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The optic nerve of rats with EAE was examined at various times to determine the integrity of the blood-brain barrier (BBB) and to assess monocyte-macrophage, T cell, and microglial responses. In naive control animals, leakage of horseradish peroxidase (HRP) and the presence of cells expressing major histocompatibility complex (MHC) class II antigen were evident in the meninges of the retrobulbar optic nerve. In rats with EAE, microglia in the region of the lamina cribrosa and in the regions adjacent to the meninges became activated from day 7 to 8 postinduction (pi). HRP leakage was also evident in the region of the lamina cribrosa from day 7 to 8 pi. On day 8 pi, infiltration of inflammatory cells and Monastral blue leakage were apparent in the myelinated region of the optic nerve. The intensity of these cellular and vascular changes peaked at day 12 pi, when signs of clinical disease became manifest. Monocytes-macrophages expressing MHC class II and the ED1 antigen, together with lymphocytes expressing the alphabetaT cell receptor, constituted the major proportion of cells associated with inflammatory lesions. Thus: (i) the inherent weakness of the BBB as well as the presence of both antigen (myelin) and MHC class II+ cells in the retrobulbar optic nerve are likely susceptibility factors for the frequent involvement of this region in EAE and multiple sclerosis; and (ii) activation of microglia occurs early in the pathogenesis of experimental optic neuritis.

The microvascular and cellular responses in the retina during acute EAE were characterized using whole-mount preparations. The earliest detectable event was the accumulation of monocytes and T cells within veins on day 7 postinduction (pi). Mild breakdown of the blood-retinal barrier (BRB), activation of microglia and infiltration of monocytes and T cells into the retinal parenchyma were first evident on days 7 to 8 pi. Monocyte adhesion to the vessel wall and breakdown of the BRB were colocalized in the same vessel segments and occurred predominantly in veins. The marked difference in response observed in the retina versus the myelinated region of the optic nerve suggests that two types of inflammatory cascades are initiated. A mild response, characterised by very low numbers of T cells and monocytes and an absence of expression of MHC class II by resident microglia, is initiated when only small amounts of the encephalitogenic antigen are present in the perivascular space or associated with perivascular antigen-presenting cells. A full blown inflammatory reaction, as observed in the optic nerve, is initiated in the presence of substantial amounts of encephalitogenic antigen. This severe response is characterised by the infiltration of large numbers of CD4+, CD8+ T cells and ED1+ monocytes, and by abundant MHC class II expression by resident microglia as well as other cell types. Thus, MHC class II expression by resident microglia may be a possible effective amplifier mechanism if the encephalitogenic antigen is encountered in the tissue parenchyma.

This study utilized the kinesiograph to assess the effect of orthognathic surgery on mandibular rest positions in a group of 27 patients exhibiting vertical facial deformities (14 long faced and 13 short faced patients). Two distinct rest positions were confirmed--clinical and physiological. Following surgery to alter the vertical dimensions, the clinical freeway space was found to adapt immediately whereas the physiological rest position partially adapted in the immediate postoperative period and continued to do so over the following year.

The external nose is generally considered to have a relatively static shape. Movement of the nose and the potential for change of external shape and the internal airway have been assessed by (1) cadaver dissection and (2) dynamic studies during a standardized series of facial expressions in 13 subjects; video recording of the movement of skin markers, electromyography, and moving magnetic resonance images. A standard description of muscle anatomy is presented. The dynamic investigations indicated the following. Video analysis showed the components of muscle action: dilatation, constriction of the nostril, depression of the tip, vertical contraction in the bridge, and elevation of the alar, in different expressions. Electromyography confirmed muscle actions during expression and phonation. Magnetic resonance imaging demonstrated large changes in the external shape of the nose and nasal aperture due to muscle actions. Muscle function should be given greater consideration in aesthetic and cleft rhinoplasty.

Expression of the relaxin-like factor (RLF) was studied at the messenger RNA (mRNA) and protein levels in the testes and ovaries of the mouse, as well as through testicular development and differentiation in the mouse testis. In situ hybridization or RT-PCR, and immunohistochemistry using a polyclonal antibody raised against a recombinant protein, provided mutually confirmatory results for a high expression of RLF in the Leydig cells of the adult testis and at a much lower level of expression in the luteal cells of the ovary through the cycle, pregnancy, and in lactation. Analysis of protein and mRNA expression, through postnatal testicular development, indicated moderate RLF expression also in the fetal population of Leydig cells, even in the hpg mutant mouse, lacking an active pituitary-gonadal axis. Prepubertal Leydig cells, however, exhibit only very low-level RLF gene expression, this phenotype persisting in the adult hpg mouse. In summary, fetal Leydig cells express RLF in an LH/human CG-independent fashion, whereas LH/human CG is essential to induce RLF expression in the adult-type Leydig cell. In cultured adult Leydig cells or in the mouse tumor MA-10 cell line, RLF mRNA is expressed in a constitutive fashion. RLF thus seems to be a useful marker of Leydig cell differentiation status.

As computer game playing is a popular activity among adolescents, a questionnaire study was undertaken with 387 adolescents (12-16 years of age) to establish their "dependence" using a scale adapted from the DSM-III-R criteria for pathological gambling. Analysis indicated that one in five adolescents were currently "dependent" upon computer games. Boys played significantly more regularly than girls and were more likely to be classified as "dependent." The earlier children began playing computer games it appeared the more likely they were to be playing at "dependent" levels. These and other results are discussed in relation to research on other gaming dependencies.

