A novel lipase with dual localisation in Trypanosoma brucei.

Phospholipases are esterases involved in lipid catabolism. In pathogenic micro-organisms (bacteria, fungi, parasites) they often play a critical role in virulence and pathogenicity. A few phospholipases (PL) have been characterised so far at the gene and protein level in unicellular parasites including African trypanosomes (AT). They could play a role in different processes such as host-pathogen interaction, antigenic variation, intermediary metabolism. By mining the genome database of AT we found putative new phospholipase candidate genes and here we provided biochemical evidence that one of these has lipolytic activity. This protein has a unique non-canonical glycosome targeting signal responsible for its dual localisation in the cytosol and the peroxisomes-related organelles named glycosomes. We also show that this new phospholipase is excreted by these pathogens and that antibodies directed against this protein are generated during an experimental infection with T. brucei gambiense, a subspecies responsible for infection in humans. This feature makes this protein a possible tool for diagnosis.

Quantifying stratosphere-troposphere transport of ozone using balloon-borne ozonesondes, radar windprofilers and trajectory models.

In a series of 10-day campaigns in Ontario and Quebec, Canada, between 2005 and 2007, ozonesondes were launched twice daily in conjunction with continuous high-resolution wind-profiling radar measurements. Windprofilers can measure rapid changes in the height of the tropopause, and in some cases follow stratospheric intrusions. Observed stratospheric intrusions were studied with the aid of a Lagrangian particle dispersion model and the Canadian operational weather forecast system. Definite stratosphere-troposphere transport (STT) events occurred approximately every 2-3 days during the spring and summer campaigns, whereas during autumn and winter, the frequency was reduced to every 4-5 days. Although most events reached the lower troposphere, only three events appear to have significantly contributed to ozone amounts in the surface boundary layer. Detailed calculations find that STT, while highly variable, is responsible for an average, over the seven campaigns, of 3.1% of boundary layer ozone (1.2 ppb), but 13% (5.4 ppb) in the lower troposphere and 34% (22 ppb) in the middle and upper troposphere, where these layers are defined as 0-1 km, 1-3 km, and 3-8 km respectively. Estimates based on counting laminae in ozonesonde profiles, with judicious choices of ozone and relative humidity thresholds, compare moderately well, on average, with these values. The lamina detection algorithm is then applied to a large dataset from four summer ozonesonde campaigns at 18 North American sites between 2006 and 2011. The results show some site-to-site and year-to-year variability, but stratospheric ozone contributions average 4.6% (boundary layer), 15% (lower troposphere) and 26% (middle/upper troposphere). Calculations were also performed based on the TOST global 3D trajectory-mapped ozone data product. Maps of STT in the same three layers of the troposphere suggest that the STT ozone flux is greater over the North American continent than Europe, and much greater in winter and spring than in summer or fall. When averaged over all seasons, magnitudes over North America show similar ratios between levels to the previous calculations, but are overall 3-4 times smaller. This may be because of limitations (trajectory length and vertical resolution) to the current TOST-based calculation.

Effects of a silica-based feed supplement on performance, health, and litter quality of growing turkeys.

Poor litter quality is a potential challenge to footpad health as well as the primary cause of ammonia volatilization. High ambient ammonia concentration is one of the most significant factors negatively affecting poultry production today. Some minerals have been reported to reduce ammonia release from poultry litter. Silicon dioxide, a highly pure and natural mineral, shows promise in decreasing ammonia volatilization and improving litter quality. The objective of the current study was to investigate the effects of feed-borne silicon dioxide on litter quality and how this impacts bird performance, general health and footpad health throughout a 12-wk posthatching turkey study. Supplementing the diet with silicon dioxide was found to significantly improve turkey BW gain and the efficiency of feed conversion. The severity of footpad dermatitis was monitored throughout the experimental period but no significant effect of diet was seen. The feeding of silicon dioxide reduced litter pH which decreased the conversion of NH4⁺ to NH3 thereby reducing nitrogen losses from litter. It was concluded that, under our study conditions, the feeding of 0.02% silicon dioxide offers potential economic benefits to turkey producers.

Extensive projections of myenteric serotonergic neurons suggest they comprise the central processing unit in the colon.

