PubMed Health⌕ Search

Biomedical subjects

R J Hinton

Publications and source records attributed to R J Hinton.

At least 19 recordsLinked to original sources
PubMed2006

Cellular response to force application at craniofacial sutures.

OBJECTIVES: To provide a comprehensive review of the literature describing research done on the responses of suture cells to force application in vitro and in vivo. DESIGN AND RESULTS: This review outlines the types of forces that can be applied, methods of applying the forces, the sutures used in experiments, and the changes in morphology, molecular biology (gene and protein expression), and cell biology (proliferation, differentiation, apoptosis) in response to these forces. CONCLUSION: The molecular response of sutures to force needs to be further investigated as these molecules can be used to enhance the way in which craniofacial sutures respond to mechanical force during orthopedic-orthodontic treatment.

Apoptosis↗
PubMed2005

Specificity of meal pattern analysis as an animal model of determining temporomandibular joint inflammation/pain.

Analyzing feeding behavior, and in particular meal duration, can be used as a biological marker for temporomandibular joint (TMJ) inflammation/pain. The present study determined the specificity of meal duration as a measure of TMJ inflammation/pain in a rodent model. The model was also used to test the efficacy of dexamethasone (DEX) as a treatment for TMJ inflammation/pain that was induced by TMJ injection of complete Freund's adjuvant (CFA). In the first study, anesthetized male Sprague-Dawley rats housed in computerized feeding modules received bilateral intra-articular knee injections of CFA or saline. The next day, CFA-injected rats had significant knee swelling and impaired mobility. Food intake in the CFA-injected group was reduced over the next two days and this was due to reduced meal number with no change in meal size. Notably, meal duration was normal in both the CFA and saline knee-injected groups. In the second study, male rats were assigned to one of four groups: Group 1, no CFA and no DEX treatment; Group 2, no CFA and treatment with DEX (0.4 mg/kg i.m. once daily); Group 3, bilateral TMJ CFA injection and no DEX treatment; and Group 4, bilateral TMJ CFA injection and treatment with DEX. CFA significantly increased TMJ swelling and stress-induced chromodacryorrhea in Group 3, but treatment with DEX attenuated these effects in Group 4. Compared to the controls, meal duration was significantly lengthened 24 and 48 h post-CFA injection in Group 3, whereas DEX treatment attenuated TMJ swelling, chromodacryorrhea and normalized meal duration. The data demonstrate that meal pattern analysis, and in particular meal duration, can be used as a non-invasive specific measure of TMJ inflammation/pain and can be used as a marker of DEX treatment efficacy.

Animals↗
PubMed2001

Direct process integration of cell disruption and fluidised bed adsorption in the recovery of labile microbial enzymes.

The practical feasibility and generic applicability of the direct integration of cell disruption by bead milling with the capture of intracellular products by fluidised bed adsorption has been demonstrated. Pilot-scale purification of the enzyme L-asparaginase from unclarified Erwinia chrysanthemi disruptates exploiting this novel approach yielded an interim product which rivalled or bettered that produced by the current commercial process employing discrete operations of alkaline lysis, centrifugal clarification and batch adsorption. In addition to improved yield and quality of product, the process time during primary stages of purification was greatly diminished. Two cation exchange adsorbents, CM HyperD LS (Biosepra/Life Technologies) and SP UpFront (custom made SP form of a prototype stainless steel/agarose matrix, UpFront Chromatography) were physically and biochemically evaluated for such direct product sequestration. Differences in performance with regard to product capacity and adsorption/desorption kinetics were demonstrated and are discussed with respect to the design of adsorbents for specific applications. In any purification of L-asparaginase (pI = 8.6), product-debris interactions commonly diminish the recovery of available product. It was demonstrated herein, that immediate disruptate exposure to a fluidised bed adsorbent promoted concomitant reduction of product in the liquid phase, which clearly counter-acted the product-debris interactions to the benefit of product yield.

Adsorption↗
PubMed2000

Age-associated changes in decorin in rat mandibular condylar cartilage.

