies. This work used a literature data set (n = 550) to evaluate predictions of total-tract digested neutral detergent fiber (NDF), fatty acid (FA), crude protein (CP), and nonfiber carbohydrate (NFC) estimated by the NRC (2001) dairy model. Mean biases suggested that the NRC (2001) lactating cow model overestimated true FA and CP digestibility by 26 and 7%, respectively, and under-predicted NDF digestibility by 16%. All NRC (2001) estimates had notable mean and slope biases and large root mean squared prediction error (RMSPE), and concordance (CCC) ranged from poor to good. Predicting NDF digestibility with independent equations for legumes, corn silage, other forages, and nonforage feeds improved CCC (0.85 vs. 0.76) compared with the re-derived NRC (2001) equation form (NRC equation with parameter estimates re-derived against this data set). Separate FA digestion coefficients were derived for different fat supplements (animal fats, oils, and other fat types) and for the basal diet. This equation returned improved (from 0.76 to 0.94) CCC compared with the re-derived NRC (2001) equation form. Unique CP digestibility equations were derived for forages, animal protein feeds, plant protein feeds, and other feeds, which improved CCC compared with the re-derived NRC (2001) equation form (0.74 to 0.85). New NFC digestibility coefficients were derived for grain-specific starch digestibilities, with residual organic matter assumed to be 98% digestible. A Monte Carlo cross-validation was performed to evaluate repeatability of model fit. In this procedure, data were randomly subsetted 500 times into derivation (60%) and evaluation (40%) data sets, and equations were derived using the derivation data and then evaluated against the independent evaluation data. Models derived with random study effects demonstrated poor repeatability of fit in independent evaluation. Similar equations derived without random study effects showed improved fit against independent data and little evidence of biased parameter estimates associated with failure to include study effects. The equations derived in this analysis provide interesting insight into how NDF, starch, FA, and CP digestibilities are affected by intake, feed type, and diet composition.



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Randomized clinical field trial on the effects of butaphosphan-cyanocobalamin and propylene glycol on ketosis resolution and milk production

Publication date: Available online 2 March 2017
Source:Journal of Dairy Science
Author(s): J.L. Gordon, S.J. LeBlanc, D.F. Kelton, T.H. Herdt, L. Neuder, T.F. Duffield
The purpose of this study was to determine the effects of a butaphosphan-cyanocobalamin combination product (B+C) and 2 durations of propylene glycol treatment (PG; 3 versus 5 d) on ketosis resolution and early lactation milk yield. Cows from 9 freestall herds (8 in Ontario and 1 in Michigan) were tested at weekly intervals between 3 and 16 d in milk. Ketosis was defined as blood β-hydroxybutyrate (BHB) ≥1.2 mmol/L. Ketotic cows were randomly assigned to treatment with 25 mL of B+C or 25 mL of saline placebo for 3 d and 3 or 5 d of 300 g of PG orally in a 2 × 2 factorial arrangement. Outcomes evaluated for all farms included ketosis cure (blood BHB <1.2 mmol/L at 1 wk after enrollment), maintenance of ketosis cure (blood BHB <1.2 mmol/L 1 and 2 wk after enrollment), and blood BHB concentrations at 1 and 2 wk after enrollment. Daily milk weights were collected in 3 herds. Poisson regression was used to evaluate cure and maintenance of cure, whereas repeated-measures ANOVA was used to evaluate blood BHB concentrations in the 2 wk after enrollment and average daily milk production in the 30 d after treatment. A total of 594 animals were enrolled in the study with 124 treated with B+C and 5 d of PG, 176 treated with B+C and 3 d of PG, 128 treated with saline and 5 d of PG, and 166 treated with saline and 3 d of PG. Animals with blood BHB >2.4 mmol/L at the time of enrollment were 1.7 times more likely [95% confidence interval (CI): 1.4 to 2.2] to cure and had a decrease of 0.25 ± 0.11 mmol/L blood BHB at 1 wk after enrollment if treated with 5 d of PG compared with 3 d, though this response was not seen in animals with BHB of 1.2 to 2.4 mmol/L at enrollment. Cows with blood glucose concentrations <2.2 mmol/L at enrollment produced 3.1 kg/d (95% CI: 1.3 to 5.0) more milk if treated with B+C and 3.4 kg/d (95% CI: 1.7 to 5.1) more milk if treated with 5 d of PG compared with their respective controls. This response was not seen in animals with blood glucose ≥2.2 mmol/L at enrollment and there was no interaction between treatments. These results indicate that extended PG treatment is beneficial in decreasing blood BHB concentrations in more severely affected animals. Additionally, both B+C treatment and extended PG treatment improved milk yield in animals with low blood glucose at the time of ketosis diagnosis.



