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Telemedicine system using computed tomography van of high-speed telecommunication vehicle.

The current medical system provides medical services to patients who visit hospitals. However, medical services can be provided at or close to the home of the patient using fully equipped mobile telemedicine systems. Such a system can identify the disease at an early stage, improve quality of life and prognosis through early diagnosis and treatment, and reduce the costs of medical service. Furthermore, the unit can provide mass screenings of the population, as well as full medical service to remote areas. The Telecommunications Advanced Organization of Japan, Matsumoto, Japan, and Shinshu University Hospital, Matsumoto, Japan, established a research center for a unique telemedicine project using a mobile system. The mobile unit consists of a van that houses a spiral computed-tomography (CT) machine and various telecommunications equipment. The unit allows medical examination, CT scanning, and on-line two-way transfer of image data/teleconferencing to a medical center for consultation with various specialists. We have used the system thus far for the early detection of lung cancer through mass screenings over a four-year period in 29 administrative districts. Mass screenings of 19117 residents resulted in the identification of 75 cases of early lung cancer who were later treated by partial pneumonectomy at Shinshu University Hospital and affiliated hospitals. We have also used the system to provide medical services to rural areas, as telemedicine support at remote areas, wintertime telemedicine support to an international sports competition, and various medical services to a home-care facility.

Humans↗

In awe of subcellular complexity: 50 years of trespassing boundaries within the cell.

In this review I describe the several stages of my research career, all of which were driven by a desire to understand the basic mechanisms responsible for the complex and beautiful organization of the eukaryotic cell. I was originally trained as an electron microscopist in Argentina, and my first major contribution was the introduction of glutaraldehyde as a fixative that preserved the fine structure of cells, which opened the way for cytochemical studies at the EM level. My subsequent work on membrane-bound ribosomes illuminated the process of cotranslational translocation of polypeptides across the ER membrane and led to the formulation, with Gunter Blobel, of the signal hypothesis. My later studies with many talented colleagues contributed to an understanding of ER structure and function and aspects of the mechanisms that generate and maintain the polarity of epithelial cells. For this work my laboratory introduced the now widely adopted Madin-Darby canine kidney (MDCK) cell line, and demonstrated the polarized budding of envelope viruses from those cells, providing a powerful new system that further advanced the field of protein traffic.

Cell Membrane↗

Chondral delamination of the knee in soccer players.

This study reports on the diagnosis and treatment of chondral delamination in the knees of soccer players. The authors attempted to provide a clinical standard for comparison with emerging cartilage repair techniques. Twenty-three consecutive chondral fractures in a homogeneous population of athletes were evaluated using physical examination, magnetic resonance imaging, and arthroscopic examination. All knees were treated with debridement to a stable border and removal of the calcified cartilage base. Results were graded using the scoring system advocated by proponents of chondral transplant. All patients reported pain that limited soccer activities. Effusions occurred in 48%, joint line tenderness in 33%, and crepitus in 19% of the knees. Results from magnetic resonance imaging correlated with arthroscopic examination in 21% of the knees. Arthroscopic examinations revealed lesions on the medial femoral condyle in eight patients, the patella in six, the lateral femoral condyle in six, and the trochlea in three. Athletes returned to play at an average of 10.8 weeks. Repeat arthroscopic procedures in eight cases revealed fibrocartilage at the initial site. There were 6 excellent, 9 good, 0 fair, and 0 poor results at their 1-year followup examinations (15 knees). The definitive diagnosis of chondral delamination relies on a thorough arthroscopic probing of the articular surface. The early functional results of this treatment compare favorably with the autologous transplantation technique. Caution, however, is recommended when treating articular cartilage injuries because no long-term data exists on whether any treatment modality can prevent the development of degenerative joint disease.

Adult↗

[Formation of plastic connections of the visual system during ontogenesis of the rabbit].

Analysis of evoked potentials and unit activity in the visual cortical projection area of rabbits revealed a definite succession of forming of interneuronal connections in ontogeny. In early postnatal period, the neuronal reactions were characterized by stable responses with one excitatory phase corresponding to initially negative surface evoked potential. Similarity of reactions of neurones situated in the same vertical column was observed and explained by functioning of a system of rigid connections of the thalamic relay nuclei afferents with cortical pyramidal neurones. Beginning from the third week of postnatal life of rabbits the neuronal reactions assumed a distinctly expressed phasic character, and variability of responses was seen along the vertical line. The changes revealed correlated with formation of a system of interneurones providing a possibility of plastic neuronal interaction. A study of the influence of preliminary cortical stimulation of the associative areas showed that intercentral cooperation mediated by cortical interneurones providing a systemic analysis of visual information began to form from the third week of postnatal life and reached the definitive level at later stages of development.

