Zymosan-induced leukocyte and cytokine changes in pigs: a new model for streamlined drug testing against severe COVID-19

Injection of 0.1 mg/kg zymosan in pigs i.v. elicited transient hemodynamic disturbance within minutes, without major blood cell changes. In contrast, infusion of 1 mg/kg zymosan triggered maximal pulmonary hypertension with tachycardia, lasting for 30 min. This change was followed by a transient granulopenia with a trough at 1 h, and then, up to about 6 h, a major granulocytosis, resulting in a 3-4-fold increase of neutrophil-to-lymphocyte ratio (NLR). In parallel with the changes in WBC differential, qRT-PCR and ELISA analyses showed increased transcription and/or release of inflammatory cytokines and chemokines into blood, including IL-6, TNF-, CCL-2, CXCL-10, and IL-1RA. The expression of IL-6 peaked at already 1.5-2.5 h, and we observed significant correlation between lymphopenia and IL-6 gene expression. While these changes are consistent with zymosans known stimulatory effect on both the humoral and cellular arms of the innate immune system, what gives novel clinical relevance to the co-manifestation of above hemodynamic, hematological, and immune changes is that they represent independent bad prognostic indicators in terminal COVID-19 and other diseases involving cytokine storm. Thus, within a 6 h experiment, the model enables consecutive reproduction of a symptom triad that is characteristic of late-stage COVID-19. Given the limitations of modeling cytokine storm in animals and effectively treating severe COVID-19, the presented relatively simple large animal model may advance the R&D of drugs against these conditions. One of these disease markers (NLR), obtained from a routine laboratory endpoint (WBC differential), may also enable streamlining the model for high throughput drug screening against innate immune overstimulation.

The COMPARE Data Hubs.

Data sharing enables research communities to exchange findings and build upon the knowledge that arises from their discoveries. Areas of public and animal health as well as food safety would benefit from rapid data sharing when it comes to emergencies. However, ethical, regulatory and institutional challenges, as well as lack of suitable platforms which provide an infrastructure for data sharing in structured formats, often lead to data not being shared or at most shared in form of supplementary materials in journal publications. Here, we describe an informatics platform that includes workflows for structured data storage, managing and pre-publication sharing of pathogen sequencing data and its analysis interpretations with relevant stakeholders.

Coding-complete sequence of a vaccine-derived recombinant porcine reproductive and respiratory syndrome virus strain isolated in Hungary.

Porcine reproductive and respiratory syndrome virus 1 is a major cause of swine morbidity and mortality in various parts of the world, including Hungary. A national elimination programme to reduce the associated economic burden was initiated in Hungary in 2012. Using extensive laboratory surveillance, we identified and isolated an unusual PRRSV strain. The complete coding sequence of this isolate was determined and analyzed. The genome of this Hungarian PRRSV1 strain, HUN60077/16, is 15,081 nucleotides in length. Phylogenetic and recombination analysis showed a mosaic structure of the genome where a large fragment of ORF1b and the genomic region coding for ORF3 to ORF7 showed a very close genetic relationship to the vaccine virus Unistrain, while the ORF1a region, the 3' end of ORF1b, and the whole ORF2 were only distantly related to this or any other PRRSV1 strain whose genome sequence is available in the GenBank database. Genomic characterization of PRRSV strains is crucial when possible vaccine-associated cases are identified. This approach not only helps to identify genetic interactions between vaccine and wild-type PRRSV1 strains but may also be needed to prevent trust in commercial vaccines from being undermined.

Equestrian-related injuries, predictors of fatalities, and the impact on the public health system in Sweden.

