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M W King

Publications and source records attributed to M W King.

At least 19 recordsLinked to original sources

POU-domain sequences from the flatworm Dugesia tigrina.

Redundant primers matching well-conserved sequences within vertebrate and invertebrate POU genes were used in the PRC (polymerase chain reaction) to detect three different POU-domain-containing genes within the genome of the flatworm, Dugesia tigrina. Two of these genes appear to be close homologues of class-III POU genes previously identified in Dugesia japonica, while the third represents a new class-III POU gene. Nucleotide sequences encoding highly related POU domains within the two species of planaria exhibited a surprisingly poor degree of shared identity. Since only class-III POU genes have been observed to date in flatworms, these genes may represent ancestral versions of the multiple classes of POU genes that exist in vertebrates, and may play important roles in the development of metazoan nervous tissue.

Amino Acid Sequence

Evaluating the Dialine vascular prosthesis knitted from an alternative source of polyester yarns.

The sudden and unilateral decision by E. I. DuPont de Nemours & Co., Inc., to withdraw its polymers for use in implantable devices has presented the medical device industry with an immediate and serious challenge to find alternative sources of biomaterials. In France, the company Cardial S.A. has already taken steps to find an alternative polyester yarn to replace Dacron by developing a new arterial prosthesis knitted from polyester yarns supplied by Rhône-Poulenc Fibres. This article describes an in vitro and in vivo study of this French device, called the Dialine prosthesis, with a view to determining its relative performance compared to current American and British prostheses, which rely on DuPont's Dacron yarn. In addition to analyses of the morphology and textile structure, and measurements of its physical and chemical properties, the Dialine graft was implanted as a thoracoabdominal bypass in dogs for periods ranging from 4 h to 6 months. In addition to our pathologic and histologic observations, we cleaned and evaluated the explanted prostheses for in vivo changes in dimensions, strength, and crystalline microstructure. The Dialine graft was found to differ structurally from other polyester prostheses because it is warp-knitted from a mixture of flat and texturized yarns with finer filaments. Its denser structure has a lower water permeability, greater flexibility and ease of handling, satisfactory strength, and dimensional stability, and it presents different textures on its luminal and external surfaces. The in vivo trial demonstrated that it has excellent biocompatibility and biostability over 6 months. With no thrombi observed on the luminal surface after 3 months, it has a faster rate of healing, generates compact external and internal capsules with a thinner neointima, and has an overall milder inflammatory response than is normally observed with Dacron-based prostheses.

Angiography

Polyvinylidene fluoride (PVDF) as a biomaterial: from polymeric raw material to monofilament vascular suture.

This study identified the effects of various manufacturing processes on the crystalline microstructure, mechanical properties, and biocompatibility of a polyvinylidene fluoride (PVDF) suture. To achieve this, changes in the crystalline microstructure and the tensile behavior of PVDF monofilaments were monitored in vitro after different thermal processing, coloration, and sterilization treatments. In addition, the in vivo biocompatibility of the manufactured and sterilized PVDF suture was assessed by using it to anastomose a preclotted polyester vascular prosthesis as a thoracoabdominal bypass in a series of dogs. The tissue response was followed by histologic and scanning electron microscopy over implantation periods ranging from 4 h to 6 months. Differential scanning calorimetry and infrared spectroscopy (FTIR-ATR) showed that thermal processing and the addition of a coloring agent had a direct effect on modifying the crystalline microstructure and hence changing the mechanical properties. For example, thermal processing converted some of the alpha phase into the beta and gamma polymorphs, whereas coloration led only to a major increase in the beta-to-alpha ratio. The tensile properties were found to be optimized when the relative proportion of the beta and gamma phases combined compared to the alpha form gave rise to an FTIR A509/A532 absorption ratio between 4.0 and 4.5. Sterilization was found to cause some modifications to the crystalline microstructure near the surface of the monofilaments, but it did not change their mechanical properties. Pathologic examination of the anastomotic regions after different periods of implantation revealed a minimal cellular response, with no mineralization, intimal hyperplasia, or excessive fibrous tissue reaction. This good biocompatibility, together with other desirable characteristics such as ease of manipulation and satisfactory mechanical strength, makes PVDF an attractive alternative monofilament suture material for cardiovascular surgery.

Animals

Carbodiimide cross-linked gelatin: a new coating for porous polyester arterial prostheses.

