Search

Abstract Background Besides being a causative agent for genital warts and cervical cancer, human papillomavirus (HPV) contributes to 40-85% of cases of anal, penile, vaginal and vulvar cancer and precancerous lesions. HPV types 16 & 18 in particular contribute to 74-93% of these cases. Overall the number of new cases of these four cancers may be relatively high implying notable health care cost to society. The aim of this study was to estimate the incidence and the health care sector costs of anal, penile, vaginal and vulvar cancer. Methods New anogenital cancer patients were identified from the Danish National Cancer Register using ICD-10 diagnosis codes. Resource use in the health care sector was estimated for the year prior to diagnosis, and for the first, second and third years after diagnosis. Hospital resource use was defined in terms of registered hospital contacts, using DRG (Diagnosis Related Groups) and DAGS (Danish Outpatient Groups System) charges as cost estimates for inpatient and outpatient contacts, respectively. Health care consumption by cancer patients diagnosed in 2004–2007 was compared with that by an age- and sex-matched cohort without cancer. Hospital costs attributable to four anogenital cancers were estimated using regression analysis. Results The annual incidence of anal cancer in Denmark is 1.9 per 100,000 persons. The corresponding incidence rates for penile, vaginal and vulvar cancer are 1.7, 0.9 and 3.6 per 100,000 males/females, respectively. The total number of new cases of these four cancers in Denmark is about 270 per year. In comparison, the total number of new cases cervical cancer is around 390 per year. The total cost of anogenital cancer to the hospital sector was estimated to be 7.6 million Euros per year. Costs associated with anal and vulvar cancer constituted the majority of the costs. Conclusions Anogenital cancer incurs considerable costs to the Danish hospital sector. It is expected that the current HPV vaccination program will markedly reduce this burden.

Abstract Background A large number of people in both developing and developed countries rely on medicinal plant products to maintain their health or treat illnesses. Available evidence suggests that medicinal plant consumption will remain stable or increase in the short to medium term. Knowledge on what factors determine medicinal plant consumption is, however, scattered across many disciplines, impeding, for example, systematic consideration of plant-based traditional medicine in national health care systems. The aim of the paper is to develop a conceptual framework for understanding medicinal plant consumption dynamics. Consumption is employed in the economic sense: use of medicinal plants by consumers or in the production of other goods. Methods PubMed and Web of Knowledge (formerly Web of Science) were searched using a set of medicinal plant key terms (folk/peasant/rural/traditional/ethno/indigenous/CAM/herbal/botanical/phytotherapy); each search terms was combined with terms related to medicinal plant consumption dynamics (medicinal plants/health care/preference/trade/treatment seeking behavior/domestication/sustainability/conservation/urban/migration/climate change/policy/production systems). To eliminate studies not directly focused on medicinal plant consumption, searches were limited by a number of terms (chemistry/clinical/in vitro/antibacterial/dose/molecular/trial/efficacy/antimicrobial/alkaloid/bioactive/inhibit/antibody/purification/antioxidant/DNA/rat/aqueous). A total of 1940 references were identified; manual screening for relevance reduced this to 645 relevant documents. As the conceptual framework emerged inductively, additional targeted literature searches were undertaken on specific factors and link, bringing the final number of references to 737. Results The paper first defines the four main groups of medicinal plant users (1. Hunter-gatherers, 2. Farmers and pastoralists, 3. Urban and peri-urban people, 4. Entrepreneurs) and the three main types of benefits (consumer, producer, society-wide) derived from medicinal plants usage. Then a single unified conceptual framework for understanding the factors influencing medicinal plant consumption in the economic sense is proposed; the framework distinguishes four spatial levels of analysis (international, national, local, household) and identifies and describes 15 factors and their relationships. Conclusions The framework provides a basis for increasing our conceptual understanding of medicinal plant consumption dynamics, allows a positioning of existing studies, and can serve to guide future research in the area. This would inform the formation of future health and natural resource management policies.

