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Biomedical subjects

D C Phillips

Publications and source records attributed to D C Phillips.

71 records · Page 4Linked to original sources
PubMedDate not supplied

X-ray crystallography of the binding of the bacterial cell wall trisaccharide NAM-NAG-NAM to lysozyme.

Hen egg white lysozyme was the first enzyme whose structure was determined by X-ray crystallography. The proposed mechanism based on this structure involves the distortion of the saccharide residue (2-acetamido-2-deoxy-D-muramic acid, NAM) in the natural substrate (an alternating beta (1 leads to 4) linked oligomer of 2-acetamido-2-deoxy-D-glucose (NAG) and NAM residues) bound to site D in the binding cleft. The importance of substrate distortion has prompted numerous enzymatic, chemical, theoretical, and physical studies, but there is little direct crystallographic evidence on the conformation of a NAM residue bound at site D. We now present the X-ray structure of the non-hydrolysed trisaccharide NAM-NAG-NAM bound in subsites B, C, D. Our interpretation of the 2.5-A resolution difference map does not involve distortion of this residue in site D. Comparison with the structure of the delta-lactone derived from tetra N-acetylchitotetraose (NAG)3NAL) bound to lysozyme suggests we may be looking at a Michaelis complex.

Binding Sites↗
PubMedDate not supplied

Tertiary structural similarity between a class A beta-lactamase and a penicillin-sensitive D-alanyl carboxypeptidase-transpeptidase.

beta-Lactam antibiotics--the penicillins, cephalosporins and related compounds--act by inhibiting enzymes that catalyse the final stages of the synthesis of bacterial cell walls. Recent crystallographic studies of representative enzymes are beginning to reveal the structural bases of antibiotic specificity and mechanism of action, while intensive efforts are being made to understand the beta-lactamase enzymes that are largely responsible for bacterial resistance to these antibiotics. It has been suggested that the beta-lactamases and beta-lactam target enzymes may be evolutionarily related and some similarity of amino-acid sequence around a common active-site serine residue supports this idea. We present here the first evidence from a comparison of three-dimensional structures in support of this hypothesis: the structure of beta-lactamase I from Bacillus cereus is similar to that of the penicillin-sensitive D-alanyl-D-alanine carboxypeptidase-transpeptidase from Streptomyces R61.

Carboxypeptidases↗
PubMedDate not supplied

Alpha-lactalbumin possesses a novel calcium binding loop.

Calcium performs a unique role in biology, achieving biological effects through highly specific interactions with and modulation of target proteins. It has been proposed that calcium-modulated proteins possess a characteristic, evolutionarily related, binding fold, known as the EF-hand. The high-resolution X-ray structure of alpha-lactalbumin reveals a Ca2+ binding fold that resembles an EF-hand only superficially and presumably has no evolutionary relationship with it. However, there is clear homology with the corresponding loop in c-type lysozyme (the 'parent' molecule of alpha-lactalbumin). This study, at 1.7 A resolution, represents one of the most accurate analyses of a calcium binding protein yet reported.

Binding Sites↗
PubMedDate not supplied

Normal and abnormal middle ear ventilation.

Studies in infants and children have suggested a functional rather than mechanical obstruction of the Eustachian tube as a predisposing factor in middle ear effusions (MEE). To simulate this condition in the laboratory, an animal model was prepared using juvenile Rhesus monkeys. The tensor veli palatini muscle was transected or expunged posterior to the hamulus of the medial pterygoid lamina. Transection of the muscle resulted in negative middle ear pressure without effusion, whereas when the muscle was expunged, the animals developed a brief episode of negative middle ear pressure followed by a persistent MEE that was sterile for bacteria. An acute bacterial MEE developed following instillation of Streptococcus pneumoniae into the nasopharynx of animals that had had a previous unilateral transection of the muscle. The condition of the middle ear was documented by impedance measurements and presence of the effusion was verified by myringotomy. Animals were periodically examined and tested for Eustachian tube ventilatory function over a period of one year. Before surgical alteration of the tensor muscle. Eustachian tube function tests demonstrated normal ventilatory function, whereas, functional Eustachian tube obstruction patterns similar to studies in children who had MEE were found during the postoperative period. Only after the development of a reliable animal model can current and future methods of management of MEE be tested under controlled laboratory conditions. These data suggest that the Rhesus monkey appears to be an excellent model for the study of normal as well as abnormal tubal function.

Animals↗
PubMedDate not supplied

Dilation of the eustachian tube by electrical stimulation of the mandibular nerve.

