In vitro and in vivo regulation of chloroplast glyceraldehydes-3-phosphate dehydrogenase isozymes from Chenopodium rubrum. I. Purification and properties of isozymes

Chloroplast glyceraldehyde-3-phosphate dehydrogenase (GPD, EC 1.2.1.13) was purified from leaves of Chenopodium rubrum L. Aggregated (≥ 10⁶) and disaggregated (165 × 10³) molecular weight forms were obtained by gel filtration in the presence of NAD⁺ and NADP⁺, respectively. The disaggregated enzyme was separated into two isozymes by inverse ammonium sulphate gradient solubilization: "NADP-GPD I" was homotetrameric (subunit molecular weight 39 × 10³); "NADP-GPD II" was heterotetrameric (subunit molecular weights 39 × 10³ and 43 × 10³). Isoelectric focusing of the isozymes, both aggregated and disaggregated, revealed two isoelectric forms in each case, at 4.3 and 7.7. Chloroplast GPD was "NADP-suppressed" in crude extracts due to partial oxidation, incubation with dithioerythritol restored full activity.     

Aspects of the ecology and energetics of the egg sac parasites of the wolf spider Pardosa lugubris (Walckenaer)

Summary The egg sacs of the wolf spider Pardosa lugubris are parasitised by two ichneumonid wasps, namely Gelis micrurus and Hidryta sordidus. The parasites are bivoltine, the first generation parasitising the summer egg sac of P. lugubris and the second generation parasitising, and overwintering in, the autumn egg sac. Parasitised egg sacs have a characteristic appearance.The degree of egg sac parasitism was 9.8% and 6.9% for the two egg sac batches in 1965 and 2.9% and 34.8% for those of 1966. The degree of parasitism varies from area to area as does the percentage parasitism attributable to the two species.The gross growth efficiency for Hidryta from the first egg sac is 31.2% and from the second 16.2%. 3.0% in 1965 and 4.3% in 1966, of the annual yield from P. lugubris to predators and parasites, goes to the egg sac parasites. This represents reductions in recruitment of 9.1% in 1965 and 10.9% in 1966.     

STUDIES ON CHLOROPLAST DEVELOPMENT AND REPLICATION IN EUGLENA : I. Vitamin B12 and Chloroplast Replication

When Euglena gracilis is grown under vitamin B₁₂ deficiency conditions, the amount of protein and of chlorophyll per cell increase with decrease of B₁₂ in the medium and consequently in the cell. The increase in cell protein is proportional to and precedes an increase in the number of chloroplasts per cell. This replication of the chloroplasts under deficiency conditions is not accompanied by nuclear or cell division. It is concluded that chloroplast replication in Euglena gracilis is independent of nuclear and cellular replication, at least under B₁₂ deficiency conditions. We established a graph of the growth of Euglena under different concentrations of vitamin B₁₂ added to the growth medium, which permitted us to calculate that at least 22,000 molecules of vitamin B₁₂ per cell are required to give normal growth.     

FEMORAL SHAFT FRACTURES—A Study of Closed Reduction and Open Treatment

A comparative study was made of 58 cases of closed femoral shaft fractures treated by skeletal traction, and 24 cases of closed femoral shaft fractures treated by open reduction with internal fixation.Although complications occurred in some cases, intramedullary nailing appeared to be the most satisfactory method, resulting in primary union, in decreased time of recumbency and time in hospital, in earlier ambulation and in less residual disability.Success of intramedullary nailing depends largely upon adequate training or experience of the surgeon in the technical operative aspects of the procedure and in postoperative management.Placing supplemental autogenous iliac bone chips at the fracture site in closed femoral fractures in which intramedullary nailing is performed appears to enhance callus formation and bony consolidation.Skeletal traction should be utilized on all patients whose general physical condition does not permit operative intervention.     

