Use of monoclonal antibodies against human HLA II antigens for the detection of bovine B lymphocytes and macrophages]

The crossreactivity of mouse monoclonal antibodies (MoAbs) (Tab. I) prepared against human HLA-DR and HLA-DP antigens was studied in various bovine cells: lymphocytes from lymph nodes and peripheral blood, adherent (B) and nonadherent (T) lymphocytes, monocytes, granulocytes and platelets. In the immunofluorescence test, MoAbs Bra13, Bra14, Bra20, Bra22, Bra30, Bra70, HL-38 reacted with bovine B lymphocytes and monocytes, but not with other tested cells (Tab. III, IV). These antibodies, except Bra22, were positive with B lymphocytes in the complement dependent cytotoxic test (Tab. II). The similarity of the bovine antigens and HLA-DR antigens determined by used MoAbs was also proved by immunoblotting. Monoclonal antibodies Bra38 and BraFB6 did not react with the bovine cells and separated antigens. The epitope (HLA-DR) recognized by the antibody Bra38 is probably absent in cattle. The presence of HLA-DP analogue determined by the antibody BraFB6 has not been confirmed. The crossreactive MoAbs could be used for the detection of B lymphocytes and macrophages in veterinary immunology.     

Purification and characterization of hepatic triacylglycerol lipase isolated from rainbow trout,Oncorhynchus mykiss

Trialcylglycerol (TG) lipase was isolated and partially purified from rainbow trout liver. Triacylglycerol lipase activity was assayed by measuring¹⁴C-oleic acid release from¹⁴C-triolein.¹⁴C-oleic acid release was linear for up to two hours. Optimal activity occurred at pH 7.0 and 15°C. Most of the lipase activity was recovered in the cytosolic fraction. A 27,000-fold purification was achieved after Sepharose (Bio-gel A 0.5 M, 200–400 mesh) chromatography of a resuspended 20% ammonium sulfate fraction. The molecular weight of the trout hepatic lipase as determined by size-exclusion chromatography and by SDS-polyacrylamide gel electrophoresis was 40–43 kD. Lipase-mediated hydrolysis of TG resulted in the production of diacylglycerols, monoacylglycerols, and fatty acids. Kinetic analysis indicated that V_max=0.016 nmol/h/mg protein and that K_m=0.28 mM triolein. Lipolytic activity was enhanced in the presence of cAMP/ATP-Mg²⁺. These results suggest that the liver of trout possesses a neutral TG lipase that is responsible for mobilizing stored TG and is catalytically activated by phosphorylation.A part of this work was presented at the Annual Meeting of the American Society of Zoologists, December 26–30, 1990, San Antonio, TX.     

Regulation of Capacitation of Canine Spermatozoa during Co-culture with Heterologous Oviductal Epithelial Cells

Progress of essential steps of the capacitation is coordinated in the oviductal isthmus, where sperm are stored in close contact with the epithelium. A crucial capacitational event is the phosphorylation of sperm membrane proteins. Regulation of the tyrosine phosphorylation by the oviduct has not been examined in dog sperm yet. The aim of this work was to study the effect of dog sperm binding to porcine oviductal epithelium on capacitation‐induced cellular and molecular changes. Epithelial cells were stripped from the oviducts of post‐puberal sows and cultured for 5–7 days at 39°C and 5% CO₂ on Biomatrix‐covered Chamber slides. Sperm washed through Percoll was co‐incubated with the oviductal epithelium cell cultures in a bicarbonate Tyrode's medium. During co‐incubation, sperm membrane changes, the state of tyrosine phosphorylation and motility were determined after 3, 30, 90, 180, 240 and 360 min. Significant increases in the percentage of capacitated and dead cells were observed in unbound sperm, while bound sperm remained uncapacitated, live and motile. An increasing tyrosine phosphorylation of tail proteins in bound, unbound and control sperm suspensions and a subsequent phosphorylation of head proteins in unbound and control sperm suspensions were observed. A significant difference regarding head phosphorylation (p < 0.05) was found between sperm bound to oviductal epithelium and unbound sperm. Binding occurred mainly in sperm with non‐ phosphorylated heads, while higher proportions of phosphorylated cells were found in unbound populations. The head phosphorylation progressed significantly during incubation in unbound spermatozoa (p < 0.05); however, it was suppressed in population of sperm attached to oviductal epithelium. Significant correlations between motility parameters related to hyperactivation and tail phosphorylation were found in unbound sperm. These observations support the hypothesis that spermatozoa with non‐phosphorylated heads preferentially attach to epithelial cells. It can be concluded that tyrosine phosphorylation of head membrane proteins and capacitation are delayed in canine spermatozoa being in closed contact with oviductal epithelium.     

