A mutant of Bacillus thuringiensis var. israelensis (B.t.i.) resistant to antibiotics

Summary PST, a spontaneous mutant of Bacillus thuringiensis var. israelensis (B.t.i.) resistant to penicillin, streptomycin and tetracycline was isolated by serial selections. In the absence of antibiotics it showed genetic stability for 16 generations. Mosquito larvicidal activity of PST was similar to that of B.t.i., its parental strain. It also maintained the specific antigenicity of B.t.i. although its rate of growth was somewhat lower, a generation time of 55 min for PST vs. 38 min for B.t.i. Cell concentration plays a major role in the phenomenon of PST resistance to penicillin.This antibiotic resistant mutant of b.t.i. provides us with an efficient tool to trace B.t.i. among the indigenous bacteria present in septic habitats in the field as well as inside the larval gut.     

From Biodiversity to Ecodiversity: A Landscape-Ecology Approach to Conservation and Restoration

An environmental revolution is urgently needed that will lead to a post-industrial symbiosis between man and nature. This can be realized only if the present unrestrained biological impoverishment and neotechnological landscape degradation are replaced by the creation of healthy and attractive landscapes. Restorationists can fulfill a vital role in this process. They must broaden their scales from biodiversity restoration in small, protected nature islands to the large-scale restoration of natural and cultural landscapes. To achieve this they must restore not only the patterns of vegetation but also the processes that create these patterns, including human land uses. Their goal should be to restore the total biological, ecological, and cultural landscape diversity, or “ecodiversity,” and its intrinsic and instrumental values of highly valuable, endangered seminatural, agricultural and rural landscapes. For this purpose it is essential to maintain and restore the dynamic flow equilibrium between biodiversity, ecological, and cultural landscape heterogeneity, as influenced by human land uses, which occur at different spatial and temporal scales and intensities. Recent advances in landscape ecology should be utilized for broader assessment of ecodiversity, including proposed indices of ecodiversity, new techniques such as Intelligent Geographical Information Systems (IGIS), and Green Books for the holistic conservation and restoration of valuable endangered landscapes. Restoration ecology can make an important contribution to an urgently needed environmental revolution. This revolution should lead to a new symbiosis between man and nature by broadening the goal of vegetation restoration to ecological and cultural landscape restoration, and thereby to total landscape ecodiversity.     

Relationships among the herbicide and functional sites of acetohydroxy acid synthase from Chlorella emersonii

The properties of acetohydroxy acid synthase (AHAS, EC 4.1.3.18) from wild-type Chlorella emersonii (var. Emersonii, CCAP-211/11n) and two spontaneous sulfometuron methyl (SMM)-resistant mutants were examined. The AHAS from both mutants was resistant to SMM and cross-resistant to imazapyr (IM) and the triazolopyrimidine sulfonanilide herbicide XRD-498 (TP). The more-SMM-resistant mutant had AHAS with altered catalytic parameters (K _m, specificity), but unchanged sensitivity to the feedback inhibitors valine and leucine. The second mutant enzyme was less sensitive to the feedback inhibitors, but had otherwise unchanged kinetic parameters. Inhibition-competition experiments indicated that the three herbicides (SMM, IM, TP) bind in a mutually exclusive manner, but that valine can bind simultaneously with SMM or TP. The three herbicide classes apparently bind to closely overlapping sites. We suggest that the results with C. emersonii and other organisms can all be explained if there are separate binding sites for herbicides, feedback inhibitors and substrates.Abbreviations AHAS acetohydroxy acid synthase - AL acetolactate - AHB acetohydroxybutyrate - IM imazapyr - TP triazolopyrimidine sulfonanilide herbicide XRD-498 - R enzyme specificity - SMM sulfometuron methyl This research was supported in part by the United States — Israel Binational Science Foundation (BSF), Jerusalem, Israel (Grant 86-00205) and the Fund for Basic Research, Israel Academy of Sciences.     

Microsite differentiation in a Mediterranean oak savanna

Abstract. Microscale variation of vegetation was studied in a Mediterranean oak savanna dominated by annual plants at Neve Ya'ar Experimental Station, in the lower Galilee of Israel. Species presence/absence data were recorded in 20 cm x 20 cm quadrats on a 500-quadrat transect. These data were ordinated with Detrended Correspondence Analysis. Soil nitrate, phosphorous, potassium, pH, soil moisture and salinity were determined at 2-m intervals on the 100-m transect, at monthly intervals from November 1981 to April 1982. Litter cover and canopy shade were surveyed in October 1981 and April 1982. A sharp rise in available nitrate occurred in November, following the first winter rains. Patterns in herbaceous vegetation, as expressed by sample ordination scores, correlated best with litter cover in October. This suggested that vegetation pattern in this community may be strongly influenced by environmental conditions at the time of seedling germination and establishment.     

