New data on the mechanism of action of immuno-adjuvants: Applications in veterinary medicine

Certain natural or synthetic peptidoglycans which act as watersoluble adjuvants represent a new family of drugs which can act selectively on certain lymphoid cells, B or T cells or macrophages. Some of these molecules can inhibit the immune response.We have developed a technique which enables either normal membranous vesicles, (cellular surface on the outside, RSO), or inverted vesicles, (cellular surface on the inside, IO), to be taken from a murine plasmacytoma. Some peptidoglycans act on the RSO vesicles, others on the IO vesicles, in every case by activating and/or inhibiting membranous enzymes such as 5-nucleotidase and Na⁺/K⁺ ATPase.It is likely that the immuno-adjuvants act selectively and that the same substance can activate or inhibit according to the dose used.

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Kurzfassung Bestimmte natürliche oder synthetisch hergestellte Peptidoglykane (wasserlösliche Adjuvanzien) sind eine neue Familie pharmakodynamischer Agenzien, die elektiv auf bestimmte Lymphoidzellen, B-Zellen, T-Zellen oder Makrophagen wirken. Bestimmte Moleküle können die Immunreaktion inhibieren. Es wurde ein Verfahren erarbeitet, mit dem aus einem Plasmozytom bei Ratten oder Mäusen Membranbläschen gewonnen werden konnten, die entweder richtig geformt (Zelloberfläche aussen, RSO) oder umgekehrt (Zelloberfläche innen, IO) sind. Manche Peptidoglykane wirken auf RSO, andere auf IO; in allen Fällen ist diese Wirkung gekennzeichnet durch die Activierung und/oder Inhibierung von Membranenzymen wie der 5-Nukleotidase und der Na+/KA—ATPase.Voraussichtlich haben die Immunitätsadjuvanzien eine Zielspezifität und derselbe Stoff kann je nach Dosierung stimulierend oder inhibierend wirken.Riassunto Gli adiuvanti idrosolubili, che sono peptido-glicani naturali o sintetici, costituiscono una nuova famiglia di agenti farmacodinamici in grado di agire selettivamente su alcune cellule linfoidi, le cellule B, T o macrofagi. Alcune molecole possono inibire la reazione immunitaria.Gli autori hanno messo a punto una tecnica che permette di ottenere, partendo da un plasmocitoma murino, vescicole membranose girate nel senso giusto (superficie cellulare rivolta all-esterno RSO) o rovesciate (superficie cellulare all'interno, IO). Alcuni peptido-glicani sono attivi sulle RSO, altrillo sono sulle IO; in ambedue i casi detta attività è caratterizzata dall-attivazione e/o dall'inibizione di enzimi membranosi come la 5 nucleotidasi e la Na⁺/K⁺ ATP-asi.Si puo ipotizzare che gli adiuvanti dell'immunita posseggano una specificita di bersaglio, e che la medesinna sostanza possa rivelarsi stimolante od inibitoria a seconda della dose impiegata.

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Resume Les adjuvants hydrosolubles que sont certains peptidoglycanes naturels ou synthétiques représentent une nouvelle famille d'agents pharmacodynamiques capables d'agir électivement sur certaines cellules lymphoides, cellules B, T ou macrophages. Certaines molécules peuvent inhiber la réponse immunitaire.Nous avons mis au point une technique permettant d'obtenir à partir d'un plasmocytome murin des vésicules membranaires tournées dans le bon sens (surface cellulaire à l'extérieur, RSO) ou inversées (surface cellulaire à l'intérieur, IO). Certains peptidoglycanes sont actifs sur les RSO, d'autres sur les IO: dans tous les cas cette activité est caractérisée par l'activation et/ou l'inhibition d'enzymes membranaires telles que la 5-nucléotidase et la Na⁺/K⁺ ATPase.Il est prévisible que les adjuvants de l'immunité ont une spécificité de cible et qu'une même substance peut se révéler stimulante ou inhibitrice selon la dose utilisée.

     

Synthesis of immunoglobulin by normal and antigenically stimulated fetal sheep.