BACKGROUND: 5-Hydroxytryptamine (5-HT, serotonin) is an important regulator of colonic motility and secretion; yet the role of serotonergic neurons in the colon is controversial. METHODS: We used immunohistochemical techniques to examine their projections throughout the enteric nervous system and interstitial cells of Cajal (ICC) networks in the murine proximal to mid colon. KEY RESULTS: Serotonergic neurons, which were mainly calbindin positive, occurred only in myenteric ganglia (1 per 3 ganglia). They were larger than nNOS neurons but similar in size to Dogiel Type II (AH) neurons. 5-HT neurons, appeared to make numerous varicose contacts with each other, most nNOS neurons, Dogiel Type II/AH neurons and glial cells. 5-HT, calbindin and nNOS nerve fibers also formed a thin perimuscular nerve plexus that was associated with ganglia, which contained both nNOS positive and negative neurons, which lay directly upon the submucosal pacemaker ICC network. Neurons in perimuscular ganglia were surrounded by 5-HT varicosities. Submucous ganglia contained nNOS positive and negative neurons, and calbindin positive neurons, which also appeared richly supplied by serotonergic nerve varicosities. Serotonergic nerve fibers ran along submucosal arterioles, but not veins. Varicosities of serotonergic nerve fibers were closely associated with pacemaker ICC networks and with intramuscular ICC (ICC-IM). 5-HT2B receptors were found on a subpopulation of non-5-HT containing myenteric neurons and their varicosities, pacemaker ICC-MY and ICC-IM. CONCLUSIONS & INFERENCES: Myenteric serotonergic neurons, whose axons exhibit considerable divergence, regulate the entire enteric nervous system and are important in coordinating motility with secretion. They are not just interneurons, as regularly assumed, but possibly also motor neurons to ICC and blood vessels, and some may even be sensory neurons.

Conjugation of deoxynivalenol by Alternaria alternata (54028 NRRL), Rhizopus microsporus var. rhizopodiformis (54029 NRRL) and Aspergillus oryzae (5509 NRRL).

Deoxynivalenol (DON, vomitoxin) is a trichothecene mycotoxin which can be considered to be an indicator of Fusarium mycotoxin contamination in grain, feed and food. Recent studies have described the presence of glucose conjugated DON, which is a product of plant metabolism, but there is a lack of information available on DON conjugation by fungi. The aim of the current study was, therefore, to investigate the ability of fungi to metabolize DON into hydrolysable conjugated DON. Alternaria alternata (54028 NRRL) and Rhizopus microsporus var. rhizopodiformis (54029 NRRL) were found to be capable of metabolizing DON into hydrolysable conjugated DON. This ranged from 13-23 % conjugation of DON in potato dextrose agar media and from 11-36 % in corn-based media. There was, however, considerable variation between fungal strains in the ability to conjugate DON as only a slight increase in hydrolysable conjugated DON (1-6 %) was observed when incubating with A. oryzae (5509 NRRL). A. oryzae (5509 NRRL) was also shown to degrade DON (up to 92 %) over 21 days of incubation on corn-based media. The current study shows that conjugation of DON can be achieved through fungal metabolism in addition to being a product of plant metabolism.

Assessment of residual active chlorine in sodium hypochlorite solutions after dissolution of porcine incisor pulpal tissue.

BACKGROUND: In previous studies, surfactant-containing Hypochlor brands of sodium hypochlorite showed better tissue solubilizing abilities than Milton; differences not explained by original active chlorine content or presence of surfactant. It was postulated that exhaustion of active chlorine content could explain differences. This study aimed to assess whether Milton's poorer performance was due to exhaustion of active chlorine. Parallel experiments assessed the influence of titration methods, and the presence of chlorates, on active chlorine measurements. METHODS: Time required to dissolve one or groups of 10 samples of porcine incisor pulp samples in Milton was determined. Residual active chlorine was assessed by thermometric titration. Iodometric and thermometric titration was carried out on samples of Milton. Chlorate content was also measured. RESULTS: Dissolution of single and 10 pulp samples caused a mean loss of 1% and 3% respectively of active chlorine, not being proportional to tissue dissolved. Thermometric ammonium ion titration resulted in 10% lower values than iodometric titration. Chlorate accounted for much of this difference. CONCLUSIONS: Depletion of active chlorine is not the reason for differences in tissue dissolving capabilities of Milton. Thermometric ammonium ion titration gives more accurate measurement of active chlorine content than iodometric titration.

