Lysine biosynthesis inMethanobacterium thermoautotrophicum is by the diaminopimelic acid pathway

Methanobacterium thermoautotrophicum, an archaebacterium, possesses the first and last enzymes of the diaminopimelic acid pathway for lysine biosynthesis, dihydrodipicolinate synthase, and diaminopimelate decarboxylase. It does not have saccharopine dehydrogenase, the last enzyme of the aminoadipate pathway for lysine biosynthesis. The dihydrodipicolinate synthase is inhibited but not repressed by lysine. We conclude that this microbe uses the diaminopimelate pathway for synthesis of lysine.Deceased.     

Rat striatal dopamine and tetrahydrobiopterin content following an intrastriatal injection of manganese chloride

Injection of manganese into the rat corpus striatum causes a rapid fall in the biopterin and dopamine (DA) content ipsilateral to the lesion. Two weeks after the lesion both biopterin and DA are partially recovered. Controls, injected with saline or magnesium, do not show alterations in their DA or cofactor levels. It is proposed that the fall in DA levels results from a rapid displacement of the amine from its storage sites by manganese followed by a decrease in the rate of DA synthesis causes by the drop in cofactor levels.     

Delimitation of morphologically similar sponge crab species of the genus Pseudodromia (Crustacea, Decapoda, Dromiidae) from South Africa

Stewart, B. A., Gouws, G., Daniels, S. R. & Matthee, C. A. (2004). Delimitation of morphologically similar sponge crab species of the genus Pseudodromia (Crustacea, Decapoda, Dromiidae) from South Africa. — Zoologica Scripta , 33 , 45–55.
Presently, three Pseudodromia sponge crab species are recognized, all of which are endemic to the continental shelf off the coast of South Africa. Two of these differ only in the morphology of their rostral teeth, making them difficult to distinguish, and can thus be considered as cryptic species. In addition they have very similar distribution ranges, thus raising doubts as to their specific status. Discriminant function analysis of morphometric data, differentiation at 10 allozyme loci, and sequence data derived from the 12S rRNA mitochondrial gene were used to test whether specimens identified as P. latens and P. rotunda are morphological forms of a single, widespread species, or represent two, distinct, reproductively isolated species, and to establish whether these two taxa are sister species, and thus form a monophyletic entity. The presence of fixed allele differences at three, and strong genetic heterogeneity at five other allozyme loci, indicating no gene flow occurring between sympatric populations, as well as the relatively high degree of 12S rRNA and allozyme genetic differentiation observed, supported the recognition of P. latens and P. rotunda as separate species. The 12S rRNA topology suggested that the genus Pseudodromia , as presently constituted, is paraphyletic, thus inferring that the morphological characters used to define this taxon might not be useful for phylogenetic inferences. It was concluded that in view of the uncertainties raised regarding the designation and composition of certain genera within the family Dromiidae, further rigorous analyses of morphological and genetic data are needed to further our understanding of the taxonomy of the sponge crabs.     

Gene expression in Brassica campestris showing a hypersensitive response to the incompatible pathogen Xanthomonas campestris pv. vitians

Xanthomonas campestris pv. vitians, a pathogen of lettuce, elicits a hypersensitive response within 12 hours of inoculation into Brassica leaves, characterized by tissue collapse, loss of membrane integrity, vein blockage and melanin production. In contrast, the compatible pathogen, X. c. pv. campestris, has no visible effects on leaves for 48 hours, after which inoculated areas show chlorosis which eventually spreads, followed by rotting.mRNA was prepared from leaves inoculated with suspensions of both pathovars or with sterile medium up to 24 hours following inoculation. In vitro translation of total and poly A⁺ RNA in rabbit reticulocyte lysate in the presence of ³⁵S methionine followed by separation of the polypeptide products by 2D-PAGE, allowed comparison of the effects of these treatments on plant gene expression. Major changes in gene expression were observed as a consequence of the inoculation technique. In addition, after inoculation with X. c. vitians, up to fifteen additional major polypeptides appeared or greatly increased by four hours. Some of these had disappeared by nine hours and several more had appeared. No major polypeptides disappeared or decreased greatly in intensity following inoculation with X. c. vitians.     

