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The framework and assumptions for the economic and efficiency evaluation of Campylobacter control in the Dutch broiler meat chain as applied in the Campylobacter risk assessment and management (CARMA) project is given. The resultant cost-utility ratio showed the relative efficiency of several policy options adopted to reduce the infections.

The impaired immune function of systemic lupus erythematosus (SLE) may be associated with reduced purine activity. SLE is an autoimmune connective tissue disorder. Researchers measured purine enzyme activity in white blood cell samples from 17 patients with SLE and 43 healthy volunteers. The activity of the 5-prime nucleotidase enzyme was more than 50% lower in blood cells from SLE patients than in those from healthy volunteers. Impaired enzyme activity may result in the accumulation of nucleotides and other substances that alter metabolic activity.

Introduction The standards relating to bathing water are a burning issue. Many studies have shown that bathing in sea or fresh waters that meet current standards may lead to illness.[1-12] [...]

The Florida Complaint and Outbreak Reporting System (FL-CORS) database is used by the Florida Department of Health's Food and Waterborne Disease Program as one of the tools to detect foodborne disease outbreaks (FBOs). We present a descriptive and spatial network analysis of FL-CORS data collected during 2015 to 2018. We also quantified FBOs that were investigated and confirmed because of a filed complaint and the etiological agents involved in these outbreaks. An increasing number of unique complaints filed in FL-CORS was observed during 2015 to 2018, with a sharp increase during 2017 to 2018 and a different seasonal pattern in 2018. The preferred mechanism of reporting varied by age group, with younger people more frequently filing complaints online and older people preferring reporting in person or by phone. Spatial network analysis revealed that 87% of complaints had the same county of residence and county of presumed exposure. Frequency of complaints was negatively associated with linear distance between place of residence and place of exposure at the zip code level. Counties located in North and Central Florida, as well as some coastal areas in South Florida, had higher incidence rates of complaints. Those counties tend to have a large population density, and some are popular vacation destinations. On average, 96 FBOs were reported in Florida annually, of which 60% were confirmed with successful identification of the causative agent. The 56% of the confirmed FBOs were triggered by a complaint. Throughout the years, 2.4 to 2.8 FBOs and 1.4 confirmed FBOs were identified per 100 complaints. Ciguatera toxin was the cause of 40% of all FBOs in Florida, and only 28% of outbreaks were detected through complaints. In contrast, complaints were the main source of identifying outbreaks of norovirus, nontyphoidal Salmonella enterica,and scombroid food poisoning, as well as rare outbreaks of Clostridium perfringens, Cryptosporidiumspp., Shigellaspp., and Vibrio vulnificus.

Live broilers contain micro-organisms as Campylobacter and Salmonella on their feathers and feet and once in the slaughterhouse may contaminate the carcasses during processing. The results can be a final [...]

Several produce-borne outbreaks have been associated with the use of contaminated water during preharvest applications. Salmonellahas been implicated in a number of these outbreaks. The purpose of this study was to evaluate the microbial quality of agricultural surface water used in preharvest production on the Eastern Shore of Virginia in accordance with the Food Safety Modernization Act's Produce Safety Rule water standards. The study also examined the prevalence, concentration, and diversity of Salmonellain those water sources. Water samples (1 L) from 20 agricultural ponds were collected during the 2015 and 2016 growing seasons (n= 400). Total aerobic bacteria, total coliforms, and Escherichia coliwere enumerated for each sample. Population levels of each microorganism were calculated per 100-mL sample and log transformed, when necessary. Samples (250 mL) were also enriched for Salmonella. Presumptive Salmonellaisolates were confirmed by PCR (invAgene) and were serotyped. In 2016, the concentration of Salmonellain each sample was also estimated by most probable number (MPN). Indicator bacteria and environmental and meteorological factors were analyzed for their association with the detection of a Salmonella-positive water sample by using logistic regression analysis. Seventeen of the 20 ponds met the Food Safety Modernization Act's Produce Safety Rule standards for production agricultural water. Three ponds did not meet the standards because the statistical threshold value exceeded the limit. Salmonellawas detected in 19% of water samples in each year (38 of 200 in 2015 and 38 of 200 in 2016). Of the 118 Salmonellaisolates serotyped, 14 serotypes were identified with the most prevalent being SalmonellaNewport. E. coliconcentration, farm, and total aerobic bacteria concentration were significantly associated with the likelihood of detecting a Salmonella-positive sample The average concentration of Salmonellain all samples was 4.44 MPN/100 mL, with the limit of detection being 3.00 MPN/100 mL. The highest concentration of Salmonellawas 93.0 MPN/100 mL. These data will assist in a better understanding of the risks that production water poses to produce contamination events.