The small proteoglycan decorin strongly binds the fibrils of collagen types I and II; this interaction is thought to play a part in the maintenance of tissue integrity and biomechanical properties. In limb articular cartilage, there is evidence that decorin synthesis increases with age and that it is elevated in response to increased loading or in osteoarthritic cartilage. The aim here was to characterize the presence and relative amount of decorin in the condylar cartilage of the temporomandibular joint (TMJ) with maturation by Western blotting, and to assess its tissue localization by immunohistochemistry. Comparative data were obtained from tibial articular cartilage, which has been extensively studied. Cartilage from the mandibular condyle and tibial plateau was harvested from 24-day-old (growing) and 161-day-old (young adult) female Sprague-Dawley rats. In growing animals, decorin appeared slightly more abundant in the mandibular condylar cartilage than in articular cartilage, whereas in young adult animals the decorin content in the TMJ cartilage was noticeably less than in limb articular cartilage. Although there was an increase in decorin abundance with age at the TMJ, the increase in decorin with age in limb articular cartilage was considerably more pronounced. These data indicate that, although decorin is present in mandibular condylar cartilage, its abundance in adults is less than in limb articular cartilage; thus, maturation-associated changes may be dissimilar in magnitude from those documented for limb articular cartilage.

Aging↗
PubMed2000

Calcitonin gene-related peptide and substance P immunoreactivity in rat trigeminal ganglia and brainstem following adjuvant-induced inflammation of the temporomandibular joint.

The immunoreactivity of two inflammatory mediators, calcitonin gene-related peptide (CGRP) and substance P, was measured in the trigeminal ganglia and brainstem to characterize an adjuvant-induced inflammation within the rat temporomandibular joint at various acute (6, 24 and 48 h) and intermediate (10 day) time intervals. Concentrations of adjuvant-related neuropeptides were compared to those in both contralateral vehicle-related tissues and non-injected controls. By 6 h, CGRP immunoreactivity in the trigeminal ganglia was significantly above that in contralateral vehicle-injected tissue. The CGRP had decreased at each of the following time-points, but remained significantly elevated at 10 days. Substance P in the ganglion on the injected side was significantly increased for all four time periods. In brainstem subnucleus caudalis, CGRP was significantly increased for all four time periods. Substance P immunoreactivity in the subnucleus caudalis was significantly increased for the initial three time periods, but by day 10 had been reduced to that of the control. These data show that the pattern of changes in neuropeptides following the induction of inflammation is different between substance P and CGRP. Moreover, the pattern of change varies between the brainstem and the trigeminal ganglion. This suggests that the two neuropeptides may have different roles in the inflammatory process, and that this process may be modulated by different mechanisms at the brainstem and ganglion.

Afferent Pathways↗
PubMed1998

Crystallization of NAD+-dependent phenylalanine dehydrogenase from Nocardia sp239.

The NAD+-dependent phenylalanine dehydrogenase from Nocardia sp239 has been crystallized by the hanging-drop method of vapour diffusion using ammonium sulfate as the precipitant. Two crystal forms were obtained in the presence and absence of the enzyme substrates phenylpyruvic acid or phenylalanine and its coenzyme NADH. Crystals of the native protein belong to the hexagonal system, with the space group being one of the enantiomorphic pair P6122 or P6522. The cell dimensions are a = b = 111.0, c = 174.5 A, alpha = beta = 90 and gamma = 120 degrees. Crystals grown from the protein co-crystallized with its substrates all belong to the trigonal system, space group P3121 or P3221, with unit-cell dimensions of a = b = 88.1, c = 112.6 A, alpha = beta = 90 and gamma = 120 degrees. Preliminary protein-sequencing experiments have established that this enzyme is related to the octameric PheDH's which are members of the wider superfamily of amino-acid dehydrogenases. However, gel-filtration studies suggest that this enzyme is active as a monomer. The full determination of the three-dimensional structure of this phenylalanine dehydrogenase will add to the understanding of the molecular basis of the differential substrate specificity within this enzyme superfamily. In turn this will contribute to the rational design of an amino-acid dehydrogenase which could be used for the diagnosis of phenylketonuria and for the chiral synthesis of high-value pharmaceuticals.

