This work reports the results of analyses of three complete mycoplasma genomes, a pathogenic (7448) and a nonpathogenic (J) strain of the swine pathogen Mycoplasma hyopneumoniae and a strain of the avian pathogen Mycoplasma synoviae; the genome sizes of the three strains were 920,079 bp, 897,405 bp, and 799,476 bp, respectively. These genomes were compared with other sequenced mycoplasma genomes reported in the literature to examine several aspects of mycoplasma evolution. Strain-specific regions, including integrative and conjugal elements, and genome rearrangements and alterations in adhesin sequences were observed in the M. hyopneumoniae strains, and all of these were potentially related to pathogenicity. Genomic comparisons

Chromobacterium violaceum is one of millions of species of free-living microorganisms that populate the soil and water in the extant areas of tropical biodiversity around the world. Its complete genome sequence reveals (i) extensive alternative pathways for energy generation, (ii) Ϸ500 ORFs for transport-related proteins, (iii) complex and extensive systems for stress adaptation and motility, and (iv) widespread utilization of quorum sensing for control of inducible systems, all of which underpin the versatility and adaptability of the organism. The genome also contains extensive but incomplete arrays of ORFs coding for proteins associated with mammalian pathogenicity, possibly involved in the occasional but often fatal cases of human C. violaceum infection. There is, in addition, a series of previously unknown but important enzymes and secondary metabolites including paraquat-inducible proteins, drug and heavy-metal-resistance proteins, multiple chitinases, and proteins for the detoxification of xenobiotics that may have biotechnological applications.T he genomes of soil-and water-borne free-living bacteria have received relatively little attention thus far in comparison to pathogenic and extremophilic organisms, yet they provide fundamental insights into environmental adaptation strategies and represent a rich source of genes with biotechnological potential and medical utility. A particularly interesting organism of this kind is Chromobacterium violaceum, a Gram-negative ␤-proteobacterium first described at the end of the 19th century (1), which dominates a variety of ecosystems in tropical and subtropical regions. This bacterium has been found to be highly abundant in the water and borders of the Negro river, a major component of the Brazilian Amazon (2) and as a result has been studied in Brazil over the last three decades. These, in general, have focused on the most notable product of the bacterium, the violacein pigment, which has already been introduced as a therapeutic compound for dermatological purposes (3). Violacein also exhibits antimicrobial activity against the important tropical pathogens Mycobacterium tuberculosis (4), Trypanosoma cruzi (5), and Leishmania sp. (6) and is reported to have other bactericidal (2, 7-10), antiviral (11), and anticancer (12, 13) activities.Some other aspects of the biotechnological potential of C. violaceum have also begun to be explored, including the synthesis of poly(3-hydroxyvaleric acid) homopolyester and other shortchain polyhydroxyalkanoates, which might represent alternatives to plastics derived from petrochemicals (14, 15), the hydrolysis of plastic films (16), and the solubilization of gold through a mercury-free process, thereby avoiding environmental contamination (17, 18). These studies, however, have been based on knowledge of only a tiny fraction of the genetic constitution of the organism. In addition, the more basic issues of the mechanisms and strategies underlying the adaptability of C. violaceum, including its observed but infrequent infection of h...

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Subacute inflammatory demyelinating polyneuropathy is a definite entity bridging the gap between Guillain-Barré syndrome and chronic inflammatory demyelinating polyneuropathy.

Electrophysiologic mapping experiments on the excitation of the A-V node of isolated rabbit heart have been performed with intracellular microelectrodes. An area of tissue showing peculiar physiologic characteristics was shown to coincide, at least in part, with the histologic A-V node and His bundle. Three functional regions have been determined in the physiologic mapping of the A-V node. The middle layer (N) shows signs of decremental conduction and is the site of the slowest propagation velocity in the A-V node. The other layers are transitional regions between the N layer and atrium (AN) or the His bundle (NH). The AN layer shows a progressively slower propagation velocity as the N region is approached. The same holds for the NH region. A precise correlation between these findings and the 3 existent histologic layers remains to be determined. The existence of an area of peculiar tissue just above the A-V valves and encircling the A-V orifice has been confirmed. This area is continuous with the AN layer of the functional A-V node and has been named the atrioventricular ring (AVR) on the assumption that it is a remnant of the embryonic A-V ring. Wenckebach cycles and A-V block of nodal origin are tentatively explained as resulting from increased decrement in the middle nodal layer (N).