Microbial contamination of fresh produce (fresh fruits and vegetables) poses serious public health concerns worldwide. This study was conducted as a comprehensive analysis of biological hazards in the global fresh produce chain. Data about produce-related outbreaks and illness were collected from the annual reports and databases of foodborne outbreak surveillance systems in different regions and countries from 2010 to 2015. The global patterns of and regional differences in documented outbreaks and cases were analyzed, and produce commodities and pathogens of greatest concern were identified. Data on sporadic illnesses were also collected through a comprehensive literature review of case-control studies. We found 988 produce-related outbreaks (with known agents) and 45,723 cases in all regions and countries. The numbers of produce-related outbreaks per million person-years were approximately 0.76, 0.26, 0.25, 0.13, 0.12, and 0.05 in New Zealand, Australia, the United States, the European Union, Canada, and Japan, respectively. The top three food categories and pathogens contributing to produce-related outbreaks were vegetables and nonfruits (i.e., food other than fruits; 27.0%), unspecified vegetables (12.2%), and vegetable row crops (11.7%) and norovirus (42.4%), Salmonella enterica(19.9%), and Staphylococcus aureus(7.9%), respectively. Produce consumption was identified as a protective factor, a risk factor, and either a protective or risk factor for sporadic illnesses in 11, 5, and 5 studies, respectively, among 21 case-control studies. Risks associated with produce consumption in the United States and the European Union have been linked to various factors such as irrigation water, cross-contamination, storage time and temperature abuse, infected food handlers, and unprocessed contaminated ingredients. The results of the current study indicate the complexity of produce products consumed across the globe and the difficulty in tracing illnesses back to specific food ingredients.

The U.S. Food and Drug Administration (FDA) has defined standards for the microbial quality of agricultural surface water used for irrigation. According to the FDA produce safety rule (PSR), a microbial water quality profile requires analysis of a minimum of 20 samples for Escherichia coliover 2 to 4 years. The geometric mean (GM) level of E. colishould not exceed 126 CFU/100 mL, and the statistical threshold value (STV) should not exceed 410 CFU/100 mL. The water quality profile should be updated by analysis of a minimum of five samples per year. We used an extensive set of data on levels of E. coliand other fecal indicator organisms, the presence or absence of Salmonella,and physicochemical parameters in six agricultural irrigation ponds in West Central Florida to evaluate the empirical and theoretical basis of this PSR. We found highly variable log-transformed E. colilevels, with standard deviations exceeding those assumed in the PSR by up to threefold. Lognormal distributions provided an acceptable fit to the data in most cases but may underestimate extreme levels. Replacing censored data with the detection limit of the microbial tests underestimated the true variability, leading to biased estimates of GM and STV. Maximum likelihood estimation using truncated lognormal distributions is recommended. Twenty samples are not sufficient to characterize the bacteriological quality of irrigation ponds, and a rolling data set of five samples per year used to update GM and STV values results in highly uncertain results and delays in detecting a shift in water quality. In these ponds, E. coliwas an adequate predictor of the presence of Salmonellain 150-mL samples, and turbidity was a second significant variable. The variability in levels of E. coliin agricultural water was higher than that anticipated when the PSR was finalized, and more detailed information based on mechanistic modeling is necessary to develop targeted risk management strategies.

In a time of increasing threats and decreasing financial resources, monitoring and controlling all possible foodborne hazards at the same time and to the same extent has become more challenging than ever. Therefore, attention is increasingly being paid to the so-called “risk ranking” methods that enable decision makers to focus on the most important foodborne hazards — even when time is limited and knowledge incomplete. In this review paper, we provide an overview of the most common quantitative methods and metrics used for ranking the risks associated with foodborne parasites and present the state of the art on risk ranking exercises for foodborne parasites.

This study aimed at identifying explanatory variables that were associated with Campylobacterand Escherichia coliconcentrations throughout processing in two commercial broiler slaughterhouses. Quantative data on Campylobacterand E. colialong the processing line were collected. Moreover, information on batch characteristics, slaughterhouse practices, process performance, and environmental variables was collected through questionnaires, observations, and measurements, resulting in data on 19 potential explanatory variables. Analysis was conducted separately in each slaughterhouse to identify which variables were related to changes in concentrations of Campylobacterand E. coliduring the processing steps: scalding, defeathering, evisceration, and chilling. Associations with explanatory variables were different in the slaughterhouses studied. In the first slaughterhouse, there was only one significant association: poorer uniformity of the weight of carcasses within a batch with less decrease in E. coliconcentrations after defeathering. In the second slaughterhouse, significant statistical associations were found with variables, including age, uniformity, average weight of carcasses, Campylobacterconcentrations in excreta and ceca, and E. coliconcentrations in excreta. Bacterial concentrations in excreta and ceca were found to be the most prominent variables, because they were associated with concentration on carcasses at various processing points. Although the slaughterhouses produced specific products and had different batch characteristics and processing parameters, the effect of the significant variables was not always the same for each slaughterhouse. Therefore, each slaughterhouse needs to determine its particular relevant measures for hygiene control and process management. This identification could be supported by monitoring changes in bacterial concentrations during processing in individual slaughterhouses. In addition, the possibility that management and food handling practices in slaughterhouses contribute to the differences in bacterial contamination between slaughterhouses needs further investigation.

