Dietary patterns and the risk of pregnancy-associated hypertension in the Danish National Birth Cohort: a prospective longitudinal study.

OBJECTIVE: To examine the association between mid-pregnancy dietary patterns and pregnancy-associated hypertension (PAH). DESIGN: A prospective longitudinal cohort study. SETTING: Denmark. POPULATION: About 55 139 Danish women with single enrolments and recorded food frequency questionnaire dates with complete information on dietary intake. METHODS: Women were eligible if they could speak Danish and were planning to carry to term. Diet was assessed using a validated semi-quantitative 360-item food frequency questionnaire and dietary patterns were derived using factor analysis. MAIN OUTCOME MEASURES: Gestational hypertension (GH) and pre-eclampsia (PE). RESULTS: Disease prevalence was 14% for GH (5491/39 362); 2% for PE (1168/54 778), and 0.4% for severe PE (234/55 086). Seven dietary patterns were characterised in the population, of which two were associated with PAH. The Seafood diet characterised by high consumption of fish and vegetables was inversely associated with the odds of developing GH [odds ratio (OR) 0.86; 95% CI 0.77-0.95)] and PE (OR 0.79; 95% CI 0.65-0.97). The Western diet characterised by high consumption of potatoes (including French fries), mixed meat, margarine and white bread increased the odds of developing GH (OR 1.18; 95% CI 1.05-1.33) and PE (OR 1.40; 95% CI 1.11-1.76). No association was seen with severe PE. CONCLUSIONS: We found protective associations of Seafood diet and harmful associations of Western diet with PAH. Dietary interventions encouraging the reduction of Western diet may contribute to a decrease of PAH. TWEETABLE ABSTRACT: Western diet increases (Seafood diet decreases) the likelihood of developing pre-eclampsia among Danish pregnant women.

Relationships between food neophobia and food intake and preferences: Findings from a sample of New Zealand adults.

Food neophobia (FN) has been shown to be a strong influence on food preferences using primarily small data sets. This has limited the explanatory power of FN and the extent to which it can be related to other factors that influence food choice. To address these limitations, we collected Food Neophobia Scale data from 1167 adults from New Zealand over a 45-month period. Participants also completed a 112-item food preference questionnaire and a self-report 24 h, a 145 item food intake recall survey, and the Food Choice Questionnaire (FCQ). As a way of providing a structure to the food intake and preference data, in each case the food items were condensed into patterns described in terms of the foods/beverages with highest factor loadings. We then determined the impact of season and participant age, gender, education and income on these factors, as well as the interaction of these variables with FN scores, divided into tertiles. FN was a strong influence on both intake frequency and preferences in the majority of the intake/preference factor patterns. When significant associations with FN were established, both frequency of intake and preference was lower among high FN individuals. Notably, the effect of FN on food preferences was evident on many commonplace foods making up the diet, suggesting that high FN individuals like food overall less than do those with lower degrees of FN. Seasonal effects in food intake were demonstrated, but with smaller impact for higher levels of FN. While associations between FN varied according to all demographic variables, these relationships varied as a function of the intake/preference patterns. Overall, the results suggest that FN is an important barrier to dietary change and addressing diet-related health problems.

Siblings Promote a Type 1/Type 17-oriented immune response in the airways of asymptomatic neonates.

BACKGROUND: Siblings have been shown to reduce the risk of childhood asthma and allergy, but the mechanism driving this association is unknown. The objective was to study whether siblings affect the airway immune response in healthy neonates, which could represent an underlying immune modulatory pathway. METHODS: We measured 20 immune mediators related to the Type 1, Type 2, Type 17, or regulatory immune pathways in the airway mucosa of 571 one-month-old asymptomatic neonates from the Copenhagen Prospective Studies on Asthma in Childhood2010 birth cohort (COPSAC2010 ). The association between airway mediator levels and presence of siblings was investigated using conventional statistics and principle component analysis (PCA). RESULTS: Neonates with siblings had an upregulated level of airway immune mediators, with predominance of Type 1- and Type 17-related mediators. This was supported by the PCA showing a highly significant difference between children with vs without siblings: P < 10(-10) , which persisted after adjustment for potential confounders including pathogenic airway bacteria and viruses: P < 0.0001. The immune priming effect was inversely associated with time since last childbirth: P = 0.0015. CONCLUSIONS: Siblings mediate a Type 1/Type 17-related immune-stimulatory effect in the airways of asymptomatic neonates, also after adjustment for pathogenic bacteria and viruses, indicating that siblings exert a transferable early immune modulatory effect. These findings may represent an in utero immune priming effect of the fetal immune system caused by previous pregnancies as the effect was attenuated with time since last childbirth, or it could relate to the presence of unidentified microbes, but further studies are needed to confirm our findings.