Waldschutz gegen Borkenkäfer: Der Beitrag von Duftstoffmeteorologie und Populationsdynamik

Zusammenfassung Mit theoretischen Modellen kann die Effektivität verschiedener Ansätze (z. B. Fallenzahl und-abstand, Ködermenge und-abgaberate) für den Einsatz von synthetischen Pheromonen in Pest-Management-Systemen verglichen werden. Das GAUSS-sche Modell, das von einer Normalverteilung der Konzentrationsabnahme um eine Lockstoffquelle ausgeht, wurde benutzt, um fürT. lineatum die lockwirksame Reichweite und die Konkurrenzsituation von Naturköder und Falle abzuschätzen. Bei 100 000 /ha und 2 Wochen Flugdauer ergeben sich für (+)-Lineatin (Reaktionsschwelle 0,1 ng/m³, Produktion 6,43 ng//d, Produktionsdauer 3 d/) mittlere Tagesraten von 128,6 g/ha bzw. eine Gesamtproduktion von 1,929 mg/ha. Die lockwirksame Reichweite x_max beträgt je nach Wetterlag für 1T. lineatum- 20–40 cm, für Fallen mit 1 CONREL-Faser (Abgaberate 10 g/d [+]-Lineatin) 13–16m. Für eine 10fach höhere lockwirksame Reichweite sind nach dem Modell etwa 100fach, in der Praxis nach verschiedenen Autoren sogar 200fach stärkere Köder erforderlich.In Freilandversuchen über die Relation von Fangleistung und Beifängen unterschiedlicher Fangsysteme waren die Coleoptera (bes. Rhizophagidae und Staphylinidae) am häufigsten. In Trichterfallen lagen die Beifänge mit 9,2% deutlich höher als in Schlitzfallen (1,2%). Das Giftpolter hatte demgegenüber das größte Beifangspektrum; hier traten auch Ordnungen auf, die von anderen Fangsystemen nicht betroffen waren.Massenfang kan prinzipiell in Holzhöfen und Waldbeständen erfolgreich durchgeführt werden. Allerdings ist das Problem der Beifänge noch ungelöst und die Relations von potentieller Vermehrung der Borkenkäfer und maximal möglicher Populationsreduzierung durch Fallenfang unbekannt. Zudem kann eine nach Massenfang geringere Populationsdichte im Folgciahr durch zunehmende Reproduktionsraten oder durch ersatzweise Einnischung (competive displacement) anderer Borkenkäferarten ausgeglichen und damit der Forstschutzeffekt zunichte gemacht werden.

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Bark beetle control: contribution of pheromone meteorology and population dynamics