Objectives:To estimate the fraction of human cases of enterically transmitted illness by five major pathways (food, environment, direct animal contact, human–human transmission, and travel) and by 11 groups within the food pathway.Methods:Food safety experts were asked to provide their estimates of the most likely range for each of the parameters. Joint probability distributions were created by probabilistic inversion (PI).Results:Sixteen experts participated in the study. PI resulted in good fits for most pathogens. Qualitatively, expert estimates were similar to earlier published studies but the estimated fraction of foodborne transmission was lower for most pathogens. Biologically less plausible pathways were given some weight by the experts. Uncertainties were smallest for pathogens with dominant transmission routes.Conclusions:Structured expert studies are a feasible method for source attribution, but methods need further development.Applications:These estimates can be combined with data on incidence, disease burden and costs to provide specific estimates of the public health impact of foodborne illness, and to identify the food groups that have the highest impact.

Flow cytometry may be used to isolate large amounts of living, fluorescently labeled cells. Certain fluorescent labels, like enhanced cyan fluorescent protein (ECFP) and enhanced yellow fluorescent protein (EYFP), allow the assessment of direct protein–protein interaction in situ, by fluorescence resonance energy transfer (FRET). However, current flow cytometric methods either require elaborate technical adaptations or, using a single laser protocol, are hampered by background signal. We optimized a single 408‐nm laser protocol to detect FRET between ECFP/EYFP‐tagged proteins.Cell lines stably expressing ECFP and/or EYFP or an EYFP‐ECFP fusion protein were used to design the settings for the flow cytometer to detect FRET‐positive cells using a single 408‐nm laser. Using these settings, interactions between the subunits of the transcription factor NF‐Y were studied.Flow cytometric analysis of the cells expressing an EYFP‐ECFP fusion protein yielded a discrete FRET‐positive population. Using the same settings, in cells expressing NF‐YB‐CFP and NF‐YC‐YFP fusion proteins, FRET could also be detected. These cells were sorted and FRET was confirmed by confocal microscopy.FRET‐positive cells, expressing ECFP‐ and EYFP‐tagged proteins, can be detected using single 408‐nm laser excitation, with low background signal. This allows high‐throughput analysis and isolation of viable FRET‐positive and ‐negative cells for subsequent biological experiments. © 2006 International Society for Analytical Cytology

Leukocyte function associated antigen 1 (LFA-1) is an adhesion molecule indispensable in immune and inflammatory reactions, but its role in hematopoiesis is poorly understood. LFA-1 is considered as a marker of late stage stem cell maturation when expressed on CD34+bone marrow cells. We observed that CD34+bone marrow cells express LFA-1, that based on LFA-1 expression several subpopulations can be distinguished, and that the level of expression appeared highly variable among different donors. Unanticipated, in time course experiments we observed that CD34+LFA-1−cells expressed LFA-1 within 24 hours upon culture. These in vitro findings support the hypothesis that once contacts with bone marrow stroma are lost, LFA-1 is upregulated by default, due to the lack of negative regulating signals from stromal cells. This might also explain the widely variable expression of LFA-1 as a result of crowding of cells in the bone marrow with subsequent loss of contact with stroma and upregulation of LFA-1, thus equipping cells with adequate adhesion receptors to migrate throughout the bone marrow. Interestingly, the expression of the LFA-1 specific activation epitope L16 on these cells is low, even after culture. This demonstrated that LFA-1 is not activated, as was confirmed by low adhesion to ICAM-1. Activation of adhesion molecules is induced by several growth factors. Indeed, we show here that an osteoblastic cell line under normal conditions does hardly produce hematopoietic growth factors but these are rapidly induced after stimulation. Such rapid induction endows the bone marrow stroma with the property to modulate the adhesive strength and enabling migration through the different environments within the stroma. Prove for such compartments within the bone marrow is provided by histological data.

Repeated exposure to morphine and amphetamine induces long-lasting sensitization of their psychomotor stimulant properties, whereas pretreatment with morphine causes cross-sensitization of the locomotor effects of amphetamine. Here, we investigated whether pre-exposure to amphetamine also results in cross-sensitization to morphine. Rats pretreated with amphetamine (5 × 2.5 mg/kg, i.p.) displayed neither short-term (3 days post-treatment) nor long-term (3 weeks post-treatment) cross-sensitization of the locomotor effects of morphine (2 or 5 mg/kg, s.c.). Two other amphetamine pretreatment protocols (1 × 5 mg/kg, i.p. and 14 × 2.5 mg/kg, i.p.) also failed to induce cross-sensitization to morphine. In contrast, all amphetamine pretreatment regimens induced sensitization of the locomotor effects of amphetamine (1 mg/kg, i.p.) and pretreatment with morphine (14 × 10 mg/kg, s.c.) induced both short- and long-term sensitization of the locomotor effects of both morphine and amphetamine. These data suggest that the expression of sensitization of the locomotor effects of morphine and amphetamine, at least partially, involves distinct neuroadaptive phenomena.Neuropsychopharmacology (1999) 21 550–559.10.1016/S0893-133X(99)00051-2