Amino Acid Oxidoreductases↗
PubMed1998

Capsaicin application to the temporomandibular joint alters calcitonin gene-related peptide levels in the trigeminal ganglion of the rat.

The aim of this investigation was to determine the temporal effect of an intra-articular injection of capsaicin to the temporomandibular joint on the levels of calcitonin gene-related peptide-like immunoreactivity (CGRP-ir) in the trigeminal ganglion of the rat. The temporomandibular joints of 26 adult female rats were injected on one side with capsaicin and contralaterally with a control vehicle. Another 8 animals served as an untreated control group and received no injections. Animals were sacrificed at time intervals of 4 hours, 48 hours, 10 days, and 21 days following treatment. The trigeminal ganglia were extirpated, and CGRP-ir levels were quantified using a radioimmunoassay. Results demonstrated that when the capsaicin-treated side and the vehicle-treated side were compared, CGRP-ir levels decreased initially at 4 hours and increased at 48 hours. At 10 days, CGRP-ir levels had again dropped below control levels, followed by an increase at 21 days. CGRP-ir levels for the first two time periods investigated, which simulate an acute inflammatory state, mimic results observed in studies using limb joints, while the other time periods, which represent an intermediate and a chronic condition, respectively, suggest a more complex interaction with capsaicin-sensitive primary afferents.

Animals↗
PubMed1997

Reactive changes in the temporomandibular joint after mandibular midline osteodistraction.

The purpose of this study was to evaluate the histologic changes within the condyle in response to mandibular widening using osteodistraction. Mandibular midline osteotomies were made in nine Macaca mulatta monkeys and tooth-borne distraction devices were bonded to the mandibular dentition. Distraction was continued until a 3-5 mm widening was achieved. The appliances were then stabilized for a period of 4 weeks. Non-decalcified sagittal sections of the lateral, middle and medial thirds of the condyles were analyzed. Although three of the seven animals showed no unusual morphology, four others exhibited morphologic differences within the fibrous layer, cartilage layer or bone/cartilage interface. Histologic changes were seen to occur in the fibrous layer, cartilaginous layer and cartilage/bone interface. The severity of these changes were correlated with the likely rotational forces directed at the condyle on the postero-lateral and antero-medial surfaces.

Analysis of Variance↗
PubMed1997

Effect of postoperative diet on condylar cartilage response to discectomy.

PURPOSE: Recent studies have shown that metabolic and structural changes occur in the condylar cartilage after surgical removal of the articular disc. There is some evidence that these structural changes are less pronounced in animals provisioned with a soft diet after disc removal. This study was initiated to assess whether a soft diet after discectomy in growing rats also results in alterations in the composition or metabolic activity of the matrix of the condylar cartilage. MATERIALS AND METHODS: In two identical experiments, 28 thirty-day-old female Sprague-Dawley rats underwent unilateral surgical removal of the articular disc (discectomy) and were then provisioned with either a soft, mushy diet or a diet of normal rat pellets. When they were killed 3 weeks later, the condylar cartilages were removed, weighed, pulse-labeled in organ culture with [35S]-sulfate for 2 hours, and analyzed for sulfated glycosaminoglycan (GAG) content and [35S]-sulfate uptake. RESULTS: As in previous studies, tissue weights and hydration were increased, and sulfated GAG content and [35S]-sulfate uptake decreased on the surgery side in animals fed a hard diet. Overall, fewer differences were present in the animals fed a soft diet, although tissue weights and hydration were still elevated on the discectomized side. Analysis of ratios of surgery/nonsurgery values in the hard diet versus the soft diet sample indicated that dietary consistency was a significant factor for tissue weights and [35S]-sulfate uptake, but not for hydration or sulfated GAG content. CONCLUSIONS: These data suggest that discectomy exerts a more pronounced effect on growing animals fed a hard diet, although some changes also persist after discectomy in animals fed a soft diet.

Animals↗
PubMed1996

Amplified expression and large-scale purification of protein L'.