Tooth periodontal ligament: Direct 3D microCT visualization of the collagen network and how the network changes when the tooth is loaded

The periodontal ligament (PDL), a soft tissue connecting the tooth and the bone, is essential for tooth movement, bone remodeling and force dissipation. A collagenous network that connects the tooth root surface to the alveolar jaw bone is one of the major components of the PDL. The organization of the collagenous component and how it changes under load is still poorly understood. Here using a state-of-the-art custom-made loading apparatus and a humidified environment inside a microCT, we visualize the PDL collagenous network of a fresh rat molar in 3D at 1 μm voxel size without any fixation or contrasting agents. We demonstrate that the PDL collagen network is organized in sheets. The spaces between sheets vary thus creating dense and sparse networks. Upon vertical loading, the sheets in both networks are stretched into well aligned arrays. The sparse network is located mainly in areas which undergo compressive loading as the tooth moves towards the bone, whereas the dense network functions mostly in tension as the tooth moves further from the bone. This new visualization method can be used to study other non-mineralized or partially mineralized tissues, and in particular those that are subjected to mechanical loads. The method will also be valuable for characterizing diseased tissues, as well as better understanding the phenotypic expressions of genetic mutants.     

Inhibition of melanoma development in the Nras(Q61K)::Ink4a−/− mouse model by the small molecule BI‐69A11

To date, there are no effective therapies for tumors bearing NRAS mutations, which are present in 15–20% of human melanomas. Here we extend our earlier studies where we demonstrated that the small molecule BI‐69A11 inhibits the growth of melanoma cell lines. Gene expression analysis revealed the induction of interferon‐ and cell death‐related genes that were associated with responsiveness of melanoma cell lines to BI‐69A11. Strikingly, the administration of BI‐69A11 inhibited melanoma development in genetically modified mice bearing an inducible form of activated Nras and a deletion of the Ink4a gene (Nras^(Q61K)::Ink4a^?/?). Biweekly administration of BI‐69A11 starting at 10 weeks or as late as 24 weeks after the induction of mutant Nras expression inhibited melanoma development (100 and 36%, respectively). BI‐69A11 treatment did not inhibit the development of histiocytic sarcomas, which constitute about 50% of the tumors in this model. BI‐69A11‐resistant Nras^(Q61K)::Ink4a^?/? tumors exhibited increased CD45 expression, reflective of immune cell infiltration and upregulation of gene networks associated with the cytoskeleton, DNA damage response, and small molecule transport. The ability to attenuate the development of NRAS mutant melanomas supports further development of BI‐69A11 for clinical assessment.     

The N-terminal domain of the regulatory subunit is sufficient for complete activation of acetohydroxyacid synthase III from Escherichia coli

We have previously proposed a model for the fold of the N-terminal domain of the small, regulatory subunit (SSU) of acetohydroxyacid synthase isozyme III. The fold is an alpha-beta sandwich with betaalphabetabetaalphabeta topology, structurally homologous to the C-terminal regulatory domain of 3-phosphoglycerate dehydrogenase. We suggested that the N-terminal domains of a pair of SSUs interact in the holoenzyme to form two binding sites for the feedback inhibitor valine in the interface between them. The model was supported by mutational analysis and other evidence. We have now examined the role of the C-terminal portion of the SSU by construction of truncated polypeptides (lacking 35, 48, 80, 95, or 112 amino acid residues from the C terminus) and examining the properties of holoenzymes reconstituted using these constructs. The Delta35, Delta48, and Delta80 constructs all lead to essentially complete activation of the catalytic subunits. The Delta80 construct, corresponding to the putative N-terminal domain, has the highest level of affinity for the catalytic subunits and leads to a reconstituted enzyme with k(cat)/K(M) about twice that of the wild-type enzyme. On the other hand, none of these constructs binds valine or leads to a valine-sensitive enzyme on reconstitution. The enzyme reconstituted with the Delta80 construct does not bind valine, either. The N-terminal portion (about 80 amino acid residues) of the SSU is thus necessary and sufficient for recognition and activation of the catalytic subunits, but the C-terminal half of the SSU is required for valine binding and response. We suggest that the C-terminal region of the SSU contributes to monomer-monomer interactions, and provide additional experimental evidence for this suggestion.