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.     

‘Action substances’ of peripheral nerve re-visited

Fifty years ago, four action substances were described as being liberated from peripheral nerve in the course of activity. An attempt has been made to update this information. Confirmatory evidence is available that acetylcholine and thiamine do play a role in the course of activity. However, the question of the relationship in time between their release and the electrical events, and the possible effect on specific ionic gating mechanisms, remains unclarified. The liberation of potassium has been found to be a transmembrane K efflux, and the question of the existence of another substance similar to thiamine has not been pursued.a member of the Editorial Board of EXPERIENTIA from 1974 to 1982     

Pineal ‘synaptic’ ribbon numbers and melatonin synthesis of rat are resistant to guanethidine sympathectomy

Chemical sympathectomy is widely used to study the impact of the noradrenergic system on neuronal and neuroendocrine circuits. We tested the effects of intraperitoneal injections of guanethidine, an adrenergic neuron blocking agent, on selected functional parameters of the rat pineal gland which are known to be under sympathetic influence. The reliability of the method was demonstrated by the clear enophthalmus developed by experimental animals. However, neither the numbers of synaptic ribbons nor melatonin synthesis differed between treated and control rats, both parameters exhibiting the nocturnal increase seen in intact animals. These results are in striking contrast to those obtained upon chemical sympathectomy with 6-hydroxydopamine or surgical superior cervical ganglionectomy. We conclude that guanethidine is not capable of sufficiently removing noradrenergic influence from the rat pineal gland, and that this substance is thus inferior to other experimental methods of sympathectomy.     

2-Methoxybenzoyl phosphate: a new substrate for continuous fluorimetric and spectrophotometric acyl phosphatase assays

A new aromatic acyl phosphate, 2-methoxybenzoyl phosphate, has been synthesized. The compound shows an intrinsic fluorescence; it displays an intense emission band at 390 nm upon excitation in the near UV region. This band practically disappears after hydrolysis of the product. On the other hand, the product displays differences in the near UV absorption spectra measured before and after hydrolysis. The at 301 nm is 2720 M^–1 cm^–1, a value that is 4.3-fold higher than that of benzoyl phosphate (the usual substrate for acylphosphatase assay) at 283 nm. The main kinetic parameters of three different acylphosphatase molecular forms (the muscular isoenzyme and two subtypes of the organ common isoenzyme) were determined using both benzoyl phosphate and 2-methoxybenzoyl phosphate as substrates, and then compared. These kinetic data and the UV absorption and fluorescence properties of 2-methoxybenzoyl phosphate sugest that this compound has better substrate features than benzoyl phosphate, and can be used for both high sensitivity continuous fluorimetric and UV absorption spectrophotometric assays of acylphosphatase.     

Population biology of bryozoans: correlates of sessile,colonial life histories in freshwater habitats

Bryozoans are common, often abundant, sessile, colonial invertebrates of freshwaters. By reviewing what is known of their general ecology and considering in further detail the population biology of specific groups, we provide evidence that 1) populations are locally ephemeral and regional persistence is attained via a dynamic equilibrium between dispersal and colonization, and 2) sex may often be infrequent and a high degree of clonality obtains at both local and regional scales. On the basis of these characteristics we discuss how the group offers exceptional opportunities for investigating a number of fundamentally important ecological and evolutionary questions.The highly curious modification of the Molluscan type which the Polyzoa [=Bryozoa] present... the great beauty of their forms, and the facility with which they can in general be observed in a living state, cannot but render them special favorites for every lover of Nature; and for the more profound student must confer on them a peculiar significance, and invest their study with a scientific interest which is scarcely surpassed by that of any other group of animals.... (Allman 1856)     

Presence of hsp65 in bacterial extracts (OM-89): A possible mediator of orally-induced tolerance?