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Mutual growth-promoting effect between Bifidobacterium bifidum WBBI03 and Listeria monocytogenes CMCC 54001

Publication date: Available online 2 March 2017
Source:Journal of Dairy Science
Author(s): Dong Yang, Xiaoli Wu, Xiaomin Yu, Lihua He, Nagendra P. Shah, Feng Xu
In this paper, Bifidobacterium bifidum WBBI03 and Listeria monocytogenes CMCC 54001 were selected to detect the changes in their growth pattern after mutual interaction between them. The proteomic changes after the interaction between the 2 bacteria were detected by the isobaric tags for relative and absolute quantitation method. The proteins related to the biosynthesis and cell reproduction were selected, and their changes at the transcriptional level were monitored by fluorescent quantitative PCR. Also, 3 other types of probiotic organisms and opportunistic pathogens were used to verify the results mentioned above. The results showed that growing the 2 organisms together could promote the growth of each other, resulting in earlier entry into the logarithmic phase. The results also showed that the expression of these proteins mostly tended to be upregulated at the translational and transcriptional level. The increase in the expression of these proteins might help promote the growth and reproduction of B. bifidum WBBI03 and L. monocytogenes CMCC 54001. One aspect of the biological significance of their presence in the normal intestine may be that the opportunistic pathogens promote the growth of the probiotics.



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Short communication: Feeding distillers dried grains in replacement of forage in limit-fed dairy heifer rations: Effects on post-trial performance

Publication date: Available online 2 March 2017
Source:Journal of Dairy Science
Author(s): A.K. Manthey, J.L. Anderson
The objective of this study was to determine the effect of increasing the inclusion rate of distillers dried grains with solubles (DDGS) in replacement of forage in diets that were limit-fed during peripubertal growth on performance during the first 3 mo of first lactation. A secondary objective was to also characterize reproductive performance of the dairy heifers. A 16-wk randomized complete block design study was conducted using 48 Holstein heifers (199 ± 2 d of age) with 3 treatments. Treatments were (1) 30% DDGS, (2) 40% DDGS, and (3) 50% DDGS, with the remainder of the diet consisting of grass hay and 1.5% mineral mix. Heifers were individually limit-fed using Calan gates at 2.65, 2.50, and 2.35% of body weight on a dry matter basis for 30, 40, and 50% DDGS, respectively. After completing the feeding study, heifers were fed a common diet according to standard herd management. Data on reproductive performance and milk production for the first 3 mo of lactation were collected for each heifer from dairy herd records. At 3 wk prepartum and at calving, body weight, frame measurements, and body condition score were recorded. We found no differences in reproductive or frame measurements taken around parturition. However, due to relatively small numbers of heifers for evaluation of reproductive parameters, results should be viewed as qualitative rather than conclusive, and more research is necessary. We noted a treatment by month effect for somatic cell count; however, there were no other differences for any of the lactation parameters measured. Results demonstrate that up to 50% of diet can be fed in limit-fed rations as DDGS, compared with 30 or 40%, to peripubertal dairy heifers without negative consequences to first-lactation performance during the first 3 mo.



http://ift.tt/2lC99NU

Adoption and consistency of application of premilking preparation in Ontario dairy herds