Divergent response profile in activated cord blood T cells from first-born child implies birth-order-associated in utero immune programming.

BACKGROUND: First-born children are at higher risk of developing a range of immune-mediated diseases. The underlying mechanism of 'birth-order effects' on disease risk is largely unknown, but in utero programming of the child's immune system may play a role. OBJECTIVE: We studied the association between birth order and the functional response of stimulated cord blood T cells. METHOD: Purified cord blood T cells were polyclonally activated with anti-CD3-/anti-CD28-coated beads in a subgroup of 28 children enrolled in the COPSAC2010 birth cohort. Expression levels of seven activation markers on helper and cytotoxic T cells as well as the percentage of CD4(+) CD25(+) T cells were assessed by flow cytometry. Production of IFN-γ, TNF-α, IL-17, IL-4, IL-5, IL-13, and IL-10 was measured in the supernatants. RESULTS: IL-10 secretion (P = 0.007) and CD25 expression on CD4(+) helper T cells (P = 0.0003) in the activated cord blood T cells were selectively reduced in first-born children, while the percentage of circulating CD4(+) CD25(+) cord blood T cells was independent of birth order. CONCLUSION: First-born infants display a reduced anti-inflammatory profile in T cells at birth. This possible in utero 'birth-order' T-cell programming may contribute to later development of immune-mediated diseases by increasing overall immune reactivity in first-born children as compared to younger siblings.

Skin reaction and regeneration after single sodium lauryl sulfate exposure stratified by filaggrin genotype and atopic dermatitis phenotype.

BACKGROUND: Filaggrin is key for the integrity of the stratum corneum. Mutations in the filaggrin gene (FLGnull) play a prominent role in atopic dermatitis (AD) pathogenesis. People with AD have increased susceptibility to irritants. However, little is known about the effect of filaggrin genotype and AD phenotype on irritant response and skin regeneration. OBJECTIVES: To investigate the role of FLGnull and AD groups for skin reaction and recovery after sodium lauryl sulfate (SLS) irritation. METHODS: This is a case-control study comprising 67 subjects, including healthy controls and patients with and without FLGnull and AD. Reactivity to different doses of SLS at 24, 48, 72 and 145 h after SLS application was measured by transepidermal water loss (TEWL) and laser Doppler flowmetry (LDF). Reactivity was assessed univariately and by pattern analysis. RESULTS: All patient groups showed a higher degree of skin-barrier disruption and inflammation than did controls in response to SLS. Assessing reactivity by the delta value of the area under the curve for both TEWL and LDF showed significant differences between healthy controls and those with the AD phenotype, irrespective of filaggrin mutation. The poorest regeneration was among those with the AD phenotype. The two AD phenotype groups were separated by multivariate technique, due to earlier inflammatory reactivity among subjects with FLGnullplus AD compared with the AD phenotype alone. CONCLUSIONS: Both skin reaction and regeneration were significantly different between the patient population and the healthy controls. Additionally, response severity and regeneration depended more on AD phenotype than on filaggrin genotype, whereas the response was more rapid among the FLGnullplus AD individuals.

Effect of high-volume systematic local infiltration analgesia in Caesarean section: a randomised, placebo-controlled trial.