The performance of a polyester arterial prosthesis impregnated with gelatin and cross-linked with carbodiimide (Uni-graft) was compared with its porous parent graft (Protegraft) using a canine thoraco-abdominal bypass model. The grafts were investigated in terms of their handling characteristics, imperviousness at implantation, surface thrombogenicity and healing behaviour. Prostheses 30 cm in length were implanted for the following periods: 4, 24 and 48 h, 1, 2 and 4 weeks, 2, 3, 4, 5 and 6 months. Both types of graft had good handling characteristics. The ready-to-use impregnated graft provided satisfactory haemostasis at implantation with no blood permeating through the wall after flow was restored. Both grafts exhibited low surface thrombogenicity, as determined by the uptake of labelled fibrin and platelets, and the healing sequence of the impregnated graft after resorption of the gelatin was equivalent to that of the preclotted control. Biodegradation of the gelatin was complete within 1 month of implantation with the subsequent development of a collagenous internal capsule at both anastomoses. Endothelial cells were observed between 4 and 6 months, but were confined to small islets distributed along the luminal surface. The prostacyclin/thromboxane A2 (PGI2/TXA2) ratio, which gives an indication of the level of endothelial cell activity, was greater than 1.0 after 1 week of implantation for the control graft. For the impregnated graft it reached 1.0 only after 3 months of implantation, but remained above 1.0 for periods of up to 6 months.(ABSTRACT TRUNCATED AT 250 WORDS)

Analysis of Variance

Removing fresh tissue from explanted polyurethane prostheses: which approach facilitates physico-chemical analysis?

Chemical, physical and structural analyses of polymers from explanted vascular prostheses are frequently jeopardized because of incomplete removal of the encroaching host tissue. In this study, microporous polyurethane arterial prostheses implanted as a canine thoraco-abdominal bypass were explanted after 1 and 12 months and were cleaned without fixation using four different digesting enzyme treatments, including collagenase, pancreatin and trypsin alone and collagenase and pancreatin in series, followed by washing in a solution of Triton X-100 detergent. By following this approach all the fresh tissue attached to the external and internal walls of the prostheses was removed with minimal damage to the underlying synthetic polymer. The morphology of the explanted and cleaned polyurethane prostheses could be obtained readily by light and scanning electron microscopy. Surface microporous features and the presence of polyurethane microfibres that had experienced in vivo biodegradation could therefore be identified easily. The surface and bulk physico-chemical properties of the polyurethane polymer were determined by electron spectroscopy for chemical analysis, attenuated total reflectance-Fourier transform infrared spectroscopy and differential scanning calorimetry. It was found that the most successful approach for removing fresh tissue and exposing a clean and uncontaminated polyurethane surface was to incubate the explanted samples first in collagenase followed by digestion in pancreatin. This particular cleaning technique has proved valuable in enabling us to monitor small in vivo changes in the surface chemistry and in the bulk microphase segmented structure of polyurethane biomaterials.

Animals

Polyvinylidene fluoride monofilament sutures: can they be used safely for long-term anastomoses in the thoracic aorta?

Polyvinylidene fluoride (PVDF) represents an attractive alternative to polypropylene as a monofilament vascular suture because of its satisfactory physicochemical properties, it ease of handling, and its good biocompatibility. However, the polymer's ability to remain mechanically and chemically stable when exposed to a mild hydrolytic environment over the long term has yet to be demonstrated. One in vitro study involved the comparison of the long-term relative resistance of PVDF and polypropylene sutures to hydrolysis for a period of 9 years. The PVDF suture showed major molecular rearrangements from the original ratio of three crystalline structures to the single beta crystalline phase. The observation of some surface oxidation and water inhibition did not significantly modify the tensile strength of the PVDF suture, which retained 92.5% of its original value. In contrast, the polypropylene sample did not undergo any recrystallization but was associated with more oxidation byproducts and more water molecules near the surface, which contributed to a 46.6% loss in initial tensile strength. An in vivo study confirmed that PVDF sutures are biocompatible and are able to maintain satisfactory biostability when used to anastomose thoracic aortic allografts for a period of 6 months in the dog. The cellular reaction of fresh allografts as well as the control autografts to PVDF sutures was minimal. In other allografts that had been preserved in a supplemented medium for 1 week prior to implantation, the PVDF sutures healed satisfactorily with the formation of neocollagen and few macrophages surrounding the monofilament. No evidence of instability at the allograft-host artery junction was observed, confirming that the PVDF sutures were able to ensure a secure anastomosis in the thoracic aorta. PVDF sutures have demonstrated superior long-term biostability in vitro and minimal tissue response in vivo. These are two essential requirements when evaluating the use of a suture for vascular surgery in general and thoracic aortic surgery in particular.

