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Biomedical subjects

C Madrid

Publications and source records attributed to C Madrid.

At least 19 recordsLinked to original sources
PubMed2002

Evidence for direct protein-protein interaction between members of the enterobacterial Hha/YmoA and H-NS families of proteins.

Escherichia coli nucleoid-associated H-NS protein interacts with the Hha protein, a member of a new family of global modulators that also includes the YmoA protein from Yersinia enterocolitica. This interaction has been found to be involved in the regulation of the expression of the toxin alpha-hemolysin. In this study, we further characterize the interaction between H-NS and Hha. We show that the presence of DNA in preparations of copurified His-Hha and H-NS is not directly implicated in the interaction between the proteins. The precise molecular mass of the H-NS protein retained by Hha, obtained by mass spectrometry analysis, does not show any posttranslational modification other than removal of the N-terminal Met residue. We constructed an H-NS-His recombinant protein and found that, as expected, it interacts with Hha. We used a Ni(2+)-nitrilotriacetic acid agarose method for affinity chromatography copurification of proteins to identify the H-NS protein of Y. enterocolitica. We constructed a six-His-YmoA recombinant protein derived from YmoA, the homologue of Hha in Y. enterocolitica, and found that it interacts with Y. enterocolitica H-NS. We also cloned and sequenced the hns gene of this microorganism. In the course of these experiments we found that His-YmoA can also retain H-NS from E. coli. We also found that the hns gene of Y. enterocolitica can complement an hns mutation of E. coli. Finally, we describe for the first time systematic characterization of missense mutant alleles of hha and truncated Hha' proteins, and we report a striking and previously unnoticed similarity of the Hha family of proteins to the oligomerization domain of the H-NS proteins.

Amino Acid Sequence↗
PubMed2000

Expression of the hemolysin operon in Escherichia coli is modulated by a nucleoid-protein complex that includes the proteins Hha and H-NS.

The Escherichia coli protein Hha is a temperature- and osmolarity-dependent modulator of the expression of the hemolysin operon. The Hha protein was purified and its DNA-binding properties analyzed. Hha binds in a non-specific manner throughout the upstream regulatory region of the hemolysin operon in the recombinant hemolytic plasmid pANN202-312. A search for interacting proteins revealed that Hha interacts with H-NS. DNA-binding studies showed that, in vitro, Hha and H-NS together form a complex with DNA that differs from those formed with either protein alone. These data, together with the effects of hha and hns mutations on the expression of the hemolysin genes, suggest that in vivo H-NS and Hha form a nucleoid-protein complex that accounts for the thermo-osmotic regulation of the hemolysin operon in E. coli.

Bacterial Proteins↗
PubMed2000

Spatiotemporal profile of synaptic activation produced by the electrical and visual stimulation of retinal inputs to the optic tectum: a current source density analysis in the pigeon (Columba livia).

The optic tectum of the pigeon is a highly organized, multilayered structure that receives a massive polystratified afference of at least five different populations of retinal ganglion cells and gives rise to various anatomically segregated efferent systems. The synaptic organization of retino-tectal circuitry is, at present, mostly unknown. To investigate the spatiotemporal profile of synaptic activation produced by differential (electrical and visual) stimulation of the retinal inputs, we performed a high-spatial-resolution current source density analysis in the optic tectum of the anaesthetized pigeon. Electrical stimuli consisted of brief pulses of different durations applied to the optic nerve head, while visual stimuli consisted of light flashes of different intensities. Electrical stimulation generated sinks confined to retinorecipient layers. The temporal structure, spatial location and thresholds of these sinks indicated that they are all due to primary tectal synapses of retinal fibers with different conduction velocities. Sinks evoked by the fastest retinal axons were more superficially located than sinks produced by slower retinal fibers. Visual stimulation, on the other hand, resulted in a more complex pattern of current sinks, with various sinks located in the retinorecipient layers and also well below. Visual stimulation induced action potentials at superficial as well as deep tectal levels. We conclude that electrical stimulation activates most of the populations of ganglion cells as well as their primary tectal synapses, but is unable to elicit a significant activation of secondary tectal synapses. Visual stimulation, on the contrary, activates just some of the incoming retinal populations, but in a way that produces noticeable secondary activation of intratectal circuits. Laminar segregation of retinally evoked tectal activity, as reported here, has also been found in other vertebrates. Similarities and differences with previous studies are discussed.