Species specificity of anti-acetylcholine receptor antibodies elicited by synthetic peptides

Two peptides corresponding to amino acid residues 351-368 of the alpha-subunits of Torpedo and human acetylcholine receptor (AChR) were synthesized. These peptides contain a segment (residues 355-364) which displays the greatest variability in amino acid sequence between the two species. Antibodies elicited against the two peptides cross-reacted with the respective native AChRs and were shown to be species specific by radioimmunoassay, immunoblotting, and immunofluorescence microscopy. Thus, antibodies against the Torpedo peptide cross-reacted with Torpedo AChR but did not bind to mammalian or chicken AChR. Antibodies against the human peptide proved to be specific probes for mammalian muscle AChR. They cross-reacted with mammalian AChR (human, calf, mouse, and rat) but not with Torpedo or chicken AChR. These antibodies were also shown to react preferentially with the extrajunctional form of muscle AChR, as compared to their reactivity with junctional muscle AChR. In immunofluorescence experiments, the anti-human peptide antibody stained AChR aggregates in sectioned or ethanol-permeabilized rat and mouse myotubes grown in culture but did not stain living myotubes. This indicates that the sequence 351-368 of the alpha-subunit of mammalian AChR is on the cytoplasmic face of muscle cell membranes, as predicted theoretically.     

Neuropeptide Y mobilizes intracellular Ca2+ and increases inositol phosphate production in human erythroleukemia cells

The intracellular concentration of free Ca2+ was monitored by measuring the fluorescence of fura-2 loaded Human Erythroleukemia Cells. Neuropeptide Y (NPY) increased intracellular Ca2+ in a dose-dependent manner and the 50% effective concentration was 2 nM. Chelation of extracellular Ca2+ by EGTA did not reduce the NPY-mediated increase in cytoplasmic Ca2+, indicating that the increase in fluorescence was due to the release of intracellular Ca2+. A second dose of NPY, after intracellular Ca2+ had returned to basal levels, failed to elicit a response, indicating that the NPY receptor had undergone desensitization. In similar experiments, NPY increased the formation of inositol phosphates, suggesting that the mobilization of Ca2+ from intracellular stores in HEL cells was secondary to the generation of inositol phosphates and stimulation of phospholipase C.     

The distribution of restriction enzyme sites in Escherichia coli.

A statistical analysis of physical map data for eight restriction enzymes covering nearly the entire genome of E. coli is presented. The methods of analysis are based on a top-down modeling approach which requires no knowledge of the statistical properties of the base sequence. For most enzymes, the distribution of mapped sites is found to be fairly homogeneous. Some heterogeneity in the distribution of sites is observed for the enzymes Pstl and HindIII. In addition, BamHI sites are found to be more evenly dispersed than we would expect for random placement and we speculate on a possible mechanism. A consistent departure from a uniform distribution, observed for each of the eight enzymes, is found to be due to a lack of closely spaced sites. We conclude from our analysis that this departure can be accounted for by deficiencies in the physical map data rather than non-random placement of actual restriction sites. Estimates of the numbers of sites missing from the map are given, based both on the map data itself and on the site frequencies in a sample of sequenced E. coli DNA. We conclude that 5 to 15% of the mapped sites represent multiple sites in the DNA sequence.     

MCA in patients with breast cancer: correlation with CEA and CA15-3

MCA serum levels were determined in 27 healthy subjects, 136 with benign pathology (42 breast) and in 289 patients with cancer (247 active). The last group includes 223 patients with breast cancer (96 without metastases, 89 with metastases and 38 no-evidence of disease). CEA and CA15-3 serum levels were determined in all the patients with breast diseases. The mean levels of MCA were 4.7 + 2.4 U/ml in the control group, considering less than 11 U/ml as normal. MCA values were abnormal in 15.4% of patients with benign pathology, mainly in those with liver cirrhosis (8/20) and lung diseases (4/20). In the majority of these cases, the rise was only moderate, lower than 15 U/ml in 97.5% of patients. In malignant diseases, important increments were found in breast cancer (19.8% Mo, 77.5% M1) and ovarian cancer stages III-IV (44.4%). When we compared MCA serum levels with CA15-3 and CEA in breast pathology, a similar specificity was observed: 92.3%, 92.3% and 100% in cases with benign pathology and 92.1%, 94.7%, and 97.4% in NED patients, respectively. MCA and CA15-3 sensitivity was similar in breast cancer without metastases (19.8%) and lower for CEA (16.7%). In patients with breast cancer without metastases, we found a relation between positivity of these tumor markers and prognostic factors (tumor size, nodal involvement). The disease free interval in patients with locoregional breast cancer was shorter in cases with abnormal presurgical levels of some of the tumor markers, but only the difference from MCA was significant (p less than 0.02).(ABSTRACT TRUNCATED AT 250 WORDS)     

Distribution of hobo transposable elements in the genus Drosophila

This study describes the distribution of hobo-hybridizing sequences in the genus Drosophila. Southern blot analysis of 134 species revealed that hobo sequences are limited to the melanogaster and montium subgroups of the melanogaster-species group. Of the hobo-bearing species, only D. melanogaster and two of its sibling species, D. simulans and D. mauritiana, were found to contain potentially complete hobo elements. The distribution of hobo sequences is one of the narrowest distributions thus far described for any Drosophila transposable element.