Publication date: Available online 2 March 2017
Source:Journal of Dairy Science
Author(s): E. Belage, S. Dufour, D.A. Shock, A. Jones-Bitton, D.F. Kelton
Milking management practices that affect udder health have been widely studied, leading to a variety of evidence-based recommendations. Lack of adoption or inconsistency in milking practices can interfere with efforts to prevent mastitis in the herd. The study objective was to assess the variation in adoption and application consistency of important milk harvest practices between and within farms over time. During the summer of 2013, 50 herds in southern Ontario were visited twice within a month, at milking time, and a single person observed and time-recorded premilking preparation procedures. A generalized mixed model was used to partition the variance for predisinfectant contact time and preparation lag time (time between the first contact with the teats and cluster attachment), and determine the proportion of variation attributable to farms, milkers, visits, and characteristics of a cow milking. Using logistic regression, models were built to assess factors affecting adequate contact time and adequate preparation lag time, respectively. Farm, the person(s) milking, and visit number were used as random effects in both instances. In both models, farm-to-farm differences and variations between cows during a specific milking accounted for the largest part of the variability seen in both contact time (47 and 44%, respectively) and preparation lag time (40 and 36%, respectively). For both outcomes, milkers were consistent in their routines over the 2 visits (only 9 and 3.1% of total variance for contact and preparation lag time, respectively). Parlors were more likely to meet the recommended contact time than tie-stalls; increased number of milkers at milking time and having contact times under 30 s had negative effects on meeting recommended preparation lag time. The majority of farms in the study complied with the recommendations for adequate milking practices; however, most did not follow a consistent timed protocol. There are several potential sources of variation in the milking routine on a dairy farm. To improve milk quality and udder health, it is important to identify whether best management practices are being implemented on each farm. Producers appeared to be consistent in the application of milking procedures across time, regardless of whether or not they were correct. Hence, with corrective education and training, improvements in these practices could be experienced and maintained to promote better udder health.



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Variants of β-casofensin, a bioactive milk peptide, differently modulate the intestinal barrier: In vivo and ex vivo studies in rats

Publication date: Available online 2 March 2017
Source:Journal of Dairy Science
Author(s): Jérémie Bruno, Aurélie Nicolas, Sandra Pesenti, Jessica Schwarz, Jean-Luc Simon, Joëlle Léonil, Pascale Plaisancié
β-Casofensin is a bioactive milk peptide that modulates the intestinal barrier, particularly through its action on goblet cells. β-Casofensin corresponds to fragment (f) 94–123 of the bovine β-casein (β-CN) A2 variant. Fifteen genetic variants of bovine β-CN (A1–3, B–G, H1–2, I–L) are known, of which the A2, A1, and B forms are the most common. These variants differ from each other by the substitution of one or more AA, some of which are localized in f94 to 123. The aim of our study was to compare the intestinal effects of β-casofensin A2 and its 3 main variants: A1, A3, and B. For this purpose, a solution (0.1 µM; 10 μL/g of body weight, postnatal d 10–20) containing β-casofensin A2, one of its variants (A1, A3, or B), or drinking water (control; CT) was administered to rat pups orally. After euthanasia (postnatal d 20), intestinal segments were collected for biochemical and histochemical analysis and also used to determine paracellular permeability to fluorescein isothiocyanate-labeled 4-kDa dextran in an Ussing chamber (Harvard Apparatus, Holliston, MA). We also studied the direct effects of β-casofensin A2 and its A1 variant on the paracellular permeability of jejunum segments of adult rats. β-Casofensin A2 and its B variant significantly increased the population of goblet cells compared with the CT, A1, and A3 groups. The mucin 2 mRNA level was significantly higher in the β-casofensin A2 group than in the CT, A3, and B groups. Our results also revealed that the protein expression of zonula occludens-1 and occludin was reduced in the jejunum of rats in the A1, A3, and B groups compared with the CT group. However, the A1 variant was the only peptide to decrease jejunal permeability compared with the CT group. This variant, tested directly in the apical compartment of an Ussing chamber at a concentration of 0.1 nM, also reduced jejunal permeability. In conclusion, the substitution of a single AA alters the effect of β-CN sequence f94 to 123 on goblet cells and on intestinal permeability. A genetic polymorphism of β-CN can affect the biological activity of peptides derived from this protein. These data should be taken into account in the production of bioactive foods.