THE CELLULAR COMPLEMENT OF THE SKELETAL SYSTEM STUDIED AUTORADIOGRAPHICALLY WITH TRITIATED THYMIDINE (H3TDR) DURING GROWTH AND AGING

An autoradiographic study has been made of mouse femora from birth to one year of age. Tritiated thymidine was used to study the proliferative rate of various skeletal tissues. The labeling index of the periosteum was found to be highest at birth (0.085). At 8 weeks of age the index fell sharply to 0.007 and remained low throughout the entire period of study. Chondro-osteogenic cells of the periosteum, especially those at the perichondrial zone, were most frequently labeled initially. The labeling index of the epiphyseal disk remained high (0.054 to 0.048) until about the 6th week of age. Following this age the index fell sharply to 0.018 at 8 weeks and 0.002 at 26 weeks of age. Autoradiographs showed that the cells of the articulating surface of the epiphysis and disk are derived at least in part from migrating chondro-osteogenic cells of the periosteum residing at the perichondrial region, and that these cells serve as the progenitor pool necessary for both circumferential growth and expansion of the epiphyseal disk.     

Enzymes Involved in Anaerobic Polyethylene Glycol Degradation by Pelobacter venetianus and Bacteroides Strain PG1

In extracts of polyethylene glycol (PEG)-grown cells of the strictly anaerobically fermenting bacterium Pelobacter venetianus, two different enzyme activities were detected, a diol dehydratase and a PEG-degrading enzyme which was characterized as a PEG acetaldehyde lyase. Both enzymes were oxygen sensitive and depended on a reductant, such as titanium citrate or sulfhydryl compounds, for optimal activity. The diol dehydratase was inhibited by various corrinoids (adenosylcobalamin, cyanocobalamin, hydroxocobalamin, and methylcobalamin) by up to 37% at a concentration of 100 μM. Changes in ionic strength and the K⁺ ion concentration had only limited effects on this enzyme activity; glycerol inhibited the enzyme by 95%. The PEG-degrading enzyme activity was stimulated by the same corrinoids by up to 80%, exhibited optimal activity in 0.75 M potassium phosphate buffer or in the presence of 4 M KCI, and was only slightly affected by glycerol. Both enzymes were located in the cytoplasmic space. Also, another PEG-degrading bacterium, Bacteroides strain PG1, contained a PEG acetaldehyde lyase activity analogous to the corresponding enzyme of P. venetianus but no diol dehydratase. Our results confirm that corrinoid-influenced PEG degradation analogous to a diol dehydratase reaction is a common strategy among several different strictly anaerobic PEG-degrading bacteria.     

Characterization of the eaeA gene from rabbit enteropathogenic Escherichia coli strain RDEC-1 and comparison to other eaeA genes from bacteria that cause attaching-effacing lesions

Abstract A number of enteric pathogens, including enteropathogenic (EPEC) and enterohemorrhagic (EHEC) Escherichia coli , Hafnia alvei , a strain of Citrobacter freundii , and rabbit EPEC strain RDEC-1 cause attaching-effacing (AE) lesions in the gut mucosa. These bacteria have a pathogenicity cassette (locus of enterocyte effacement or LEE) containing the eaeA gene. This gene encodes intimin, an outer membrane protein required for production of AE lesions. RDEC-1, a non-invasive enteropathogen in young rabbits, produces AE lesions morphologically indistinguishable from lesions caused by human AE bacterial strains. The RDEC-1 example of E. coli diarrhea in rabbits is an important model for studying the pathogenesis of AE bacteria in a natural infection and for analyzing specific roles of the components of LEE. In order to better understand the role of intimin in the development of AE lesions, a portion of DNA within RDEC-1 LEE, containing the eaeA gene and an upstream open reading frame (ORF), was sequenced. The RDEC-1 eaeA gene shared 87%, 92%, and 93% DNA sequence identity and > 80% amino acid sequence identity with the eaeA genes of C. freundii biotype 4280, EHEC O157:H7, and EPEC O127:H6, respectively. The carboxy-terminal 280 amino acid residues of intimin has 80%, 56%, and 54% identity with C. freundii , EHEC O157:H7, and EPEC O127:H6 intimins, respectively. The predicted protein encoded by the upstream ORF (156 amino acids) shares 95%, 97%, and 99% amino acid identity with predicted proteins from C. freundii , EHEC O157:H7, and EPEC O127:H6, respectively. The high degree of sequence homology of the ORF and the eaeA gene of RDEC-1 with those of other AE bacteria suggests an evolutionary relationship of LEE and supports and facilitates the use of the RDEC-1 model for studying the role of LEE in pathogenesis.