Glucocorticoid receptor (GR)‐mediated facilitation of striatal dopaminergic (DA) neurotransmission has been proposed to play a role in behavioral sensitization induced by intermittent exposure to drugs of abuse or stressors. Searching for possible common neuronal substrates acted upon by drugs of abuse and corticosterone, we addressed the question as to whether such a facilitatory effect is apparent (i.e., persists) in primary cultures of rat striatum subsequent to intermittent (prenatal) morphine administration. As previously observed in striatal slices of morphine‐treated rats, intermittent morphine exposure in vivo caused a long‐lasting increase in DA D1 receptor‐stimulated adenylyl cyclase activity, that appeared to persist in primary cultures of rat striatal γ‐aminobutyric acid (GABA) neurons. Subsequent in vitro exposure of these striatal neurons to corticosterone or dexamethasone, simultanously activating GR and mineralocorticoid receptors (MR), about doubled this adaptive effect of previous in vivo morphine administration. The selective MR agonist aldosterone was ineffective in this respect. Prior in vivo morphine treatment also enhanced the stimulatory in vitro effect of corticotropin releasing hormone (CRH) on adenylyl cyclase in cultured GABA neurons. However, the enhanced CRH receptor functioning was not potentiated by in vitro corticosterone exposure. Activation of GR by corticosterone did not facilitate the increase in D1 receptor efficacy induced by sustained activation of muscarinic receptors in cultured striatal neurons. These data indicate that previous intermittent morphine administration induces a long‐lasting synergistic effect of corticosterone on enhanced striatal DA neurotransmission at the level of postsynaptic D1 receptors. Moreover, the induction of this neuroadaptation seems to display opioid receptor selectivity and its long‐term expression may be confined to D1 receptors. Since exposure to drugs of abuse or stressors not only increase striatal DA release but also plasma corticosterone levels, we hypothesize that this adaptive phenomenon in DA‐sensitive GABA neurons is involved in the expression of morphine‐induced long‐term behavioral sensitization to drugs of abuse and stressors. Synapse 25:381–388, 1997. © 1997 Wiley‐Liss, Inc.

Abstract: The role of release-inhibitory α2-adrenoceptors on or near noradrenergic nerve terminals was investigated in slices of rat nucleus accumbens and medial prefrontal cortex, representing major projection areas of mesocorticolimbic dopamine neurons. As expected, the electrically evoked release of [3H]noradrenaline from superfused medial prefrontal cortex slices was strongly inhibited (to about 25% of control values) by exogeneous noradrenaline and by the α2-adrenoceptor agonist oxymetazoline, while blockade of α2-adrenoceptors with phentolamine or rauwolscine caused a large increase (to almost 250% of control values) in neurotransmitter release. In striking contrast and much to our surprise, these drugs did not at all affect the electrically evoked release of [3H]noradrenaline from nucleus accumbens slices. These results indicate that α2-adrenoceptor autoregulation of noradrenaline release may not be a major feature of noradrenergic varicosities in rat nucleus accumbens. Thus, the functioning or presence of these receptors may vary with the brain region innervated by locus coeruleus neurons and may not represent a ubiquitous feature of noradrenergic neurons.

No abstract.

WE evaluated the incentive motivational properties of MK-801 by determining its priming effect on drug-seeking behaviour following extinction of cocaine self-administration, an animal model for drug craving. Rats were allowed to self-adminstrate cocaine (0.5mg/kg) or saline during 10 daily sessions. MK-801 (0.1–0.52mg/kg, i.p.) dose-dependently reinstated responding for cocaine following an extinction period of 3 weeks. Responding was selectively enhanced in the previously drug-paired hole and was completely absent in rats with a history of saline self-administration. These data provide evidence for a possible role of NMDA receptors in the incentive motivation underlying cocaine-seeking behaviour. In addition, the ability of MK-801 to elicit drug-seeking behaviour may prove to be a serious drawback for the proposed use of NMDA antagonists in the treatment of drug addiction.