OBJECTIVES: Horse riding is a popular activity but has also been found to lead to many injuries and even fatalities. No reduction in the numbers of those being admitted to hospital for equestrian-related injuries have been seen in Sweden in recent years. The aim of this work was to examine injuries, fatalities, and predictors of fatalities in equestrian-related activities and to investigate the cost of these injuries to the public health system. STUDY DESIGN: The study is a retrospective analysis of hospital data. METHODS: National Swedish hospital and mortality registers were retrospectively examined, inclusive of the years 1997-2014. Logistic regression was used to examine the predictors of fatal injuries, and cost of hospital treatment was considered. RESULTS: Over the study period, there were an observed 29,850 injured cases and 51 fatalities. Women comprized almost 90% of those injured and 70% of fatalities. The average age was 26.8 years (range 0-91, standard deviation [SD] = 16.1) for injured and 43 years (range 7-78 years, SD = 20.5) for fatal cases. Men dominated both injured and fatal samples in the older age ranges (Fatal: >50 years; Injured: >70 years), although overall numbers were small. Injuries to the head contributed more than any other body region for both injured and fatal cases, and fractures were the most frequently seen injury type. A chi-squared analysis confirmed that injury type and injured body region were not independent of age. Logistic regression examining the association between fatality and age, gender, home region, and year of injury, indicating trends over time, found that there was an increase of 5.1% in the odds of fatality for every year increase in age of the patient and men had 2.2 times higher odds to be in the fatal sample than women. The conservative estimated cost of injury was 1800 Euro per injury event, equating to over 3 million Euro per year. CONCLUSION: Equestrian-related injury events present a major public health concern. Observed decreases in fatalities suggest improved health care, yet head injury and fatality rates are still high, indicating a need for further intervention. The type of injury changes with the age group, and a better understanding of injury patterns with age is needed to identify protective measures for the different user groups.

Prevention of the development of heart failure with preserved ejection fraction by the phosphodiesterase-5A inhibitor vardenafil in rats with type 2 diabetes.

AIMS: Heart failure with preserved ejection fraction (HFpEF) has a great epidemiological burden. The pathophysiological role of cyclic guanosine monophosphate (cGMP) signalling has been intensively investigated in HFpEF. Elevated levels of cGMP have been shown to exert cardioprotective effects in various cardiovascular diseases, including diabetic cardiomyopathy. We investigated the effect of long-term preventive application of the phosphodiesterase-5A (PDE5A) inhibitor vardenafil in diabetic cardiomyopathy-associated HFpEF. METHODS AND RESULTS: Zucker diabetic fatty (ZDF) rats were used as a model of HFpEF and ZDF lean rats served as controls. Animals received vehicle or 10 mg/kg body weight vardenafil per os from weeks 7 to 32 of age. Cardiac function, morphology was assessed by left ventricular (LV) pressure-volume analysis and echocardiography at week 32. Cardiomyocyte force measurements were performed. The key markers of cGMP signalling, nitro-oxidative stress, apoptosis, myocardial hypertrophy and fibrosis were examined. The ZDF animals showed diastolic dysfunction (increased LV/cardiomyocyte stiffness, prolonged LV relaxation time), preserved systolic performance, decreased myocardial cGMP level coupled with impaired protein kinase G (PKG) activity, increased nitro-oxidative stress, enhanced cardiomyocyte apoptosis, and hypertrophic and fibrotic remodelling of the myocardium. Vardenafil effectively prevented the development of HFpEF by maintaining diastolic function (decreased LV/cardiomyocyte stiffness and LV relaxation time), by restoring cGMP levels and PKG activation, by lowering apoptosis and by alleviating nitro-oxidative stress, myocardial hypertrophy and fibrotic remodelling. CONCLUSIONS: We report that vardenafil successfully prevented the development of diabetes mellitus-associated HFpEF. Thus, PDE5A inhibition as a preventive approach might be a promising option in the management of HFpEF patients with diabetes mellitus.

Pre-registration efficacy study of a novel marker vaccine against classical swine fever on maternally derived antibody positive (MDA+) target animals.