Abstract Background A recent qualitative study in Denmark showed that genital warts (GWs) can considerably lower the quality of life of heterosexual patients. In this follow-up study, we interviewed men having sex with men (MSM) suffering from GWs to obtain an in-depth understanding of their perception of GWs and determine the extent to which minority (homosexual) cultural issues affect these patients' experiences. Qualitative interviews with six MSM were performed using a semi-structured interview guide. Questions were formulated on the basis of the earlier qualitative study in heterosexual patients with GWs along with a literature review. Data were analysed using a medical anthropological approach. Findings Many MSM worried about being stigmatised and excluded from the small homosexual 'scene', their key social group, thereby lowering their chances of finding sex and love. Most participants had suffered from GWs for several years which added to the negative psycho-sexual and social effects of the disease. Participants' fears of developing anal cancer were similar to those expressed about cervical cancer by females with GWs. Conclusions Ano-genital human papillomavirus (HPV) infection is common and has a serious psychological and sexual impact among MSM. However, they do not benefit to the same extent as heterosexual men from the herd immunity effect of HPV vaccination of girls. The pathological profile and concerns specific to MSM should be addressed when communicating with these patients, and should be taken into account when considering HPV vaccination of boys.

Abstract Background Genital warts, which are caused by infection with human papillomavirus (HPV), are one of the most common sexually transmitted diseases in Europe. Although genital warts are commonly perceived as a non-serious condition, treatment is often long, of varying effectiveness and the recurrence rate is high. Very few studies have been performed on the personal consequences of genital warts. The aim of this qualitative study, set in Denmark, was to examine the ways in which genital warts may affect patients' quality of life. Methods To obtain an in-depth understanding of patients' perceptions of genital warts, we used qualitative focus-group interviews with five men and five women aged between 18 and 30 years who had genital warts. The interview guide was based on a literature review that identified important issues and questions. The data were analysed using a medical anthropological approach. Results Patients' experiences were related to cultural conceptions of venereal diseases and the respective identities and sexuality of the sexes. The disease had negative psychological and social effects both for men and for women and it affected their sex and love lives, in particular. The psychological burden of the disease was increased by the uncertain timeline and the varying effectiveness of treatment. We identified a need for more patient information about the disease and its psycho-sexual aspects. Conclusions The men and women participating in this study considered their quality of life to be significantly lowered because of genital warts. The experiences described by the participants give insights that may be valuable in treatment and counselling. The quadrivalent HPV vaccine that has now been added to the childhood vaccination programme for girls in Denmark for the prevention of cervical cancer can also prevent 90% of cases of genital warts. Our results suggest that HPV vaccination could considerably reduce the largely unacknowledged psychological and social burden associated with genital warts, in men as well as women.

Abstract Background Sodium retention and ascites are serious clinical problems in cirrhosis. Urodilatin (URO) is a peptide with paracrine effects in decreasing sodium reabsorption in the distal nephron. Our aim was to investigate the renal potency of synthetic URO on urine sodium excretion in cirrhosis patients with sodium retention and ascites. Methods Seven cirrhosis patients with diuretics-resistant sodium retention received a short-term (90 min) infusion of URO in a single-blind, placebo-controlled cross-over study. In the basal state after rehydration the patients had urine sodium excretion < 50 mmol/24 h. Results URO transiently increased urine sodium excretion from 22 ± 16 μmol/min (mean ± SD) to 78 ± 41 μmol/min (P < 0.05) and there was no effect of placebo (29 ± 14 to 44 ± 32). The increase of URO's second messenger after the receptor, cGMP, was normal. URO had no effect on urine flow or on blood pressure. Most of the patients had highly elevated plasma levels of renin, angiotensin II and aldosterone and URO did not change these. Conclusion The short-term low-dose URO infusion increased the sodium excretion of the patients. The increase was small but systematic and potentially clinically important for such patients. The small response contrasts the preserved responsiveness of the URO receptors. The markedly activated systemic pressor hormones in cirrhosis evidently antagonized the local tubular effects of URO.

Abstract Background A depressed Apgar score at 5 minutes is a marker for perinatal insults, including neurologic damage. We examined the association between 5-minute Apgar score and the risk of epilepsy hospitalization in childhood. Methods Using records linked from population registries, we conducted a cohort study among singleton children born alive in the period 1978–2001 in North Jutland County, Denmark. The first hospital discharge diagnosis of epilepsy during the follow-up time was the main outcome. We followed each child for up to 12 years, calculated absolute risks and risk differences, and used a Poisson regression model to estimate risk ratios for epilepsy hospitalization. We adjusted risk ratio estimates for birth weight, gestational age, mode of delivery, birth presentation, mother's age at delivery, and birth defects. Results One percent of the 131,853 eligible newborns had a 5-minute Apgar score Conclusion An Apgar score