1The effects of selective opioid receptor agonists and antagonists on N‐methyl‐D‐aspartate (NMDA, 10 μM)‐induced release of [3H]‐dopamine and [14C]‐acetylcholine (ACh) from superfused neostriatal slices were studied to investigate the possible occurrence of functional κ‐opioid receptor subtypes in rat brain.2The κ receptor agonists (−)‐ethylketocyclazocine ((−)‐EKC), U69593 and the endogenous opioid peptide dynorphin A1–13caused a naloxone‐reversible inhibition of NMDA‐induced [3H]‐dopamine release, with pD2values of about 9, 8.5 and 8.2, respectively, whereas both the μ agonist Tyr‐D‐Ala‐Gly‐(NMe)Phe‐Gly‐ol (DAMGO) and theδ agonist D‐Pen2‐D‐Pen5‐enkephalin (DPDPE) were ineffective in this respect. The inhibitory effect of submaximally effective concentrations of dynorphin A1–13, U69593 and (−)‐EKC on NMDA‐induced [3H]‐dopamine release were not changed by the δ1/δ2‐opioid receptor antagonist naltrindole (up to a concentration of 1 μM), but reversed by the κ receptor antagonist nor‐binaltorphimine (nor‐BNI), with an IC50as low as 0.02 nM, indicating the involvement of U69593‐sensitive κ1‐opioid receptors.3NMDA‐induced [14C]‐ACh release was reduced in a naloxone‐reversible manner by DPDPE (pD2about 7.2), dynorphin A1–13(pD26.7) and EKC (pD26.2), but not by U69593 and DAMGO. The inhibitory effect of a submaximally effective concentration of DPDPE, unlike those of dynorphin A1–13and (−)‐EKC, on NMDA‐induced [14C]‐ACh release was antagonized by naltrindole with an IC50of 1 nM, indicating the involvement of δ‐opioid receptors in the inhibitory effect of DPDPE. On the other hand, the inhibitory effects of dynorphin A1–13and (−)‐EKC on [14C]‐ACh release were readily antagonized by nor‐BNI with an IC50of about 3 nM. A 100 fold higher concentration of nor‐BNI also antagonized the inhibitory effect of DPDPE, indicating the involvement of U69593‐insensitive κ2‐opioid receptors in the inhibitory effects of dynorphin A1–13and (−)‐EKC.4Although naloxone benzoylhydrazone (NalBzoH), displaying high affinity towards the putative κ3‐opioid receptor, antagonized the inhibitory effects of dynorphin A1–13and (−)‐EKC on [3H]‐dopamine and [14C]‐ACh release as well as that of U69593 on [3H]‐dopamine release, it displayed a low apparent affinity (IC50about 100 nM) in each case.5In conclusion, whereas activation of κ1‐opioid receptors causes presynaptic inhibition of NMDA‐induced dopamine release, κ2receptor activation results in inhibition of ACh release in rat neostriatum. As such, this study is the first to provide unequivocal in vitroevidence for the existence of functionally distinct κ‐opioid receptor subtypes in the brain.

The release of 14C-ACh from rat nucleus accumbens slices, induced by 15 mM [K+], was inhibited by the µ- and d-opioid agonists DAMGO and DPDPE, respectively, whereas only the ? agonist U50,488 reduced the release of 3H-DA.

3H-noradrenaline release from rat neocortical slices induced by 15 mM K+ was concentration-dependently inhibited by morphine, [D-Ala2-D-Leu5] enkephalin (DADLE) and the calcium entry blocker Cd2+. Blockade of presynaptic a2-adrenoceptors with phentolamine, almost doubling K+-induced 3H-noradrenaline release, slightly enhanced the relative inhibitory effects of morphine and DADLE, whereas that of Cd2+ remained unaffected. In contrast, activation of presynaptic a2-adrenoceptors with clonidine (1 µM) or TL-99 (1 µM), inhibiting release by about 50%, completely abolished the inhibitory effects of morphine and DADLE without affecting that of Cd2+. When in the presence of 1 µM clonidine adenylate cyclase was activated with forskolin (10 µM), which restored release to the drug-free control level, the opioids still did not display their inhibitory effects. Therefore, µ-opioid receptor efficacy appears to be dependent on the degree of activation of a2-adrenoceptors in central noradrenergic nerve terminals, probably through a local receptor interaction within the nerve terminal membrane.

Possible local interactions between noradrenergic and serotonergic systems in the dorsal raphe region of the rat were investigated by studying the effects of various drugs on depolarization (20 mmol/l K+)-induced release of [3H]5-hydroxytryptamine (5-HT) and [3H]noradrenaline (NA) in vitro using a superfusion method. Exogenous 5-HT did not influence the release of [3H]NA. However, NA (in the presence of 10 µmol/l desipramine) as well as the selective a2-adrenoceptor agonists clonidine and oxymetazoline strongly inhibited [3H]5-HT release. The selective a1-adrenoceptor agonists phenylephrine and methoxamine did not affect the release of either [3H]5-HT or [5H]NA. The inhibition by NA of both [3H]5-HT and [5H]NA release was not affected by the ß-adrenoceptor antagonist sotalol nor by the selective a1-adrenoceptor antagonist prazosin. However, phentolamine and the selective a2-adrenoceptor antagonists yohimbine and rauwolscine competitively antagonized the inhibitory effect of NA on [3H]NA release (respective pA2-values 7.5 and 8.3) and on [3H]5-HT release (respective pA2-values 7.7 and 8.2). Moreover, the release of [3H]NA and also, but to a lesser extent, that of [3H]5-HT were increased by the antagonists. It is concluded that the release of both 5-HT and NA in the dorsal raphe region may be subject to presynaptic inhibition by NA via activation of a2-adrenoceptors.

The effects of oxymetazoline and noradrenaline (in the presence of desipramine) on the release of 3H-noradrenaline from rat brain cortex synaptosomes were studied using a superfusion technique. Both drugs (at 1µM concentrations) were found to reduce the depolarization-induced (15 mM K+) release of 3H-noradrenaline. The release-modulating effect of noradrenaline was antagonized by phentolamine and yohimbine.

The characteristics of 3H-DA release from striatal slices by electrical stimulation were analyzed and the effects of a number of neuroleptics thereon were examined under different experimental conditions. The butyrophenones, haloperidol and spiroperidol, already at low concentrations (0.1–1 µM) increased basal tritium efflux in a dose-dependent manner. The phenothiazines, chlorpromazine and fluphenazine, were much less effective in this respect.

