Rejection of worm load through singly and repeatedly sensitized peritoneal exudate cells during experimental ancylostomiasis

Sensitized peritoneal exudate cells from Swiss albino mice donors infected with a single dose of 1000 A. caninum larvae could expel a challenge dose of 500 larvae from recipients at a faster rate when compared to cells from repeatedly infected (250 + 250 + 500) donors. However, at 36 h after challenge, the larval expulsion was almost the same in both the groups. Because of the bowel sensitization by the cells, some larvae (not expelled) in the 1st group, readily migrated into muscles where they met allergic immobilization and death due to infiltration of inflammatory cells and their exudates at these sites.     

Studies on experimental ancylostomiasis: Transfer of acquired immunity toAncylostoma caninum in mice through sensitized thymus and bone marrow cells

Summary An attempt has been made to transfer acquired immunity toAncylostoma caninum infective larvae from infected Swiss albino female mice to nonimmune, isologous recipients of same sex, through immunized thymus and bone marrow cells. Immunized cells from donors infected with a single high dose of 1000 larvae were found to be more immunocompetent than cells from donors infected with a single, but low dose of 500 larvae.Acknowledgment. We are indebted to Professor B. M. Sinha for providing facilities and to Council of Scientific and industrial Research, New Delhi for funds.     

Studies on experimental ancylostomiasis: transfer of acquired immunity to Ancylostoma caninum in mice through sensitized thymus and bone marrow cells.

An attempt has been made to transfer acquired immunity to Ancylostoma caninum infective larvae from infected Swiss albino female mice to nonimmune, isologous recipients of same sex, through immunized thymus and bone marrow cells. Immunized cells from donors infected with a single high dose of 1000 larvae were found to be more immunocompetent than cells from donors infected with a single, but low dose of 500 larvae.     

Protective effect of Shigyaku-to,a traditional Chinese herbal medicine,on the infection of herpes simplex virus type 1 (HSV-1) in mice

The antiviral activity of Shigyaku-to (TJS-109), a traditional Chinese herbal medicine, was investigated in mice infected with herpes simplex virus type 1 (HSV-1). TJS-109 is a combination of the medicinal plant extracts fromZingiberis siccatum rhizoma,Aconiti tuber andGlycyrrhizae radix in a specific proportion. Mice infected with a 10 LD₅₀ dose of HSV-1 were treated with TJS-109 orally at doses of 1.25 to 20 mg/kg 2 days before, and 1 and 4 days after the infection. The treated groups had 80% (1.25 mg/kg), 40% (5 mg/kg) and 23% (20 mg/kg) mortality rates 25 days after the infection as compared with a 100% mortality rate in control mice treated with saline. When HSV-1 infected mice (recipients) received CD8⁺T cell fractions derived from spleens of mice treated with TJS-109 (donors), 70% of recipients survived, as compared with 0% survivors in the groups of mice treated with saline, B cell fractions, CD4⁺ T cell fractions or macrophage-enriched fractions prepared from the same donors. TJS-109 did not show any virucidal activities against HSV-1 or any virostatic activities on the growth of HSV-1 in Vero cells. These results suggest that TJS-109 protected mice exposed to lethal amounts of HSV-1 through the activation of CD8⁺ T cells.     

Experimental ancylostomiasis in chickens: effect of various dose of infective Ancylostoma caninum larvae on their migration and distribution

White leghorn chickens, when infected with repeated doses of Ancylostoma caninum larvae, expel the larvae at a faster rate than when infected with a single dose. This suggests that the initial dose induces some resistance in the host. An initial dose of 1000 and 2000 larvae, followed by a 2nd dose of the same order, induces resistance in the alimentary tract causing the entire larval burden either to migrate to other tissues (organs) or to be expelled in 24 h.     

Diuretic and antifeedant actions byManduca sexta diuretic hormone in lepidopteran larvae

Neuroendocrine disruption of water balance in insect larvae was evaluated as the basis for a new approach to pest insect control. Effects on water balance and food consumption were measured for larvae ofHeliothis virescens (cotton budworm) treated with syntheticManduca sexta diuretic neurohormone (MasDH). Synthetic MasDH (50–250 nM) caused dose-dependent increases in fluid secretion by larval Malpighian tubules in vitro; higher concentrations resulted in lower fluid secretion. Last instar larvae injected with 20–100 pmol MasDH exhibited increasing weight loss. Larvae injected with 500 pmol MasDH dramatically reduced both water excretion and food consumption by 70%. These latter results indicate that high titers of diuretic hormone can suppress feeding damage by larval insects by depressing water excretion and food consumption.     

Brain factor fromGalleria mellonella (Lepidoptera) stimulating silk gland activity

Brain extracts from day 1–4 last instar larvae ofGalleria mellonella (Lepidoptera) stimulate RNA synthesis in cultured silk glands from day 3 last instar larvae. When the fibroin-synthesizing posterior parts of silk glands were incubated for 3 h in vitro in the presence of brain extract (0.1 brain equivalent), [³H]-uridine incorporation into RNA was stimulated more than twofold. The stimulating effect of brain extract showed a dose response relationship. It is suggested that the heat-resistant and protease-sensitive brain factor is a peptide.     