Maternally Derived Antibodies (MDA) can have a negative effect on the efficacy of live attenuated vaccines against classical swine fever (CSF). For this reason, a marker vaccine candidate CP7_E2alf was tested for its efficacy in the presence of MDA. Pregnant sows were vaccinated four weeks before farrowing with CSF virus (CSFV) strain "Thiverval". A total of 40 piglets with MDAs were included in this study. At six weeks of age the piglets were allocated into three treatment groups using generalized randomized block design (GRBD) blocking on serological status and pen location. Of the 40 piglets with MDAs, 30 piglets were vaccinated either orally (n = 15) or intramuscularly (n = 15) with a single dose of vaccine candidate produced under Good Laboratory Practice (GLP) conditions. The ten remaining piglets were allocated into the untreated control group. All 40 piglets were oronasally challenged with 2 ml of the highly virulent CSFV strain "Koslov" 14 days after vaccination. It was revealed that presence of MDAs negatively influences the efficacy of the live marker vaccine candidate, however, the extent of this negative impact depends on the route of vaccine administration. Based on our observations, intramuscular vaccination is recommended during CSF control programs in order to develop superior immune protection.

Novel technologies applied to the nucleotide sequencing and comparative sequence analysis of the genomes of infectious agents in veterinary medicine.

Next-generation sequencing (NGS), also referred to as deep, high-throughput or massively parallel sequencing, is a powerful new tool that can be used for the complex diagnosis and intensive monitoring of infectious disease in veterinary medicine. NGS technologies are also being increasingly used to study the aetiology, genomics, evolution and epidemiology of infectious disease, as well as host-pathogen interactions and other aspects of infection biology. This review briefly summarises recent progress and achievements in this field by first introducing a range of novel techniques and then presenting examples of NGS applications in veterinary infection biology. Various work steps and processes for sampling and sample preparation, sequence analysis and comparative genomics, and improving the accuracy of genomic prediction are discussed, as are bioinformatics requirements. Examples of sequencing-based applications and comparative genomics in veterinary medicine are then provided. This review is based on novel references selected from the literature and on experiences of the World Organisation for Animal Health (OIE) Collaborating Centre for the Biotechnology-based Diagnosis of Infectious Diseases in Veterinary Medicine, Uppsala, Sweden.

Le séquençage de nouvelle génération (également désigné « séquençage à très haut débit ¼ ou « séquençage massivement parallèle ¼) est un nouvel outil extrêmement puissant permettant de procéder à des diagnostics sophistiqués et d'assurer un contrôle vétérinaire intensif de maladies infectieuses complexes. Les technologies du séquençage de nouvelle génération sont également d'un grand secours pour étudier l'étiologie, la génomique, l'évolution et l'épidémiologie des maladies infectieuses ainsi que les interactions hôtes­agents pathogènes et bien d'autres aspects de la biologie des maladies infectieuses. Pour présenter les progrès et les accomplissements les plus récents dans ce domaine, les auteurs décrivent d'abord une série de techniques innovantes puis quelques exemples d'applications du séquençage de nouvelle génération dans le champ de la biologie des maladies animales infectieuses. Ils exposent un certain nombre d'étapes et de processus opérationnels régissant la sélection et la préparation des échantillons, l'analyse séquentielle et les études de génomique comparative, ainsi que ceux qui permettent d'améliorer la justesse prédictive de la génomique ; les exigences particulières de la bio-informatique sont également évoquées. Cette analyse est complétée par quelques exemples d'applications de l'analyse séquentielle et de la génomique comparative en médecine vétérinaire. Cette synthèse est basée sur une sélection de références bibliographiques récentes ainsi que sur l'expérience acquise par le Centre collaborateur de l'Organisation mondiale de la santé animale (OIE) pour le diagnostic basé sur la biotechnologie des maladies infectieuses en médecine vétérinaire, situé à Uppsala (Suède).

La secuenciación de próxima generación, también denominada secuenciación profunda, de alto rendimiento o masivamente paralela, es una nueva y poderosa herramienta para efectuar diagnósticos complejos y vigilar muy de cerca enfermedades infecciosas complejas en el ámbito de la medicina veterinaria. Estas técnicas también se utilizan cada vez más para estudiar la etiología, genómica, evolución y epidemiología de las enfermedades infecciosas, así como las interacciones entre patógeno y anfitrión y otros aspectos de la biología de las infecciones. Los autores resumen brevemente una serie de logros y adelantos obtenidos últimamente en este ámbito, presentando en primer lugar un conjunto de técnicas novedosas y ofreciendo después ejemplos de aplicaciones de la secuenciación de próxima generación a la biología de las infecciones veterinarias. También describen varios protocolos y procesos de trabajo para la obtención y preparación de muestras, el análisis de secuencias y las labores de genómica comparada, explican cómo mejorar la exactitud de la predicción genómica y examinan las herramientas bioinformáticas necesarias para ello. A continuación presentan ejemplos de aplicaciones basadas en técnicas de secuenciación y en la genómica comparada en medicina veterinaria. Este artículo está basado en referencias muy recientes, tomadas de publicaciones científicas, y en la experiencia del Centro Colaborador de la Organización Mundial de Sanidad Animal (OIE) para el Diagnóstico de las enfermedades infecciosas de la medicina veterinaria basado en la biotecnología, sito en Upsala (Suecia).

