Inhibition of the cytotoxicity of protein toxins by a novel plant metabolite,Mansonone-D

We have studied the effect of several structurally related mansonones on the cytotoxicity of plant and bacterial toxins in Vero and BER-40, a brefeldin A-resistant mutant of Vero cells. Mansonone-D (MD), a sesquiterpenoid ortho-naphthoquinone, inhibited the cytotoxicity of ricin, modeccin, Pseudomonas toxin, and diphtheria toxin in Vero cells to different extents. The inhibition of ricin cytotoxicity was dose dependent and reversed upon removal of the drug. Protection of ricin cytotoxicity was also observed in the presence of cycloheximide, indicating that de novo protein synthesis is not required for the protective effect. Although MD inhibited the degradation and excretion of ricin, the binding and internalization of ricin was not affected. In contrast, MD strongly reduced the specific binding of diphtheria toxin in Vero cells. Fluorescence microscopic studies show that MD treatment dramatically alters the morphology of the Golgi apparatus in Vero cells. The kinetic studies reveal that the protection of ricin cytotoxicity is the consequence of decreased toxin translocation to the cytosol in MD-treated cells. The reactive ortho-quinone moiety of MD is important for the protective effect as thespesone, a para-naphthoquinone with a heterocyclic ring structure identical to that of MD, did not inhibit the cytotoxicity of toxins. Thespone, a dehydromansonone-D, lacking two hydrogens from the heterocyclic dihydrofuran ring of MD, inhibited the cytotoxicity of ricin, but was albeit less potent than MD. Neither mansonone-E nor mansonone-H with reactive ortho-quinone moiety, but with a different heterocyclic structure, had any effect on the cytotoxicity of ricin indicating that the protective effect of MD is specifically related to the overall structure of the metabolite. J. Cell. Physiol. 176:40–49, 1998. Published 1998 Wiley-Liss, Inc.

1 This article was prepared by a group of United States government employees and non-United States government employees, and as such is subject to 17 U.S.C. Sec. 105.

     

Effect of nitrogen on growth and water relations of radiata pine families

The effect of nitrogen fertilisation on growth, foliar nutrients and water relations of four families of radiata pine (Pinus radiata, D. Don) currently in the Australian breeding program was examined from age six to 11 years. At this stage, the stand was ready for commercial thinning. The annual rainfall at the site varied from 563 to 733 mm.Application of nitrogen fertiliser resulted in stem wood volume at age nine years of 178 m³ ha^-1 in the controls, compared with 228 m³ ha^-1 in plots treated with 600 kg N ha^-1. Pre-dawn needle water potential () measured in three consecutive summers (when rainfall ranged from 53 to 106 mm) were consistently higher (less water stress) in nitrogen fertilised than in control trees. Similarly, the water stress integral (S) decreased consistently with increasing levels of nitrogen, although total water use in fertilised trees would have been substantially higher because fertiliser application increased the leaf area index. The relationship between S and basal area was strong and paralleled that of foliar nitrogen concentration and basal area growth. Therefore, nitrogen application increased growth rates of trees by improving the nutrient status of trees and lowering the water stress on trees in summer.Families showed markedly different responses of basal area growth to nitrogen, ranging from an increase of 9.4% over three years for the least responsive family to 99.0% for the most responsive. There was no nitrogen × family interaction on or S suggesting that the large genetic variation in the growth response to nitrogen is mediated by factors other than water relations. These results have implications for managing highly productive plantations grown in an environment where rainfall is low compared to potential evapotranspiration.     