The rate of appearance, quantity and immunochemical character of serum immunoglobulins which appear normally during the development of fetal sheep and following the injection of antigens at different stages of gestation have been studied. After 70 days' gestation a percentage of normal fetal sheep synthesise IgM. Although the concentration of IgM in the circulation of these animals was very low, the precentage with IgM increased with fetal age. A few late term fetuses were detected which also had IgG1 in their circulation, although none were detected with IgG2 or IgA. Fetuses injected with antigens before 71 days' gestation only synthesised IgM, while fetuses injected after 79 days' gestation synthesised both IgM and IgG1. Neither IgG2 nor IgA were detected by single-radial immunodiffusion analyses during the first 14 days of primary immune responses to a variety of antigens, although trace amounts of IgG2 were detected late in responses occurring in older fetuses. The immunoglobulins synthesised by antigenically stimulated fetal sheep appeared identical to adult sheep 19S IgM, 7S IgG1 and 7S IgG2 respectively, when analysed by immunoelectrophoresis, isoelectric focusing and G200 Sephadex column chromatography.     

S-methylcysteine sulphoxide,the brassica anaemia factor (a valuable dietary factor for man?)

Ruminant animals fed mainly or exclusively on brassica crops may develop a severe Heinz body anaemia. Clinical signs of the disease include haemoglobinuria, jaundice, increase in pulse rate and fall in milk production; there may also be growth retardation. Nitrate and thiocyanates were excluded as primary haemolytic factors in kale which contains up to 2% in DM of S-methylcysteine sulphoxide (SMCO). In test goats, daily intakes of SMCO in the range 15 to 19 g/100 kg liveweight, whether given as kale or the pure synthetic compound, elicited comparable haemolytic responses. SMCO was thus implicated as the major primary toxin in brassica poisoning. Rumen organisms converted SMCO into dimethyl disulphide which mimicked the haemolytic action of kale and SMCO in goats and is thus considered to be the secondary haemolysin. The practical consequence of this disulphide mechanism is that, because there is no simple way of counteracting the systemic action of dimethyl disulphide, prophylactic measures are restricted to limiting the intake of SMCO or to inhibiting disulphide production. Manipulation of rumen flora by dietary means to exclude organisms with SMCO-lyase is seen as a possible way of minimising the problem which in the long term however calls for developing brassica varieties with low SMCO content. The SMCO content of leafy brassicas increases as the plants mature; within a single plant highest SMCO levels are found in young leaves and growing shoots. SMCO is a heritable character and its selection will probably not lead to marked changes in protein or carbohydrate contents. In steers acute haemolytic response with growth inhibition was obtained with daily SMCO intakes, given as cabbage, of 18 to 35 g/100 kg liveweight. At the lower daily intake of 10–15 g the response was sub-acute, but the low grade anaemia produced would probably be incompatible with maximum productivity. Daily intakes below 10 g/100 kg generally lead to mild disturbances only in blood picture; and it is speculated that, on balance, at this level the effects of SMCO may even be beneficial to the animal. The possible prophylactic action of the S-alkylcysteine sulphoxides in coronary heart disease in man is briefly discussed.Resume Les ruminants nourris principalement ou exclusivement de crucifères peuvent contracter une grave anémie à corpuscules endoglobulaires. Les signes cliniques de la maladie comprennent l'hémoglobinurie, la jaunisse, l'élévation du pouls et la baisse de la production de lait et éventuellement aussi un retard de la croissance. Le nitrate et les thiocyanates ont été exclus comme facteurs hémolytiques primaires du chou à feuilles, qui contient jus qu'à 2% en matières sèches du sulfoxyde de s-méthylcystéine (SMCO). Chez les chèvres utilisées comme sujets d'expériences, des doses quotidiennes de SMCO de l'ordre de 15 à 19 g par 100 kg de poids corporel, absorbée sous la forme de chou ou sous celle de composé synthétique pur ont donné des réactions hémolytiques comparables. On a donc conclu que le SMCO était la principale toxine primaire responsable de l'empoisonnement brassicaire. Les organismes du rumen ont transformé le SMCO en disulphide de diméthyle qui a reproduit l'action hémolytique du chou et du SMCO chez les chèvres, et qui est donc considéré comme l'hémolysine secondaire. Étant donné qu'il n'y a pas de moyens simples de contrecarrer l'action systémique du disulphide de diméthyle, les seules mesures prophylactiques sont la limitation de l'absorption de SMCO ou l'inhibition de la production de disulphide. L'action sur la flore du rumen par des moyens diététiques, à fin d'exclure les organismes à SMCO-lyase, semble un moyen possible d'atténuer le problème. Mais à long terme, il faudra développer des variétés de crucifères à faible teneur en SMCO. La teneur en SMCO des crucifères feuillus augmente au fur et à mesure de la maturation. Sur une même plante, les teneurs les plus élevées se rencontrent dans les jeunes feuilles et dans les pousses en croissance. Le SMCO est un caractère héréditaire et une sélection operée en fonction de lui n'entraînera probablement pas de modifications importantes des teneurs en protéines ou en hydrates de carbone. Une réaction hémolytique aiguë avec inhibition de la croissance a été provoquée chez les bouvillons au moyen de doses quotidiennes de SMCO de 18 à 35 g par 100 kg de poids corporel, administrées sous forme de chou. A la dose quotidienne plus faible de 10 à 15 g la réaction a été sub-aiguë mais l'anémie faible qui en résulte serait probablement incompatible avec une productivité optimale. Les doses quotidiennes inférieures à 10 g par 100 kg ne provoquent généralement que de légères perturbations de la formule hématologique. On estime qu'à ce taux les effets du SMCO peuvent même, dans l'ensemble, être bénéfiques à l'animal. L'action prophylactique possible des sulfoxydes de s-alkylcysteine dans les cas de maladies coronaires chez l'homme est examinée brièvement.