BACKGROUND: Pain after Caesarean section is often treated with opioids with a risk of side effects. Wound infiltration with local anaesthetics is effective and has few side effects, but volume vs. dose concentration has not been examined. METHODS: Ninety patients scheduled for elective Caesarean section included in a randomised, double-blinded, placebo-controlled trial receiving infiltration with 50 ml ropivacaine 0.5% or 125 ml ropivacaine 0.2% or 50 ml 0.9% saline (placebo) during surgery. Surgery was performed under lumbar spinal anaesthesia. Primary endpoint was post-operative pain. Secondary endpoints were rescue analgesic, post-operative nausea and vomiting, time spent in the postanesthesia care unit (PACU) and time to first mobilisation. RESULTS: No difference in pain response between groups, but time until maximum pain score was prolonged in the ropivacaine 0.5% group compared with the placebo group (P = 0.0493). The administration of ketobemidone at 24 h post-operatively in the ropivacaine 0.5% group was reduced compared with the placebo group (P = 0.020), and between the ropivacaine 0.2% group and the ropivacaine 0.5% group (P = 0.044). No significant differences between groups were found concerning time spent in the PACU, to first mobilisation or in number of women with nausea/vomiting (P ≥ 0.05). No complications related to ropivacaine were observed. CONCLUSIONS: Systematic infiltration with a high concentration, low volume compared with low concentration, high volume showed no significant effect on post-operative pain intensity. However, a statistically significant, but clinically limited opioid sparing effect was demonstrated compared with placebo in the high concentration, low volume group.

Quantification of individual fatty acids in bovine milk by infrared spectroscopy and chemometrics: understanding predictions of highly collinear reference variables.

Predicting individual fatty acids (FA) in bovine milk from Fourier transform infrared (FT-IR) measurements is desirable. However, such predictions may rely on covariance structures among individual FA and total fat content. These covariance structures may change with factors such as breed and feed, among others. The aim of this study was to estimate how spectral variation associated with total fat content and breed contributes to predictions of individual FA. This study comprised 890 bovine milk samples from 2 breeds (455 Holstein and 435 Jersey). Holstein samples were collected from 20 Danish dairy herds from October to December 2009; Jersey samples were collected from 22 Danish dairy herds from February to April 2010. All samples were from conventional herds and taken while cows were housed. Moreover, in a spiking experiment, FA (C14:0, C16:0, and C18:1 cis-9) were added (spiked) to a background of commercial skim milk to determine whether signals specific to those individual FA could be obtained from the FT-IR measurements. This study demonstrated that variation associated with total fat content and breed was responsible for successful FT-IR-based predictions of FA in the raw milk samples. This was confirmed in the spiking experiment, which showed that signals specific to individual FA could not be identified in FT-IR measurements when several FA were present in the same mixture. Hence, predicted concentrations of individual FA in milk rely on covariance structures with total fat content rather than absorption bands directly associated with individual FA. If covariance structures between FA and total fat used to calibrate partial least squares (PLS) models are not conserved in future samples, these samples will show incorrect and biased FA predictions. This was demonstrated by using samples of one breed to calibrate and samples of the other breed to validate PLS models for individual FA. The 2 breeds had different covariance structures between individual FA and total fat content. The results showed that the validation samples yielded biased predictions. This may limit the usefulness of FT-IR-based predictions of individual FA in milk recording as indirect covariance structures in the calibration set must be valid for future samples. Otherwise, future samples will show incorrect predictions.

OCT4 expression in outgrowth colonies derived from porcine inner cell masses and epiblasts.

The present study was conducted to test different methods for porcine inner cell mass (ICM) and epiblast isolation and to evaluate the morphology and expression of pluripotency genes in ICM- and epiblast-derived outgrowth colonies (OCs) and passages thereof with particular attention on the relationship between OCT4 expression and embryonic stem cell (ESC)-like morphology. A total of 104 zona pellucida-enclosed and 101 hatched blastocysts were subjected to four different methods of ICM and epiblast isolation, respectively: Manual isolation, immunosurgery, immunosurgery with manual cleaning, or whole blastocyst culture. OCs were established on mouse embryonic fibroblast (MEF) cells and categorized according to morphology and OCT4 staining. Although all isolation methods resulted in ESC-like OCs, immunosurgery with manual cleaning yielded significantly higher rates of ICM/epiblast attachment and subsequent ESC-like morphology, whereas no significant difference was found between ICM and epiblasts with respect to these characteristics. All ESC-like OCs showed nuclear OCT4 staining and expression of OCT4, NANOG and SOX2 as evaluated by RT-PCR. Upon initial passages, the expression of pluripotency markers was, however, gradually lost in spite of maintained ESC-like morphology. In conclusion, we have established a robust system for derivation of ESC-like OCs from porcine ICM and epiblasts and we have shown that localization of OCT4 is associated with an ESC-like morphology although this relationship is lost during early passages.