The use of semiochemicals in pest management systems of bark beetles is viewed in relation to optimization of resources investment, comparing beetle numbers in univoltine, requisitegoverned woodboringTrypodendron lineatum (Oliv.) with multigeneration bark inhabitingIps typographus (L.). The efficiency of different numbers of and distance between traps and pheromone load of dispensers was analysed using steady state models and comparing results with published and own field data. Competition between traps and beetle-infested logs, attractive distance and overlap of pheromone plume was compared forT. lineatum using models which assume a GAUSS distribution of concentration decrease around a pheromone source. For (+)-lineatin (reaction threshold 0.1 ng/m³, production 6.43 ng//d, duration of production 3 d/), 100,000 /ha and a flight period of 2 weeks, daily mean production is 128.6 g/ha. Attractive distance x_max is 20–40 cm for 1T. lineatum- and 13–16 m for a trap with 1 CONREL-dispenser (mean release rate 10 g/d (+)-lineatin). A 10-fold increase of attractive distance x_max needs a 100-fold increase of pheromone release rate according to the model, and 200-fold according to field experiments.In field experiments, Coleoptera (esp. Staphylinidae and Rhizophagidae) were the most abundant non-target trap catches. Insecticide-treated trap logs had the widest range of non-target insects including orders that were not affected by the non-insecticide trapping systems. Mass trapping can be successful in logging areas and/or forests. However, the exclusion of non-target insects still is an unsolved problem and we still have only limited knowledge on potential population increase of beetles and maximum reduction that can be achieved by trapping. Also, population levels reduced by mass trapping can be compensated by increasing reproductive effort of the succeeding generation and/or competitive displacement by secondary bark beetle species.

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Mit 5 Abbildungen und 2 Tabellen

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Mit Unterstützung der Deutschen Forschungsgemeinschaft.

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Nach einem Vortag bei der Fortbildungstagung des Hessischen Forstvereins in Solms/Lahn am 23. Mai 1991.

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Herrn Prof. Dr. Dr.G. Wellenstein zum 85. Geburtstag.     

Direct electron microscopy and serology with plant viruses in leaf material dried and stored over calciumchloride

Most of 43 viruses could easily be detected directly in 53 out of 66 leaf samples dried and stored over CaCl₂ for varying periods of time up to 20 1/2 years. Detection usually was with PTA pH 6.5, but alfalfa mosaic, cucumber mosaic and tomato aspermy viruses required PTA pH 3.0 to 4.0. Bean common mosaic, cowpea aphid-borne mosaic and cowpea mosaic viruses were also easily observed in newly dehydrated samples obtained for diagnosis from Morocco and Tanzania.Broad bean wilt virus, cowpea mosaic virus and cucumber mosaic virus were detected with agar gel-diffusion tests in dry leaf material ground in buffer. This serological assay demonstrated a high concentration of cucumber mosaic virus in leaf material dried over CaCl₂ 20 years ago. This paper further coroborates the value of the CaCl₂ method of dehydration and storage of plant viruses in leaf material.Samenvatting Zeer uiteenlopende plantevirussen kunnen goed worden bewaard in bladmateriaal van geïnfecteerde planten dat is gedroogd en bewaard boven CaCl₂. Bij rechtstreekse elektronenmicroscopische toetsing van 66 monsters met verschillende isolaten van 43 virussen konden in 53 monsters gemakkelijk virusdeeltjes worden waargenomen (Tabel 1 en Fig. 1 en 2). Meestal lukte dit met fosforwolfraamzuur pH 6,5 dat doorgaans voor in ruw plantesap voorkomende virussen wordt gebruikt. Bij luzernemozaïekvirus en komkommermozaïek virus gelukte dit alleen maar bij lagere pH (3,0 en 4,0). Ook het tomate-aspermievirus was dan veel gemakkelijker aantoombaar. Methylaminewolfraamzuur gaf geen beter resultaat.Draden van bonerolmozaïekvirus en van cowpea aphid-borne mosaic virus en bolletjes van cowpea mosaic virus waren snel waarneembaar in elektronenmicroscopische preparaten gemaakt van bladmateriaal van recent uit Marokko en Tanzania ontvangen monsters.Cowpea mosaic virus, tuinboneverwelkingsvirus en komkommermozaïekvirus konden eveneens gemakkelijk en snel serologisch worden aangetoond in met bufferoplossing vermalen droog blad van respectievelijk cowpea, erwt en tabak. Het laatstgenoemde, reeds 20 1/2 jaar geleden gedroogde bladmateriaal, bleek in vergelijking met pas geïnoculeerd vers blad van komkommer enChenopodium quinoa zelfs zeer veel serologisch actief virusmateriaal te bevatten (Fig. 3).De beschreven waarnemingen bevestigen nogmaals de waarde van de toegepaste methode van virusbewaring en tonen aan dat het mogelijk is vele virussen te herkennen in van elders ontvangen gedroogde bladmonsters, zonder het risico te lopen van virusontsnapping, zoals altijd aanwezig bij werk met toetsplanten in de kas.Guestworker from Istituto Sperimentale per la Patologia Vegetale, Rome, Italy, participating in the investigations from September 21 to December 21, 1977.     