Cooperativity in the interactions among proteins subunits and DNA is crucial for DNA recognition. LexA repressor was originally thought to bind DNA as a monomer, with cooperativity leading to tighter binding of the second monomer. The main support for this model was a high value of the dissociation constant for the LexA dimer (micromolar range). Here we show that the protein is a dimer at nanomolar concentrations under different conditions. The reversible dissociation of LexA dimer was investigated by the effects of hydrostatic pressure or urea, using fluorescence emission and polarization to monitor the dissociation process. The dissociation constant lies in the picomolar range (lower than 20 pM). LexA monomers associate with an unusual large volume change (340 ml/mol), indicating the burial of a large surface area upon dimerization. Whereas nonspecific DNA has no stabilizing effect, specific DNA induces tightening of the dimer and a 750-fold decrease in the K d . In contrast to the previous model, a tight dimer rather than a monomer is the functional repressor. Accordingly, the LexA dimer only loses its ability to recognize a specific DNA sequence by RecA-induced autoproteolysis. Our work provides insights into the linkage between protein-protein interactions, DNA recognition, and DNA repair.DNA-binding proteins recognize specific sequences of DNA, and the molecular basis for the interactions is currently the focus of intense research (1-3). X-ray crystallography and nuclear magnetic resonance spectroscopy have revealed that the reading of a DNA sequence is achieved by a combination of van der Waals', electrostatic, and hydrogen bonds (2, 3). The cooperative interactions among protein subunits, regulatory proteins, enzymes, and DNA are responsible for many biological functions, such as transcription, DNA replication, DNA repair, and differentiation (2, 4 -8). Most sequence-specific DNA-binding proteins are dimers or tetramers, and in many cases, the crystal structure of the protein-DNA complex reveals that each subunit of a dimer contacts each half of the operator (2, 3). In several cases dimerization only occurs upon DNA binding, as for example with the DNA-binding domains of the steroidnuclear receptors (9). Dimerization upon binding to DNA was also shown to be the relevant pathway for some basic region leucine zipper and basic region helix-loop-helix zipper transcription factors (10). However, in many DNA-binding proteins the subunits associate with high affinity in solution in the absence of DNA, with K d values in the range of 10 Ϫ7 -10 Ϫ10 M (2, 3, 11-13). In these latter cases, a free energy linkage between folding/oligomerization and DNA binding is still crucial for the recognition process (14 -18). LexA repressor is a 202-residue protein that regulates transcription in Escherichia coli of the SOS system, with about 17 genes that encode proteins involved in DNA repair (19,20). LexA has two domains, and it undergoes autocleavage at residues 84 -85 in a flexible hinge between the two domains (21). The so...

ABSTRACT. Growth by serial transfers of the trypanosomatid Crithidia deanei in culture medium containing 1 mg/ml of the β‐lactam antibiotics ampicillin or cephalexin resulted in shape distortion of its endosymbiont. The endosymbiont first appeared as filamentous structures with restricted areas of membrane damage. An increase of electron lucid areas was also observed in the endosymbiont matrix. The continuous treatment with β‐lactam antibiotics, resulted in endosymbiont membranes fragmentation; and later on the space previously occupied by the symbiont was identified as an electron lucid area in the host cytoplasm. The putative targets of β‐lactam antibiotic were two membrane‐bound penicillin‐binding proteins (PBPs) detected in the Sarkosyl‐soluble fraction of purified symbionts labeled with [3H]‐benzylpenicillin. The apparent molecular weight of the proteins were 90 kDa (PBP1) and 45 kDa (PBP2). PBP2 represented 85% of the total PBP content in the membrane fraction of the endosymbionts. Competition experiments using the tested antibiotics and [3H]‐benzylpenicillin showed that ampicillin and cephalexin have half saturating concentrations considerably higher than [3H]‐benzylpenicillin and indicated that PBP1 is the probable lethal target of the antibiotics tested. These results suggest that a physiologically active PBP is present in the cell envelope of C. deanei endosymbionts and may play important roles in the control of processes such as cell division and shape determination.

A new temperature-sensitive mutant strain of Escherichia coli K-12 which forms filaments at 42 C has been described. The mutant, Y16, maintained growth and deoxyribonucleic acid synthesis at 42 C. The resulting multinucleate filaments gradually lost their viability at 42 C but could be recovered, even after 240 min of incubation, upon return to 30 C. Septation was resumed and growth was promptly re-established at normal rates. Recovery still took place in the presence of chloramphenicol added to the culture at the time of temperature shift from 42 to 30 C. A study has been made of the effects of adenine and various nucleosides on cultures of strain Y16 as compared with another This paper K-12 (A) Wild type N. Zinder Hfr H(AB259) Hfr thi-1 rel-1 A-K. B. Low Ra-2 Hfr supE42 mal-28XA X K. B. Low KL25 Hfr supE42 X-K. B. Low KL14 Hfr thi-1 rel-1 A-K. B. Low PK191 Hfr A(proB-lac) X111 sup-36A