Effects of feed-borne Fusarium mycotoxins on performance, serum chemistry, and intestinal histology of New Zealand White fryer rabbits.

The objective of the present study was to determine the effects of feeding diets containing grains naturally contaminated with Fusarium mycotoxins to fryer rabbits. The efficacy of a glucomannan mycotoxin adsorbent (GMA) was also examined. Thirty 5-wk-old male New Zealand White rabbits were fed a control diet, a contaminated diet, or a contaminated diet + 0.2% GMA for 21 d. Experimental diets contained deoxynivalenol (DON; vomitoxin) at a concentration of 0.25, 4.3, and 4.9 µg/g, respectively. Feed intake was measured daily and water intake was measured every 3 d. At the end of the feeding period, blood was collected for evaluation of serum chemistry and hematology. Visceral organs were excised, weighed, and processed for histopathological examination. Body weight gain and water intake were greater in rabbits fed the contaminated diet (P = 0.075 and 0.020, respectively) and those fed the contaminated + GMA diet (P = 0.026 and 0.002, respectively) compared with controls. Rabbits fed the contaminated + GMA diet had significantly increased serum urea concentrations (P = 0.023) and decreased serum alkaline phosphatase activity (P = 0.020) compared with controls. Increase in BW gain of rabbits fed the contaminated diets was caused by increased water consumption. There was no effect (P > 0.05) of diet on relative organ weights, but decreased infiltrations with eosinophilic granulocytes were observed in different regions of the intestine in rabbits fed the contaminated or the contaminated + GMA diet. It was concluded that rabbits could be adversely affected by feed-borne Fusarium mycotoxins but appear to be less sensitive than other mammalian species. Supplementation with GMA did not reduce many of the effects of feed-borne mycotoxins.

Ca(2+) transients in submucous neurons during the colonic migrating motor complex in the isolated murine large intestine.

BACKGROUND: The colonic migrating motor complex (CMMC) is a spontaneous, rhythmic, and neurally mediated motor pattern generated by myenteric neurons, which can propel fecal pellets in mice. Our aim was to determine whether submucous neurons were also activated during the CMMC. METHODS: :The isolated murine colon was opened and sections of mucosa were removed to expose the submucous ganglia, which were then loaded with Fluo-4. KEY RESULTS: Colonic migrating motor complexes, which occurred spontaneously or by mechanically stimulating the mucosa, were identified by displacement of the tissue (duration = 23.3 s). Between CMMCs, spontaneous Ca(2+) transients (frequency = 0.9 Hz) were observed in 55% (n = 8) of submucous neurons. During the CMMC, 98% (seven ganglia, n = 7) of submucous neurons within the same ganglion exhibited rapid Ca(2+) transients (1.6 Hz) superimposed on a sustained rise in Ca(2+) (duration ∼23 s) that occurred 1.7 s following the mucosal stimulus; whereas other neurons exhibited a similar, but delayed response that occurred either at 7 or 13 s following the stimulus. The activity in submucous neurons was correlated with activity in adjacent nerve varicosities. Ondansetron (1 mm; 5-HT(3) antagonist) significantly reduced the frequency and duration of the Ca(2+) transient responses. CONCLUSIONS & INFERENCES: Activity in the submucous neurons appears to be secondary to that in the myenteric plexus and appears to be generated largely by activity in myenteric descending (serotonergic) interneurons. During the CMMC, there is likely to be an increase in secretion to lubricate and facilitate fecal pellet propulsion.

Activity in varicosities within the myenteric plexus between and during the colonic migrating motor complex in the isolated murine large intestine.