Thoracic aortic strain can affect endograft sizing in young patients.

OBJECTIVE: Aortic computed tomography angiography (CTA) examination with electrocardiography gating is becoming the clinical routine image acquisition protocol for diagnosis and intervention planning. To minimize motion artifact, the images are reconstructed in the diastolic phase of the cardiac cycle. The aim of our study was to quantify aortic strain in an elderly nonaneurysmatic patient cohort and to identify the phases of the R-R cycle that correspond to the minimal and maximal aortic diameters. The quantification of aortic strain may enable the improvement of intervention planning and the introduction of more effective dose-saving protocols for CTA scans. METHODS: We assessed CTA images of 28 patients (14 men; mean age, 74 years). Aortic calcium score was calculated on native images. Angiography images were reconstructed in equally spaced 10 phases of the R-R cycle. After semiautomatic centerline analysis, we measured the cross-sectional areas in each of the 10 phases at 9 specific segments between the ascending aorta and the common iliac bifurcation representing the attachment sites of thoracic and abdominal stent grafts. Area-derived effective diameter, pulsatility (Amax - Amin), and strain [(Amax - Amin)/Amin] were calculated. Repeated measurements were taken to evaluate inter-reader and intrareader reproducibility (10-10 patients each). RESULTS: A total of 4320 measurements were performed. We found significant difference between diastolic and systolic diameters (DD,Z0 = 33.2, DS,Z0 = 34.4; P < .001). Pulsatility values of the vessel diameters were 1.0 to 1.1 mm in the thoracic aorta, 0.7 to 0.9 mm in the abdominal aorta, and 0.5 to 0.6 mm in the common iliac arteries. Negative, moderate correlations were found between aortic strain and age (r = -0.586; P = .001), aortic strain and plaque area (r = -0.429; P = .026), and age and body mass index (r = -0.412; P = .029). We found positive, moderate correlation between age and plaque area (r = 0.594; P = .001). The aortic pulsatility curve has a positive extreme at 30% and a negative extreme at 90% of the R-R cycle throughout the aorta. Lin concordance coefficients were 0.987 for inter-reader and 0.994 for intrareader correlations. CONCLUSIONS: Aortic strain can be reliably quantified on electrocardiography-gated CTA images. Pulsatility of the aorta can be substantial in the thoracic aortic segments of young patients; therefore, the routine use of systolic images is not recommended. In addition, we demonstrated that images at 30% of the heart cycle correspond to the largest diameter of the aorta.

Pre-registration efficacy study of a novel marker vaccine against classical swine fever on Maternally Derived Antibody negative (MDA-) target animals.

Classical swine fever (CSF) marker vaccine candidate CP7_E2alf produced under Good Laboratory Practice (GLP) conditions by Pfizer was tested on 40 six-week-old MDA-piglets according to the European Pharmacopoeia (Ph.Eur.) requirements. Single doses of CP7_E2alf were given to 15 piglets orally, while 15 other piglets were intramuscularly vaccinated. Ten additional animals were included as unvaccinated controls. All piglets were oronasally challenged with the highly virulent CSF virus (CSFV) strain "Koslov" 14 days after vaccination. CP7_E2alf administered i.m. provided a complete protection, while p.o. administratrion triggered only partial protection. The level of protection was determined by the development of clinical signs, viraemia and rate of mortality. The vaccine candidate met the criteria of Ph. Eur Monograph 0065, "Swine-fever vaccine (live, prepared in cell cultures), classical" 7th Edition, which claims the efficacy test is invalid if fewer than 50 per cent of the control piglets display typical signs of serious infection of CSF or die, and if fewer than 100 per cent of the control piglets show clinical signs of disease within 21 days following challenge. Fulfilling these validity criteria is a key step in the registration procedure for a vaccine candidate to become openly available.