Background and Aims: Cardiovascular disease (CVD) poses significant health challenges globally. While substantial data exists for most populations, the Arctic Inuit's CVD incidence rates remain understudied. This research aimed to change this by estimating CVD incidence and mortality rates in Greenland from 1994 to 2021., Methods: Using nationwide registers, a retrospective observational study was conducted, focusing on individuals born in Greenland to Greenlandic-born parents. Data were sourced from the Greenlandic Hospital Discharge Register and the nationwide electronic medical record., Results: A total of 65,824 individuals were included. the age- and sex-specific incidence rates (IR) of ischemic heart disease, stroke, and heart failure (HF) declined from 1994 to 2021, with the most substantial decline observed for HF among women. Conversely, the IR of atrial fibrillation/flutter increased in both men and women, while the IR of myocardial infarction rose among men. The IR for stroke was particularly elevated compared to other CVD subgroups. Mortality rates for those diagnosed with CVD were 2.4 times higher than those without. Men exhibited a 40 % elevated mortality risk relative to women., Conclusion: The study provides pivotal insights into CVD trends within the Arctic Inuit population, highlighting both positive developments and areas of concern. Given the increasing elderly demographic in Greenland, proactive health strategies are crucial. Emphasizing primary prevention and addressing specific CVD risks, particularly the elevated stroke IR, is imperative for future public health efforts., Competing Interests: The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: HEL reports financial support was provided by Karen Elise Jensen Foundation. HEL reports financial support was provided by Novo Nordisk Foundation. HEL reports travel was provided by A. P. Møller Foundation. HEL reports travel was provided by Ketty and Ejvind Lyngsbæks Foundation. FG reports a relationship with Bayer that includes: consulting or advisory. FG reports a relationship with Pfizer that includes: consulting or advisory. FG reports a relationship with Astra-Zeneca that includes: consulting or advisory. FG reports a relationship with Ionis that includes: consulting or advisory. FG reports a relationship with Alnylam that includes: consulting or advisory. FG reports a relationship with Pharmacosmos that includes: consulting or advisory. FG reports a relationship with Abbott that includes: consulting or advisory. FG reports a relationship with Novartis that includes: speaking and lecture fees. FG reports a relationship with Orion Pharma that includes: speaking and lecture fees. MEJ reports a relationship with Astra-Zeneca that includes: funding grants. MEJ reports a relationship with Sanofi Aventis that includes: funding grants. MEJ reports a relationship with Boehringer Ingelheim that includes: funding grants. MEJ reports a relationship with Novo Nordisk that includes: funding grants. MEJ reports a relationship with Novo Nordisk that includes: equity or stocks. If there are other authors, they declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (© 2024 The Authors.)

Heart Failure (HF) constitutes a significant burden for healthcare around the world. In Greenland, risk factors like smoking, diabetes, and obesity are prevalent. Yet, the prevalence of HF remains unexplored. This register-based cross-sectional study uses data from the national medical record in Greenland to estimate the age- and gender-specific prevalence of HF and to describe the characteristics of patients with HF in Greenland. A total of 507 patients (26% women) with a mean age of 65 years were included based on a diagnosis of HF. The overall prevalence was 1.1% and higher among men compared to women (1.6% vs. 0.6%, p < 0.05). The highest prevalence was among men above 84 years (11.1%). More than half (53%) had a body mass index above 30 kg/m2 and 43% were current daily smokers. The proportion diagnosed with ischaemic heart disease (IHD) was 33%. The overall prevalence of HF in Greenland is consistent with that in other high-income countries, yet high among men in some age groups, compared to Danish men. Almost half the patients were obese and/or smokers. A low prevalence of IHD was observed indicating that other factors may play a role in developing HF among Greenlanders.

We demonstrate a continuous-wave (CW) coherent detection lidar (CDL) capable of detecting micro-Doppler (propeller) signatures and acquiring raster-scan images of small unmanned aerial systems/vehicles (UAS/UAV). The system uses a narrow-linewidth 1550 nm CW laser and takes advantage of mature and low-cost fiber-optics components from the telecommunications industry. Using either collimated or focused probe beam geometry, lidar based detection of characteristic periodic motions of drone propellers up to a remote distance of 500 m has been achieved. Furthermore, by raster scanning a focused CDL beam with a galvo-resonant mirror beamscanner, two-dimensional images of flying UAVs up to 70 m range were obtained. Each pixel of the raster-scan images provides both lidar return signal amplitude and target radial speed information. The raster-scan images obtained up to 5 frames per second make it possible to discriminate various UAV types based on their profile and even resolve the presence of payloads. With feasible improvements, the anti-drone lidar is a promising alternative to expensive EO/IR and active SWIR cameras used in counter-UAV systems.