Hemolymph proteins in an ascitic condition,induced by lethal1(3)gl tumorous tissue inDrosophila hydei

Summary Drosophila hydei larvae homozygous for the gene1(3)gl may survive to the age of 3 weeks, become bloated and be incapable of metamorphosis. Wild-type flies after the injection of a fragment of the1(3)gl disc, which has been previously culture in vivo for 40–500 days, also become bloated (ascitic reaction). In the hemolymph of both the advanced1(3)gl larvae and ascitic adult hosts the accumulation of a specific protein is observed.Acknowledgments. This work was supported by the Swiss National Science Foundation under Grant No.3.792-0.76 and the Georges und Antoine Claraz-Schenkung. We thank Prof. H. Gloor, University of Geneva, and Prof. P.S. Chen, University of Zürich, for advice and criticism.     

Detection of toxic metabolites in the hemolymph ofBeauveria bassiana infectedSpodoptera exigua larvae

Highly active metabolites have been detected in the hemolymph of the lepidopteranSpodoptera exigua infected with the mycopathogen,Beauveria bassiana. A combination of phenyl sepharose and CM ion exchange chromatography was utilized to extract the active metabolites from infected hemolymph samples. The active in vivo metabolites, having a molecular mass greater than 10 KDa, were thermolabile and were inactivated by proteinase K. These metabolites were characterized by their ability to disrupt metamorphosis, killing treated larvae at the wandering or pupal stage. Additionally, injection ofS. exigua larvae with active samples caused a reduction in the number of filopodial-producing hemocytes. The biological activities and biochemical properties suggest that novel compounds are produced duringB. bassiana mycosis.     

Analysis of the relationship between age of larvae at mutagen treatment and frequency and size of spots in the wing somatic mutation and recombination test inDrosophila melanogaster

The relationship between the induction of mutant clones and the time of mutagen treatment was studied in the somatic mutation and recombination test (SMART) in wing cells ofDrosophila melanogaster. Larvae trans-heterozygous for the recessive marker mutations multiple wing hairs (mwh) and flare (flr) were produced. Batches of these larvae were then treated with mutagen at different ages spanning all three larval instars. Methyl methanesulfonate was fed acutely for 2 h by immersing the larvae in a solution of the mutagen mixed with powdered cellulose. Wings of the surviving adult flies were mounted and scored for the presence of spots. The frequency and size of single and twin spots were recorded separately. Twin spots are produced exclusively by mitotic recombination, whereas single spots can results from various types of mutational and exchange events. There exists a clear correlation between time of induction and frequency as well as size of the single spots. In young larvae only few but very large spots are induced, whereas in older larvae the frequencies are considerably increased but the sizes are smaller. The twin spots show a different relationship. Practically no twin spots are found in very young and in very old larvae. The results demonstrate that in the wing spot test the optimal age of the larvae for mutagen treatment is 72 h.     

Leishmania donovani in hamsters: Stimulation of non-specific resistance by some novel glycopeptides and impact on therapeutic efficacy

Summary Several glycopeptides structurally related to muramyl dipeptide (MDP) have been synthesized and evaluated for their ability to stimulate the non-specific resistance of hamsters againstL. donovani infection. These compounds have been named CDRI compounds. The synthetic procedure used for compounds 86/448 and 84/212 is described.MDP and its synthetic congeners were administered as immunostimulants at a prophylactic dose of 3 mg/kg at two weeks interval. The challenge infection (1×10⁷ amastigotes i.c./hamster) was given in between two doses of the compounds. One of the glycopeptides, CDRI comp. 86/448, has been found to be significantly more potent than MDP, effecting 92% inhibition of the challenge dose, whereas MDP produced only 26.5% inhibition. The effect of comp. 86/448 lasted until day 7 of challenge. The efficacy of sodium stibogluconate was appreciably improved in hamsters treated with comp. 86/448.     

Nosiheptide,a sulfur-containing peptide antibiotic isolated fromStreptomyces actuosus 40037

Summary Nosiheptide (9671 R.P.) isolated fromStreptomyces actuosus 40037 (NRRL 2954) is a sulfur-containing polypeptidic antibiotic, quite different from all the other members of this family. Very active in vitro against gram-positive bacteria, it is inactive in vivo in experimentally infected mice. Not toxic, even at high dose, it may be used as a feed additive for chickens and pigs and it shows a favourable effect on the growth and conversion index.     