Animals↗
PubMed1999

Reversible mechanical fixation of eye position in awake head-restrained pigeons (Columba livia).

Here we describe a method to fix gaze positions and to significantly reduce saccadic oscillations in pigeons. The procedure consists of a mechanical immobilization of the eye through the use of an electromagnet that exerts a radial force upon a small metal rectangle glued to the dorsal part of the eye. The method can be used in avian visual neurophysiology in order to hold the eye immobilized for periods of time, long enough to map the properties of visual receptive fields and investigate the possible functions of saccadic oscillations.

Animals↗
PubMed1999

[The architectural balance of the face: a 3D cephalometric concept].

The use of the c2000 software led us to develop a new 3D cephalometric construction, based on the selection on the axial CT Scans of 8 anatomical landmarks and of teeth, all of which were situated along the trigeminal neuro matriciel facial growth axes. The analysis of this construction is based on the use of an original mathematical tool in biology: the axes of inertia. Using the selection of both mental foramen, both infra and supra orbital foramen and the head of both malleus, the C 2000 software creates a geometrical construction called: "the maxillo-facial frame", as well as, a 3D cephalometric analysis: angles, distances, areas, volumes center of gravity and axes of inertia. Using the selection of teeth, the C 2000 software calculates the axes of inertia of each tooth or of groups of teeth. The use of the axes of inertia allow us to create of a hierarchy of anatomical levels the teeth, the half arches, the arches, both arches and the maxillo-facial frame. In addition, for each of these anatomical levels, the axes of inertia create a 3D landmark which allows the calculation of the orientation of each of these elements in relation to the others. The study of 28 orthomorphic people using this analysis revealed the existence of a maxillo-facial balance that is unique for each individual.

Adolescent↗
PubMed1998

Osmolarity modulates the expression of the Hha protein from Escherichia coli.

The effect of the osmolarity of the culture medium on the expression of the hha gene of Escherichia coli was investigated. When cells were grown in LB medium, expression reached a maximum in the exponential phase of growth and decreased in the stationary phase. Increasing the osmolarity of the LB medium had no significant effect on the expression of the hha gene, but depletion of NaCl led to a significant decrease in expression. Expression of the hha gene is thus sensitive to the osmolarity of the growth medium. High levels of expression of the hha gene when cells are grown at medium to high osmolarity are consistent with the finding that the Hha protein appears to play its main modulatory role when cells grow under these conditions.

Bacterial Proteins↗
PubMed1997

Construction of a double hha hns mutant of Escherichia coli: effect on DNA supercoiling and alpha-haemolysin production.

A double hha hns Escherichia coli mutant was constructed. The effect of the single hns mutation and of the double hha hns mutation on the expression of the alpha-haemolysin determinant of plasmid pANN202-312 was assessed. Whereas the hns mutant moderately increased expression of the toxin, the double hha hns mutant strongly enhanced transcription of the hly operon and hence expression of the toxin. This suggests that both Hha and H-NS proteins participate in the modulation of the expression of the toxin. The enhancement of haemolysin expression in the double mutant could not be correlated to a global alteration of DNA topology: DNA preparations of a reporter plasmid isolated from this mutant gave a topoisomer distribution similar to that of the parental strain.

Bacterial Proteins↗
PubMed1997

Distribution and characterization of faecal verotoxin-producing Escherichia coli (VTEC) isolated from healthy cattle.