BACKGROUND: Neuronal communication within the myenteric plexus occurs when action potentials along nerve fibers produce Ca(2+) transients in varicosities leading to exocytosis of vesicles and neurotransmitters release. We used Ca(2+) transients in varicosities to monitor action potential activity in myenteric nerve pathways both between and during the colonic migrating motor complex (CMMC) in the isolated murine colon. METHODS: Strips of longitudinal muscle were removed to reveal the myenteric ganglia, which were then loaded with Fluo-4. KEY RESULTS: Many varicosities, including synaptotagmin 1 labeled varicosities, exhibited ongoing Ca(2+) transients (duration of unitary Ca(2+) transient 3.9 s). Between CMMCs, varicosities fired at a frequency of 0.6 Hz, which correlated with spontaneous inhibitory junction potentials in the circular muscle, suggesting they were mainly in inhibitory nerve pathways. During a CMMC other previously quiescent varicosities fired at 1.3 Hz (max. 2.0 Hz) for the duration (24 s) of the CMMC, suggesting they were on excitatory nerve pathways. Activity in varicosities was correlated with Ca(2+) transient responses in a number of neurons. Some varicosities appeared to release an inhibitory neurotransmitter that reduced activity in nNOS-positive neurons. Varicosities along the same nerve fiber exhibited identical patterns of activity that allowed nerve fibers to be traced throughout the myenteric plexus and internodal strands. Activity in varicosities was reduced by hexamethonium (100 µmol L(-1) ), and blocked by ω-conotoxin GVIA (200 nM) and tetrodotoxin (1 µmol L(-1) ; TTX). CONCLUSIONS & INFERENCES: Ca(2+) imaging of varicosities allows for a determination of activity in neural pathways within the enteric nervous system.

Validation of the Jamaican Maternal Sexual Role Modelling Questionnaire.

OBJECTIVE: Urban Jamaican adolescent girls face significant risk for sexually transmitted infections including HIV Studies from the United States ofAmerica have found that parents influence adolescents' sexual risk attitudes and behaviours through parent-child sexual communication and monitoring/supervision. Data from an ongoing mother-daughter HIVrisk reduction intervention study in Kingston, Jamaica identified an additional influence of adolescent girls' sexual risk - maternal sexual role modelling (MSRM). As no reliable and valid questionnaires existed to measure MSRM, one was developed. The objective of the current study was to evaluate the psychometric properties of the Jamaican Maternal Sexual Role Modelling questionnaire. METHOD: Data were collected from 209 Jamaican female adolescents recruited from Kingston, St Andrew and St Catherine parishes. RESULTS: The final 19-item Jamaican MSRM questionnaire was found to have excellent internal reliability (Cronbach's alpha = 0.89). Content validity expert ratings and modified kappa statistics were all 1.0. Principal component analysis identified a three-factor structure that accounted for 53.7% of the variance. Greater MSRM scale scores, indicating more positive and protective maternal sexual role modelling, were associated with less sexual experience, lower intentions to have sex, greater intentions to use condoms if having sex and greater condom use self-efficacy among adolescent girls. CONCLUSION: The MSRM scale was found to be a reliable and valid measure of Jamaican adolescent females'perceptions of their mothers'sexual role modelling. Further research is needed to assess the reliability and validity of the instrument with other populations.

Effects of dietary Fusarium mycotoxins on intestinal lymphocyte subset populations, cell proliferation and histological changes in avian lymphoid organs.

An experiment was conducted to investigate the effects of dietary Fusarium mycotoxins on gut immunity, cell proliferation, and histology of avian lymphoid organs. The efficacy of a polymeric glucomannan mycotoxin adsorbent (GMA) was also determined. Seventy-two one-day-old male turkey poults were fed corn, wheat, and soybean meal-based diets for 21 days. Diets included control grains, contaminated grains and contaminated grains +0.2% GMA. The major contaminant was deoxynivalenol (3.9 µg/g) with lesser amounts of zearalenone (0.67-0.75 µg/g), 15-acetyl-deoxynivalenol (0.34 µg/g) and HT-2 toxin (0.078-0.085 µg/g). T- and B-lymphocyte populations and crypt cellular proliferation in duodenum, jejunum, ileum and cecal tonsil were measured immunohistochemically on day 14 and 21. Histological changes were recorded after 14 and 21 days of feeding. Feeding contaminated grains significantly increased the percentage of B-lymphocytes in ileum on day 14, and reduced (P<0.05) the percentages of CD8(+)-lymphocytes in cecal tonsil on day 21. GMA supplementation prevented these effects. The feeding of contaminated diets also caused a reduction (P<0.05) in ileal crypt proliferating cells and a significant increase in spleen secondary follicle on day 21. It was concluded that the feeding of grains naturally contaminated with Fusarium mycotoxins results in adverse effects on gut immunity and mucosal cell proliferation.

Morphologic changes in the intestine of broiler breeder pullets fed diets naturally contaminated with Fusarium mycotoxins with or without coccidial challenge.