Anastomosis, Surgical

[Macroscopic, histologic and ultrastructural study of 89 prostheses of anterior cruciate ligament excised because of prosthesis failure].

PURPOSE OF THE STUDY: This study concerns the etiology of failed synthetic anterior cruciate ligament (ACL) prostheses, and attempts to identify the primary mechanisms that lead to their premature rupture. MATERIAL AND METHODS: A total of 89 failed and surgically excised ACL prostheses were retrieved from young and active patients (27 +/- 7 years) at various orthopaedic centres in France. Their average duration of implantation was 34 +/- 24 months. They were examined macroscopically, histologically and by scanning electron microscopy (SEM) to determine the model, the manufacturer, the surgical technique used at implantation, the extent of healing, the site of rupture, as well as the morphology of the damage fibers. RESULTS: Seventy two of these explants represented 6 different models. While all 6 were fabricated from polyester fibres, each had a different textile construction, and each were associated with a unique healing and mechanical response in vivo. SEM observations confirmed that abrasion of the textile fibres were a phenomenon common to all models, and were the primary cause of prosthesis failure. Such wear zones were particularly prevalent at the exit of the tibial tunnel and around the femoral condyle. Collagenic infiltration into the synthetic ACL was poorly organized and unpredictable. It did not increase with the duration of implantation. In fact in certain models, it appeared to have caused deterioration and fraying of the textile structure rather than serving as a reinforcing matrix around the prosthesis. DISCUSSION: A synthetic ACL prosthesis is to be preferred for patients who do not have tissue available for autologous ligamentoplasty. Yet none of the synthetic devices examined in the present study were capable of stabilizing the knee over the long term. Among the factors that influenced their failure we found that the three most common mechanisms were flat abrasion against an osseous surface, flexural and rotational fatigue of the fibres, and loss of integrity of the textile structure due to unpredictable tissue infiltration during healing. CONCLUSION: The results of the present study show that none of the current models succeed in replacing the natural ACL. Future improvements may be achieved by developing surgical procedures for implantation combined with a prosthesis made from fibres and textile structures which are more abrasion resistant and promote predictable and controlled tissue infiltration.

Adult

Novel HOX, POU and FKH genes expressed during bFGF-induced mesodermal differentiation in Xenopus.

Cells from the cap of the animal hemisphere of the early Xenopus embryo are determined to form ectodermal lineages. When these cells are explanted and cultured in the presence of various growth factors a change in fate to cells of mesodermal lineage can be observed. Proteins of the fibroblast growth factor (FGF) family belong to this class of fate altering compounds. The ability of FGFs to change animal cap cell fate is in part due to an alteration in the program of genes expressed in these explanted cells. Several genes that are known to be pattern regulating in other systems have been shown to be induced by FGFs in the animal cap assay. We have utilized a PCR-based sib-selection and cloning protocol to identify a large number of cDNAs of the HOX, POU and FKH families that are present in animal caps very early during bFGF-induced mesodermal differentiation. A total of 11 different HOX, 7 POU and 4 FKH cDNAs were identified in an induced animal cap cDNA library. In several cases, pairs of highly related sequence variants were identified that presumably represent expression from the duplicated alleles of the ancestrally tetraploid Xenopus genome. In this report we characterize the temporal and spatial expression of three novel Xenopus genes during early development as well as during bFGF-induced mesodermal differentiation.

Amino Acid Sequence

Morphological, physical and chemical evaluation of the Vascugraft arterial prosthesis: comparison of a novel polyurethane device with other microporous structures.

In this study the morphology, physical properties, surface chemical characteristics and microstructure of the Vascugraft arterial prosthesis have been investigated. This is a novel microporous polyurethane device, recently developed by the company Braun-Melsungen AG in Germany for use as a small calibre arterial substitute. This comparative study included two other synthetic grafts: the Mitrathane prosthesis, a hydrophilic prototype polyetherurethane urea graft with closed internal pores, and the commercially successful expanded polytetrafluoroethylene reinforced Goretex prosthesis with an open microporous structure. The Vascugraft prosthesis contains a network of fused microfibres of varying thickness and orientation which provide open and communicating pores similar in size to those in the Goretex material. In addition, they extend from one side of the graft wall to the other. As well as having superior longitudinal and radial compliance to the reinforced Goretex device, the Vascugraft prosthesis has more than adequate bursting and suture retention strengths. Through the use of contact angle measurements, electron spectroscopy for chemical analysis, Fourier transform infrared spectroscopy, differential scanning calorimetry and molecular weight analysis by size exclusion chromatography, the surface of the Vascugraft prosthesis has been shown to be uniquely hydrophobic, as well as containing carbonate groups within an aliphatic polyesterurethane polymer. In addition, variations in micro-phase separation structure of hard and soft segment domains between different sizes and batches of product are marginal. Because of the interesting physical and chemical properties, it is recommended that in vitro biocompatibility and biostability studies be undertaken prior to using the prosthesis in animal or clinical trials.