The aim of these studies was to investigate the ability of cysteamine and its congeners to arrest the proliferation of leukemic cells and to determine the physico-chemical properties responsible for this ability. Fifteen low molecular weight thiol-bearing compounds were shown to arrest the proliferation of CCRF-CEM cells and a methotrexate-resistant subline, with IC50 values between 10(-5) and 10(-4) M. Cysteamine arrested proliferation by slowing the passage of cells through S phase. These cells subsequently resumed cycling, although a proportion went on to die by apoptosis. The antiproliferative action of cysteamine was shown to depend, in part, on H2O2 production. This ability to generate peroxide is shared by many thiol compounds, and molecular modeling indicated that thiol groups were required for the antiproliferative actions of the congeners of cysteamine. Molecular modeling also revealed that the most efficacious antiproliferative agents were those that had their amino acid and thiol moieties separated by an intramolecular distance of 3.17 to 5.9 A, as exemplified by WR 1065 and the aminothiophenols. These findings indicate that thiol-bearing compounds may have some efficacy in the treatment of drug-naive and -resistant leukemia cells.

Objective: The effectiveness of lithium in preventing recurrences of bipolar disorder was examined prospectively for 2 years in two representative samples of bipolar I patients being treated in a comprehensive program following recommended guidelines., Method: One hundred and twenty patients were recruited from consecutive admissions to two catchment area psychiatric services (one in the United Kingdom, the other in New Zealand). They were seen at 3-monthly intervals by a member of the research team. Treatment was adjusted according to clinical needs., Results: Overall, two-thirds of the patients had a recurrence. Of the 57 on lithium as sole treatment, 39 (68%) had a further episode; 17 after stopping lithium. The 42 on other mood stabilisers and/or an antipsychotic, with or without lithium, did no better. By contrast, only eight (38%) of the 21 who were taking no prophylactic medication had a recurrence., Conclusions: Lithium is much less effective in clinical practice than would be expected from clinical trial results. A major reason for this is poor compliance. Alternative treatment strategies are needed to improve the outcome for bipolar disorder patients.

Exposure of plasma from apolipoprotein E gene knockout (apoE-/-) and control (CBA or C57BL/6J) mice plasma to a constant rate of aqueous peroxyl radicals (ROO.) resulted in the depletion of ascorbate, urate and alpha-tocopherol (alpha-TOH), with substantial and little lipid peroxidation, respectively. Alpha-TOH levels were 3-times higher in plasma from apoE-/- than control mice and its addition enhanced the oxidizability of control mouse plasma. In apoE-/- mouse plasma, alpha-TOH was associated primarily with very low density lipoprotein (VLDL), whereas in plasma from control mice, the vitamin was located largely in high density lipoproteins. Oxidation of isolated lipoproteins by ROO. resulted in the accumulation of lipid hydroperoxides to an extent that reflected the plasma concentration and alpha-TOH content of the different lipoprotein fractions. Oxidation of 'plasma' reconstituted from components of apoE-/- mice and/or human plasma showed that human and apoE-/- mouse lipoproteins peroxidized with similar kinetics, although the initiation of lipid peroxidation was greater in the presence of mouse than human lipoprotein-deficient plasma. Also, the chain length of lipid peroxidation in apoE-/- mouse plasma after ascorbate depletion appeared to be independent of the rate of ROO. generation. Together, these results show that the ROO. induced peroxidation of plasma lipoproteins in atherogenesis-susceptible apoE-/- mice exhibits some, though not all, features of tocopherol-mediated peroxidation (TMP). Therefore, apoE-/- mice may represent a suitable animal model to test a role for TMP in atherogenesis and the prevention of this disease by anti-TMP agents.

The pathogenesis of human cerebral malaria (CM) remains unresolved. In the most widely used murine model of CM, the presence of T lymphocytes and/or interferon (IFN)-gamma is a prerequisite. IFN-gamma is the key inducer of indoleamine 2,3-dioxygenase (IDO), which is the catalyst of the first, and rate-limiting, step in the metabolism of tryptophan (Trp) along the kynurenine (Kyn) pathway. Quinolinic acid (QA), a product of this pathway, is a neuro-excitotoxin, like glutamic acid (Glu) and aspartic acid (Asp). Kynurenic acid (KA), also produced from the Kyn pathway, antagonizes the neuro-excitotoxic effects of QA, Glu, and Asp. We therefore examined the possible roles of IDO, metabolites of the Kyn pathway, Glu, and Asp in the pathogenesis of fatal murine CM. Plasmodium berghei ANKA infection was studied on days 6 and 7 post-inoculation (p.i.), at which time the mice exhibited cerebral symptoms such as convulsions, ataxia, coma, and a positive Wooly/White sign and died within 24 hours. A model for noncerebral malaria (NCM), P. berghei K173 infection, was also studied on days 6 and 7 and 13 to 17 p.i. to examine whether any changes were a general response to malaria infection. Biochemical analyses were done by high-pressure liquid chromatography and gas chromatography/mass spectrometry/mass spectrometry (GC/MS/MS). IDO activity was low or absent in the brains of uninfected mice and NCM mice (days 6 and 7 p.i.) and was induced strongly in the brains of fatal murine CM mice (days 6 and 7 p.i.) and NCM animals (days 13 to 17 p.i.). This induction was inhibited greatly by administration of dexamethasone, a treatment that also prevented CM symptoms and death. Furthermore, IDO induction was absent in IFN-gamma gene knockout mice, which were also resistant to CM. Brain concentrations of Kyn, 3-hydroxykynurenine, and the neuro-excitotoxin QA were significantly increased in both CM mice on days 6 and 7 p.i. and NCM mice on days 13 to 17 p.i., whereas an increase in the ratio of brain QA to KA occurred only in the CM mice at the time they were exhibiting cerebral symptoms. Brain concentrations of Glu and Asp were significantly decreased in CM and NCM mice (days 13 to 17 p.i.). The results imply that neuro-excitation induced by QA may contribute to the convulsions and neuro-excitatory signs observed in CM.