D-1 dopamine receptor-stimulated cyclic AMP efflux from rat neostriatal slices (induced by 30 µM dopamine + 10 µM (-)sulpiride) was concentration-dependently reduced by morphine, [abetd-Ala-abetd-Leu]-enkaphalin (DADLE), [d-Pen-abetd-Pen]enkephalin (DPDPE) and bremazocine. Naloxone (0.1 µM) selectively antagonized the inhibitory effect of (a submaximally effective concentration of) morphine, whereas ICI 174864 (0.75 µM) completely blocked the inhibitory effects of DADLE, DPDPE and bremazocine without affecting that of morphine, indicating a role of µ- as well as d-opioid receptors. Upon simultaneous activation of D-1 dopamine receptors and b-opioid receptors the (µ-receptor-mediated) inhibitory effect of morphine was abolished, while it was not changed following simultaneous activation of D-1 and (inhibitory) D-2 dopamine receptors. Cyclic AMP efflux induced by isoprenaline or adenosine was not affected by the opioids and that induced by vasoactive intestinal peptide (VIP) was inhibited by morphine and DADLE only. In the latter case naloxone, but not ICI 174864, antagonized the inhibitory effects.

In Ca2+-free EGTA-containing medium veratrine (3–25 µM) concentration-dependently enhanced the efflux of 3H-noradrenaline from (radiolabelled) rat neocortical slices. Clonidine (1 µM) inhibited and phentolamine (3 µM) enhanced veratrine-induced 3H-noradrenaline release and the modulatory effects were inversely related to the veratrine concentration used. Dibutyryl-cyclic AMP, 8-Bromo-cyclic AMP (10 µM-3 mM) and the adenylate cyclase activators NaF (2 mM) and forskolin (10 µM) enhanced 3H-noradrenaline release induced by 3 µM veratrine, but had no effect on spontaneous tritium efflux. In the presence of these drugs the modulatory effects of clonidine and phentolamine on 3H-noradrenaline release were reduced as expected from the enhanced efficacy of veratrine. In contrast to these drugs the selective cyclic AMP-phosphodiesterase inhibitor ZK62771 reduced veratrine (3 µM)-induced 3H-noradrenaline release in Ca2+-free medium. In the presence of 1.2 mM Ca2+, 3H-noradrenaline release induced by 13 mM K+ was also inhibited. However, when 3H-noradrenaline release was effected in the presence of tetrodotoxin (0.3 µM) or by electrical field-stimulation (1 Hz), ZK 62771 slightly but significantly enhanced the release.

Brain cortical slices were superfused with Krebs-Ringer media and the effects of oxymetazoline (an a-adrenoceptor agonist) and phentolamine (an a-antagonist) on depolarization-induced 3H-NA release were examined. Depolarization was effected by various K+-concentrations or by electrical pulses.

The effects of various drugs on the distribution of 3H-dopamine in rat striatal synaptosomes were studied after superfusion with this radiolabelled neurotransmitter. By compartmental analysis of the efflux of radioactivity, drug effects on the total accumulation of 3H-dopamine, on the “bound fraction” and on the rate constant of the efflux of radioactivity from the tissue were determined. Subsequently, the synaptosomes were exposed to 40 mM K+, in order to assess also the effects of these drugs on depolarization-induced 3H-dopamine release.

In the present paper we report an accurate method to quantify the accumulation of radiolabelled dopamine in striatal synaptosomes after superfusion of these particles with low concentrations of 3H-dopamine. Compartmental analysis of the efflux of radioactivity following 3H-dopamine accumulation was carried out. Moreover, the effects of some drugs and the uptake kinetics were studied in order to confirm that 3H-dopamine was accumulated in dopaminergic synaptosomes. The data show that:1.Extracellular 3H-dopamine disappears from the superfusion chambers within 10 min, after which a steady state of efflux is rapidly established.2.Accumulated 3H-dopamine distributes in two compartments: an efflux compartment, and a bound fraction which does not contribute to the efflux.3.Extrapolation from the steady state to the start of the efflux accurately estimates the initial accumulation in the efflux compartment.4.Radioactivity extracted from synaptosomes, previously superfused with 3H-dopamine, consisted for about 90% of unmetabolized dopamine and this percentage slightly increased with time.5.Both cocaine and tyramine at a concentration of 10 µM inhibited the accumulation of 3H-dopamine by 85%, although their different mechanisms of action could be clearly distinguished by compartmental analysis.6.At least 90% of the 3H-dopamine retained had been transported via the high affinity membrane carrier for dopamine, as concluded from the uptake kinetic data.7.The maximal uptake rate was found to be 100±2.1 pmol·min-1·mg-1 protein (n=4), which is higher than the values reported in the literature.8.This method makes possible a reliable differentiation between uptake inhibiting and releasing actions of drugs.