Ecological succession in the vaginal microbiota during pregnancy and birth.

The mother's vaginal microbiota represents the first microbes to which a child is exposed when delivered vaginally. However, little is known about the composition and development of the vaginal microbiota during pregnancy and birth. Here, we analyzed the vaginal microbiota of 57 women in pregnancy week 24, 36 and at birth after rupture of membranes but before delivery, and further compared the composition with that of the gut and airways of the 1-week-old child. The vaginal community structure had dramatic changes in bacterial diversity and taxonomic distribution, yet carried an individual-specific signature. The relative abundance of most bacterial taxa increased stepwise from week 24 of pregnancy until birth, with a gradual decline of Lactobacillus. Mother-to-child vertical transfer, as suggested by sharing, was modest, with the strongest transfer being for Clostridiales followed by Lactobacillales and Enterobacteriales. In conclusion, late gestation is associated with an increase in maternal vaginal microbiota diversity, and vaginal bacteria at birth only modestly predict the composition of the neonatal microbiota.

Prevotella bryantii 25A used as a probiotic in early-lactation dairy cows: effect on ruminal fermentation characteristics, milk production, and milk composition.

Ingestion of high levels of rapidly fermented carbohydrates after parturition often leads to the production of excessive quantities of organic acids that may exceed the buffering capacity of the rumen and cause pH to drop. Ruminal acidosis results in animal discomfort, anorexia, depression, decreased digestibility, and decreased milk production. In the present study, we examined the effects of daily addition of cells of a newly isolated strain of Prevotella bryantii (25A) to the rumen of 12 ruminally cannulated cows in early lactation. This strain was selected based on earlier in vitro studies that indicated its ability to grow rapidly, compete for starch, and produce organic acids other than lactate. After calving, all cows received increasing amounts of an energy-dense diet containing barley grain, corn silage, and grass silage in a 40:60 forage-to-concentrate ratio. Animals were blocked according to milk production from their previous lactation. Treatments (control and P. bryantii) were distributed among cows within the same block. Cows were fed once a day. Six cows were given a daily dose of P. bryantii (2 x 10(11) cells/dose), administered directly with a syringe through the rumen cannula, from 3 wk prepartum up to 7 wk postpartum. Rumen fluid was sampled before feeding and at 2 and 3 h postfeeding on wk 1, 2, 3, 4, 6, and 7 postpartum. Feed intake and milk yield were recorded daily and milk composition was recorded 2 d/wk, up to wk 7 of lactation. Feed intake was similar between control and treated cows. Prevotella bryantii did not change milk production, but milk fat tended to be greater in treated cows compared with control cows (3.9 vs. 3.5%). Rumen pH was similar between the 2 groups and differed across sampling times, being higher before feeding (6.3) as opposed to 2 h (5.9) and 3 h (5.7) postfeeding. Rumen lactate concentration was similar before feeding between control and treated cows; however, 2 to 3 h after feeding, lactate concentration was lower in cows receiving P. bryantii compared with control cows (0.7 vs. 1.4 mM). This difference was maintained throughout the experimental period. Concentration of NH(3)-N was greater in treated cows than in control cows (174 vs. 142 mg/L). Acetate (65.5 vs. 57.8 mM), butyrate (12.7 vs. 10.5 mM), and branched-chain C4 fatty acid (0.90 vs. 0.75 mM) concentrations were greater in postfeeding samples of treated cows compared with control cows. Supplementing early-lactating cows with P. bryantii 25A increased ruminal fermentation products and milk fat concentration. Because signs of subacute ruminal acidosis were not observed in either treated or control cows, no conclusions can be made about possible protection against acidosis by P. bryantii.

De electricitatis catholici musculari - Concerning the electrical properties of muscles, with emphasis on meat quality.