Animals

In vivo performance of the polyesterurethane Vascugraft prosthesis implanted as a thoraco-abdominal bypass in dogs: an exploratory study.

Among the various prototype vascular prostheses that have been developed over recent years as small vessel substitutes, the Vascugraft polyurethane device produced by Braun-Melsungen AG has a number of attractive features. As well as having high mechanical compliance similar to that of the arterial tree, it has been manufactured from a specially synthesized poly(ester urethane) with improved biostability and its microfibrous structure provides a highly porous wall with open communicating pores. With a view to evaluating the in vivo biofunctionality and biostability of this prosthesis in the dog, 10 mm diameter grafts were implanted as thoraco-abdominal bypasses for prescheduled periods of 1 months and 12 months, and their performance monitored in terms of gross morphology, histology and the measurement of the chemical and physical properties of the explanted and cleaned specimens. Both grafts were patent at retrieval. Each had a smooth and glistening flow surface without organized mural thrombi and showed the development of a thin collagenous internal capsule with the presence of endothelial-like cells. Both grafts were well encapsulated externally and revealed a small distal bend or kink which is frequently observed by any thoraco-abdominal bypass in dogs. The fresh explanted prostheses were cleaned by a new enzyme treatment which provided specimens for microscopic, mechanical and thermal analyses, as well as studies of the surface and bulk chemistry. By comparing the results from the explanted and cleaned material with those of the virgin prosthesis, we have observed some deterioration in the integrity of the microfibrous structure, some loss in mechanical performance, marginal changes in molecular weight, and an apparent microphase separation of the hard and soft segment domains at a depth of a few microns. While the biofunctionality of a 10 mm calibre device has been demonstrated, additional in vivo studies are recommended to assess the biofunctionality at different diameters and the biostability over longer periods of implantation.

Abdomen

Quantitative analysis of the surface morphology and textile structure of the polyurethane Vascugraft arterial prosthesis using image and statistical analyses.

The surface morphology and textile structure of the Vascugraft polyurethane arterial prosthesis were investigated. Novel methods of image analysis and the presentation of statistical data were used to obtain quantitative results of the surface morphology of the non-woven microfibrous structure of prostheses of three different sizes. These techniques have identified apparent differences in the distributions of thickness and orientation of the microfibres between the internal and external surfaces and between the three prostheses investigated.

Biocompatible Materials

Functional analysis of the AUG- and CUG-initiated forms of the c-Myc protein.

Activation of the c-myc proto-oncogene by chromosomal translocation or proviral insertion frequently results in the separation of the c-myc coding region from its normal regulatory elements. Such rearrangements are often accompanied by loss or mutation of c-myc exon 1 sequences. These genetic alterations do not affect synthesis of the major c-myc protein, p64, which is initiated from the first AUG codon in exon 2. However they can result in mutation or loss of the CUG codon located in exon 1 that normally serves as an alternative translational initiation codon for synthesis of an N-terminally extended form of c-Myc (p67). It has been hypothesized that p67 is a functionally distinct form of c-Myc whose specific loss during c-myc rearrangements confers a selective growth advantage. Here we describe experiments designed to test the functional properties of the two c-Myc protein forms. We introduced mutations within the translational initiation codons of a normal human c-myc cDNA that alter the pattern of Myc protein synthesis (p64 vs. p67). The functions of each of these proteins were experimentally addressed using co-transformation and transcriptional activation assays. Both the p64 and p67 c-Myc proteins were independently able to collaborate with bcr-abl in the transformation of Rat-1 fibroblasts. In addition, both the exon 1- and exon 2-initiated forms of the c-Myc protein stimulated transcription of a Myc/Max-responsive reporter construct to a similar level. Given the apparent absence of functional differences between p64 and p67, we conclude that the basis for c-Myc oncogenic activation lies primarily in the overall deregulation of its expression and not in alterations in the protein. The existence of the CUG translational initiator may reflect a mechanism for the continued synthesis of c-Myc protein under conditions where AUG initiation is inhibited.

Amino Acid Sequence

Characterization and expression of the Xenopus c-Myb homolog.