Faecal swabs obtained from a random sample of 268 cows and 90 calves on 19 Lugo farms were examined for verotoxin-producing Escherichia coli (VTEC). We found VTEC on 95% of the farms. The prevalence rates of VTEC infection in asymptomatic cows and calves were estimated to be 35 and 37%, respectively. The proportion of animals infected on each farm ranged from 0 to 100%. VTEC strains isolated from healthy cattle belonged to 27 O serogroups; however, 57% (85 of 149) were of one of 8 serogroups (O2, O8, O22, O77, O82, O105, O113 and O171). Nearly 60% of the bovine VTEC strains belonged to serogroups that cause haemorrhagic colitis and haemolytic uraemic syndrome in humans. The VTEC were all non-O157:H7; 91% were eae-negative and 86% produced VT2 or VT1 and VT2. These characteristics are different from those of VTEC isolated from calves with diarrhoea.

Animals↗
PubMed1997

[A new 3-dimensional cephalometry model. A new 3-dimensional parameter analysis: the axis of inertia. A new idea: maxillofacial equilibrium].

The authors propose a biomensurativ 3 D method based on the plotting of 8 anatomical landmarks situated on the trigeminal axis and the selection of teeth. In the analysis of this construction, we have employed mathematical tools unusual in developmental biology, the axis of inertia. The 3 D analysis allows to evidence the architectural, structural and functional balance of the dentomaxillofacial complex. This balance must be substituted to the normality concept to directly linked to cultural criterias and to the selection of mean parameters among culturally and ethnically homogeneous populations. Therapy does not have to lead to the normality of mean parameters but to the return or to the discovery of a real balance.

Cephalometry↗
PubMed1996

Complementation of the hha mutation in Escherichia coli by the ymoA gene from Yersinia enterocolitica: dependence on the gene dosage.

The Hha protein from Escherichia coli is highly similar (82%) to the YmoA protein from Yersinia enterocolitica. Both are members of a new class of proteins that modulates gene expression, probably by influencing DNA topology. In this paper, complementation of the hha mutation in E. coli by the ymoA gene from Y. enterocolitica has been studied. We show that the ymoA gene complements one of the phenotypic properties of hha mutants (high level of haemolysin production when they carry the recombinant plasmid pANN202-312) when cloned in a medium-copy-number plasmid but not when carried in a low-copy-number plasmid. Western blot analysis of the expression of YmoA in E. coli rules out inefficient expression of the protein. Surprisingly, the hha gene itself fails to complement the hha mutation when cloned in a medium-copy-number vector and causes genetic rearrangements of the E. coli chromosome as a consequence of insertion sequences mobilization.

Bacterial Proteins↗
PubMed1996

The Hha protein as a modulator of expression of virulence factors in Escherichia coli.

We constructed hha derivatives from both a clinical uropathogenic Escherichia coli isolate (strain FVL4) and a wild E. coli strain causing bovine diarrhea (strain CCB21) and analyzed the effect of the hha allele on the expression of the different virulence factors exhibited by these strains. Expression of hemolysin and of the Vir antigen was altered in hha mutants. Whereas production of hemolysin by strain FVL4 was repressed both at a low temperature and at high osmolarity, the hha allele accounted for a significant increase of hemolysin production under these conditions. Also, the low temperature-sensitive expression of the Vir adhesin was modified in hha mutants, which were able to express this adhesin at a low temperature. Expression of other virulence factors, such as cytotoxic necrotizing factor type 1 and 2 toxins, remained unmodified in hha derivatives of strains FVL4 and CCB21.

Adhesins, Bacterial↗
PubMed1994

Environmental regulation of alpha-haemolysin expression in Escherichia coli.

The effect of osmolarity, temperature and anaerobiosis on the expression of Escherichia coli alpha-haemolysin was investigated. Low osmolarity of the culture medium, growth at high temperature (37 degrees C) and anaerobiosis increase haemolysin production in E. coli cells harbouring the haemolytic plasmid pHly152 as well as in haemolytic strains isolated from urinary-tract infections.

Anaerobiosis↗
PubMed1994

Isolation and characterization of a Tn5-induced tolQ mutant of Escherichia coli.