This study aims to explore the potential of evoked non-invasive surface electromyography (SEMG) analysis, in predicting meat quality traits in livestock. Evoked SEMG is a system that records, transdermally, electrical signals generated in muscle fibres upon external stimulation. These signals are reported as compound muscle action potentials (CMAP). CMAP parameters of LD correlated negatively and significantly to ultimate pH (pH 24h) at day 61, but not at day 153 after birth, and a similar albeit positive correlation was observed for muscle glycogen content. Muscle glycogen content and pH 24h correlated negatively in LD and BF. Negative significant correlations between CMAP parameters and shear force were found in LD at day 153 after birth, which might, in the range of the recording electrodes, reflect the combined effect of large cross-sectional area fibres and reduced perimysium content per unit volume of muscle. The fact that correlations between CMAP characteristics and quality traits of both metabolic and non metabolic origin could be established, warrants a fuller investigation of this method in terms of its potential as a predictive tool for meat quality traits in live animals.

Klebsiella to Salmonella gene transfer within rumen protozoa: implications for antibiotic resistance and rumen defaunation.

The rumen has long been thought to be a site of gene transfer for microorganisms. Rumen protozoa (RPz) are active predators of bacteria that can harbor antibiotic resistance genes. In this study, RPz were assessed as sites of gene transfer between two bacterial species, Salmonella and Klebsiella. One Klebsiella isolate carried a plasmid bearing bla(CMY-2), encoding an extended-spectrum beta-lactamase conferring ceftriaxone resistance, while the Salmonella was susceptible to ceftriaxone yet capable of thriving within protozoa. In vitro studies revealed that ceftriaxone-resistant Salmonella could be isolated following co-incubation of Salmonella and Klebsiella with RPz obtained from adult cattle and goats. Ceftriaxone-resistant Salmonella were not recovered in the presence of an inhibitor of protozoa engulfment or when a protozoa-sensitive Salmonella was part of the co-incubation. This transfer event was additionally observed in vitro for protozoa-independent stressors although at a significantly lower frequency. The gene transfer event was related to bacterial conjugation since a conjugation inhibitor, nalidixic acid, perturbed the phenomenon. Ceftriaxone-resistant Salmonella were recovered from calves, sheep, and goats co-challenged with ceftriaxone-resistant Klebsiella and ceftriaxone-sensitive Salmonella. However, the transfer event was not observed in calves and sheep that were defaunated prior to the co-challenge. Moreover, Salmonella transconjugants were isolated from separate bovine in vivo studies involving a Klebsiella donor carrying a plasmid conferring colicin activity while no such transconjugants were obtained from defaunated calves. These results provide an important basis for evaluating and preventing the spread of antibiotic resistance and other selective advantages for pathogens present in ruminants.

Evaluation of corn furan fatty acid putative endocrine disruptors on reproductive performance in adult female chickens.

Based on evidence from rodent models, it was hypothesized that furan fatty acids found in corn would inhibit reproduction in the laying hen. An isomeric mixture of furan fatty acids [9, (12)-oxy-10,13-dihydroxystearic acid and 10, (13)-oxy-9,12-dihydroxystearic acid] was administered for a period of 3 wk via the diet (1 and 3 ppm) at levels greater than those in corn to 20-wk-old pullets. There were no overt indications of acute or chronic toxicity (no effects on mortality, feed intake, or average daily gain). Similarly, there was no dose-dependent effect on reproductive parameters [egg production, egg weight, shell thickness, ovarian weight, number or weight of large yolky preovulatory follicles, and number of small yellow follicles (4-8 mm in diameter)]. The present data do not suggest that furan fatty acids are a cause of concern to the poultry industry.

Rumen contents as a reservoir of enterohemorrhagic Escherichia coli.

We investigated the role of the rumen fermentation as a barrier to the foodborne pathogen, Escherichia coli O157:H7. Strains of E. coli, including several isolates of O157:H7, grew poorly in media which simulated the ruminal environment of a well-fed animal. Strains of E. coli O157:H7 did not display a superior tolerance to ruminal conditions which may facilitate their colonization of the bovine digestive tract. Unrestricted growth of E. coli was observed in rumen fluid collected from fasted cattle. Growth was inhibited by rumen fluid collected from well-fed animals. Well-fed animals appear less likely to become reservoirs for pathogenic E. coli. These results have implications for cattle slaughter practices and epidemiological studies of E. coli O157:H7.

Steady-state and time-resolved spectroscopy of F420 extracted from methanogen cells and its utility as a marker for fecal contamination.