Predictors of relapse in patients with Crohn's disease in remission after 1 year of biological therapy.

BACKGROUND: Some of the most important questions relating to the use of biological therapy in inflammatory bowel diseases concern the duration of maintenance therapy. AIM: To assess the disease course and frequency of relapse of Crohn's disease (CD) following discontinuation of biological therapy, and to determine predictive factors for relapse. METHODS: One hundred twenty-one CD patients who had achieved clinical remission following 1 year of biological therapy and for whom biological therapy was then discontinued participated in this prospective observational study. Eighty-seven CD patients had received infliximab and 34 adalimumab. The definition of relapse was an increase of >100 points in CDAI to at least a CDAI of 150 points. RESULTS: Biological therapy was restarted within 1 year of treatment cessation in 45% of patients. Logistic regression analysis revealed that previous biological therapy (P = 0.011) and dose intensification during the 1-year course of biological therapy (P = 0.024) were associated with the need for and the time to the restarting of biological therapy. Smoking was observed to have an effect that was not statistically significant (P = 0.053). CONCLUSIONS: Biological therapy was restarted a median of 6 months after discontinuation in almost half of Crohn's disease patients in who had been in clinical remission following 1 year of biological therapy. These results suggest that, in the event of the presence of certain predictive factors, biological therapy should probably be continued for more than 1 year by most patients.

Detection of Mycoplasma bovis with an improved pcr assay.

A Mycoplasma bovis species-specific PCR assay has been developed with improvement of a previously described method (Ghadersohi et al., 1997). This test and its semi-nested version (Hayman and Hirst, 2003) did not function at all in our hands. A new reverse primer (Mbr2) was designed using previously published sequence data. For testing specificity, DNA was extracted from the most frequently occurring mycoplasma species and bacteria of bovine origin. The new PCR detected only Mycoplasma bovis. Moreover, no cross-reaction was observed with the genetically closest relative species, M. agalactiae. The target organism could be detected in a dose as low as 150 CFU ml(-1) in broth cultures using ethidium-bromide-stained agarose gels.

Cytopathogenicity markers in the genome of Hungarian cytopathic isolates of bovine viral diarrhoea virus.

Since genetic recombination is a major factor in the evolution of the cytopathogenic (cp) bovine viral diarrhoea virus (BVDV) biotypes, in this study the cytopathogenicity markers were investigated in the genomes of two cp BVDV strains recently isolated from mucosal disease (MD) cases in Hungary. In the genome of strain H4956, a Jiv-like insertion was found similar to those described in reference strain NADL and in other BVDV 1, BVDV 2 and border disease virus (BDV) strains. The 133 amino acid Jiv-like sequence is inserted at nucleotide position 4984 (amino acid position 1533), 9 nucleotides upstream of that of strain NADL. The insertion showed 96% amino acid sequence identity with the cellular Jiv protein. In the genome of cp BVDV strain H115/PCR, an ubiquitin-containing duplication was found. The duplicated sequence started at nucleotide position 7978 (amino acid 2531) in the NS4B gene. The duplication contained a complete ubiquitin monomer of 76 amino acids and the complete NS3 gene starting at nucleotide position 5153 (amino acid 1589), which corresponds to the first N-terminal amino acid of NS3. The duplication was located further downstream of the known ubiquitin-containing genomic regions of cp BVDV strains, and it consisted of the shortest inserted nucleotide sequence. The insertions and duplication of strains H4956 and H115/PCR further confirmed that recombinations occurring at positions A and B are the most common mechanisms leading to the development of BVDV cytopathogenicity.

[Results of laparoscopic repair of abdominal wall hernias using an ePTFE-polypropylene composite mesh]. / Ergebnisse der laparoskopischen Versorgung von Bauchwandhernien mit einem PTFE-Polypropylen-Komposite-Netz.