The effects of feeding diets containing grains naturally contaminated with Fusarium mycotoxins on intestinal histology were studied in chickens raised to 10 wk of age in the absence or presence of coccidial challenge. Experimental diets included the following: controls, diets containing grains naturally contaminated with Fusarium mycotoxins, and diets containing contaminated grains + 0.2% polymeric glucomannan mycotoxin adsorbent. Contaminated diets contained up to 3.8 microg/g deoxynivalenol (DON), 0.3 microg/g 15-acetyl DON, and 0.2 microg/g zearalenone. An optimized mixture (inducing lesions without mortality) of Eimeria acervulina, Eimeria maxima, and Eimeria tenella was used to challenge birds at 8 wk of age. Intestinal tissues were collected from duodenum, jejunum, and ileum prior to challenge; at the end of the challenge period (7 days postinfection; PI); and at the end of the recovery period (14 days PI). Mean villus height (VH) in the duodenum of birds fed the contaminated diets in the absence of coccidial challenge was significantly lower than that of the controls. Mean VH in the jejunum and ileum of the same birds was significantly higher compared to controls, indicating a compensatory mechanism. Fusarium mycotoxins retarded duodenal recovery from coccidial lesions, as indicated by lower duodenal VH and apparent villus surface area comparing challenged birds fed the contaminated diets to challenged controls of the same age. Increased VH was frequently associated with cryptal hyperplasia and increased numbers of mitotic figures in crypts. It was concluded that diets contaminated with Fusarium mycotoxins below levels that negatively affect performance could alter intestinal morphology and interfere with intestinal recovery from an enteric coccidial infection.

ICC-MY coordinate smooth muscle electrical and mechanical activity in the murine small intestine.

BACKGROUND: Animals carrying genetic mutations have provided powerful insights into the role of interstitial cells of Cajal (ICC) in motility. One classic model is the W/W(V) mouse which carries loss-of-function mutations in c-kit alleles, but retains minimal function of the tyrosine kinase. Previous studies have documented loss of slow waves and aberrant motility in the small intestine of W/W(V) mice where myenteric ICC (ICC-MY) are significantly depleted. METHODS: Here, we used morphological and electrophysiological techniques to further assess the loss of ICC around the circumference of the small intestine and determine consequences of losing ICC-MY on electrical activity, Ca(2+) transients and contractions of the longitudinal muscle (LM). KEY RESULTS: In wild-type mice, there was coherent propagation of Ca(2+) transients through the ICC-MY network and spread of this activity to the LM. In short segments of small intestine in vitro and in exteriorized segments, slow waves coordinated smoothly propagating Ca(2+) waves and contractions in the LM of wild-type mice. In W/W(V) mice, Ca(2+) waves were initiated at variable sites along and around intestinal segments and propagated without constraint unless they collided with other Ca(2+) waves. This activity resulted in abrupt, uncoordinated contractions. CONCLUSIONS & INFERENCES: These results show how dominance of pacemaking by ICC-MY coordinates propagating con-tractions and regulates the spontaneous activity of smooth muscle.

Effects of feeding diets naturally contaminated with Fusarium mycotoxins on protein metabolism in late gestation and lactation of first-parity sows.

A study was conducted to assess the effects of feeding a blend of grains naturally contaminated with Fusarium mycotoxins to sows on the capacity for protein synthesis in skeletal muscle, the protein content per cellular unit, and the efficacy of a polymeric glucomannan adsorbent (GMA) to prevent these effects in late gestation and in lactation. Thirty-two Yorkshire sows were assigned to 4 treatment groups (8 per treatment) from 91 +/- 3 d of gestation up to weaning on d 21 after farrowing. Diets included 1) control, 2) contaminated grains, and 3) contaminated grains + 0.2% GMA. A fourth treatment of feeding sows the control diet at a restricted feed allowance was also included. The variables measured include ADFI, average daily BW change, serum total protein, urea, and ammonia, and skeletal muscle DNA, RNA, and protein. To assess the capacity for protein synthesis, ratios of RNA:DNA, and RNA:protein were compared among dietary treatments. To assess the degree of muscle protein mobilization in gestation and lactation, ratios of protein:DNA were compared among dietary treatments. Muscle samples were obtained from the triceps brachii. Blood and muscle samples were obtained 3 times: the first was obtained 1 d before the sows began to receive the experimental diets (90 +/- 3 d of gestation), a second sample was obtained 14 d later (104 +/- 3 d of gestation), and the third sample was obtained 10 d after farrowing. Serum ammonia concentrations were similar in sows fed the contaminated feed and sows fed the restricted feed compared with controls, but serum ammonia concentrations were greater in sows fed contaminated feed (P = 0.02) and restricted-fed sows (P = 0.008) compared with sows fed the contaminated grains plus GMA on 104 +/- 3 d of gestation. There were no reductions in the capacity for protein synthesis caused by mycotoxins or restricted feeding compared with controls. A reduction in ADFI (P = 0.003) was observed in sows fed the 2 contaminated diets in lactation. Muscle protein mobilization was not affected by diet, but a reduction (P = 0.04) in the content of protein per cellular unit was observed in lactation compared with gestation. Reduction in protein:DNA could be caused by the catabolic state in lactation, which was augmented by a low ADFI. The rate of muscle mobilization could be the result of the indirect effect of the reduction in ADFI in lactation rather than a direct effect of Fusarium mycotoxins in the capacity for protein synthesis.