Synaptosomal preparations, obtained from rat corpus striatum, were superfused with Krebs-Ringer medium. During superfusion the synaptosomes were exposed to various concentrations of 3H-dopamine for a few minutes. Depolarization was effected by superfusion with medium containing elevated K+-concentrations. Both the accumulation and the subsequent K+-induced release of 3H-dopamine were quantified in the same preparation. This method was used to detect the presence of 3H-dopamine in different pools in isolated dopaminergic nerve terminals. The data obtained can be summarized as follows:1.The percentage of 3H-dopamine released and, hence, the distribution of 3H-dopamine into intrasynaptosomal transmitter pools, was independent of the amount of 3H-dopamine accumulated in the synaptosomes.2.Upon repeated depolarization, 3H-dopamine release was markedly depressed. Supplying the synaptosomes with 3H-dopamine between two stimulations fully restored the release in response to depolarization. Therefore, the depression of 3H-dopamine release upon repeated depolarization is probably due to the selective depletion of a releasable pool and not to an impairment of stimulus-secretion coupling processes.3.When synaptosomes were subjected twice to K+-stimulation and exposed to 3H-dopamine either before the first stimulation or between both stimulations, the most recently accumulated 3H-dopamine was released (by the second stimulation) in preference to 3H-dopamine accumulated previously.4.14C-Dopamine synthesized in the synaptosomes from 14C-tyrosine was not released in preference to 3H-dopamine that entered synaptosomal pools through the high affinity uptake mechanism.5.Our results strongly support the view that dopamine is stored heterogeneously in the varicosities of nigro-striatal neurones in the rat. The remarkably slow redistribution of stored 3H-dopamine into synaptosomal pools indicates that differences in vesicle localization with respect to the nerve terminal membrane might explain the occurrence of different dopamine pools.

A comparison is made between interlaboratory studies in which individual chlorobiphenyls (CBs) are measured in fish and sludge samples. The target coefficient of variation (CV(R)) between laboratories at CB levels of 0.01–1.0 mg/kg should be in the order of 20–30%. An improvement in the CV(R) is clearly shown in studies in which a learning program, paying attention to the optimization of gas Chromatographic operating conditions, is included. For standard solutions, where no interference problems of the matrix occur, excellent CV(R)s are obtained in the order of 6%. In fish oil CV(R)s of 10–30% and in sludge CV(R)s of 20–30% are reached.

To further define the role of CD66 glycoproteins in the regulation of neutrophil function, we analysed the effects of a CD66 monoclonal antibody, Kat4c, which recognizes an epitope present on AB domains of CD66a, CD66b and CD66c. Intact Kat4c and F(ab′)2 fragments were found to augment fMLP-induced oxidation of 1,2,3-dihydrorhodamine (oxidant species production) and β2 integrin-mediated adhesion to fibrinogen but did not promote β2 integrin-mediated binding of albumin coated latex beads. Since the latter assay is a sensitive indicator of neutrophil CD11b/CD18 functional activation, these results imply CD66 may exert differential effects upon β2 integrin activity. Neutrophil oxidant species production and spreading on fibrinogen substrates were further potentiated by cross-linking of Kat4c F(ab′)2, in keeping with the suggestion that ligation of CD66 regulates neutrophil function. However, although intact Kat4c promoted β2 neutrophil adhesion, F(ab′)2 fragments were without effect, implying a role for Fc receptors in this effect which has previously been attributed to CD66. Together these data define more clearly the role of CD66 in regulation of neutrophil function and further suggest that augmented β2 integrin-mediated adhesion following CD66 ligation occurs independently of affinity regulation.

We studied the incidence and potential prognostic value of thyroid abnormalities after allogeneic bone marrow transplantation (BMT) without total body irradiation (TBI) conditioning. 77 consecutive patients who received a chemotherapy-alone-based conditioning regimen pretransplant were included. Free serum thyroxine (FT4), free serum triiodothyronine (FT3) and serum thyrotropin (TSH) levels were assayed before and 3 and 14 months after BMT. Patients were classified in three categories: normal thyroid profile if FT3 and FT4 were within the normal range and TSH was normal or low, peripheral thyroid insufficiency (PTI) if TSH was >4 mIU/l, or an ‘euthyroid sick syndrome’ (ETS) if FT3 and/or FT4 were low and TSH was normal or low. The incidence of thyroid dysfunction at 3 months was 57%, and 29% at 14 months. This was mostly due to the occurrence of ETS which was more frequent at 3 months (48%, 29/61) than at 14 months (19%, 9/48). Furthermore, at 3 months, survival was significantly lower in the ETS group (34.5%) than in the euthyroid group (96.2%), or in the PTI group (83.3%) (P  <  0.0001). PTI was observed even in the absence of TBI in 11 patients (14%) and was equally distributed at 3 months (n  =  6) and 14 months (n  =  5).

There has been a steady production of Dutch missionary literature during the years since the end of World War II. This literature and the thinking which it expresses are generally unknown in America. The Missionary Research Library has sought someone for the last four years to report on the Dutch literature of missions. Through the kindness of Dr. Oussóren this survey statement has been made available.–Ed.