The c-Myb protein is a sequence specific DNA-binding transcriptional regulator that is critically involved in the regulation of hematopoietic differentiation. Its role in these processes suggests that the function of c-Myb may be important early in the establishment of the hematopoietic lineage. We have isolated cDNA and partial genomic clones representing the Xenopus c-Myb homolog (Xc-Myb) in order to examine the role this gene plays in early mesodermal patterning in the frog embryo. The establishment of these clones as c-Myb homologs, as opposed to Myb-related sequences, is based upon both predicted amino acid sequences and the location of the exon-intron boundaries within the Xc-Myb gene. Maternally derived Xc-Myb RNA is degraded following fertilization then, beginning at midblastula, re-accumulates throughout early development. Xc-Myb RNA is localized to the animal cap region of the early blastula. Following the onset of gastrulation expression predominates in the ventral half of the embryo. During neurulation expression of Xc-Myb is observed in both the anterior dorsal and ventral vegetal regions of the embryo. Expression of Xc-Myb occurs in several adult tissues, the highest levels of which are in the intestine, heart, liver, lung and ovary. Xc-Myb encodes a protein of 624 amino acids and exhibits a mobility in SDS-PAGE of approximately 75 kDa, identical with that of the murine c-Myb protein. Xc-Myb protein exhibits 70% identity with avian and 67% identity with mammalian c-Myb proteins.

Amino Acid Sequence

A rapid and efficient, nonradioactive method for screening recombinant DNA libraries.

In this report we present a rapid and inexpensive PCR-based method to screen recombinant DNA libraries. The efficiency of this method was demonstrated by the isolation of clones of interest from three different libraries using different vector systems. This method is nonradioactive and makes it easier to handle a large number of samples since there is no need for DNA extraction. The advantages and applications of the method are discussed.

Animals

Breast implants.

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Consumer Product Safety

Histopathological and immunological investigations of synthetic fibres and structures used in three prosthetic anterior cruciate ligaments: in vivo study in the rat.

Three types of prosthetic anterior cruciate ligaments were investigated by enzymatic and histological analysis of the tissue surrounding each implant and immunologically by a cytofluorometric analysis of T-cell populations in the peripheral blood of rats. Two of the prostheses had a braided construction, one made from polyester and the other from high performance polyethylene fibres. The third type also contained high performance polyethylene fibres, but had been manufactured in a knitted construction (Raschel high performance polyethylene). Five specimens from each prosthesis were implanted intraperitoneally in rats by a trocar for different periods of time up to 4 wk. A control group of rats underwent the surgery, but not the implant. No modification in peripheral T-cell populations was induced by the presence of any implant. Whilst the levels of acid phosphatase and esterase activity appeared to have increased slightly following implantation of any of the prostheses, such increases were not highly significant. Histologically, all three materials induced an intense acute inflammatory reaction at 3 d which gave way to a typical chronic response after 4 wk. The only major difference between the prostheses was that after 4 wk the polyester fibres exhibited less inflammation, and the surrounding tissue was more mature, more vascularized and more densely infiltrated with collagen than with the two high performance polyethylene implants. In conclusion, all three devices provided satisfactory biocompatibility in terms of cellular and healing response.

Acid Phosphatase

Expression of two distinct homologues of Xenopus Max during early development.

The Max protein belongs to the basic region-helix-loop-helix-leucine zipper family of transcriptional regulators. Max heterodimerizes with Myc family proteins to form sequence-specific DNA-binding complexes. In order to elucidate the potential role of Myc and Max during amphibian embryogenesis, we have isolated and analyzed the expression of two Xenopus Max complementary DNAs: XMax1 and XMax2. Comparison of XMax1 and XMax2 with their mammalian counterparts demonstrates a strikingly high degree of conservation at both the nucleotide and amino acid levels, with the exception of a 24-residue deletion in both XMax proteins within their COOH termini. In addition, the two Xenopus Max proteins differ in that XMax2 contains a unique 27-amino acid insertion that interrupts the COOH-terminal end of the zipper domain; XMax1 lacks this insertion. Despite these differences, both XMax1 and XMax2 can form complexes with either Xenopus or human c-Myc proteins. Analysis of XMax expression during embryogenesis reveals that both mRNA and protein are expressed throughout early development, including the egg, 32-cell stage, and midblastula transition. Although the expression of XMax1 RNA appears to predominate at all stages examined, the ratios of XMax1 to XMax2 protein vary during development as well as between different tissue culture cell lines, suggesting a potential for cell type-specific regulation. Our results demonstrate the presence of Xenopus Max throughout frog development, raising the possibility that Myc and Max could function as a complex even during early embryogenesis.

Amino Acid Sequence