The origin of meiotic bridges by chiasma formation in heterozygous inversions in Prosimulium multidentatum (Diptera: Simuliidae)

Prosimulium multidentatum (Twinn) has three metacentric pairs in its chromosome complement. All six arms are individually identifiably in polytene nuclei. XY₁ males are heterozygous for a small basal non-conformity in section 59 of the non-pairing sex differential segments which extends from sextion 58 to section 62 of the IIL arm. XY₂ males carry an additional large heterozygous inversion in the center of this same arm. Meiosis is chiasmate in both kinds of male. In XY₂ individuals 14.2% of the pachytene nuclei show reverse loop pairing and 12.5% of the anaphase I cells form bridge-fragment configurations. A majority of these bridges persist into second division and 7.1% double sized spermatids are formed. No pachytene loops or anaphase bridges were found in XY₁ males. It is concluded therefore that the bridges and fragments of XY₂ males result from chiasma formation within the Y₂ inversion.     

Uptake of Zn65 from dry soil by plants

Summary Absorption of Zn⁶⁵ by plant roots from a layer of dry top soil (suction > 15 bar) was measured using a technique that obviated water stress in the plant.Roots growing through the dry layer absorbed significant amounts of Zn⁶⁵ provided they absorbed water from lower depths. Mucilage accumulated around the roots in the dry soil and may have facilitated ion transfer in the immediate vicinity of the root. The ability of roots to grow in dry soil and to absorb Zn⁶⁵ varied between plant species.     

Staphylococcal meningoencephalitis, nematodiasis, and typhlocolitis in a squirrel monkey (Saimiri sciureus)

Background  Seizures were observed in a 16-year old male Guyanese squirrel monkey with a history of inappetence and weakness.
Methods and results  Complete blood count, biochemical profile, and urinalysis indicated systemic disease. Nematode larvae were detected in the feces. Cerebrospinal fluid (CSF) analysis revealed leukocytes and gram-positive cocci. Staphylococcus aureus was isolated from the CSF. Histopathological evaluation revealed systemic lesions with inflammation and nematodes in the small and large intestine.
Conclusion  This is the first report describing spontaneous staphylococcal CNS infection in a squirrel monkey.     

Efficient hydrolysis of the chemical warfare nerve agent tabun by recombinant and purified human and rabbit serum paraoxonase 1

Paraoxonase 1 (PON1) has been described as an efficient catalytic bioscavenger due to its ability to hydrolyze organophosphates (OPs) and chemical warfare nerve agents (CWNAs). It is the future most promising candidate as prophylactic medical countermeasure against highly toxic OPs and CWNAs. Most of the studies conducted so far have been focused on the hydrolyzing potential of PON1 against nerve agents, sarin, soman, and VX. Here, we investigated the hydrolysis of tabun by PON1 with the objective of comparing the hydrolysis potential of human and rabbit serum purified and recombinant human PON1. The hydrolysis potential of PON1 against tabun, sarin, and soman was evaluated by using an acetylcholinesterase (AChE) back-titration Ellman method. Efficient hydrolysis of tabun (100 nM) was observed with ∼25-40 mU of PON1, while higher concentration (80-250 mU) of the enzyme was required for the complete hydrolysis of sarin (11 nM) and soman (3 nM). Our data indicate that tabun hydrolysis with PON1 was ∼30-60 times and ∼200-260 times more efficient than that with sarin and soman, respectively. Moreover, the catalytic activity of PON1 varies from source to source, which also reflects their efficiency of hydrolyzing different types of nerve agents. Thus, efficient hydrolysis of tabun by PON1 suggests its promising potential as a prophylactic treatment against tabun exposure.     

Inhibition of Helicobacter hepaticus-induced colitis by IL-10 requires the p50/p105 subunit of NF-kappa B