Fibrochondrogenesis is a very rare form of lethal short-limb dwarfism, with 8 cases described since it was first reported in 1978. It is becoming clear that this condition has certain radiological and histological characteristics that distinguish it from other skeletal dysplasias. We herein present a further case of fibrochondrogenesis diagnosed in a fetus of 17 weeks, which is the youngest patient reported so far. In addition, the fetus showed severe micrognathia and a bifid tongue. These are not previously described manifestations, which extend the phenotype of this rare condition.

The polycythemic strain of the Friend spleen focus-forming virus (SFFVp) is a replication-defective, acutely transforming retrovirus inducing a bistage erythroleukemia in susceptible mice. The first stage of the disease is an acute polyclonal erythroblastosis induced by the proliferation-promoting effect of gp55. gp55 is expressed from a spliced subgenomic message of SFFVp and stimulates the cellular receptor for erythropoietin. Using a selectable SFFVp that otherwise mimics the specificity of the disease, we demonstrate that the kinetics of the polyclonal expansion depends on the transcriptional strength of the retroviral cis-active elements. By exchanging gp55 for apathogenic genes, we show that SFFVp enhancer and splice signals can be successfully utilized for the development of retroviral vectors mediating very efficient transgene expression in hematopoietic cells. Apathogenic selectable SFFVp-based vectors carrying distinct enhancer alterations are a valuable tool to analyze transcriptional control of leukemia viruses in the absence of oncogenic proteins. Moreover they might have therapeutic potential.

This retrospective cephalometric study examined the stability of mandibular incisor decompensation following orthognathic treatment. It also studied the relationship between the mandibular incisor-mandibular plane angle (LIMnp) and the maxillary-mandibular planes angle (MMA). Two groups of 22 subjects each underwent either mandibular advancement only or bimaxillary surgery with mandibular setback. Serial radiographs of each subject were digitized. Twenty-seven Class I subjects who were part of the Leighton Growth Study served as the control. The control group showed a significant association between LIMnp and MMA. For the surgical groups, the only significant relationship between LIMnp and MMA was seen in the mandibular advancement group at debond. No definite indicator was found to suggest that the final MMA dictates the amount of mandibular incisor relapse following debond. Only the bimaxillary group showed an association between changes in LIMnp and changes in MMA during the review period.

This paper presents two cases in which supplemental premolars were an incidental finding. Although anomalies of tooth number and form are quite common, both cases demonstrate late forming supernumeraries with one case illustrating their development in a more unusual site. It is not routine practice to screen for late development of teeth during orthodontic treatment. Therefore the possibility of their interference with occlusal development or orthodontic mechanics such as space closure, should always be kept in mind.

In 1992, a virus (DPP2209) isolated from sentinel cattle located at Coastal Plains Research Station, latitude 12 degrees 39'S, longitude 131 degrees 20'E, approximately 60 km east of Darwin, Northern Territory. This virus was identified as a serotype of epizootic haemorrhagic disease (EHD) of deer virus previously undescribed in Australia. An additional 17 isolation of this virus were made from eight animals during the period February to May. Electron microscopic studies showed the presence of orbivirus-like structures. Serogrouping ELISA, indirect immunofluorescence assay and the serogrouping plaque reduction neutralisation test indicated the virus was a member of the epizootic haemorrhagic disease serogroup. Serotype specific plaque reduction neutralisation tests, indicated the virus was a member of the epizootic haemorrhagic disease serogroup not previously isolated in Australia. Analysis of the VP3 gene confirmed this observation. Cross neutralisation testing of the isolate with known epizootic haemorrhagic disease serotype viruses including endemic Australian and exotic strains identified isolate DPP2209 as epizootic haemorrhagic disease virus serotype 1.

The post mortem demonstration continues to play an important role in medical education at both undergraduate and postgraduate levels. However, for a variety of reasons its format needs to be adaptable in order to maximize the educational benefit and increasingly to comply with legislation. We have developed a method of demonstration based upon a still video camera system which we have found is acceptable to our students and overcomes some of the problems associated with other formats. In this paper we describe the system itself, compare it with other systems, discuss its potential applications and present the results of a questionnaire-based survey of the first groups of medical students to be taught in this way. These results confirm that the technique is popular with students and that it appears to be a useful teaching modality worthy of further development.

The human thyrotropin receptor (hTSHR) represents an autoantigen that plays a central role in the hyperthyroidism of Graves' disease (GD). hTSHR transcripts have recently been detected by reverse transcription (RT)-PCR in various extrathyroidal tissues, suggesting that the hTSHR may be more widely distributed than previously thought, and that it may serve as a common antigen in the thyroidal and extrathyroidal manifestations of GD. Using techniques other RT-PCR, we examined whether RNA encoding hTSHR and hTSHR-specific immunoreactivity can be detected and localized in cultured orbital fibroblasts (OF), orbital connective tissue, extraocular muscle and various extraorbital tissues derived from both patients with Graves' ophthalmopathy (GO) and normal individuals. Using in situ hybridization with a digoxigenin-labeled antisense oligonucleotide probe specific for the extracellular domain of hTSHR, specific perinuclear and cytoplasmic hTSHR gene expression was detected in OF of patients with GO and, to a lesser degree, normal individuals. Using a highly sensitive immunostaining technique and a panel of monoclonal and polyclonal antibodies directed against different epitopes of recombinant hTSHR, distinct cytoplasmic hTSHR-like immunoreactivity was detected in methanol-fixed OF and orbital connective tissue, which was absent in abdominal fibroblasts, or when using isotype-matched non-immune immunoglobulins. Mouse monoclonal and pig polyclonal hTSHR antibodies detected cytoplasmic hTSHR-like immunoreactivity in perimysial fibroblasts within extraocular muscle, but not in extraocular muscle fibers. Immunocytochemical staining with rabbit polyclonal hTSHR antibody revealed, in addition, distinct cell surface-associated immunoreactivity in paraformaldehyde-fixed OF, but not in abdominal fibroblasts. Taken together, our results suggest that RNA encoding hTSHR is present and actively processed to immunoreactive hTSHR-like protein in OF residing within orbital connective tissue and extraocular muscle. These data support the concept that OF expressing intact or variant hTSHR may act as extrathyroidal targets for sensitized T-cells and immunoglobulins in GD.