BACKGROUND: The standard technique for laparoscopic ventral hernioplasty (peritoneal onlay) reduces the recurrence rate and the rate of other complications, in particular the risk of infections. However, the procedure requires a special mesh to mitigate the risk of visceral adhesions. For this purpose an ePTFE mesh is generally used. There are limited reported clinical data regarding the long-term results and benefits of other biomaterials. STUDY DESIGN: Bard Composix mesh was used for laparoscopic ventral hernia repair. This biomaterial combines two different clinically proven materials--polypropylene with a thin layer of ePTFE--to maximize ingrowth of surrounding tissue, while mitigating the risk of visceral adhesions. Demographic, operative and postoperative data were collected and analyzed. Follow-up clinical evaluations were conducted 6-10 days, 4 weeks, and every 6 (th) month after surgery in all patients. RESULTS: Fifteen patients were involved in the study. The mean operating time was 101.2 min (64-190 min). There were no intraoperative complications or conversion to open procedure. Postoperative complications were 1 seroma, 4 cases of prolonged ileus, one tacks-site pain requiring second-look laparoscopy. Hospital discharge occurred on average 6 days (3-10 days) after surgery. The mean follow-up was 12.4 months (range 5-29 months). No late complications or hernia recurrence were observed. CONCLUSIONS: Laparoscopic prosthetic ventral hernioplasty using the composite polypropylene/ePTFE mesh offers short hospital stay and acceptable complications for primary and recurrent ventral hernias. Disadvantage of the composite mesh is the difficulty to roll this product into a sufficiently small size to permit its easy introduction into the abdominal cavity.

[Antireflux procedures at our unit (1990-1999)]. / Refluxgátló mútétek klinikánk gyakorlatában (1990-1999).

UNLABELLED: A retrospective analysis was carried out to evaluate the changing attitude of antireflux surgical practice of the authors. Between 1990 and 1994 24 antireflux procedures were performed. Laparoscopic antireflux surgery was introduced in 1995. Since then 41 laparoscopic and 23 open repairs have been performed. Indication for surgery was always based on thorough preoperative examination protocol including functional tests of the esophagus. Each patients had undergone endoscopic and X-ray examinations. The patients are under continuous follow-up surveillance. During the postoperative follow-up studies more than 2/3 of the patients (61 patients) agreed to carry out the functional tests postoperatively. RESULTS: The demographic data of patients of the two study periods proved to be comparable. The length of history was 10.5 years in the first group, while it was 8.9 years in the second group. In both groups the surgical repair caused significant reflux control confirmed by decrease of reflux index and number of reflux episodes furthermore by increase of resting lower esophageal sphincter pressure. Actually 4 recurrences are known in the first group and 2 in the second group. One of the 4 recurrences observed in the first period was treated by redo surgery. The patients having recurrences in the second period can easily be managed by medical therapy. There was no mortality and severe morbidity during the study periods. Few patients reported mild temporary dysphagia. CONCLUSIONS: In a given proportion of reflux patients the individually planned and perfectly performed antireflux surgery offers an acceptable definitive treatment option. The availability of endoscopic surgery enhances this trend, which can be seen all over the world. An increase can be seen in the number of procedures: nearly three times more patients were operated on during the second period. The early clinical results of laparoscopic antireflux procedures are good enough. Good long-term results can be expected if we accept and apply the basic technical principles determined during the open surgery era.

Functional and immunocytochemical evidence that galanin is a physiological regulator of human jejunal motility.