Background: The clonal diversity underpinning trends in multidrug resistant Escherichia coli causing bloodstream infections remains uncertain. We aimed to determine the contribution of individual clones to resistance over time, using large-scale genomics-based molecular epidemiology., Methods: This was a longitudinal, E coli population, genomic, cohort study that sampled isolates from 22 512 E coli bloodstream infections included in the Norwegian surveillance programme on resistant microbes (NORM) from 2002 to 2017. 15 of 22 laboratories were able to share their isolates, and the first 22·5% of isolates from each year were requested. We used whole genome sequencing to infer the population structure (PopPUNK), and we investigated the clade composition of the dominant multidrug resistant clonal complex (CC)131 using genetic markers previously reported for sequence type (ST)131, effective population size (BEAST), and presence of determinants of antimicrobial resistance (ARIBA, PointFinder, and ResFinder databases) over time. We compared these features between the 2002-10 and 2011-17 time periods. We also compared our results with those of a longitudinal study from the UK done between 2001 and 2011., Findings: Of the 3500 isolates requested from the participating laboratories, 3397 (97·1%) were received, of which 3254 (95·8%) were successfully sequenced and included in the analysis. A significant increase in the number of multidrug resistant CC131 isolates from 71 (5·6%) of 1277 in 2002-10 to 207 (10·5%) of 1977 in 2011-17 (p<0·0001), was the largest clonal expansion. CC131 was the most common clone in extended-spectrum β-lactamase (ESBL)-positive isolates (75 [58·6%] of 128) and fluoroquinolone non-susceptible isolates (148 [39·2%] of 378). Within CC131, clade A increased in prevalence from 2002, whereas the global multidrug resistant clade C2 was not observed until 2007. Multiple de-novo acquisitions of both bla CTX-M ESBL-encoding genes in clades A and C1 and gain of phenotypic fluoroquinolone non-susceptibility across the clade A phylogeny were observed. We estimated that exponential increases in the effective population sizes of clades A, C1, and C2 occurred in the mid-2000s, and in clade B a decade earlier. The rate of increase in the estimated effective population size of clade A (N e =3147) was nearly ten-times that of C2 (N e =345), with clade A over-represented in Norwegian CC131 isolates (75 [27·0%] of 278) compared with the UK study (8 [5·4%] of 147 isolates)., Interpretation: The early and sustained establishment of predominantly antimicrobial susceptible CC131 clade A isolates, relative to multidrug resistant clade C2 isolates, suggests that resistance is not necessary for clonal success. However, even in the low antibiotic use setting of Norway, resistance to important antimicrobial classes has rapidly been selected for in CC131 clade A isolates. This study shows the importance of genomic surveillance in uncovering the complex ecology underlying multidrug resistance dissemination and competition, which have implications for the design of strategies and interventions to control the spread of high-risk multidrug resistant clones., Funding: Trond Mohn Foundation, European Research Council, Marie Skłodowska-Curie Actions, and the Wellcome Trust., Competing Interests: Declaration of interests NJC reports grants from GlaxoSmithKline and personal fees from Antigen Discovery, outside the submitted work. JP reports grants from Wellcome Trust, during the study. All other authors declare no competing interest. All authors had full access to all the data in the study and had final responsibility for the decision to submit for publication., (Copyright © 2021 The Author(s). Published by Elsevier Ltd. This is an Open Access article under the CC BY-NC-ND 4.0 license. Published by Elsevier Ltd.. All rights reserved.)

Cardiac stimulation via sympathetic neurons can potentially trigger arrhythmias. We present approaches to study neuron-cardiomyocyte interactions involving optogenetic selective probing and all-optical electrophysiology to measure activity in an automated fashion. Here we demonstrate the utility of optical interrogation of sympathetic neurons and their effects on macroscopic cardiomyocyte network dynamics to address research targets such as the effects of adrenergic stimulation via the release of neurotransmitters, the effect of neuronal numbers on cardiac behavior, and the applicability of optogenetics in mechanistic in vitro studies. As arrhythmias are emergent behaviors that involve the coordinated activity of millions of cells, we image at macroscopic scales to capture complex dynamics. We show that neurons can both decrease and increase wave stability and re-entrant activity in culture depending on their induced activity-a finding that may help us understand the often conflicting results seen in experimental and clinical studies., Competing Interests: Declaration of Interests The authors declare no competing interests., (Copyright © 2020 The Authors. Published by Elsevier Inc. All rights reserved.)