Pathophysiology of colonic causes of chronic constipation.

Colonic sensorimotor dysfunction is recognized as the principal pathophysiological mechanism underpinning chronic constipation. This review addresses current understanding derived from both human and animal studies, with particular reference made to methods of investigation.

Effects of feed naturally contaminated with Fusarium mycotoxins on metabolism and immunity of dairy cows.

A previous study in dairy cows showed some effect of feed contaminated with Fusarium mycotoxins on metabolism and immunity. A subsequent experiment investigated the effect of feedborne Fusarium mycotoxins on some immune functions in more detail. A total mixed ration (TMR) containing a blend of feedstuffs naturally contaminated with Fusarium mycotoxins was fed for 63 d to 12 mid-lactation Holstein cows with an average milk production of 36 kg/d in a completely randomized design with repeated measures including 1) control TMR and 2) contaminated TMR. Wheat, corn, hay, and corn silage were the contaminated feedstuffs. Deoxynivalenol was the major contaminant and was found in TMR at 3.5 mg/kg of dry matter. The parameters measured were 1) performance: body weight, body condition score, dry matter intake, milk production, composition and somatic cell count; 2) health: blood serum chemistry, hematology, coagulation profile, and rumen fluid ammonia levels; 3) immune function: total serum immunoglobulins (IgA, IgG, IgM), specific antibody response to ovalbumin, and neutrophil phagocytosis. Dry matter intake, body weight, milk production, and milk composition were not affected by diet. Neutrophil phagocytosis was depressed throughout the experiment in cows fed the contaminated diet. Serum sodium concentrations and osmolality were significantly elevated throughout the experiment in cows fed the contaminated diet. Primary antibody response to ovalbumin immunization was higher in cows fed the contaminated diet compared with controls. It was concluded that feed naturally contaminated with Fusarium mycotoxins can affect metabolic parameters and immune function of dairy cows.

Signal transduction and gene expression in cultured accessory olfactory bulb neurons.

Glutamate and norepinephrine (NE) are believed to mediate the long-lasting synaptic plasticity in the accessory olfactory bulb (AOB) that underlies pheromone recognition memory. The mechanisms by which these neurotransmitters bring about the synaptic changes are not clearly understood. In order to study signals that mediate synaptic plasticity in the AOB, we used AOB neurons in primary culture as a model system. Because induction of pheromone memory requires coincident glutamatergic and noradrenergic input to the AOB, and requires new protein synthesis, we reasoned that glutamate and NE must induce gene expression in the AOB. We used a combination of agonists that stimulate alpha1 and alpha2 adrenergic receptors in combination with N-methyl-d-aspartic acid and tested expression of the immediate-early gene (IEG) c-Fos. We found that the glutamatergic and noradrenergic stimulation caused significant induction of c-Fos mRNA and protein. Induction of c-Fos was significantly reduced in the presence of inhibitors of protein kinase C, mitogen-activated protein kinase (MAPK) and phospholipase C. These results suggest that glutamate and NE induce gene expression in the AOB through a signaling pathway mediated by protein kinase C and MAPK.

Bves directly interacts with GEFT, and controls cell shape and movement through regulation of Rac1/Cdc42 activity.