Apparent independence of glucose tolerance and body fat content in 8‐week‐old broilers

1. A possible relationship between glucose tolerance and body‐fat content was examined in broilers selected at 2 and 4 weeks of age for fast or slow glucose disposal.

2. At 8 weeks of age, selected chickens were different in glucose tolerance but similar in body weight, food conversion efficiency, carcass composition and glucose‐induced insulin release.

3. Therefore, variations in glucose regulation and insulin sensitivity which are detectable at an early age, do not appear to be related to body composition in 8‐week‐old broilers.     

Effects of atmospheric ammonia on pulmonary bacterial clearance in the young pig.

Young pigs were exposed to an aerosol of a nonpathogenic strain of Escherichia coli and then were retained in air-pollutant exposure chambers for a 2-hour clearance period. In series 1 (n = 80 pigs), 40 exposed young pigs (principals; 15.5 days of age) were placed in an atmosphere of filtered room air + 50 ppm of atmospheric NH3 during the clearance period; control pigs were exposed to filtered room air without added NH3. In series 2 (n = 24 pigs), 12 exposed young pigs (principals; 6.2 days of age) were similarly maintained, but at a lower concentration of atmospheric NH3 (75 ppm). At the end of the clearance period pigs were killed and pulmonary bacterial clearance was determined. Pigs kept in the NH3-contaminated atmospheres (either concentration) harbored more bacteria, on the average, in their lungs than did the controls. If series 1 and 2 data were combined, pigs kept in the NH3-contaminated atmospheres had 51% more bacteria in their lungs than did the controls. Pulmonic weight and ratio of pulmonic weight to body weight of pigs kept in the NH3-contaminated atmosphere were greater than those of the controls in series 1, but not in series 2. Gross and histopathologic examinations of lung tissue generally revealed no differences between controls and principals in either series 1 or 2.     

Gesunde Pflanzen unter zukünftigem Klima

Predicted changes in average values of global climate variables (increased temperatures, altered precipitation patterns, increased concentrations of atmospheric CO₂) and changes in the frequency, duration, and degree of extremes (frost, heat, drought, hail, storms, floods, etc.) will affect agricultural crops, agroecosystems, and agricultural productivity. Although forecasts of regional climate changes are still imprecise, mean temperature increases in Europe are expected to be greater in the north (2.5–4.5°C) than in the south (1.5–4.5°C). Regional forecasts for precipitation changes are also very far from precise; however, problems with drought are expected to increase, especially in Mediterranean countries. Overall, shortage of water will be the predominant factor affecting plant growth. As higher temperatures are known to enhance plant development and especially the grain-filling duration of cereals, grain yield losses are possible in a warmer climate. On the other hand, elevated atmospheric CO₂ concentrations are known to stimulate photosynthesis and enhance growth and yield (“CO₂ fertilization”); concomitantly, leaf transpiration is reduced, resulting in improved water use efficiency. Total biomass and yield were enhanced by 20–30% in experiments with elevated CO₂ exposure (550–700 ppm) under more or less ideal growth conditions. Elucidating the interactions between positive and negative effects of climate change is of crucial importance for any prediction of future crop yields. The present paper is a brief summary mainly of the potential effects of elevated temperatures and atmospheric CO₂ on crop growth, quality, and yield. Also, adaptation measures, possible interactive effects of different climate variables, and interactions of climate change components with other growth variables (pathogens, air pollutants) are briefly described.