The gene fragment (PPL') encoding the functional unit of peptostreptococcus protein L was isolated using PCR and expressed in E. coli. As the gene fragment lacked its own promoter, the 5' PCR primer was designed to incorporate an Nde1 restriction site (CATATG) into the gene. This enabled the gene to be cloned in frame into an Nde1 restriction site immediately downstream of a trp promoter. To prevent read through, a stop codon was introduced into the 3' primer. Expression of PPL' was up to 27% total cell protein which compares favourably to the 0.1% total soluble cell protein obtained from the original clone of peptostreptococcus. Following a heat step homogeneous PPL' was recovered by a single anion-exchange chromatography step on Q-Sepharose FF in yields of 90%.

Bacterial Proteins↗
PubMed1995

Cloning, expression and purification of Ppl-1, a kappa-chain binding protein, based upon protein L from Peptostreptococcus magnus.

Protein L is a multi-domain, cell wall constituent of certain strains of Peptostreptococcus magnus, which binds to the variable domain of the L-chains of Ig. A gene fragment which codes for a single Ig-binding domain of protein L (Ppl-1) was cloned into a modified pKK223-3 vector and over-expressed in E. coli JM103. A rapid protein purification protocol is described. In these studies, purified Ppl-1 was immobilised on to an agarose gel and tested against an array of Igs and Ig fragments. It was found that Ppl-1-bound Igs from a number of different sources via interactions with the L kappa-chain. An enzyme linked immunosorbant assay has been developed to assay the binding of Ppl-1 to IgG. The incubation of Ppl-1 with human serum does not produce an immunoprecipitate, thus suggesting one unique interaction per binding domain which has been confirmed by ELISA. These experiments demonstrate the potential value of Ppl-1 as an immunological tool and as an affinity chromatography ligand for the purification of Igs.

Amino Acid Sequence↗
PubMed1993

Effect of dietary consistency on matrix synthesis and composition in the rat condylar cartilage.

Little is known of how the matrix of the condylar cartilage of the mandible changes in response to an alteration in local biomechanical circumstances, although this has been a habitual focus of studies of articular cartilage in the orthopedic literature. This study was formulated to investigate (1) whether matrix changes would occur in the condylar cartilage of rats fed a diet of soft consistency, a circumstance which would presumably reduce articular forces at the mandibular joint; (2) whether these changes, once established, could be reversed by restoring a diet of hard rat pellets, and (3) if these changes were localized to a particular region(s) of the cartilage. In the first experiment, male Sprague-Dawley rats were provisioned with either a soft, mushy diet or hard rat pellets beginning at weaning and subsequently sacrificed at 44 days of age. In the second experiment, all animals were initially given the soft diet, but in half a normal hard diet was reinstituted for 7 days prior to sacrifice at 44 days of age. In the third experiment, cartilages from rats fed soft and hard diets were subdivided into a superior fraction (that portion directly opposite the cranial articulation) and a posterior fraction. The results of experiment 1 showed the water content of the condylar cartilage to be significantly reduced in the soft-diet group, as well as one measure of [35S]uptake into sulfated glycosaminoglycans ([35S]sulfate dpm/micrograms uronic acid). Both wet and dry tissue weights of the condylar cartilage were greatly reduced in rats fed a soft diet.(ABSTRACT TRUNCATED AT 250 WORDS)

Animal Feed↗
PubMed1992

Alterations in rat condylar cartilage following discectomy.

The purpose of this study was to examine the effect of unilateral extirpation of the articular disc of the temporomandibular joint on selected metabolic and compositional characteristics of the condylar cartilage. Unilateral discectomy was performed in one group of growing (28-day-old) rats, while another group received only unilateral arthrotomy. The effect of the discectomy procedure was assessed by comparison of ratios of the operated to unoperated sides in the discectomy and arthrotomy groups. Total uronic acid content (micrograms/mg dry tissue wt) of the condylar cartilage and [35S]-sulfate incorporation (dpm/micrograms uronic acid) into the condylar cartilage were decreased in discectomy-group animals at nine, 16, and 28 days following surgery, while hydration of the cartilage was increased in discectomy animals at both 16 and 28 days post-surgery. Wet and dry tissue weights of the cartilage were dramatically increased in discectomy-group animals at all post-operative intervals. These metabolic and compositional changes were accompanied by pronounced structural alterations in the cartilage, including chondrocyte clustering and tears parallel to the articular surface. With the exception of [35S]-sulfate incorporation, the changes demonstrated in the condylar cartilage following discectomy were similar to early osteoarthritic alterations reported in limb articular cartilage following partial meniscectomy or cruciate ligament resection.