The complete protein-coding region of the human relaxin-like factor (RLF; formerly Ley-I-L) was cloned by reverse transcription-polymerase chain reaction from human testis and subcloned into a bacterial expression plasmid for the production of recombinant human RLF in Escherichia coli. Polyclonal antibodies were raised against the recombinant RLF, as well as against a peptide epitope from the B-domain of the RLF polypeptide. Antibodies were used for immunohistochemistry of Bouin-fixed, paraffin-embedded samples of human testis tissues. Specific immunoreactivity was located exclusively in the Leydig cells with a consistent high intensity of staining, showing similar spatial distribution to other Leydig cell markers, such as the luteinizing hormone (LH) receptor and 3 beta-hydroxysteroid dehydrogenase (3 beta-HSD), and to the pattern of RLF mRNA shown by in-situ transcript hybridization. In biopsy samples from patients with severe disturbances of spermatogenesis, RLF staining intensity was consistently high in all cases, unlike staining for 3 beta-HSD which varied considerably between patients. Immunostaining for RLF would thus appear to offer an interesting new marker for Leydig cells in human testis samples.

Cerebral expression of c-fos protein was studied by immunocytochemistry in murine cerebral malaria (CM) and malaria without cerebral involvement (non-CM). c-fos expression, low in the brains of uninfected mice, increased in frequency, intensity, and distribution during the course of fatal CM (e.g., a 70-fold increase on day 7 after inoculation). These changes paralleled the timing and degree of the neurologic complications and histopathologic changes. Only a slight increase in c-fos expression was detectable in non-CM mice on day 7 after inoculation. Dexamethasone treatment (days 0 and 1 after inoculation) of the CM mice largely prevented the increased cerebral c-fos expression, histopathologic changes, cerebral complications, and death. Increased c-fos expression may indicate the specific neuronal pathways activated by the immunopathologic process of fatal murine CM and could be associated with the behavioral changes and neurologic complications in this model.

The mouse macrophage cell-line RAW264.7, stimulated with lipopolysaccharide, was used as a model for the study of the production of tumor necrosis factor (TNF) isoforms. TNF is synthesised initially as a 26 kDa transmembrane precursor, which is then processed enzymatically by a protease to release a mature molecule of 17 kDa. Dose-dependent production of transmembrane TNF was assessed by fractionation of cell membranes and Western blot analysis followed by autoradiography and densitometry. Isoforms of both the precursor and mature molecules were separated using two-dimensional (2-D) electrophoresis with immobilised pH gradient 3-10 linear gels as the first dimension. After radiolabelling of cells with 35S, both cell-associated and supernate-associated TNF isoforms were immunoprecipitated. A large number of protein spots were visualised on the 2-D gel map, for both the transmembrane and mature TNF species, more than have been detected previously using one-dimensional sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE). The likelihood that these putative isoforms were the result of differential glycosylation was tested by preincubating the cells with tunicamycin. This had the effect of reducing the number of protein spots, notably the higher molecular weight species. There were a number of precursor TNF isoforms that were unchanged upon tunicamycin treatment and these presumably reflect protein modifications other than glycosylation.

The National Breast Screening Programme affords the opportunity to study breast carcinomas at an early stage in their development. E-cadherin is a calcium-dependent, intercellular adhesion molecule whose loss of expression may facilitate the processes of invasion and metastasis of some human tumours. From a group of screen-detected ductal carcinomas less than or equal to 10 mm in diameter, 16 with lymph node metastasis were identified and matched for grade, size and patient age with node negative tumours. The level of expression of E-cadherin (detected by immunocytochemistry) was compared in the matched pairs using a simple semi-quantitative intensity distribution scoring system. The results showed a significant (P = 0.05 Wilcoxon paired rank test) reduction of E-cadherin expression in tumours with lymph node metastases compared to those without. In the context of the small size of these tumours it is proposed that these results support the hypothesis that reduction in E-cadherin expression is an early event in the development of metastases.

Background: The quest for a drug that would provide analgesia with minimal motor deficiency, through the selective inhibition of impulses in small-diameter fibers, was brightened by a previous report of veratridine's C-fiber-selective actions on the isolated rabbit vagus nerve. The goal of the present research was to demonstrate the same actions on rat sciatic nerve in vitro and to observe the functionally differential blockade in the rat in vivo., Methods: Sciatic nerves were removed from rats, mounted in a recording chamber, wherein a 1-cm length of the ensheathed nerve was superfused with the plant alkaloid veratridine (2 microM) in bicarbonate-buffered Liley's solution, and the compound action potential (CAP) was stimulated supramaximally to give A- and C-fiber elevations. Onset, steady-state, and recovery from veratridine effects were assayed for a range of stimulus frequencies. Open-field behavior and quantitative neurological assessments of proprioception, motor function, and nociception were tested in 15 trained rats after injection near the sciatic nerve of 0.1 ml veratridine at 0.5, 0.7, and 1.0 mM each plus epinephrine (1:200,000)., Results: Veratridine inhibited the C-fiber component of the CAP in a frequency-dependent manner. At 0.1 Hz the CAP was 65% of the control amplitude, 50% at 0.5 Hz, and 40% at 5 Hz. A-fiber elevations were unattenuated at stimulus frequencies as high as 50 Hz. Steady-state inhibition was reached 5 min after drug administration, and recovery from the effects was 30% complete by 15 min of drug washout. Proprioception, measured as a "hopping" or "placing" reaction, was inhibited dose dependently by maximum degree and for durations of, respectively, 0.5 mM, 61%, 180 min; 0.7 mM, 100%, 360 min; and 1 mM, 100%, 420 min. Extensor postural thrust, as a measure of motor function, was inhibited by and for 0.5 mM, 77%, 240 min; 0.7 mM, 99%, 390 min; and 1 mM, 100%, 420 min. Analgesia, as a prolonged withdrawal latency to a noxious thermal stimulus, had the following profile: 0.5 mM, 10%, 30 min; 0.7 mM, 52%, 150 min; and 1 mM, 66%, 150 min., Conclusions: Despite the fact that veratridine gave a C-fiber preferential blockade in the isolated sciatic nerve, heightened analgesia over motor block was not achieved in vivo. Indeed, just the opposite occurred. If preferential C-fiber blockade also occurs in vivo, then its traditionally expected correlation with analgesia must be re-examined.