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Kurzfassung Bei Wiederkäuern, die hauptsächlich oder ausschliesslich Brassica-Futter erhalten, kann es zu einer schweren Innenkörperanämie kommen. Klinische Anzeichen hierfür sind u.a. Bluthernen, Gelbsucht, erhöhte Pulsfrequenz und Rückgang der Milcherzeugung, ausserdem unter Umständen ein verlangsamtes Wachstum. Nitrat und Thiocyanat wurden als primäre hämolytische Faktoren im Brassica oleracea, das bis zu 2% S-Methylcystein sulfoxid (SMCO) in der Trockenmasse enthält, ausgeschlossen. Bei Test-Ziegen führten Tagesdosen von SMCO im Bereich von 15 bis 19 g/100 kg Lebendgewicht in Form von Brassica oleracea oder dem reinen, synthetisierten Bestandteil zu vergleichbaren hämolytischen Reaktionen. SMCO war also als wichtigstes Primärtoxin bei Brassica-Vergiftungen beteiligt. Die Pansenorganismen setzten SMCO in Dimethyldisulfid um, das die selbe hämolytische Wirkung wie Brassica oleracea und SMCO bei Ziegen hatte und so als sekundäres Hämolysin gilt. Die praktische Folge dieses Disulfidmechanismus ist, dass vorbeugende Massnahmen auf die Begrenzung der SMCO-Aufnahme oder die Verhinderung der Disulfidproduktion beschränkt sind, da es keine einfache Möglichkeit gibt, der systematischen Aktion von Dimethyldisulfid entgegenzuwirken. Eine Manipulation der Pansenflora durch entsprechende Ernährung zur Ausschaltung von Organismen mit SMCO-Lyase gilt als Möglichkeit zur Minimalisierung des Problems, das langfristig gesehen jedoch die Entwicklung von Brassica-Arten mit geringem SMCO-Gehalt erforderlich macht. Der SMCO-Gehalt von Blattkohl nimmt mit zunehmender Reife der Pflanze zu; innerhalb einer Pflanze findet sich der grösste SMCO-Gehalt in jungen Blättern und wachsenden Trieben. SMCO ist ein erbliches Merkmal, und seine Selektion wird vermutlich nicht zu grösseren Aenderungen im Protein-oder Kohlehydratgehalt führen. Bei Ochsen wurde eine akute hämolytische Reaktion mit Wachstumsverzögerung bei täglichem SMCO-Dosen von 18 bis 35 g/100 kg Lebendgewicht in Form von Kohl erzielt. Bei der geringeren Tagesdosis von 10–15 g war die Reaktion subakut, die entwickelte geringfügige Anämie wäre jedoch vermutlich mit maximaler Produktivität unvereinbar. Tagesdosen unter 10 g/100 kg führen im allgemeinen nur im Blutbild zu geringen Störungen; man vermutet, dass alles in allem SMCO bei dieser Dosierung sogar eine positive Auswirkung auf das Tier haben kann. Die mögliche prophylaktische Wirkung der S-Alkylcystein sulfoxide bei Erkrankungen der Herzkranzgefässe beim Menschen wird kurz erörtert.