Heat shock proteins (HSP) have been implicated in rodent models of autoimmunity, particularly arthritis, and there is suggestive though inconclusive evidence that they may also play a role in human autoimmune disease. The simplest hypothesis is based on molecular mimicry due to the amino-acid sequence homology between mammalian and microbial HSP. Recently OM-89, an extract of several strains ofEscherichia coli, has shown some efficacy in the treatment of rheumatoid arthritis (RA) when taken orally. Using species-specific antibodies, we show here that OM-89 contains the 65 kDa HSP (hsp65), while hsp65 was not detected in another bacterial extract containing other microorganisms, includingStaphylococcus aureus (OM-85). We suggest that if the human homologue of hsp65 is a relevant target antigen in the human disease, the efficacy of the preparation could be due to induction of oral tolerance or to switching the Th1 response towards Th2. Alternatively, even if the human hsp65 is not a target molecule in RA joints, OM-89 may evoke bystander suppression of joint inflammation via induction of TGF-secreting effector cells. These hypotheses should be tested in further studies.     

Occurrence of 3-epi-22-deoxy-20-hydroxyecdysone and its phosphoric ester in diapause eggs of the silkworm,Bombyx mori

Ecdysteroids in diapause eggs of the silkworm,Bombyx mori, were analyzed using high-performance liquid chromatography (HPLC) combined with radioimmunoassay (RIA). A relatively large amount of an unidentified free ecdysteroid and its phosphoric ester (conjugated form) were detected. These two compounds were isolated by a combination of column chromatography on silicic acid, thin-layer chromatography (TLC), and HPLC using a reverse-phase (RP) column. The purified compounds were identified as 3-epi-22-deoxy-20-hydroxyecdysone (22d20E) and 3-epi-22-deoxy-20-hydroxyecdysone 2-phosphate (22d20E2P) by means of mass spectrometry and nuclear magnetic resonance spectroscopy. to our knowledge, this is the first report of 22d20E and 22d20E2P.     

Energy flow and community structure in freshwater ecosystems

The goal of this article is twofold: 1) It aims at providing an overview on some major results obtained from energy flow studies in individuals, populations, and communities, and 2) it will also focus on major mechanisms explaining community structures. The basis for any biological community to survive and establish a certain population density is on the one hand energy fixation by primary producers together with adequate nutrient supply and the transfer of energy between trophic levels (bottom-up effect). On the other hand, predator pressures may strongly control prey population densities one or more trophic levels below (top-down effect). Other interpopulation effects include competition, chemical interactions and evolutionary genetic processes, which further interact and result in the specific structuring of any community with respect to species composition and population sizes.     

Thrombospondin co-localises with TGFβ and IGF-I in the extracellular matrix of human osteoblast-like cells and is modulated by 17β estradiol

Thrombospondin (TSP) is a multifunctional glycoprotein which is synthesised by several cell types including osteoblasts, and incorporated into the extracellular matrix (ECM) of these cells. The function and regulation of TSP in bone is not clear. In this study, using a long term culture model of human osteoblast-like cells, we examined the distribution of TSP in the ECM and its modulation by added estradiol. In this model the osteoblast-like cells form a regular multilayer which continues to increase in depth up to 50 days post confluence. In the ECM of these cultures and in 19-week fetal bone, the bone markers osteocalcin and alkaline phosphatase were diffusely distributed in the matrix. In contrast, labelling for TSP was concentrated, confined to the banded collagen and its immediately adjacent ECM. This pattern of labelling resembled that of the growth factors transforming growth factor-I (TGF), and insulin-like growth factor-I (IGF-I), with which TSP label co-localised. Labelling intensities were comparable between fetal bone and the in vitro material for TSP, TGF and IGF-I. TSP label was present by 10 days post confluence, reached a maximum by 20 days, and declined slowly thereafter, a time course which was similar to that of IGF-I. Incubation of osteoblast-like cell cultures with 17 estradiol resulted in an increase in multilayer depth and a maximal 3-fold increase in TSP labeling at 30 days as well as approximately 2-fold increases for TGF and IGF-I. The dose-response relationship for these responses to estradiol treatment was biphasic with maximal increases at 10^–10 M–10^–11 M of added estradiol. Treatment with 17 estradiol produced labelling intensities that were not significantly different from controls. Studies with other cell types have suggested that TSP may be involved in modulation of growth factor activity. The similarities between TSP, TGF and IGF-I, in terms of their distribution and regulation by 17 estradiol treatment, may indicate a role for TSP in modulating bone cell proliferation and function through interaction with local growth factors.