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Transcranial LED therapy on amyloid-β toxin 25–35 in the hippocampal region of rats

Abstract

Excessive Aβ deposition in the brain is associated with the formation of senile plaques, and their diffuse distribution is related to Alzheimer's disease. Thirty rats (EG) were irradiated with light-emitting diode (photobiomodulation (PBM)) in the frontal region of the skull after being inoculated with the Aβ toxin in the hippocampus; 30 rats were used as the control group (CG). The analysis was conducted at 7, 14, and 21 days after irradiation. We observed a decreased in Aβ deposits in treated animals compared with animals in the CG. The behavioral and motor assessment revealed that the EG group covered a larger ground distance and explored the open field than the CG group on days 14 and 21 (p < 0.05). The EG group was statistically significant in the spatial memory test compared to the CG group on day 14. The use of PBM significantly reduced the presence of Aβ plaques and improved spatial memory and behavioral and motor skills in treated animals on day 21.

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DIAGNOSIS OF ENDOCRINE DISEASE: Endocrine late-effects of childhood cancer and its treatments

Endocrine complications are frequently observed in childhood cancer survivors (CCS). One of two CCS will experience at least one endocrine complication during the course of his/her lifespan, most commonly as a late-effect of cancer treatments, especially radiotherapy and alkylating agent chemotherapy. Endocrine late-effects include impairments of the hypothalamus/pituitary, thyroid and gonads, as well as decreased bone mineral density and metabolic derangements leading to obesity and/or diabetes mellitus. A systematic approach where CCS are screened for endocrine late-effects based on their cancer history and treatment exposures may improve health outcomes by allowing the early diagnosis and treatment of these complications.

http://ift.tt/2m2STcB

Depressive-like effect of prenatal exposure to DDT involves global DNA hypomethylation and impairment of GPER1/ESR1 protein levels but not ESR2 and AHR/ARNT signaling

Publication date: Available online 2 March 2017
Source:The Journal of Steroid Biochemistry and Molecular Biology
Author(s): Malgorzata Kajta, Agnieszka Wnuk, Joanna Rzemieniec, Ewa Litwa, Wladyslaw Lason, Agnieszka Zelek-Molik, Irena Nalepa, Zofia Rogóż, Adam Grochowalski, Anna K. Wojtowicz
Several lines of evidence suggest that exposures to Endocrine Disrupting Chemicals (EDCs) such as pesticides increase the risks of neuropsychiatric disorders. Despite extended residual persistence of dichlorodiphenyltrichloroethane (DDT) in the environment, the mechanisms of perinatal actions of DDT that could account for adult-onset of depression are largely unknown. This study demonstrated the isomer-specific induction of depressive-like behavior and impairment of Htr1a/serotonin signaling in one-month-old mice that were prenatally exposed to DDT. The effects were reversed by the antidepressant citalopram as evidenced in the forced swimming (FST) and tail suspension (TST) tests in the male and female mice. Prenatally administered DDT accumulated in mouse brain as determined with gas chromatography and tandem mass spectrometry, led to global DNA hypomethylation, and altered the levels of methylated DNA in specific genes. The induction of depressive-like behavior and impairment of Htr1a/serotonin signaling were accompanied by p,p'-DDT-specific decrease in the levels of estrogen receptors i.e. ESR1 and/or GPER1 depending on sex. In contrast, o,p'-DDT did not induce depressive-like effects and exhibited quite distinct pattern of biochemical alterations that was related to aryl hydrocarbon receptor (AHR), its nuclear translocator ARNT, and ESR2. Exposure to o,p'-DDT increased AHR expression in male and female brains, and reduced expression levels of ARNT and ESR2 in the female brains. The evolution of p,p'-DDT-induced depressive-like behavior was preceded by attenuation of Htr1a and Gper1/GPER1 expression as observed in the 7-day-old mouse pups. Because p,p'-DDT caused sex- and age-independent attenuation of GPER1, we suggest that impairment of GPER1 signaling plays a key role in the propagation of DDT-induced depressive-like symptoms.