Globally, two billion people suffer from micronutrient deficiencies, 151 million children under five suffer from stunting, and millions more have impaired cognitive development related to poor nutrition. This is partly due to insufficient consumption of animal-sourced foods (ASF), which supply multiple bioavailable nutrients that are lacking in the cereal-based diets of the poor. Yet, reports like the one recently published by the EAT-Lancet Commission, solely focus on the threat of ASF consumption on sustainability and human health, overestimate and ignore the tremendous variability in the environmental impact of livestock production, and fail to adequately include the experience of marginalized women and children in low- and middle-income countries whose diets regularly lack the necessary nutrients. Yet animal-source foods have been described by the World Health Organization as the best source of high-quality nutrient-rich food for children aged 6–23 months. Livestock and ASF are vital to sustainability as they play a critical role in improving nutrition, reducing poverty, improving gender equity, improving livelihoods, increasing food security, and improving health. The nutritional needs of the world's poor, particularly women and children, must be considered in sustainability debates.

The adenylate cyclase activator forskolin as well as 8-bromo-cyclic AMP enhanced the electrically evoked release of3H-noradrenaline and3H-5-hydroxytryptamine from superfused rat neocortical slices and that of3H-dopamine from neostriatal slices with comparable EC50's of about 0.5 and 50 µM, respectively, without affecting spontaneous tritium efflux. The phosphodiesterase inhibitor ZK 62771 (3–100 µM) also enhanced3H-noradrenaline and3H-dopamine release but slightly reduced3H-5-hydroxytryptamine release. However, this drug profoundly enhanced spontaneous tritium release in the latter case. The facilitatory effect of forskolin (0.3 µM) on the release of the amine neurotransmitters was potentiated in the presence of ZK 62771 (30 µM). Therefore, cyclic AMP appears to exert a general facilitatory effect on the release of these biogenic amines from central nerve terminals.

At least three different families of endogenous opioid peptides, the enkephalins, endorphins and dynorphins, are present in the mammalian central nervous system (CNS). Immuno-cytochemical studies have demonstrated their localization in neurones1–8, which supports the view that these peptides may have a role as neurotransmitters or neuromodulators. However, the target cells and cellular processes acted upon by the opioid peptides are still largely unknown. One possible function of neuropeptides, including the opioid peptides, may be presynaptic modulation of neurotransmission in certain neuronal pathways, for example, by inhibition or promotion of neurotransmitter release from the nerve terminals9–12. Here we report that dynorphin and some benzomorphans potently and selectively inhibit the release of (radiolabelled) dopamine from slices of rat corpus striatum, by activating κ-opioid receptors. In contrast, [Leu5]enkephalin and [D-Ala2, D-Leu5]enkephalin selectively inhibit acetylcholine release by activating δ-opioid receptors.

Various malignant and benign neoplastic lesions have been noted in the past to appear on tattoos (Auger, 1943; Bashir, 1976; McQuarrie, 1966). The development of malignant melanoma in a tattoo has been noted to occur in only a few cases (Kirsch, 1969; Sharlit, 1938; Wolfort et al., 1974).We are reporting hereunder two patients with malignant disease previously treated by surgery and radiotherapy, one developing a primary and the other a metastatic melanoma in tattoo sites used for marking a radiation field. This possible result of radiation and tattooing appears to be, to the best of our knowledge, the first cases in the literature.Case No. 1:A 52-year-old caucasian female with a previous history of tonsillectomy, psoriasis and fibroadenoma of her right breast underwent, in August 1975, left radical mastectomy for intraductal schirrus adenocarcinoma.Three of the eight axillary lymph nodes were invaded by the tumour. Subsequently, the patient received radiotherapy by 60Co ? rays to a total of 3600 cGy (rad) to the suprasternal, axillary, internal mammary and tangential fields. Indian ink was used for marking the radiation fields.The patient was without evidence of disease until June 1977, when a bleeding nevus 0.5 × 1.0 cm was observed on a tattoo at the border of the left tangential chest-wall field. The lesion was excised and on histology a malignant melanoma Clark Level II was found.

BackgroundThe public health impact of foodborne diseases globally is unknown. The WHO Initiative to Estimate the Global Burden of Foodborne Diseases was launched out of the need to fill this data gap. It is anticipated that this effort will enable policy makers and other stakeholders to set appropriate, evidence-informed priorities in the area of food safety.

BackgroundFoodborne diseases are an important cause of morbidity and mortality in populations across the globe. Estimates of the burden of these diseases are scarce. As infectious diseases are highly dynamic, the trends in food-related disease incidence over a 6-year period and the associated variation in disease burden are examined.

BackgroundProton pump inhibitors (PPIs) are prescribed to patients requiring prevention or treatment of gastric and duodenal ulcers. PPIs inhibit gastric acid secretion by specific action on proton pumps of parietal cells. The resulting pH increase is favourable for gastrointestinal pathogen survival. Indeed, PPIs are reported repeatedly as potential risk factors for infections, such as Campylobacterand Salmonellainfections. The current study examined national trends in PPI use, reported campylobacteriosis cases between 2004 and 2011, and estimated the excess disease burden potentially related to PPI use.

BackgroundMost estimates of disease burden due to infectious diseases focus on the impact of acute infections and do not fully capture the effects of (long-term) sequelae. To provide evidence-based and comparable estimates, the European Centre for Disease Prevention and Control (ECDC) initiated the Burden of Communicable Diseases in Europe (BCoDE) project.