Defects within the innate immune system sensitize NF-kappaB-deficient (p50(-/-); p65(+/-)) mice to Helicobacter hepaticus (Hh)-induced colitis. Because IL-10 plays a central role in the inhibition of Hh-induced colitis, we hypothesized that the ability of IL-10 to inhibit the innate inflammatory response to Hh may be compromised in NF-kappaB-deficient mice. To test this hypothesis, we evaluated the ability of an IL-10-Ig fusion protein with IL-10-like properties to inhibit Hh-induced colitis in RAG-2(-/-) (RAG) and p50(-/-); p65(+/-); RAG-2(-/-) (3X/RAG) mice. As expected, IL-10-Ig efficiently inhibited the development of colitis in RAG mice. In contrast, the ability of IL-10-Ig to inhibit colitis was compromised in 3X/RAG mice. The defect in response to IL-10-Ig appeared to be primarily the result of the absence of the p50/p105 subunit, because the ability of IL-10-Ig to inhibit colitis was also compromised in p50(-/-); RAG-2(-/-) (p50/RAG) mice. Radiation chimeras demonstrated that the presence of p50/p105 within hemopoietic cells of the innate immune system was necessary for efficient inhibition of colitis by IL-10-Ig. Consistent with a defect in the suppressive effects of IL-10 in the absence of p50/p105, we found that the ability of IL-10 to control LPS-induced expression of IL-12 p40 was significantly compromised in macrophages lacking p50/p105. These results suggest that the absence of the p50/p105 subunit of NF-kappaB within hemopoietic cells of the innate immune system interferes with the ability of IL-10 to suppress inflammatory gene expression and Hh-induced colitis.     

Marine mycoflora of south India with special emphasis on lignicolous marine fungi

A study dealing with the marine fungi associated with decaying wood samples in the brackish water mangrove ecosystem and shoreline ecosystem was carried out in south India. A total of 19 marine fungi were isolated from the brackish water mangrove ecosystem. They included 13 Ascomycetes, one Basidiomycete and five Mitosporic fungi. In terms of percent frequency of occurrence, the most frequent species obtained from the brackishwater were the Lignincola longirostris (16.60%) and Savoryella lignicola (12.09%). Nine species were found frequently. Five species were occasionally encountered. Aigialus mangrovei, Aniptodera mangrovei and Halosarpheia marina were the rare species recorded. The average number of isolates per wood sample was 1.53. A total of 27 marine fungi including 15 ascomycetes, one basidiomycete and ten mitosporic fungi were recorded from the shoreline ecosystem. In terms of percent frequency of occurrence, the most frequent species obtained from Kanyakumari were the Arenariomyces trifurcates (13.66%), Corollospora maritima (12.44%), and Cirrenalia pygmea (10.98%). Seven species were found frequently. Fourteen species were occasionally encountered. Three species were found to be rare in occurrence. The average number of isolates per wood sample was 1.21.     

STR/ort mice, a model for spontaneous osteoarthritis, exhibit elevated levels of both local and systemic inflammatory markers

Osteoarthritis is a common joint disease that currently lacks disease-modifying treatments. Development of therapeutic agents for osteoarthritis requires better understanding of the disease and cost-effective in vivo models that mimic the human disease. Here, we analyzed the joints of STR/ort mice, a model for spontaneous osteoarthritis, for levels of inflammatory and oxidative stress markers and measured serum cytokines to characterize the local and systemic inflammatory status of these mice. Markers of low-grade inflammatory and oxidative stress-RAGE, AGE, S100A4, and HMGB1-were evaluated through immunohistochemistry. Of these, AGE and HMGB1 levels were elevated strongly in hyperplastic synovium, cartilage, meniscus, and ligaments in the joints of STR/ort mice compared with CBA mice, an osteoarthritis-resistant mouse strain. These increases (particularly in the synovium, meniscus, and ligaments) correlated with increased histopathologic changes in the cartilage. Serum analysis showed higher concentrations of several cytokines including IL1β, IL12p70, MIP1β, and IL5 in STR/ort mice, and these changes correlated with worsened joint morphology. These results indicate that STR/ort mice exhibited local and systemic proinflammatory conditions, both of which are present in human osteoarthritis. Therefore, the STR/ort mouse model appears to be a clinically relevant and cost-effective small animal model for testing osteoarthritis therapeutics.