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Chemical profiles and pharmacological activities of Chang-Kang-Fang, a multi-herb Chinese medicinal formula, for treating irritable bowel syndrome

Publication date: Available online 2 March 2017
Source:Journal of Ethnopharmacology
Author(s): Qian Mao, Lei Shi, Zhi-Gang Wang, Yu-Hui Luo, Yin-Yu Wang, Xue Li, Min Lu, Jian-Min Ju, Jin-Di Xu, Ming Kong, Shan-Shan Zhou, Min-Qin Shen, Song-Lin Li
Ethnopharmacological relevanceChang-Kang-Fang formula (CKF), a multi-herb traditional Chinese medicinal formula, has been clinically used for treatment of irritable bowel syndrome (IBS). The mechanisms of CKF for treating IBS and the components that are responsible for the activities were still unknown.Aim of the studyTo investigate the chemical profiles and effects of CKF on IBS model.Materials and methodsThe chemical profiles of CKF were investigated by ultra performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry (UHPLC-Q/TOF-MS/MS). On colon irritation induced rat neonates IBS model, the influence of CKF on neuropeptides, including substance P (SP), calcitonin gene-related peptide (CGRP), vasoactive intestinal polypeptide (VIP) and 5-hydroxytryptamine (5-HT), were measured by ELISA, and the effect on intestinal sensitivity was assessed based on the abdominal withdrawal reflex (AWR) scores. In addition, the activities of CKF against acetic acid-induced nociceptive responses and prostigmin methylsulfate triggered intestinal propulsion in mice were also evaluated.Results80 components were identified or tentatively assigned from CKF, including 11 alkaloids, 20 flavanoids, 4 monoterpenoids, 9 iridoid glycoside, 9 phenylethanoid glycosides, 10 chromones, 7 organic acid, 3 coumarins, 2 triterpene and 5 other compounds. On IBS rat model, CKF was observed to reduce AWR scores and levels of SP, CGRP, VIP and 5-HT. Moreover, CKF reduced the acetic acid-induced writhing scores at all dosages and reduced the intestinal propulsion ration at dosage of 7.5 and 15.0 g/kg/d.ConclusionsCKF could alleviate the symptoms of IBS by modulating the brain–gut axis through increasing the production of neuropeptides such as CGRP, VIP, 5-HT and SP, releasing pain and reversing disorders of intestinal propulsion. Berberine, paeoniflorin, acteoside, flavonoids and chromones may be responsible for the multi-bioactivities of CKF.

Graphical abstract

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Surgical nodulectomies can heal in patients with lymphoedema secondary to Podoconiosis in resource poor settings

Abstract

In Ethiopia severe lymphoedema of the legs resulting from podoconiosis affects approximately 1 million people¹. Woody-hard fibrous nodules which complicate this renders the condition resistant to conventional treatment. We present a series of nodulectomies performed in a rural, resource limited setting.

This article is protected by copyright. All rights reserved.

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Lipocalin-2 is expressed by activated granulocytes and keratinocytes in affected skin and reflects disease activity in acne inversa / hidradenitis suppurativa

Summary

Background

Acne inversa (AI)/Hidradenitis suppurativa is a chronic inflammatory disease characterized by painful axillary, inguinal, and perianal skin lesions with deep-seated nodules, abscesses, and fistulae.

Objectives

This study aimed at the identification and characterization of key players in AI pathogenesis.

Methods

Epidemiologic and anamnestic data as well as blood and skin samples of AI patients were collected. Healthy participants and psoriasis patients served as controls. Assessment of samples and cultures of primary cells was performed by ELISA, qRT-PCR, and immunohistochemistry.