The neuropeptide galanin has species-dependent effects on intestinal motility. It has a contractile effect on rat jejunal muscle while it relaxes guinea-pig ileum by inhibiting cholinergic transmission. Its effect on human gut motility has been unknown. Extensive work led to the discovery of selective galanin analogues such as M15 [galanin(1-12)-Pro-substance-P(5-11)], M35 [galanin(1-12)-Pro-bradykinin(2-9)-amide] that competitively inhibit various actions of galanin in the central nervous system. The present study was designed to examine the effect of galanin, M15 and M35 on longitudinal jejunal smooth muscle strips isolated from humans and rats, and to localize galanin-immunoreactivity in human jejunum. Galanin and ACh were equally effective in stimulating contractions of the isolated jejunal muscle: sigmoid curve fitting showed that maximal contractile response to galanin and ACh were 25.7+/-11.1 mN and 23.7+/-9.7 in humans, while 8.0+/-0.6 and 8.1+/-0.3 mN in rats, respectively. These effects of galanin were not inhibited by either atropine (5 x 10(-6) M) or tetrodotoxin (3 x 10(-6) M). The potency of galanin inducing the contractile actions were similar in humans and rats. Interestingly, neither M15 nor M35 (up to 10(-7) M) were able to inhibit the responses of the smooth muscle to galanin. However, both putative galanin receptor antagonists showed agonist effects in our experimental models. In accordance with the functional studies, both the longitudinal and the circular muscle layers were abundant in nerve fibers and varicosities showing galanin immunoreactivity. Our data suggest that galanin is a potent physiological regulator of jejunal contractions in humans. Its action on the jejunum, however, is mediated by galanin receptors that are different from those located in the central nervous system.

Effects of putative galanin antagonists M35 and C7 on rat exocrine pancreas.

Galanin is a neuropeptide having a wide range of biological actions. Recently selective galanin receptor antagonists such as M35 [galanin(1-12)-Pro-bradykinin(2-9)-amide] and C7 [galanin(1-12)-Pro-spantide-amide] have been described. These antagonists have blocked the actions of galanin on flexor reflex, glucose-induced insulin secretion, and acetyicholine release from hippocampus. Our present aim was to investigate whether M35 and C7 can affect galanin-induced inhibition of pancreatic enzyme secretion in rats. Pancreatic enzyme secretion was studied in urethane-anesthetized rats supplied with jugular vein catheter and pancreatic cannula. Amylase secretion evoked by submaximal CCK-8 stimulation was inhibited dose-dependently by galanin in anesthetized rats. Surprisingly, neither M35 nor C7 was able to inhibit the responses of the exocrine pancreas to galanin. However, both putative galanin receptor antagonists behaved as agonists in our experimental models. Our data suggest that the effects of galanin on pancreatic enzyme secretion are not mediated by M35- or C7-sensitive galanin receptors. Therefore, these galanin receptors are different from those described in the central nervous system.

[Hand-assisted laparoscopic technique (HALS) in colorectal surgery (Case reports and review of the literature)]. / Kézzel asszisztált laparoszkópos technika (HALS) a colorectalis sebészetben (Esetismertetések és irodalmi áttekintés).

Hand-assisted laparoscopic surgery (HALS) is a new technique in which the surgeon's hand is introduced to the abdominal cavity through a mini-laparotomy, while pneumoperitoneum is maintained. We describe the technique of HALS used successfully in the treatment of two patients of colorectal disease. We describe current literature and our initial clinical experience. Our conclusion is that, besides colorectal surgery, HALS technique may prove useful for other abdominal procedures as well; however its real value should be estimated in prospective randomized trials.

Sequence analysis of the membrane protein gene and nucleocapsid gene of porcine reproductive and respiratory syndrome virus isolated from a swine herd in Hungary.

Porcine reproductive and respiratory syndrome virus (PRRSV) was isolated from blood samples taken at a pig farm in Hungary from pigs showing clinical signs of the disease. The virus (ABV 32) was identified as belonging to the European genotype by using type-specific monoclonal antibodies. This was confirmed by comparing the sequence of the membrane protein gene (ORF 6) and the nucleocapsid gene (ORF 7) with the American VR2332 and the European LV genotype reference strain, respectively. Analysis of the amino acid sequence of the ORF 6 and ORF 7 of ABV 32 revealed five amino acid changes in both ORFs when compared with LV, of which two changes in ORF 7 were only found in the Spanish isolates. Additionally, the ORF 7 sequence was compared with corresponding sequences of a total of 21 other European strains. Phylogenetic analysis using the PHYLIP package confirmed the close relationship between the Hungarian and the Spanish isolates. Of all the isolates analysed, ABV 32 and LV were the least related.