Many therapeutic interventions in disease states of heightened cardiac sympathetic activity are targeted to the myocytes. However, emerging clinical data highlights a dominant role in disease progression by the neurons themselves. Here we describe a novel experimental model of the peripheral neuro-cardiac axis to study the neuron's ability to drive a myocyte cAMP phenotype. We employed a co-culture of neonatal ventricular myocytes and sympathetic stellate neurons from normal (WKY) and pro-hypertensive (SHR) rats that are sympathetically hyper-responsive and measured nicotine evoked cAMP responses in the myocytes using a fourth generation FRET cAMP sensor. We demonstrated the dominant role of neurons in driving the myocyte ß-adrenergic phenotype, where SHR cultures elicited heightened myocyte cAMP responses during neural activation. Moreover, cross-culturing healthy neurons onto diseased myocytes rescued the diseased cAMP response of the myocyte. Conversely, healthy myocytes developed a diseased cAMP response if diseased neurons were introduced. Our results provide evidence for a dominant role played by the neuron in driving the adrenergic phenotype seen in cardiovascular disease. We also highlight the potential of using healthy neurons to turn down the gain of neurotransmission, akin to a smart pre-synaptic ß-blocker.

Unlabelled: Hypertension is associated with impaired nitric oxide (NO)-cyclic nucleotide (CN)-coupled intracellular calcium (Ca(2+)) homeostasis that enhances cardiac sympathetic neurotransmission. Because neuronal membrane Ca(2+) currents are reduced by NO-activated S-nitrosylation, we tested whether CNs affect membrane channel conductance directly in neurons isolated from the stellate ganglia of spontaneously hypertensive rats (SHRs) and their normotensive controls. Using voltage-clamp and cAMP-protein kinase A (PKA) FRET sensors, we hypothesized that impaired CN regulation provides a direct link to abnormal signaling of neuronal calcium channels in the SHR and that targeting cGMP can restore the channel phenotype. We found significantly larger whole-cell Ca(2+) currents from diseased neurons that were largely mediated by the N-type Ca(2+) channel (Cav2.2). Elevating cGMP restored the SHR Ca(2+) current to levels seen in normal neurons that were not affected by cGMP. cGMP also decreased cAMP levels and PKA activity in diseased neurons. In contrast, cAMP-PKA activity was increased in normal neurons, suggesting differential switching in phosphodiesterase (PDE) activity. PDE2A inhibition enhanced the Ca(2+) current in normal neurons to a conductance similar to that seen in SHR neurons, whereas the inhibitor slightly decreased the current in diseased neurons. Pharmacological evidence supported a switching from cGMP acting via PDE3 in control neurons to PDE2A in SHR neurons in the modulation of the Ca(2+) current. Our data suggest that a disturbance in the regulation of PDE-coupled CNs linked to N-type Ca(2+) channels is an early hallmark of the prohypertensive phenotype associated with intracellular Ca(2+) impairment underpinning sympathetic dysautonomia., Significance Statement: Here, we identify dysregulation of cyclic-nucleotide (CN)-linked neuronal Ca(2+) channel activity that could provide the trigger for the enhanced sympathetic neurotransmission observed in the prohypertensive state. Furthermore, we provide evidence that increasing cGMP rescues the channel phenotype and restores ion channel activity to levels seen in normal neurons. We also observed CN cross-talk in sympathetic neurons that may be related to a differential switching in phosphodiesterase activity. The presence of these early molecular changes in asymptomatic, prohypertensive animals could facilitate the identification of novel therapeutic targets with which to modulate intracellular Ca(2+) Turning down the gain of sympathetic hyperresponsiveness in cardiovascular disease associated with sympathetic dysautonomia would have significant therapeutic utility., (Copyright © 2016 Larsen et al.)

Genome-wide association studies implicate a variant in the neuronal nitric oxide synthase adaptor protein (CAPON) in electrocardiographic QT variation and sudden cardiac death. Interestingly, nitric oxide generated by neuronal NO synthase-1 reduces norepinephrine release; however, this pathway is downregulated in animal models of cardiovascular disease. Because sympathetic hyperactivity can trigger arrhythmia, is this neural phenotype linked to CAPON dysregulation? We hypothesized that CAPON resides in cardiac sympathetic neurons and is a part of the prediseased neuronal phenotype that modulates calcium handling and neurotransmission in dysautonomia. CAPON expression was significantly reduced in the stellate ganglia of spontaneously hypertensive rats before the development of hypertension compared with age-matched Wistar-Kyoto rats. The neuronal calcium current (ICa; n=8) and intracellular calcium transient ([Ca(2+)]i; n=16) were significantly larger in the spontaneously hypertensive rat than in Wistar-Kyoto rat (P<0.05). A novel noradrenergic specific vector (Ad.PRSx8-mCherry/CAPON) significantly upregulated CAPON expression, NO synthase-1 activity, and cGMP in spontaneously hypertensive rat neurons without altering NO synthase-1 levels. Neuronal ICa and [Ca(2+)]i were significantly reduced after CAPON transduction compared with the empty vector. In addition, Ad.PRSx8-mCherry/CAPON also reduced (3)H-norepinephrine release from spontaneously hypertensive rat atria (n=7). NO synthase-1 inhibition (AAAN, 10 μmol/L; n=6) reversed these effects compared with the empty virus alone. In conclusion, targeted upregulation of CAPON decreases cardiac sympathetic hyperactivity. Moreover, dysregulation of this adaptor protein in sympathetic neurons might further amplify the negative cardiac electrophysiological properties seen with CAPON mutations., (© 2015 American Heart Association, Inc.)

