Identification of two novel arginine binding DNAs.

RNA tertiary structure is known to play critical roles in RNA-protein recognition and RNA function. To examine how DNA tertiary structure might relate to RNA structure, we performed in vitro selection experiments to identify single-stranded DNAs that specifically bind arginine, and compared the results with analogous experiments performed with RNA. In the case of RNA, a motif related to the arginine binding site in human immunodeficiency virus TAR RNA was commonly found, whereas in the case of DNA, two novel motifs and no TAR-like structures were found. One DNA motif, found in approximately 40% of the cloned sequences, forms of hairpin structure with a highly conserved 10 nucleotide loop, whereas the second motif is especially rich in G residues. Chemical interference and mutagenesis experiments identified nucleotides in both motifs that form specific arginine binding sites, and dimethylsulfate footprinting experiments identified single guanine residues in both that are protected from methylation in the presence of arginine, suggesting possible sites of arginine contact or conformational changes in the DNAs. Circular dichroism experiments indicated that both DNAs undergo conformational changes upon arginine binding and that the arginine guanidinium group alone is responsible for binding. A model for the G-rich motif is proposed in which mixed guanine and adenine quartets may form a novel DNA structure. Arginine binding DNAs and RNAs should provide useful model systems for studying nucleic acid tertiary structure.     

Influence of available aluminium on soil micro-organisms

P. ILLMER, K. MARSCHALL AND F. SCHINNER. 1995. Forest soils were selected which covered a wide range of aluminium concentrations (7 to μmol g^-1dry matter), but which differed as little as possible from one another in their soil chemical characteristics, including pH. These soils were examined with respect to microbial biomass and respiration, activity of cellulase, N-mineralization, colony-forming units of bacteria and fungi, and the concentrations of several inorganic soil components. The influences of altitude, climate, vegetation and, especially, of soil acidity could be kept to a minimum and so differences between the soil microfloras could clearly be attributed to Al concentration.
Al concentration was recognized to be the main inhibiting factor for the microbial biomass in soil. While N-mineralization was severely inhibited by aluminium, cellulase activity was hardly affected by increasing Al concentrations.
By taking the Al concentration along with various other soil chemical parameters a linear model could be developed that allowed more than 98% of the variability of the microbial biomass in soil to be explained.     

Methods and Errors in Measurements of Synovial Fluid Volume in Stifles with Low Volume and High Viscosity Synovial Fluid. An Experimental Study in Goats

Synovial fluid (SF) volume was calculated using various methods in the stifles of goats, in which the cranial cruciate ligament had been transected on one side. Measurements were performed prior to surgery and again 4,8, and 18 weeks following surgery, by measuring the dilution of an injected radioactive tracer diluted by the SF. Later, 7 months following surgery, SF volume measurements using simple arthrocentesis were performed on stifles in 9 of the goats, and the SF that could not be aspirated, was calculated using 2 indirect methods simultaneously on identical fluids in 3 of these goats. SF was also collected directly during staged arthrotomy of the stifles in 4 goats. There were conflicting results between methods, but the resulting calculated SF volumes seemed to be larger in the operated stifles compared to the controls for all the methods at about the same degree. The 2 indirect methods used to calculate the fluid remaining in the joints following arthrocentesis gave disparate volume calculations. The experiments revealed sources of error in all methods. Direct methods failed to acquire the total fluid volume, and indirect methods were subject to improper mixing and escape of the injected fluid or synovial fluid or both. It was concluded that none of the methods could be used to measure the “true” volume of SF, if such a concept exists and can be defined. None of the methods were considered reliable to compare volumes in different type of joints containing this type of fluid. It was, however, concluded that all the methods gave indication of increased SF volume present on a relative basis when paired joints were compared.     

Apical Chlorosis Disease of Chickpea (Cicer arietinum) Caused by Tomato Spotted Wilt Virus in Brazil

A severe new disease was observed in field-grown chickpea (Cicer arietinum L.) plants in Brasilia-DF, central Brazil. Symptoms were mainly general chlorosis accompanied by necrosis of the new growth and plant stunting, but pods were symptomless. Host range studies, serology, particle morphology, and cytopathology showed that tomato-spotted wilt tospovirus (TSWV) was the cause of the disease. This is apparently the first report of a chickpea disease caused by natural infection of a tospovirus.     

Muscarinic receptors in the prenatal mouse embryo. Comparison of M35-immunohistochemistry with [3H]quinuclidinyl benzylate autoradiography

Muscarinic cholinergic receptors are widespread in nervous tissue and smooth muscsle or paracrine epithelial cells of various organs. In the embryo, muscarinic receptors are transitorily expressed in the early blastoderm and later on in blastemic tissues during morphogenesis. Recently, a monoclonal antibody (M35) against muscarinic receptor from calf brain became available. In the present study the use of M35-immunohistochemistry is compared to autoradiographic localization of muscarinic binding sites in the mouse embryo. The aim of the study is to test the suitability of the antibody for localization of muscrinic receptors in embryonic tissues. For autoradiography whole-body sagittal cryostat sections of the 17- and 18-day mouse embryo were covered with LKB-Ultrofilm after incubation with the radioactive ligand [³H] quinuclidinyl benzylate (QNB). For immunohistochemistry cryostat sections of formalin fixed tissues were used. In general, all tissues exhibiting ligand binding were also recognized by the antibody. M35-immunohistochemistry resulted in higher spatial resolution of receptor localization than [³H]QNB autoradiography. Definitive muscarinic receptors were observed in smooth muscle and the epithelial lining of the vascular, intestinal, respiratory and urinary system, in the brain, spinal cord and peripheral nerves. The embryonic type of the muscarinic receptor was detected in the mesothelium of lung and liver, in the nephrogenic blastema of the metanephros, and in lung mesenchyme. A large amount of embryonic muscarinic receptors was found in the remnants of the notochord and in the nucleus pulposus of the developing vertebral column. A function in morphogenesis is discussed of the embryonic muscarinic receptor.     

Virulence factors (aerobactin and mucoid phenotype) in Klebsiella pneumoniae and Escherichia coli blood culture isolates

Abstract We examined the presence of two virulence factors in 241 blood isolates of Klebsiella pneumoniae from patients hospitalized during 1989 and 1990 in 7 French hospitals, and 125 blood isolates of Escherichia coli from one hospital. Aerobactin was scored phenotypically and genotypically with an intragenic DNA probe of 2 kb. The mucoid phenotype was assessed by culture on trypticase soy agar and by genotypic analysis (intragenic DNA probe of 235 bp). Only 6% K. pneumoniae isolates were aerobactin-positive with no significant variation according to geographical location while 20% of K. pneumoniae isolates displayed the mucoid phenotype, with a significant variation according to hospital. Aerobactin was always associated with the mucoid phenotype. The frequency of aerobactin production but not mucoid phenotype (14%) was higher among E. coli isolates (48%). They harbored two types of large plasmids. Intraperitoneal injection into mice of 10³ cfu of K. pneumoniae producing both virulence factors demonstrated that capsular serotype K2 was the more virulent K23 and K28.