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Riassunto I ruminanti la cui dieta è in gran parte o esclusivamente rappresentata da cavoli (Brassica spp.) possono essere colpiti da una grave forma di anemia (Heinz body anaemia). I sintomi clinici della malattia comprendono emoglobinuria, ittero, aumento della frequenza cardiaca e caduta della produzione lattea. A volte si può anche avere ritardo della crescita. I nitrati e i tiocianati sono stati esclusi come fattori emolitici primari nei cavoli che contengono fino al 2% in DM di S-metilcisteina sulfossido (SMCO). In experimenti condotti sulle capre, la somministrazione giornaliera di SMCO in dosi di 15–19 g/100 kg di peso vivo, sia sotto forma di vegetale che di prodotto sintetico puro, ha provocato fenomeni emolitici relativamente simili. L'SMCO è stato pertanto indicato come la principale tossina che ha un ruolo primario nell'intossicazione da cavoli. La flora ruminale converte infatti l'SMCO in dimetil bisolfuro, il quale svolge un'azione emolitica simile a quella osservata nelle capre dopo l'assunzione di cavoli e di SMCO; il dimetil bisolfuro, pertanto, e ritenuto costituire l'emolisina secondaria. Le conseguenze pratiche di tale produzione di dimetil bisolfuro è che, non esistendo un metodo semplice per contrastare l'azione sistemica del dimetil bisolfuro, le misure profilattiche non possono essere volte ad altro se non a limitare l'assunzione di SMCO o ad inibire la produzione di bisolfuro. Gli interventi sulla flora del rumine mediante una dieta capace di eliminare i microrganismi provvisti di SMCO-liasi vengono considerati come un possibile mezzo per minimizzare il problema, il quale, tuttavia, se visto a lungo termine, richiede la creazione di varietà di cavoli a basso contenuto di SMCO. Il contenuto di SMCO delle foglie dei cavoli aumenta col maturare della pianta: nella pianta singola i livelli piu alti di SMCO si riscontrano nelle foglie giovani e nei germogli. La presenza di questa sostanza costituisce un carattere ereditario ed un intervento selettivo in tal senso non dovrebbe condurre a significative alterazioni del contenuto in proteine e carboidrati della pianta. Nei vitelloni e stato possibile indurre una risposta emolitica di tipo acuto somministrando giornalmente, sotto forma di cavoli, 18–35 g di SMCO per 100 kg di peso vivo. Un' assunzione giornaliera minore, pari a 10–15 g, ha dato luogo ad una forma emolitica subacuta, ma e tuttavia da ritenere che la lieve anemia cosi indotta non sia compatibile con una produttività ottimale. Livelli giornalieri al di sotto dei 10 g/100 kg hanno causato soltanto lievi alterazioni a carico del quadro ematico, ed è stata avanzata l'ipotesi che, tutto sommato, gli effetti dell'SMCO possano, a questi livelli, essere addirittura di beneficio all'animale. Viene discussa brevemente la possibile azione preventiva degli S-alchilcisteina sulfossidi sulle malattie cardio-coronariche dell'uomo.

     

Effects of chlordimeform on mitochondria from eggs of Spodoptera littoralis

Mitochondria were isolated from eggs of Spodoptera littoralis. With succinate (+pyruvate) as substrate, respiratory control ratios between 1.70 and 2.54 were obtained. Uncouplers and the energy transfer inhibitor oligomycin influenced these mitochondria in the well-known manner. The uncoupling activity of chlordimeform in vitro was very weak and decreased with increasing age of the eggs. Electron transport in mitochondria from chlordimeform-treated eggs was not uncoupled. Therefore, it is concluded that the ovicidal effect of this pesticide is not due to its uncoupling effect.     

Relative persistence of di-and tri-chlorophenoxyalkanoic acid herbicides in Saskatchewan soils