Currently there is a lack of targeted therapies that lead to long-term attenuation or regression of disease in patients with advanced clear cell renal cell carcinoma (ccRCC). Our group has implemented a high-throughput genetic analysis coupled with a high-throughput proliferative screen in order to investigate the genetic contributions of a large cohort of overexpressed genes at the functional level in an effort to better understand factors involved in tumor initiation and progression. Patient gene array analysis identified transcripts that are consistently elevated in patient ccRCC as compared to matched normal renal tissues. This was followed by a high-throughput lentivirus screen, independently targeting 195 overexpressed transcripts identified in the gene array in four ccRCC cell lines. This revealed 31 'hits' that contribute to ccRCC cell proliferation. Many of the hits identified are not only presented in the context of ccRCC for the first time, but several have not been previously linked to cancer. We further characterize the function of a group of hits in tumor cell invasion. Taken together these findings reveal pathways that may be critical in ccRCC tumorigenicity, and identifies novel candidate factors that could serve as targets for therapeutic intervention or diagnostic/prognostic biomarkers for patients with advanced ccRCC.

The possibility of including bacterial indicators in programmes for monitoring the disinfection of animal waste treated in biogas plants was examined. Five full-scale plants receiving waste from several farms were monitored. Of these, two used thermophilic and one mesophilic digestion exclusively whereas two employed a two stage process involving thermophilic pretreatment followed by mesophilic digestion. For each plant the reducing effect on bacterial pathogens present in the incoming waste and on E. coli and enterococci indigenous to the waste was followed over long periods of time (six to 16 months). It was found that both E. coli and enterococci could be used as indicators to monitor the reduction of vegetative bacterial pathogens, but the use of enterococci would give the broadest "safety margin" in this respect. It is concluded that thermophilic as well as mesophilic digestion with thermophilic pretreatment will result in a sufficient reduction of both vegetative pathogenic bacteria and intestinal parasites occurring in concentrations usually found in animal waste to allow for unrestricted use of the de-gassed material in agriculture. This cannot be said for mesophilic digestion used alone. To achieve a sufficient reduction in the number of pathogens during the digestion process, a decrease in the enterococcal concentration of at least 3-4 log10 units should be accomplished, which for normal waste would correspond to a maximum concentration of these bacteria in treated material in the order of 10(2) cfu/ml. It is suggested that enterococcal counts in newly digested material should be one of the parameters in a monitoring programme aimed at controlling the hygienic state of the end-product from large biogas plants.

A group of Atlantic salmon (Salmo salar) was followed through their first year of maturation and spawning. At monthly intervals, starting with juvenile fish in December, 5-7 fish of each sex were killed, and liver and plasma were sampled. The last sampling point was of spawning fish in November a year later. Variables in the cytochrome P450 (P450) system were studied in hepatic microsomes, and estradiol 17β was measured in the plasma of females to assess the maturational status. The P450 1A1-mediated 7-ethoxyresorufin O-deethylase (EROD) started at high levels in winter, but decreased to non-detectable activities in pre-spawning females. Decreases, but not to the same extent, were also observed during this period in total cytochrome P450, cytochrome b5, NADPH-cytochrome P450 reductase, and in the content of two immunochemically determined P450 isozymes. At the same time, LSI levels increased in maturing females (starting in July), and GSI levels increased in both sexes (starting in May). Sex specific differences were observed in pre-spawning fish in September and October, with levels of total P450, b5, NADPH-cytochrome P450 reductase, EROD and P450 isozymes significantly lower in females. At the same time, plasma estradiol-17β levels reached peak values in females. The results point to the important role of sex steroids such as estradiol-17β as major factors in the regulation of final sexual maturation. However, this study also indicates that there may be estradiol-17β independent events of equal importance in the early stages of gonadal maturation that may involve the P450 system. The changes observed in the P450 system (as a major drug and steroid metabolizing system) of Atlantic salmon during sexual maturation may be of importance both in the endogenous transduction of hormonal signals, and as a pharmacological basis for designing therapeutic treatment of diseases in the aquaculture industry.