The recent studies of the anatomy of the eustachian tube and related structures in the Rhesus monkey (Macaca mulatta) have shown that the monkey tubal system is similar to the human. This investigation in Rhesus monkeys was an attempt to verify previous studies in other animals that the tensor veli palatini muscle was the only dilator of the eustachian tube. Two unipolar stimulating electrodes were introduced into the foramen ovale, and the mandibular branch of the trigeminal nerve was electrically stimulated. Simultaneously, pressure-flow recordings through the eustachian tube were monitored. Stimulus-response relationships were obtained for five Rhesus monkeys. The degree of tubal dilation by the tensor veli palatini muscle contraction was shown to be a function of stimulating current levels. Artifically induced dilations were quite similar to the physiological dilations during swallowing when these animals were tested alert. Following complete transection of the tensor muscle, regardless of the stimulus level, no tubal dilations were observed. Stimulation of the nerve to the internal pterygoid and stimulation of the levator veli palatini muscle induced only constrictions of the tube. The tensor veli palatini muscle is the only paratubal muscle responsible for active dilation of the eustachian tube in the Rhesus monkey, and its motor innervation is the mandibular division of the trigeminal nerve.

Animals↗
PubMedDate not supplied

Nonhuman primate model of cleft palate and its implications for middle ear pathology.

The present study is a preliminary report on the development of a nonhuman primate model of cleft palate and middle ear (ME) disease. The causal relationship between a surgical cleft of the soft palate only or a cleft of the hard and soft palate and otitis media with effusion (OME) was investigated in rhesus monkeys. Prior to clefting, ME status was documented by pneumatic otoscopy or otomicroscopic examination and tympanometry over a period of at least five months. A minimum of four preoperative eustachian tube (ET) function evaluations were performed employing the inflation-deflation and the forced-response tests. These procedures were repeated following surgery and during a long-term follow-up. Seventeen of the 18 ears developed a recurrent OME. Postoperative ME pressures were initially high negative values. After the first two postoperative months, high positive ME pressures were recorded. The forced-response test showed little to no long-term changes in passive and active tubal resistances or in the efficiency of tubal dilation as a result of surgery. The inflation-deflation test showed higher opening and closing pressures and a limited and more variable ability to equilibrate applied positive and negative ME pressures following surgery. Both ME status and ET function appeared to improve with time. These findings indicated that the pathogenesis of recurrent OME in this animal model may have been due to changes in ET function associated with an abnormal nasopharynx rather than aberrant tensor veli palatini (TVP) muscle function.

Animals↗
PubMedDate not supplied

Effect of surgical alterations of the tensor veli palatini muscle on eustachian tube function.

Previous studies in humans have indicated that functional obstruction of the eustachian tube (ET) is an important factor in the pathogenesis of otitis media with effusion (OME). This type of obstruction appears to be related to the structural properties of the tube, or to an inefficient active tubal opening mechanism, or both. In this study, functional ET obstruction was created in 22 rhesus monkeys (Macaca mulatta) by surgically altering the tensor veli palatini (TVP) muscle using three different procedures: 1) complete excision of the muscle; 2) transection of the superficial muscle bundle; or 3) transposition of the muscle tendon medial to the hamular process. Prior to surgery, weekly tympanometry, pneumatic otoscopy, and otomicroscpic examinations were performed for a period of at least six months to document middle ear (ME) status. A minimum of four ET function tests were performed on each animal using the inflation-deflation and forced-response tests. Following surgery, these tests and examinations were continued for periods of up to one year. Postoperatively, the animals in which the TVP had been excised developed a sterile ME effusion which proved to be a chronic condition which persisted throughout the follow-up period. Eustachian tube function tests showed a complete absence of any active tubal dilation by swallowing. Animals that had the muscle transected developed abnormal ME pressures , or effusions, or both, which returned to normal in some ears, but which were recurrent or chronic in others. Eustachian tube function tests in these animals showed an initial loss of active tubal function which gradually improved, but not to normal levels, presumably as a result of healing of the muscle. In cases in which the muscle was transposed the ME pathology and ET dysfunction were similar after the surgery, but improved within a short period of time. These data suggest that alteration of the TVP muscle can create functional obstruction of the ET. The severity of ET obstruction depends upon the surgical procedure undertaken. The results of postoperative ET function tests were similar to those recorded from children with recurrent and chronic OME.

Animals↗
PubMedDate not supplied

Gas absorption in the middle ear.

The middle ear (ME) pressure of alert and anesthetized rhesus monkeys was monitored by serial tympanograms in order to determine the gas absorption process in the ME cavity. During the four-hour observation period the ME pressure showed small, random fluctuations around zero pressure in the alert animals, whereas in the anesthetized animals, following an initial positive pressure phase, the ME pressure reached a plateau at an average pressure of -60 mm H2O. The ME gas composition was changed by air or oxygen gas politzerization in anesthetized animals to determine the effect of different initial gas compositions on the rate of absorption of the gas and on the magnitude of ME pressure. Politzerization with air and with oxygen gas resulted in average maximum ME pressures of -400 and -730 mm H2O, respectively. The experimental evidence suggested that the physiological composition of ME gases is nearly that of the surrounding environment and that the rate of gas absorption is low. When the gas composition of the ME is changed, the ME pressure decreases considerably. Based on these findings, a possible mechanism for the development of negative ME pressure is proposed.

Absorption↗