Oenocyte and prothoracic gland activity inManduca sexta under varying photoperiod and light conditions

Summary Manduca sexta larvae were subjected to diapause-inducing and diapause-preventing photoperiods, using two types of fluorescents (Indorsun and Blacklight-blue). The oenocytes, prothoracic glands (PTG) and ecdysone levels were examined in 3-day-old 5th instar larvae, 2-day-old and 10-day-old pupae. Our results indicate that oenocytes and PTG cells tend to be more active under long photoperiods while oenocytes only are active under short photoperiods in pupae in diapause. UV light has a definite effect on oenocytes while PTG cells seem to be unaffected. Ecdysone and ecdysterone levels vary with PTG and oenocyte activity at the pupal stage. The significance of these findings is discussed.This work was supported by a research associateship to L.M. by the People's Republic of China and a grant from NSERC to B.J.R.P.     

Free amino acids of the haemolymph of the cotton leaf-worm,Spodoptera littoralis Boisduval full-grown larvae,infected with nuclear-polyhedrosis virus

Summary Free amino acids in the haemolymph ofSpodoptera littoralis full-grown larvae infected with a nuclearpolyhedrosis virus were compared with those in the haemolymph of normal insects. Amino acids were separated by 2-dimensional paper chromatography and quantified colorimetrically. Most of the amino acids in the haemolymph of diseased full-grown larvae decreased markedly in concentration but proline, lysine, aspartic acid and histidine occurred in greater concentration in the haemolymph of diseased full-grown larvae than in the haemolymph of healthy insects.     

Multiple overwintering mechanisms inChymomyza amoena larvae (Diptera: Drosophilidae) and laboratory induction of freeze tolerance

Summary Chymomyza amoena larvae in apples in summer were found to be either potentially freeze tolerant or to supercool. By late September only potentially freeze tolerant larvae were recovered from apples. Larvae from walnut husks in winter supercooled to avoid freezing. However, freeze tolerance could be induced in laboratory grown larvae by placing them in apples around the seed area and maintaining them at chilling temperatures for several weeks. Overwintering mechanisms employed byC. amoena larvae in Michigan appear to depend upon larval feeding site.Prof. Davis at Michigan Agricultural College placed 9C. amoena emerging from apples in August and Sept. 1891 along with codling moth in the MAC (=MSU) Entomological Museum collection.     

Allatotropic activity of the median neurosecretory cells ofGalleria mellonella (lepidoptera) larval brain

Summary The brain allatotropic hormone (ATTH) is released inGalleria mellonella from the Median neurosecretory cells located in the pars intercerebralis. These cells show the ability to elicit supernumerary larval molts upon implantation into sensitive host larvae, and the ability to in vitro stimulate the juvenile hormone synthesis in corpora cardiaca-corpora allata glands ofG. mellonella.11 November 1986Acknowledgments. I am greatly indebted to Wesleyan University (Connecticut, USA) for supporting this study.     

Crystal cell pattern modification in a melanotic tumor strain ofDrosophila melanogaster

Summary 3rd instar larvae of the melanotic tumortu-pb strain ofDrosophila melanogaster hold a lower number of free-circulating crystal cells in their hemolymph than the wild type ones. This pattern could result from an abnormal retention of mature crystal cells in the hematopoietic organs, as the strong hemocyte melanization inside the lymph glands of heat-treatedtu-pb larvae seems to demonstrate. Melanotic tumor formation and modification of the crystal cell pattern may be related.     

Detection of human cytomegalovirus (HCMV) DNA from cell-free plasma of immunosuppressed patients by nested PCR: influence of nucleic acid extraction method

Conclusions Blood cells and plasma preparations from HCMV-seropositive healthy blood donors were all nPCR negative. Detection of HCMV DNA from PBMC and granulocytes (DNAemia) of immunosuppressed patients by nPCR did not correlate with the isolation of infectious virus from these cell populations in cell culture (viremia). However HCMV could be isolated in 60% of cases from other materials of the same patient. HCMV DNA detected in blood cells persisted for up to one year in an asymptomatically infected individual after NTX. The sensitivity of HCMV DNA detection in cell-free plasma (up to 5 fg) depended on the method used for DNA isolation. The rate of HCMV DNA detection in plasma was lower than in leukocytes. In all cases of positive plasma PCR infectious virus could be isolated from any other material of the symptomatically infected patients. Therefore HCMV DNA PCR from plasma of immunosuppressed patients seems to be a suitable and easy alternative to HCMV RT/PCR for routine diagnosis of HCMV disease.     

Free amino acid composition of the intestinal contents,intestinal cells and hemolymph ofPhilosamia cynthia larvae

Summary Free amino acid composition of the intestinal contents, intestinal cells and hemolymph has been determined in larvae of the mothPhilosamia cynthia. From the hemolymph/lumen concentration ratio, an active transport could be inferred for neutral and basic amino acids. The values of cell/lumen and hemolymph/cell ratios suggested that the active step in the transport mechanism could be localized at the luminal pole of the enterocyte for neutral amino acids (except aromatic amino acids) and at the basolateral pole of the enterocyte for basic amino acids (except arginine).This work was supported by grants from Italian Consiglio Nazionale delle Ricerche and from Ministero della Pubblica Istruzione, Rome. The authors are indebted to Prof. V. Capraro for helpful discussion.