Transposon mutagenesis was used to isolate an Escherichia coli mutant that released into the external medium a heterologous protein, the Aeromonas hydrophila aerolysin. Genetic mapping and phenotypic characterization of E. coli CM209 showed that Tn5 is inserted in the tolQ gene. This mutant strain released a significant amount of unprocessed (proaerolysin) protein into the external medium, together with other periplasmic proteins. Similar levels of the toxin were detected in the intracellular compartments of the parental and mutant strains. Whereas inactivation of the tolQ gene itself was responsible for some of the phenotypic properties of strain CM209, such as tolerance to group A colicins or to filamentous bacteriophages, the leaky phenotype was associated with a polar effect exerted by Tn5 on distal genes of the tolQRA cluster.

Aeromonas hydrophila↗
PubMed1994

Excision and transposition of Tn5 upon insertion in the hha gene of Escherichia coli.

We report the occurrence of Tn5 secondary transpositions in Escherichia coli HB101 hha::Tn5(pANN202-312). Tn5-induced hha mutants (hemolysin overproducers) segregated at high frequency either hemolysin-negative clones or clones showing the parental hemolytic phenotype (reduced hemolysin production). Secondary transpositions of Tn5 appeared to be responsible for the phenotypes of both types of derivatives. The hemolysin-negative clones no longer harboured Tn5 in the hha gene, but rather Tn5 was in the hly genes of the hemolytic plasmid pANN202-312. The derivatives that recovered the parental hemolytic phenotype contained a single copy of Tn5 in a chromosomal location different from that of hha. Both kinds of transpositional events appeared to restore the function of the hha gene.

Base Sequence↗
PubMed1994

Orthopedic case management in a collaborative practice setting.

An orthopedic practice that consists mainly of total joint replacements is an ideal setting for a case manager. While fixed reimbursement demands that care be cost-effective, an older patient population presents with more complex health problems. Key factors towards achieving successful patient outcomes are thorough preoperative assessment, patient and family education, and the development of critical pathways.

Group Practice↗
PubMed1993

A tridimensional study using cuts at a low temperature of the infratemporal region.

Although the infratemporal region is well defined anatomically, its complex topography has been the subject of numerous, and sometimes, opposite works. That is the reason why it appeared necessary for the authors to re-evaluate this topic using the original method of Combelles and Boyer, allowing to define three referential planes, and thereby, a tridimensional shape and the volume of this region. This study allows to conclude that the infratemporal region is a triangular prism with an horizontal main axis 47 mm long. Its anterior base has a mean area of 733 mm2. The posterior top consists of the Juvara slot and has a mean area of 490 mm2. This infratemporal prism contains another one, the pterygomandibular space, prismatic too. It widens out from coronoïd plane (93 mm2) to mandibular foramen plane (169 mm2) before ending as a narrow groove between the neck of the mandibular condyle and the interpterygoïd fascia. The volume of the pterygomandibular space is quite superior to the value usually reported in the dental literature. It is of 4.8 ml to 5.8 ml according to denture. These results point out the opportunity to accomodate more important volumes of anesthesic solutions, than the 1.8 ml usually performed, without any leak out of the infratemporal region.

Aged↗
PubMed1992

[A study of the cranio-pharyngo-mandibular space by sections at low temperature].

After acrylic injections into the vessels and mandibular block anaesthesia the pterygo-mandibular space is studied in 38 anatomical pieces. 538 colour slides have been obtained from anatomical cuts at low temperature with the scannigraph system. The results point out that a cranio-pharyngo-mandibular space is clearly distinguished laterally, from a medial maxillo-pharyngeal space. There is no transversal communication between them. On the contrary, the cranio-pharyngo-mandibular space communicates, forward and upward, with the temporal space; forward and downward, with the sub-mandibular space. The cranio-pharyngo-mandibular space contains an interpterygoid space communicating with the oval foramen by a "chimney" largely filled by the emissary pterygoid veins. A pericondilar plexus of veins is also observed. The cranio-pharyngo-mandibular space is characterised by the cellulo-adipose tissue by which it is filled and which guides the diffusion of injected solutions to the oval foramen.

Aged↗