The major components of the cytochrome P450 (P450) system in liver microsomes of Atlantic salmon were studied using spectrophotometric, catalytic and immunochemical techniques. In juvenile fish sampled during the winter season, high basal activities of 7-ethoxyresorufin O-deethylase (EROD) were found. The Km for 7-ethoxyresorufin was 0.4 µM, and Vmax 1.23 nmol/min/mg protein in juvenile fish. In mature fish sampled from the same group of fish in December, EROD activity was barely detectable (20-30 pmol/min/mg protein). Treatment with the P450 1A1 inducer β-naphthoflavone (BNF) resulted in almost 2-fold induction of total P450, and 30-40-fold induction of EROD activity in immature fish. A similar fold increase was seen in mature fish. The differences in EROD activity between untreated and BNF-treated fish, was accompanied by similar differences in a P450 1A1 cross-reacting protein (Mr=58,000 D) in immunochemical studies using rabbit anti-cod P450 1A1 IgG. However, judging from these studies, the levels of P450 1A1-protein in mature salmon far exceeded those accounted for by the measured EROD activity in comparison to immature fish (both before and after BNF-treatment), indicating inhibiting effects of sex steroids on the measured activity. This effect was not seen on 7-ethoxycoumarin O-deethylase activity. A long-term storage experiment indicated that Atlantic salmon liver microsomes can be stored for 2 years at -80°C in 20% glycerol without losing more than 20-40% of its catalytic activity.

The levels of several environmental contaminants, including selected polyaromatic hydrocarbons (PAH), organochlorines (DDT/DDE, hexachlorobenzene), 15 polychlorinated biphenyl (PCB) congeners, and polychlorinated dibenzofurans and dibenzo-p-dioxins, PCDF/PCDD), and heavy metals (Cd, Hg, Pb, and As) were analyzed in muscle and liver of three different flatfish species (dab, Limanda limanda; flounder, Platichthys flesus; plaice, Pleuronectes platessa) caught by gill netting at different sites in the Hvaler Archipelago. Indices of biochemical effects in liver S9-fractions were studied by measuring cytochrome P450-dependent monooxygenase and UDP-glucuronyl transferase activities, and by immunoquantitating cytochrome P450 1A1 using an indirect enzyme-linked immunosorbent assay (ELISA). Only low levels of PCDD/PCDF, Cd, and Pb were observed, whereas PCB levels were significantly elevated in fish from the inner sites of the Archipelago compared to a reference site. The contaminant gradient toward the Glomma estuary was correlated with increased cytochrome P450 1A1 activity, measured as 7-ethoxyresorufin O-deethylase (EROD), and with immunoquantitated P450 1A1. In contrast, fish from the site at Idefjorden, although containing elevated contaminant levels, did not show elevated EROD activity, but apparently elevated P450 1A1 protein. These findings may reflect different pollution histories of the sites, and indicate the applicability of biochemical effect indices (i.e., EROD and P450 1A1 immunoquantitation) to monitoring studies. The integrated chemical-biochemical approach employed in this study can obviously be expanded to give fruitful information about cause-effect relationships in other contaminant situations.

1. An enzyme-linked immunosorbent assay (ELISA) based on polyclonal rabbit anti-cod cytochrome P-450 IA1 IgG has been developed in our laboratory. 2. The antibodies employed in the ELISA demonstrate good cross-reactivity with the homologous protein in a number of other fish species, giving cross-reacting protein bands o 54-59 kDa in liver microsomes, as determined by Western blotting. 3. The ELISA technique has been used in numerous experiments with both field collected and laboratory exposed fish of different species, showing good correlation with contaminant exposure. 4. In some instances of PCB exposure where the classical P-450 IA1 monooxygenase assay 7-ethoxyresorufin O-deethylase (EROD) failed to reveal any induction, the ELISA technique demonstrated increased levels of P-450 IA1 protein, indicating inhibiting effects of the PCBs on EROD measurement. 5. In tissues like gill filaments, and in whole cod larvae, where EROD activity is barely detectable, if at all, the ELISA technique showed induction after exposure to a water soluble fraction (WSF) of North Sea crude oil. 6. The results reviewed indicate the usefulness of the ELISA technique to allow rapid screening of a large number of samples, and especially when catalytic measurement is difficult or impossible due to (a) small sample or tissue size, (b) loss of activities in bad storage conditions, or (c) presence of compounds inhibiting activity.