Fatal murine cerebral malaria (FMCM) is an immunopathological process. The depletion of CD4+ T cells, or the administration of antioxidants or antibodies against certain cytokines, protect the mice against cerebral complications. We previously have shown that astrocytes, microglia, and monocytes play a role in the development of FMCM, suggesting that an active immune response occurs locally within the central nervous system. We now have investigated the functional involvement of glia and monocytes in FMCM by assessing 1) the production, 2) the temporal appearance, and 3) the cellular source of cytokine mRNA and protein in the brain. Brain sections from uninfected and FMCM mice were analyzed for the presence of cytokine mRNA and protein by in situ hybridization and immunohistochemistry. Tumor necrosis factor (TNF)-alpha mRNA and protein were associated with microglia and astrocytes, monocytes, and the cerebral vascular endothelium in FMCM mice but not uninfected animals. TNF-alpha mRNA was first detected several days before the animals showed cerebral symptoms and died. Interleukin (IL)-1 beta mRNA was found in the brains of both uninfected and FMCM mice. However, IL-1 beta protein was associated only with monocytes, the meningeal vascular endothelium, and neurons in the fronto-parietal cortex in the FMCM brains. No IL-4 or IL-6 mRNAs were detected in either group. These results provide the strongest evidence to date that cytokines, in particular TNF-alpha, produced locally in the central nervous system play a role in the pathogenesis of FMCM.

This study utilized bite-force measurements to determine the effect of various orthognathic surgery procedures on occlusal force generation in 42 patients with vertical facial deformities. The results showed that orthognathic surgery produced marked alterations in occlusal force levels which continued to occur up to a year after surgery. Measurements indicated that advancement of the mandible may result in weaker force levels, while bimaxillary surgery for the treatment of a "long face" brings the previously weaker bite force to a more "normal" level.

We have previously demonstrated an obligatory requirement for intracellular reactive oxygen species generation during T lymphocyte activation, and have proposed that intracellular reactive oxygen species may act as signalling agents in the regulation of certain cellular processes, for example, during cell cycle entry. To test this hypothesis, we have been interested to determine which, if any, cell cycle entry events are affected by oxidative signalling. In earlier studies, we have identified the transcription factors NF-kappa B and AP-1 as molecular targets for oxidative signalling processes during cell cycle entry, and have shown that oxidative signalling is involved in the regulation of early changes in gene expression during the G0 to G1 phase transition. To extend these initial observations, we have examined the effect of antioxidant treatment on the activity of the mitogen-activated protein kinases erk1 and erk2, as members of a signal transduction pathway known to directly regulate transcription factor function. Using as a probe cysteamine, an aminothiol compound with both antioxidant and antiproliferative activity, we have identified erk2, a key element of the MAP kinase pathway, as being responsive to oxidative signalling during lymphocyte activation. These observations provide further evidence to suggest a role for intracellular oxidant generation as a regulatory mechanism during cell cycle entry, and establish a link between oxidative signalling and other aspects of the intracellular signalling network that is activated in response to mitogenic stimulation.

We examined the optic nerve, as an analogous tissue to brain white matter, to assess possible relationships between changes in the blood-nerve barrier, axonal integrity, and astrocyte morphology in the central nervous system during fatal murine cerebral malaria (FMCM). In the FMCM model, namely, CBA mice infected with Plasmodium berghei ANKA, neurological symptoms begin around day 5 post-inoculation (p.i.) and mice become increasingly ill by day 7 p.i., at which time they lapse into coma and die. Using intravascular perfusion with horseradish peroxidase combined with light and electron microscopy, and GFAP immunohistochemistry, the optic nerves in malaria-infected mice were found to display i) breakdown of the blood-nerve barrier, detectable as early as day 3 p.i. (about 2 days before the onset of neurological symptoms) increasing to peak severity by day 7 p.i.; ii) monocytosis, vascular congestion, and monocyte adherence to the endothelium in the microvasculature during the later stages of the disease process; iii) an increased incidence of patchy axonal demyelination and degeneration, mostly associated with vascular changes and astrogliosis, beginning at day 5 p.i. and more evident by day 7 p.i.; and iv) an increased intensity of GFAP immunostaining, detectable from day 3 p.i. and peaking at day 7 p.i. These optic nerve changes were always seen in the infected individuals, though they varied in intensity. The temporal and anatomical coincidence between the compromised blood-nerve barrier, monocyte adherence to the vascular endothelium, astrocyte changes, neuronal degeneration, and demyelination in the optic nerve in FMCM suggests that these factors are mechanistically inter-related. These findings are consistent with the proposed immunopathological nature of FMCM and provide further evidence for the pivotal role of the CNS microvasculature in the disease process. This is the first investigation of involvement of the optic nerve in FMCM and the first demonstration, to our knowledge, of loss of axonal viability in this condition in any CNS tissue. The observed demyelination is consistent with reports by other workers on such changes in the brain in human cerebral malaria.