Bves is an integral membrane protein with no determined function and no homology to proteins outside of the Popdc family. It is widely expressed throughout development in myriad organisms. Here, we demonstrate an interaction between Bves and guanine nucleotide exchange factor T (GEFT), a GEF for Rho-family GTPases. This interaction represents the first identification of any protein that has a direct physical interaction with any member of the Popdc family. Bves and GEFT are shown to colocalize in adult skeletal muscle. We also demonstrate that exogenous expression of Bves reduces Rac1 and Cdc42 activity levels while not affecting levels of active RhoA. Consistent with a repression of Rac1 and Cdc42 activity, we show changes in speed of cell locomotion and cell roundness also result from exogenous expression of Bves. Modulation of Rho-family GTPase signaling by Bves would be highly consistent with previously described phenotypes occurring upon disruption of Bves function in a wide variety of model systems. Therefore, we propose Bves as a novel regulator of the Rac1 and Cdc42 signaling cascades.

Effects of feedborne fusarium mycotoxins on brain regional neurochemistry of turkeys.

An experiment was conducted to investigate the effects of feeding grains naturally contaminated with Fusarium mycotoxins on brain regional neurochemistry of turkeys. The possible preventative effect of a poly-meric glucomannan mycotoxin adsorbent (GMA) was also determined. Forty-five 1-d-old male turkey poults were fed wheat-, corn-, and soybean meal-based diets up to wk 6, formulated with control grains, contaminated grains, or contaminated grains + 0.2% GMA. Deoxynivalenol was the major contaminant, and the concentrations were 2.2 and 3.3 mg/kg of feed during starter and grower phases, respectively. Concentrations of brain monoamine neurotransmitters and metabolites were measured in discrete regions of the brain including the pons, hypothalamus, and cortex by HPLC with electrochemical detection. Neurotransmitters and metabolites analyzed included norepinephrine, dopamine, 3,4-dihydroxyphenylacetic acid, serotonin (5-hydroxytryptamine, 5-HT), and 5-hydroxyindoleacetic acid (5-HIAA). The concentration of 5-HIAA and the 5-HIAA:5-HT-ratio were significantly decreased in pons after feeding contaminated grains. Dietary supplementation with GMA prevented these effects. In the pons, a significant positive correlation (r = 0.52, P < 0.05) was observed between the concentration of 5-HT and BW gain after feeding contaminated diets. The feeding of contaminated diet had no significant effects on the concentrations of neurotransmitters and metabolites in hypothalamus and cortex. It was concluded that consumption of grains naturally contaminated with Fusarium mycotoxins adversely altered the pons serotonergic system of turkeys. Supplementation with GMA partially inhibited these effects.

Effects of feeding blends of grains naturally contaminated with Fusarium mycotoxins on small intestinal morphology of turkeys.

An experiment was conducted to investigate the effects of feeding grains naturally contaminated with Fusarium mycotoxins on morphometric indices of duodenum, jejunum, and ileum in turkeys. The possible preventative effect of a polymeric glucomannan mycotoxin adsorbent (GMA) was also determined. Three hundred 1-d-old male turkey poults were fed wheat, corn, and soybean meal-based starter (0 to 3 wk), grower (4 to 6 wk), developer (7 to 9 wk), and finisher (10 to 12 wk) diets formulated with control grains, contaminated grains, and contaminated grains + 0.2% GMA. Morphometric indices were measured at the end of each growth phase and included villus height (VH), crypt depth, villus width, thicknesses of submucosa and muscularis, villus-to-crypt ratio, and apparent villus surface area (AVSA). At the end of the starter phase, feedborne mycotoxins significantly decreased the VH in the duodenum, and supplementation of the contaminated diet with GMA prevented this effect. The feeding of contaminated grains also reduced (P < 0.05) VH and AVSA in jejunum, whereas none of the variables were affected in the ileum. Villus width and AVSA of duodenum, VH, and AVSA of jejunum and submucosa thickness of ileum were significantly reduced when birds were fed contaminated grains at the end of the grower phase, and supplementation with GMA prevented these effects in jejunum and ileum. No effects of diets were seen on morphometric variables at the end of the developer and finisher phases. It was concluded that consumption of grains naturally contaminated with Fusarium mycotoxins results in adverse effects on intestinal morphology during early growth phases of turkeys, and GMA can prevent many of these effects.