Animals↗
PubMed1992

Amplified expression and large-scale purification of protein G'.

PCR was used to isolate the gene fragment coding for Protein G' (SpG'), a truncated bacterial cell surface protein from Streptococcus G148 which binds to the Fc region of IgG and expressed in E. coli [Goward et al. (1990) Biochem. J. 267: 171-177]. The PCR primer was designed to change the TTG initiation triplet to ATG and to incorporate it into an NdeI restriction site (CATATG), allowing the gene to be cloned in frame into an NdeI restriction site immediately downstream of a trp promoter. Expression of SpG' was estimated as about 30% total soluble cell protein which compares very favourably to the less than 1% total soluble cell protein obtained from the original system [Goward, et al. (1990) Biochem. J. 267: 171-177]. Homogeneous SpG' was recovered by a single anion-exchange chromatography step on Q-Sepharose FF in a process which avoided use of an affinity adsorbent. Even though SpG' consists of almost identical repetitive domains from amino acid sequence analysis, different proteolytic sensitivity of each domain was observed indicating their structural dissimilarity.

Amino Acid Sequence↗
PubMed1991

Histologic examination of the temporomandibular joint after mandibular advancement with and without rigid fixation: an experimental investigation in adult Macaca mulatta.

This study evaluated the histologic response of the temporomandibular joint (TMJ) following mandibular advancement using rigid and nonrigid fixation in monkeys. Twelve adult female rhesus monkeys underwent sagittal ramus osteotomies with advancement. Six of them were placed into maxillomandibular fixation (MMF); six underwent bicortical bone-screw fixation without MMF. Changes in condylar position were quantified using lateral cephalograms with the aid of bone markers. The animals were killed at 6 weeks and the TMJs were prepared for histologic analysis. Three measures of condylar cartilage thickness were obtained for each animal and were correlated to changes in position of the condyle. Animals who underwent MMF showed a tendency for anterior movement of the condyles; animals who underwent rigid fixation showed a tendency for posterior condylar position. Thicker cartilage layers were found in the MMF animals. Animals who had posterior displacement of the condyles showed evidence of resorption of the posterior surface of the condyle and anterior surface of the postglenoid spine. There was a significant correlation between a change in the horizontal position of the condyle and the thickness of the posterior aspect of the condylar cartilage. The results of this study indicate that alterations in condylar position may induce remodeling changes within the TMJ.

Animals↗
PubMed1991

Jaw protruder muscles and condylar cartilage growth in the rat.

Many studies have explored the role of the protrusive musculature in promoting growth at the condylar cartilage and the overall lengthening of the lower jaw, with emphasis on the lateral pterygoid muscle (LPM). The largely anteroposterior orientation of the superficial part of the masseter muscle (SM) in the rat suggests that it may also function as a protruder of the lower jaw. Accordingly, it is possible that the action of the SM may play a part in the regulation of growth of the condylar cartilage and the lower jaw. To examine this hypothesis, bilateral resection of the superficial portion of the masseter muscle was performed in male Sprague-Dawley rats at 26 days of age. At 5 days after surgery, [3H]-thymidine incorporation in the condylar cartilage was increased (F = 6.93, p less than or equal to 0.01) in the SM myectomy group relative to the surgical control and unoperated control groups. However, by 20 days after surgery no differences were present. At this sacrifice interval, lower jaw dimensions relating to areas of muscle attachment, as well as ramus height, were significantly reduced in the SM myectomy group, but overall jaw length (mental foramen to condyle) was unaffected. In contrast, myotomy of the LPM resulted in a significant decrease in mitotic activity of the cartilage 4 days after surgery. This decrease was present, but not more pronounced, in animals subjected to both SM myectomy and LPM myotomy. Hence, myotomy or myectomy of these two muscles, each with a protrusive orientation, produces opposite effects on proliferative activity at the condylar cartilage.

Animals↗