Microglia are pluripotent members of the macrophage/monocyte lineage that can respond in several ways to pathological changes in the central nervous system. To determine their role in the pathogenesis of fatal murine cerebral malaria (FMCM) we have conducted a detailed study of the changes in morphology and distribution of retinal microglia during the progression of the disease. Adult CBA/T6 mice were inoculated with Plasmodium berghei ANKA. These mice died 7 days post inoculation (p.i.) with the parasite while exhibiting cerebral symptoms, increased permeability of the blood-brain barrier, and monocyte adherence to the vascular endothelium. Mice were injected i.v. with Monastral blue 2 h prior to sacrifice to identify "activated" monocytes, and their isolated retinae were incubated with the Griffonia simplicifolia (GS) lectin or reacted for the nucleoside diphosphatase enzyme to visualize microglia and the vasculature. Changes in microglial morphology were seen within 2-3 days p.i., that is, at least 3 days prior to the onset of cerebral symptoms and 4 days before death. Morphological changes included retraction of ramified processes, soma enlargement, an increasingly amoeboid appearance, and vacuolation. There was also increased staining intensity and redistribution of "activated" microglia toward retinal vessels, but no increase in density of NDPase-positive cells. The GS lectin only labeled a small population of microglia in the uninfected adult mouse retina. However, there was a striking increase in the focal density of GS-positive microglia during the progression of the disease. Extravasation of monocytes also was observed prior to the onset of cerebral symptoms. These results provide the first evidence that microglial activation is a critical component of the pathological process during FMCM.

The purpose of this study was to determine the corrosive effects of multi-species biofilms on intra-oral magnets in the presence and absence of sucrose. Using pooled human saliva as an inoculum, biofilms were grown on the surfaces of 90 neodymium-iron-boron (Nd2Fe14B) magnets in a constant depth film fermentor under aerobic conditions at 37 degrees C. The fluid phase was a mucin-containing artificial saliva (delivered at a rate of 0.72/litres day-1), and, after 15 days, 100 ml of 10% (w/v) sucrose was added (as three pulses of 33.3 ml) each day for a further 15 days. Six magnets with attached biofilms were removed periodically. On each sampling occasion the numbers of aerobes, anaerobes, streptococci, veillonellae and actinomyces in each biofilm, the pH of the fermentor effluent and the dry mass of the magnets were determined. Addition of sucrose to the fermentor resulted in a fall in pH (from a mean of 6.94 to a mean of 4.96), an increase in the proportion of streptococci and a decrease in the proportion of veillonellae comprising the biofilms. The decrease in mass of the magnets was 28-fold greater in the presence of sucrose than in its absence. The results of this study have shown that the presence of sucrose affects the microbial composition of multi-species biofilms growing on Nd2Fe14B magnets and results in a marked increase in corrosion of the magnets.

Relaxin-like factor (RLF) is a new member of the insulin/insulin-like growth factor family of hormones and growth factors, which appears to be predominantly expressed in the Leydig cells of the testis. An analysis of male and female bovine tissues indicated that in the cow the RLF gene is also highly expressed in the female, mainly in the follicular theca interna and in the corpus luteum, with a pattern of gene expression very similar to that for the related peptide relaxin in other species. Sequence analysis of bovine testicular and luteal RLF and cDNA shows that the same gene is expressed in both male and female gonads. Because the bovine RLF sequence, like those from pigs, humans, and mice, retains, the putative receptor binding motif described for relaxin, it seems plausible that RLF might functionally substitute for relaxin in the cow, in which the latter peptide appears not to be significantly expressed.

We previously have demonstrated an obligatory requirement for intracellular reactive oxygen species (ROS) generation during T lymphocyte activation, and have proposed that ROS may act as signalling agents in the regulation of certain cellular processes, for example, during cell cycle entry. In order to test this hypothesis, we have been interested to determine which, if any, cell cycle entry events are affected by oxidative signalling. Given the requirement for both oxidative signalling and altered gene expression during the G0 to G1 phase transition, we have attempted to establish the extent to which oxidative signalling affects global gene expression patterns during cell cycle entry, and to isolate and characterize mRNAs whose expression patterns are responsive to oxidative signalling during this process. Using differential display in a phenotypic screening approach, we have identified 10 mRNA species whose expression patterns were altered in response to inhibition of oxidative signalling during cell cycle entry. The expression patterns of 4 of these 10 mRNAs were unaffected during cell cycle arrest caused by a different mechanism, cyclosporin A-induced interference with calcineurin-mediated signalling events, implying that the altered expression patterns seen were not simply a consequence of cell cycle arrest. This suggests that the expression of these 4 mRNAs is regulated by a mechanism both necessary for cell cycle entry and sensitive to oxidative signalling. RNAse protection assays confirmed that 2 of these 4 mRNAs were indeed responsive to redox regulation. These observations strongly suggest an involvement for oxidative signalling in the regulation of gene expression during the G0 to G1 phase transition, in peripheral blood mononuclear cells at least.

The relationships between increased vascular permeability to protein, monocyte adherence to the endothelium, and expression of the cell adhesion molecules, intercellular adhesion molecule-1 (ICAM-1) and vascular cell adhesion molecule-1 (VCAM-1) in the central nervous system microvasculature were studied during the progression of fatal murine cerebral malaria. CBA mice were inoculated with Plasmodium berghei ANKA, and changes in the retinal microvasculature were examined on days 3, 5, and 7 postinoculation (p.i.). Evans blue dye and horseradish peroxidase (HRP) were administered intravenously to assess vascular permeability to macromolecules macroscopically and by light and electron microscopy. ICAM-1 and VCAM-1 expression were examined by immunohistochemistry. HRP leakage into the retinal parenchyma was seen macroscopically at a low level on day 3 p.i., increasing progressively at day 5 (the earliest time at which cerebral symptoms were observed) and day 7 (the day on which animals showed severe behavioral abnormalities and died). The inner retinal vascular plexus showed a slight increase in vascular permeability to intravenous Evans blue at day 3 p.i. and congestion, monocyte adherence to the endothelium, and increased vascular permeability to both Evans blue and HRP at day 7 p.i. Electron microscopic observations were consistent with these findings and also revealed disrupted light junctions and the coating of monocytes and endothelium with HRP at day 7 p.i. Immunohistochemical staining and densitometry showed a progressive increase from day 3 to day 7 p.i. in the densities of ICAM-1 and VCAM-1 on the venular endothelium of the inner retinal vascular plexus, with the appearance of adherent ICAM-1+ monocytes at the terminal stage of the disease. None of the pathological changes associated with the inner retinal plexus were seen at any stage in the outer retinal plexus. These results suggest the following sequence of events in the inner retinal vessels, particularly the venules, during the progression of fatal murine cerebral malaria: 1) a mild increase in vascular permeability at approximately day 3 p.i., 2) a progressive increase in endothelial expression of the cell adhesion molecules ICAM-1 and VCAM-1, commencing at approximately day 3 p.i., 3) monocyte adhesion to the endothelium starting at approximately day 5 p.i., and 4) frank disruption of endothelial integrity at the terminal stage (day 7 p.i.), leading to edema and hemorrhage. Similar changes in cerebral vessels may underlie the neurological complications of the disease.