The persistence of 2,4-D, 2,4-DB, dichlorprop, 2,4,5-T, and fenoprop at the 2 ppm level was studied in the laboratory on three prairie soils at 85% of field capacity and 20°C. Following extraction of the soils with aqueous acetonitrile containing acetic acid, the herbicidal acids remaining were analysed gas chromatographically. Breakdown was rapid on all soils and the average half-lives for 2,4-D, 2,4-DB, dichloroprop, 2,4,5-T, and fenoprop were < 7, < 7, 10, 12, and 12 days respectively. Degradation on air-dried soils (15% of field capacity) was negligible with over 85% of the applied herbicides being recoverable after incubation periods during which the herbicides remaining in the moist soils accounted for less than 30% of the original treatments. Persistance relative des acides di et tri-chlorophénoxy-alkanoï-ques herbicides dans des sols du Saskatchewan. La persistance du 2,4-D, du 2,4-DB, du dichlorprop, du 2,4,5-T et du fénoprop, à la concentration de 2 ppm, a étéétudiée au laboratorire, sur trois sols de prairie, a 85% de la capacité au champ et a 20°C. Après leur extraction des sols par l'acétonitrile aqueux contenant de l'acide acétique, les acides herbicides restants ont été analysés par chromatographie en phase gaseuse. La dégradation a été rapide pour tous les sols et les demi-vies moyennes du 2,4-D, du 2,4-DB, du dichlorprop, du 2,4,5-T, et du fénoprop ont été respectivement de <7, <7, 10, 12 et 12 jours. La dégradation sur des sols séchés a l'air (15% de la capacité au champ), a été négligeable, plus de 85% des quantités d'herbicides appliquées étant récupérables après des périodes d'incubation durant lesquelles les herbicides restant dans les sols humides ne représentaient plus que moins de 30% des quantités apportées à l'origine. Relative Persistenz von Di-und Trichlorphenoxyalkansäure-Her-biziden in Böden Saskatchewan In Laborversuchen wurde die Persistenz von 2,4-D, 2,4-DB, Dichlorprop, 2,4,5-T und Fenoprop in drei Prärieböden festgestellt. Die Versuche wurden bei 20°C, 85 % der Feldkapazität und einem anfänglichen Herbizidgehalt der Böden von 2 ppm durchgeführt. Die Extraktion der Böden erfolgte mit wässerigem Acetonitril mit einem geringen Anteil an Essigsäure. Die Herbizide wurden gaschromatographisch nachgewiesen. In allen Böden wurde ein schneller Abbau festgestellt. Die Halbwertszeiten betrugen für 2,4-D, 2,4-DB, Dichlorprop, 2,4,5-T und Fenoprop < 7, < 7,10, 12 bzw. 12Tage. Der Abbau im lufttrockenen Boden (15% der Feldkapazität) war zu vernachlässigen. Hier waren noch mehr als 85% der ausgebrachten Herbizidmenge vorhanden, wenn in den feuchten Böden die Konzentration bereits weniger als 30% betrug.     

An Evaluation of the USDA Preshipment Clearance Program for Propagative Material1)

One of the programs utilized by PPQ-APHIS-USDA2) to prevent the introduction of hazardous pests and pathogens is known as preclearance. Preclearance relies more on inspection at origin than it does on inspection upon arrival or entry into the United States. Two such programs have been developed by PPQ for plant materials. One, established in 1951, is for bulbs, while the other, established in 1975, is for a few ornamental crops. The USDA, in co-operation with the Plant Protection Services of the Netherlands, Belgium, France, Italy, Fed. Rep. of Germany, Israel and South Africa, has been performing preclearance for bulbous plant propagative materials. Flower bulbs are being precleared in all of the above countries at the request of the exporter's groups of the respective countries. The objective of this clearance is to facilitate the entry of this material, reduce the chances of introducing plant pests and diseases into the United States, and save APHIS manpower at US ports of entry. This is achieved by having USDA personnel working cooperatively with foreign plant protection services in the inspection of material in the field and packing houses in the country of origin. Detection and elimination of plant pests and diseases is a co-operative effort for mutual benefit of both parties.     

Nerine latent virus: some properties and serological detectability in Nerine bowdenii

Nerine latent virus (NeLV), first found inNerine bowdenii, may occur also in the otherNerine species investigated so far:N. sarniensis, N. flexuosa Alba, andN. Mansellii. Chenopodium amaranticolor, C. quinoa, andGomphrena globosa sometimes reacted with local lesions after mechanical inoculation with NeLV.Nicotiana clevelandii andHippeastrum were symptomless hosts. In this respect NeLV resembled the incompletely describedHippeastrum latent virus (HLV).Serologically NeLV was closely related to HLV and to carnation latent virus (CaLV), but differed from the latter in host plant reaction. A more distant relationship was observed with some other carlaviruses, wheareas NeLV also reacted with an antiserum to potato virus X.Depending on the lot, NeLV could be detected rather reliably with the micro-precipitin test inN. bowdenii Van Roon, but less well in 63. Better results were obtained with the microplate method of enzyme-linked immunosorbent assay (ELISA).The average particle length was 664 nm, the sedimentation coefficient 155 S and the buoyant density 1.298 g/cm³.NeLV can be considered as a member of the carlavirus group. On basis of priority HLV may be considered as NeLV.