Results

Among 35 mediators quantified in blood of AI patients, lipocalin(LCN)-2 appeared as one of the most significantly upregulated parameters compared to healthy participants [85.8±12.2 (n=18) versus 41.8±4.2 (n=15); P=0.000]. Strongly elevated LCN2 expression was present in AI lesions, with granulocytes and, to a lower extent, keratinocytes being sources of this expression. In vitro, these cells upregulated LCN2 production in response to TNF-α, and a positive relationship between systemic TNF-α and LCN2 levels (r_s=0.55; P=0.011; n=20) was evident for AI. LCN2 blood levels correlated with AI disease severity (r_s=0.65; P=0.000, n=29), but not with disease duration, age, sex, BMI, or smoking habit. Detailed analyses revealed a link with the number of skin regions containing nodules and fistulae, but not scars.

Conclusions

LCN2 might serve as a blood biomarker for objective assessment of inflammatory activity in AI. Additionally, we suggest a self-amplification loop comprising TNF-α, neutrophilic granulocytes, and LCN2, that contributes to the recurrent skin neutrophil infiltration in AI, clinically evident as pus.

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Recent advances in the understanding of severe cutaneous adverse reactions

Abstract

Severe cutaneous adverse reactions (SCARs) encompass a heterogeneous group of delayed hypersensitivity reactions, which are most frequently caused by drugs. Our understanding of several aspects of SCAR syndromes has evolved considerably over the previous decade. This review explores evolving knowledge on the immunopathogenic mechanisms, pharmacogenomic associations, in-vivo and ex-vivo diagnostics for causality assessment and medication cross-reactivity data related to SCAR syndromes. Given the rarity and severity of these diseases, multidisciplinary collaboration through large international, national and/or multicentre networks to collect prospective data on patients with SCAR syndromes should be prioritized. This will further enhance a systematised framework for translating epidemiological, clinical, and immunopathogenetic advances into preventive efforts and improved outcomes for patients.

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The Oncogenic role of miR-155 in Mycosis Fungoides: an in vitro and xenograft mouse model study

Summary

Background

miR-155 contributes to the proliferation of mycosis fungoides (MF) in vitro and is upregulated in tumours of MF compared to early MF lesions.

Objectives

To investigate the contribution of miR-155 to the cancerous phenotype and drug resistance of MF/Sezary cell lines.

Methods

miR-155 was inhibited in MF cell lines (MyLa and MJ) by transduction of miRZip anti-miR-155 and overexpressed in Hut78 cells by transduction of miRVec-miR-155; empty plasmids served as a control. Cells were analysed for response to inducers of apoptosis and cell cycle arrest using FACS. The transduced MyLa cells were subcutaneously injected into SCID mice, and tumours were analysed immunohistochemically and for final size.

Results

MyLa and MJ cells expressed a high level of miR-155 whereas Hut78 cells expressed a low level. MF cell lines stably expressing miR-155 inhibitor showed increased G2/M arrest in response to SL111, a cell-cycle-arrest inducer, followed by increased apoptosis. Additionally, they showed increased apoptosis in response to SAHA, a histone deacetylase inhibitor. Tumours formed in mice from injected anti-miR-155-expressing MyLa cells had a significantly lower volume and more apoptosis than controls. Stable overexpression of miR-155 in Hut78 cells had no effect.

Conclusions

Oncogenic miR-155 appears to contribute to the cancerous phenotype of MyLa and MJ cells, but not of Hut78 cells, by interrupting activation of the G2/M checkpoint in response to SL111 and decreasing apoptosis in response to SL111 and SAHA, thereby facilitating tumor growth. These findings have implications for the potential development of novel therapeutic modalities for MF incorporating miR-155 inhibitors.

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The mTOR inhibitor everolimus suppresses proliferation, metabolic activity and collagen synthesis of human fibroblasts in vitro and exerts antifibrogenic effects in vivo

Abstract

Fibrosclerotic and fibroproliferative diseases remain a therapeutic challenge in dermatology. Everolimus, an inhibitor of the mammalian target of rapamycin (mTOR), is approved for prophylaxis against allograft rejection, treatment of renal cell carcinoma, and use as a drug-eluting stent¹. Interestingly, rapamycin improved skin stiffness and mobility in a patient with systemic sclerosis (SSc)². A randomized, single-blind pilot study confirmed that rapamycin has beneficial effects on the modified Rodnan skin thickness score and on the patient's global assessment in SSc³.

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