Recent changes in the classification of psychiatric illnesses have resulted in the term dysmorphophobia being replaced by that of body dysmorphic disorder (BDD). This paper attempts to alert the clinician to the presenting features of the condition and discusses its management, with particular emphasis on the role of surgery and current concepts of pharmacological treatment. A number of case reports are included to illustrate the diversity of this interesting disorder and the difficulties involved in treatment.

The use of high temperature antigen retrieval methods has been of major importance in increasing the diagnostic utility of immunocytochemistry. However, these techniques are not without their problems and in this report attention is drawn to a loss of nuclear morphological detail, including mitotic figures, following microwave antigen retrieval. This was not seen with an equivalent autoclave technique. This phenomenon was quantified using image analysis in a group of B cell lymphomas stained with the antibody L26. Loss of nuclear morphological detail may lead to difficulty in identifying cells accurately, which is important in the diagnostic setting-for example, when trying to distinguish a malignant lymphoid infiltrate within a mixed cell population. In such cases it would clearly be wise to consider the use of alternative high temperature retrieval methods and accept their slightly lower staining enhancement capability compared with the microwave technique.

A cDNA clone encoding a novel endozepine-like peptide (ELP) was isolated from mouse testes, sequenced, and its mRNA expression characterized by northern and in situ hybridization. ELP mRNA was found exclusively in the late spermatid stages of spermatogenesis in the testes of sexually mature mice and in no other tissue or cell type examined. It was also expressed in rat, bovine, porcine and sheep testes. Mouse ELP-encoding cDNA was used to construct expression vectors for the production of ELP in bacteria, and the purified bacterial protein used to raise polyclonal antibodies in rats. These antibodies identified the predicted endogenous ELP in extracts of mouse testis and epididymis and in no other tissue. Immunohistochemistry confirmed that the ELP antigen was present only in late spermatids and spermatozoa, particularly within the cytoplasmic droplet which is retained by the mature spermatozoa during their transit into the epididymis. We conclude that ELP is an intracytoplasmic peptide exclusively expressed in post-meiotic spermatozoa and which may be involved in the energy metabolism of the mature sperm.

Coordinated patterns of lower facial muscle activity have been observed using 8-channel electromyography (surface electrodes in a bipolar configuration) on 11 healthy volunteers to provide a picture of the patterns of muscle activity during function. Measurements of integrated electrical activity were made to allow comparison of different muscle groups during active, active-against-resistance, and passive movements. A contraction reflex was assessed by an electromechanical device and noted in 8 of 11 subjects. Lower facial movements were found to involve simultaneous bilateral activity in all the muscle groups tested bilaterally and to require a balance between dilator and constrictor forces. The significance for facial reanimation surgery is that it is desirable to reconstruct balanced constrictor and dilator forces, the latter having vector pulls upwards, downwards and laterally to reconstruct the normal mechanism of lower facial movement.

The definition, evaluation and assurance of quality of health care are becoming increasingly important with health care moving into purchaser/provider mode. This questionnaire based study investigated patient satisfaction and changes in their quality of life following joint orthodontic/surgical treatment for the correction of facial deformity. Questionnaires were distributed to 83 pre-operative and a separate group of 100 postoperative patients with data analysis involving comparison of pre- and postoperative mood states and opinions about various aspects of appearance and personality. The results indicated that the majority of respondents were happy with the outcome of treatment. There was significant improvement in appearance, as well as in self-confidence, overall mood states and the ability to mix socially. The majority of respondents felt that the technical aspects of the operation had been well explained but almost a quarter felt that the effects following surgery were badly explained. Pre-operative counselling, therefore, needs to be improved.

This investigation examined the magnetic flux and repulsive force of neodymium-iron-boron (Nd-Fe-B) magnets based on Nd2Fe14B with acrylic coatings in different orientations. The flux was measured with a Hall probe and the force measured by electronic scales with the magnets mounted on a laboratory jack. Results show that there were no magnetic losses after embedding the magnets in acrylic bite-blocks, although there were significant flux losses when the magnet blocks were directly heated. The alignment of the magnets over each other was of the utmost importance and significantly affected the repulsive force between the magnets. When the magnets were mounted in an articulator to simulate jaw movement, the force levels between the magnets was further reduced as a result of the effects of the rotation of the articulator. It is concluded that Nd-Fe-B type magnets embedded in acrylic can be used to give predictable repulsive forces in the mouth. The dramatic reduction in force levels when the magnets are not in optimal alignment, however, makes their effectiveness in aiding intrusion of teeth doubtful.