Methanogenic bacteria, which are common inhabitants of the animal digestive tract, contain the fluorescent compound F420 (coenzyme 420), a 7,8-didemethyl-8-hydroxy-5-deazariboflavin chromophore. F420 was characterized as an initial step in determining if this compound would be useful as a fluorescent marker for the detection of fecal and ingesta contamination. Using a single anion exchange chromatographic process, F420 was separated from other cell components of a Methanobrevibacter sp. cell culture. The extent of separation was determined spectroscopically. To aid in the development of possible techniques for the detection of fecal contamination using F420 as a marker, further spectroscopic investigation of F420 was conducted using steady-state and time-resolved fluorescence methods. The fluorescence lifetime of F420 in an elution buffer of pH 7.5 was found to be 4.2 ns. At higher pH values, the fluorescence decay, F(t), was best described by a sum of two exponentials: at pH 13, F(t) = 0.31 exp(-t/4.20 ns) + 0.69 exp(-t/1.79 ns). Further investigation using front-faced fluorescence techniques has shown that emission from F420 can be collected efficiently from samples of methanogen cell cultures as well as from fecal material.

Animals as a source of Escherichia coli pathogenic for human beings.

Symptoms of SLT E coli-induced enteric disease in human beings include watery diarrhea, hemorrhagic diarrhea, and, in some cases, HUS. The most frequent serotype associated with HUS is O157:H7, although several other serotypes have also been implicated. These organisms produce SLT-I, SLT-II, or both toxins. Factors other than SLT are implicated as virulence attributes, such as adhesins and enterohemolysins, but roles for these factors in the pathogenicity of these organisms have not been defined. Colonization mechanisms for enterohemorrhagic E coli have not been defined, nor is there a defined set of characteristics by which enterohemorrhagic E coli pathogenic for human beings can be identified. Because virulence attributes are ill-defined, experimental animal models are useful in studies of pathogenicity. Gnotobiotic pigs, infant rabbits, streptomycin-treated mice, and one-day-old chickens have been used. Although the epidemiologic evidence implicating cattle as a source of zoonotic SLT E coli is strong, there is a paucity of direct evidence documenting this relationship. Until we have a better set of criteria with which to identify SLT E coli that are human pathogens, we are probably limited to epidemiologic criteria. Cattle excrete a variety of SLT E coli that includes many serotypes, in addition to O157:H7, that have been associated with disease in human beings. Surveys of the incidence of O157:H7 indicate a low incidence of these organisms in healthy cattle. However, much of these data have been derived from surveys of clinically normal cattle in daries.(ABSTRACT TRUNCATED AT 250 WORDS)

Involvement of a putative intercellular signal-recognizing G protein-coupled receptor in the engulfment of Salmonella by the protozoan Tetrahymena.

In an effort to investigate the molecular basis of protozoa engulfment-mediated hypervirulence of Salmonella in cattle, we evaluated protozoan G protein-coupled receptors (GPCRs) as transducers of Salmonella engulfment by the model protozoan Tetrahymena. Our laboratory previously demonstrated that non-pathogenic protozoa (including Tetrahymena) engulf Salmonella and then exacerbate its virulence in cattle, but the mechanistic details of the phenomenon are not fully understood. GPCRs were investigated since these receptors facilitate phagocytosis of particulates by Tetrahymena, and a GPCR apparently modulates bacterial engulfment for the pathogenic protozoan Entamoeba histolytica. A database search identified three putative Tetrahymena GPCRs, based on sequence homologies and predicted transmembrane domains, that were the focus of this study. Salmonella engulfment by Tetrahymena was assessed in the presence of suramin, a non-specific GPCR inhibitor. Salmonella engulfment was also assessed in Tetrahymena in which expression of putative GPCRs was knocked-down using RNAi. A candidate GPCR was then expressed in a heterologous yeast expression system for further characterization. Our results revealed that Tetrahymena were less efficient at engulfing Salmonella in the presence of suramin. Engulfment was reduced concordantly with a reduction in the density of protozoa. RNAi-based studies revealed that knock-down of one the Tetrahymena GPCRs caused diminished engulfment of Salmonella. Tetrahymena lysates activated this receptor in the heterologous expression system. These data demonstrate that the Tetrahymena receptor is a putative GPCR that facilitates bacterial engulfment by Tetrahymena. Activation of the putative GPCR seemed to be related to protozoan cell density, suggesting that its cognate ligand is an intercellular signaling molecule.