Nine different plasmid profiles were demonstrated among 135 Danish isolates of Salmonella dublin, with 58% of the strains enclosed in the major profile group. Two strains did not carry any plasmids. The plasmid profiles of 129 strains (96%) were made up of one or more of only four plasmids of approximate sizes 98 kb, 80 kb, 4.0 kb, and 3.8 kb. There was no significant difference in the prevalence of different plasmid profile groups between isolates from cattle and man. The serovar-specific and virulence-associated 80 kb plasmid could be demonstrated in all plasmid carrying strains, but one strain carried this plasmid at a very low copy number. Five of 47 strains tested showed resistance to antibiotics (2 to sulphathiazole, 2 to streptomycin, and 1 to streptomycin and tetracycline). Only the combined streptomycin/tetracycline resistance was transferable by a 48 kb conjugative plasmid.

An outbreak of listeric meningo-encephalitis occurred in a population of 1800 fallow deer (Dama dama) in a park during the winter and early spring of 1985 to 1986. Listeriosis was diagnosed in 41 of 42 fallow deer that showed the typical central nervous system signs of circling disease or were found dead. The diagnosis was verified by bacteriological examination of the brains of 35 animals. In five of the seven remaining cases listeriosis was diagnosed by histological examination, and in one animal by clinical signs alone. Listeria monocytogenes was isolated in three of 23 soil samples taken from the park. In addition, L monocytogenes was isolated from the intestinal contents of apparently normal fallow deer. Fifty isolates from animals and soil were serotyped and all of them belonged to serovar 4b except one from brain (serovar 1/2b) and three from intestinal contents (serovar 1/2a). In phage typing of 54 isolates, the 35 isolates from the brain and spleen of diseased animals belonged to the same lysovar, as did most isolates from other sources, but strains from intestinal contents belonged to three other phage types. No external source of L monocytogenes was demonstrated in the outbreak and stress due to the poor beech-mast crop, an increased stocking rate and a sudden change in the weather are suspected as predisposing factors.

In an experimental station for calf feeding the occurrence and distribution of drug resistant E. coli in faeces have been investigated among selected groups of calves. Samples were obatined just after the calves arrived 1--2 weeks old in January--February and at intervals until an age of 7--11 months. Daily from arrival to the age of 8 weeks all calves at the station received antibiotics in their feed. In 1971 it was chlortetracycline and in 1972 zincbacitracin. In addition, from 3.5 percent to 60 percent of all calves were every month given therapeutic doses of antibiotics, mainly penicillin + streptomycin and neomycin. Both years the highest number of treatments were administered in the months of February and March (cf. fig. 1). In each of the years studied, a high prevalence of calves harbouring resistant E. coli (total resistance: greater than 88 percent) and R-+E. coli (1971: 83 percent and 1972: 50 percent) was demonstrated at the first sampling. As the calves grew older a general tendency to exchange the resistant strains with sensitive ones was recognized. The prevalence of calves with R-+ E. coli generally devreased at a faster rate than that of total resistance. A discussion is given of the epidemiological significance of cross infection and of selection pressure of antibiotics for the establishment of a resistant flora in individuals as well as for replacement of this flora with sensitive strains.

In the period 1971-73 various extra enteral habitats have been examined for the occurrence of resistant E. coli. Of 172 isolates of E. coli from retail butcher's meat products 9.9% were resistant and 3.5% R+ E. coli. Of 203 isolates of E. coli from bakery products 7.9% were resistant and 3.5% R+ E. coli. Of 52 isolates of E. coli from freshly killed carcases of chickens 40.4% contained weak non-transferable monoresistance against sulphonamides (Table I). Of 92 isolates of E. coli from seawater receiving human sewage 15.2% were resistant and 4.4% R+ E. coli (Table III). All but one samples of sewage and crude sludge contained resistant E. coli of which 11% harboured R factors (Table II). Of 117 Salmonella isolates from human sewage 11.1% were resistant and in 4.2% resistance were transferable (Table VI). Quantitative examinations of untreated and treated sewage did not indicate any selective reduction of resistant E. coli compared to sensitive (Table IV). However, more di- and multiresistant E. coli were found in sewage than in seawater (Fig. 1). The food hygienic implications of resistant E. coli are discussed.