This investigation examined the magnetic flux and repulsive force of neodymium-iron-boron (Nd-Fe-B) magnets used in repulsion in the treatment of anterior open bite. Three different grades of magnets each of five different dimensions were used. Magnetic flux was measured by a Hall probe and the repulsive force by an Instron mechanical testing machine. Results showed that the different grades of magnet displayed similar magnetic flux with a relationship between thickness and flux. There was a significant difference between the flux measured at the end as opposed to the center of the magnet. The force between two magnets and the flux above each magnet was directly related. Using magnets in groups did not produce higher forces than using single magnets of an equivalent size. It was concluded that Nd-Fe-B magnets can be used to give predictable repulsive forces in the mouth.

Graves' disease is attributed to the presence of autoantibodies with agonist activity which interact with the TSH receptor causing thyroid hyperstimulation and hyperthyroidism. The degree of TSH-binding inhibition (TSH-BI) caused by a Graves' serum in a TSH radioligand receptor assay is considered to be an index of the prevalence of anti-TSH receptor autoantibodies in that serum. We have previously shown that the specific inhibition by Graves' serum of hCG-stimulated steroidogenesis by Leydig cells was at a site distal to receptor binding and second messenger activation. In this report, we have investigated whether the effect of Graves' serum upon Leydig cells is a property of the constitutive antibodies. Immunoglobulin-enriched fractions were obtained from Graves' and normal sera using three increasingly rigorous procedures; ammonium sulphate precipitation, caprylic acid treatment and Protein A or G-affinity purification. The TSH-BI was determined for untreated and extracted sera in two radioreceptor assays developed for use with serum, one using human thyroid membranes and the other using HeLa cells transfected with the human TSH receptor, and the results were compared with effects in the Leydig cell steroidogenesis bioassay. The specific inhibition of hCG-stimulated Leydig cell steroidogenesis by Graves' sera was not retained in the antibody fraction causing TSH-BI. Thus, the inhibitory factor appears not to be an antibody and we are now attempting to purify and identify the responsible factor from Graves' serum.

Three methods of bonding orthodontic brackets to acrylic teeth were investigated for use with a typodont. One-hundred-and-twenty Dentaurum 0.018 x 0.025 inch standard Edgewise brackets were bonded to Crylopax cross-linked acrylic teeth using three cements: Concise composite, Concise with acrylic primer, and GAC acrylic cement on a previously roughened tooth surface. Half the specimens in each group was immersed in a thermostatically controlled water bath for 15 min on 20 occasions over a 5-day period, to simulate the conditions of a typodont, prior to measuring tensile bond strength on an Instron machine. Under dry conditions Concise composite was significantly stronger than either Concise with acrylic primer or GAC cold cure acrylic resin. Under wet conditions, however, the mean bond strength of the acrylic resin increased significantly compared to the other cements. The bond strength of both Concise and Concise composite with primer deteriorated following water immersion. It is suggested that when setting up typodont teeth with brackets bonded with GAC acrylic resin, immersion of the bonded teeth in hot water prior to positioning of the teeth in the typodont may greatly reduce the risk of subsequent bracket loss.

With the aid of recombinant DNA technology (PCR/site directed mutagenesis, sequencing) the full length coding region of the human TSH receptor was manipulated to place a specific epitope peptide tag (FLAG epitope sequence) at the carboxyl end of the protein. The resulting construct was cloned into a eukaryotic expression vector and stably transfected into HeLa cells. The expression/translation of the tagged TSH receptor molecule was monitored by immune-precipitation and western blotting of protein lysates, and was found to be expressed at considerable levels using the commercially available antibodies directed towards the FLAG epitope. This analysis revealed two discrete specific bands 90-120 KDa representing, presumably, differently glycosylated forms of the receptor. TSH radio receptor assays demonstrated that the FLAG tagged TSH receptor bound TSH comparable with the wild type receptor. Furthermore TSH stimulated cAMP response in these transfected cells were comparable to the wild type receptor, thus demonstrating that the tagged receptor was functionally identical to the transfected wild type receptor. These cell lines will be of great value when analysing TSH/receptor or receptor/autoantibody interactions considering the availability of well characterized experimental anti-TSH receptor sera.

Follow-up of patients who have undergone orthognathic surgery is well documented in the literature, usually in the form of questionnaire-based studies or clinical interviews. However, there have been few long-term follow-up investigations. This study aimed to look at the long-term outcomes of such operative procedures. Questionnaires were sent to 68 patients who had undergone joint orthodontic and orthognathic surgery procedures 5 to 16 years previously, after the patients were contacted by telephone to learn the nature of the study. The 49 responses were compared with a previously analyzed preoperative group and a short-term postoperative group (1 to 3 years following surgery) from the same institution. Respondents showed high levels of satisfaction overall, with the majority believing they had made the correct decision to undergo surgery. The main problem areas identified were those concerning presurgical explanations and lack of advice regarding side-effects. This area requires improvement as postsurgical dissatisfaction can be minimized by accurate descriptions of the treatment. A number of respondents reported persistent lip paresthesia and continued limitation of mouth opening. However, the majority of patients commented that these persistent symptoms had no effect on their daily life.

The extracellular domain of the TSH receptor (TSHR-561, amino acids #78-389) was expressed as a hexa-histidine fusion protein in bacteria. The recombinant protein was purified to homogeneity and used to immunize porcine and ovine species. High titre antibodies were obtained from both species that recognized the recombinant protein in Western blot analysis but failed to interfere with the TSH radio receptor assay. An epitope library was constructed and screened with affinity purified ovine and porcine antisera and detected a number of positive clones. Sequence analysis revealed that all of the epitopes contained sequences derived from the carboxyl terminus of the recombinant immunogen. One clone defined an epitope covering 16 amino acids from the carboxyl terminus and was the common epitope found in all of the other clones. Western blot screening of a large panel of Graves' sera with recombinant TSH receptor protein identified one patient sera that also recognized linear epitopes in the TSHR-561 protein. Experimentation demonstrated that the linear epitope recognized by this human sera was identical to the sequence recognised by the animal antisera. This sequence is unique to the TSH receptor and will be useful in further studies to analyze the TSH receptor protein.