Systematic review: are lymphocytic colitis and collagenous colitis two subtypes of the same disease - microscopic colitis?

BACKGROUND: Despite similar clinical symptoms, collagenous colitis (CC) and lymphocytic colitis (LC) are considered two distinct disease entities. AIM: To compare pathoanatomical findings, clinical presentations, risk factors, course of diseases and response to treatment in CC and LC to establish whether they could be subtypes of the same disease, microscopic colitis (MC). METHODS: The MEDLINE was searched for CC, LC and MC, and clinical studies of >20 patients were included. Pooled results with 95% confidence intervals were calculated based on the number of patients. RESULTS: An abnormal number of intraepithelial lymphocytes are found in 45% (40-50%) with CC, and an abnormal subepithelial collagen band in 16% (13-20%) with LC suggesting a histological overlap. The incidence of CC and LC has increased in parallel. Mean age (CC 63 years; LC 60 years) and clinical presentation are indistinguishable, and females are predominant in CC (77%; 75-79%) as well as LC (68%; 66-70%). Risk factors such as nonsteroid anti-inflammatory drugs consumption CC 39% (36-42%); LC 32% (29-35%) are similar and prevalence of concomitant autoimmune diseases such as coeliac disease (CC 5%; CI: 4-6% and LC 7%; CI: 6-9%) do not differ. Bile acid diarrhoea is highly prevalent in CC (41%; 37-45%) and LC (29%; 24-34%). The effect of budesonide is identical. CONCLUSIONS: CC and LC could be considered histological subtypes of the same disease, MC. To facilitate recruitment to clinical trials, all MC patients could be included in future trials and stratified for subtypes.

Directed differentiation of porcine epiblast-derived neural progenitor cells into neurons and glia.

Neural progenitor cells (NPCs) are promising candidates for cell-based therapy of neurodegenerative diseases; however, safety concerns must be addressed through transplantation studies in large animal models, such as the pig. The aim of this study was to derive NPCs from porcine blastocysts and evaluate their in-vitro differentiation potential. Epiblasts were manually isolated from expanded hatched blastocysts and cultured on MEF feeder cells. Outgrowth colonies were passaged to MS5 cells and rosettes were further passaged to Matrigel-coated dishes containing bFGF and EGF. Three NPC lines were established which showed expression of SOX2, NESTIN and VIMENTIN. One line was characterised in more detail, retaining a normal karyotype and proliferating for more than three months in culture. Following differentiation, TUJI was significantly up-regulated in protocol 2 (RA and SHH; 58% positive cells) as were NF and TH. In contrast, MBP was significantly up-regulated in protocol 3 (FGF8 and SHH; 63% positive cells), whereas, GFAP was significantly up-regulated in protocols 1-4 (33%, 25%, 43% and 22%). The present study provides the first report of a porcine blastocyst-derived NPC line capable of differentiating into both neurons and glia, which may be of paramount importance for future transplantation studies in large animal models of neurodegenerative diseases.

Exposure of sheep scrapie brain homogenate to rumen-simulating conditions does not result in a reduction of PrP(Sc) levels.

AIMS: Experiments were designed to evaluate the potential of rumen-simulating conditions to reduce PrP(Sc) levels. METHODS AND RESULTS: Scrapie-positive brain material was incubated under rumen-simulating conditions. Time points were taken over a 24-h period and PrP(Sc) levels were analysed by Western blot. No loss of PrP(Sc) was observed over a 24-h time period. CONCLUSIONS: Our results indicate that a fully developed rumen fermentation does not provide significant protection against prion infection via the oral route. Developmental changes including senescence of immune system function or other developmental changes in the gastrointestinal tract are potential mechanisms by which relative bovine spongiform encephalopathy (BSE) susceptibility might vary with age. SIGNIFICANCE AND IMPACT OF THE STUDY: Epidemiology of the BSE outbreak in the United Kingdom indicates that younger animals were at higher risk of infection. The rumen undergoes pronounced developmental changes early in life, coinciding with the introduction of fibre into the diet. The timeframe of highest risk of infection overlaps the time in life prior to full rumen development. This work indicates that a fully developed rumen does not provide significant protection against prion infection via the oral route of infection. This result implicates other developmental changes that are responsible for the age-dependent susceptibility of cattle to BSE.