Animals↗

New characters for the functional interpretation of primate scapulae and proximal humeri.

The study of muscle function in nonhuman primates through the technique of electromyography (EMG) has facilitated the identification of specific functional roles for muscles in particular behaviors. This has led to a more complete understanding of the biomechanics of certain regions of the musculoskeletal system, and should facilitate our ability to identify morphological features useful in the functional interpretation of fossil material. The current paper represents one such investigation of a new set of morphometric characters of the scapula and proximal humerus suggested by EMG analyses of shoulder muscle function. A set of new metric variables were examined on the scapulae and proximal humeri of 25 species of extant anthropoid primates, as well as on casts of scapulae and humeri of three fossil primate taxa. The variables are primarily related to the line of action and attachments of the rotator cuff muscles. The position of the scapular spine, the degree of lateral expansion of the subscapular fossa, the size and shape of the subscapularis insertion facet on the lesser tubercle, and the orientation of the infraspinatus insertion facet on the greater tubercle all appear to successfully sort the extant taxa into locomotor groups. Their appearance on the fossil specimens generally supports previous functional interpretations of each taxon's locomotor abilities based on a variety of other characters, suggesting that these traits are equally applicable to fossil material.

Animals↗

Stimulus symmetry affects the bisection of figures but not lines: evidence from event-related fMRI.

Many geometric shapes retain their symmetry when bisected, but appear asymmetrical when misbisected. We have previously shown that this correspondence can guide the accuracy and speed of perceptual bisection (Landmark) judgments. Using event-related fMRI, here we examined whether the behavioural effects of symmetry are also evident at the neural level. The data showed that the presence/absence of symmetry modulates the activity of right anterior cingulate gyrus, an area associated with a variety of higher level attentional functions. A previous visual half-field study also showed that bisected lines are apprehended more quickly and accurately than misbisected lines in right, but not left, visual field. We were able to localise this advantage to right superior temporal gyrus. Significantly, we found no evidence that symmetry played a role in apprehending the midpoint of the line stimuli traditionally used to assess visual neglect. The data clarify the effects of visual symmetry on bisection behaviour, and highlight novel dissociations within the neural systems thought to underline Landmark performance.

Adult↗

Rescue of measles virus using a replication-deficient vaccinia-T7 vector.

A system which allows the reconstitution of measles virus (MV) from cloned cDNA is described. The severely host cell restricted vaccinia vector MVA-T7 expressing bacteriophage T7 RNA polymerase was used to generate full-length antigenomic MV RNA and simultaneously the mRNAs encoding the viral N, P and L proteins in order to produce replicationally and transcriptionally active nucleocapsids. The functionality of the N, P and L proteins was demonstrated first by their ability to rescue MV specific subgenomic RNAs. Assembly and budding of reconstituted MV was shown by syncytia formation and subsequently by virus isolation. The inability of MVA-T7 to produce progeny virus in most mammalian cells circumvents the necessity to separate the reconstituted MV from the MVA-T7 helper virus. Since all components are expressed transiently, this system is especially suitable for studying the functions of N, P and L. Furthermore, it is useful for investigating later steps in the MV life cycle.

Animals↗

Control pathways of the 67 kDa laminin binding protein: surface expression and activity of a new ligand binding domain.

A number of papers have been published on the clinical correlation of the expression of the 67 kDa laminin binding protein (LBP) with the metastatic potential of solid tumors. Both mRNA and protein expression levels have been reported, but both the relationship between them and the molecular nature of the 67 kDa surface product remain unclear. We have utilized a homotypic overexpression system to investigate the cell surface presentation of the 67 kDa LBP and the contribution of this protein to the invasive phenotype of cultured cell lines. We report here that the cellular mRNA levels do not directly reflect the levels of the 67 kDa LBP observed on the cell surface in this overexpression system. Methotrexate amplification of transfected plasmids expressing the 67 kDa LBP leads to an initial elevation of both the LBP mRNA and surface protein levels. This is accompanied by an altered, more flattened, cell morphology. Later, apparent adaptation of the cells to methotrexate is accompanied by a down-regulation of the surface expression of the protein. mRNA levels, however, remain elevated. A nine amino acid sequence, CDPGYIGSR (peptide 11), within the beta chain of laminin 1 has been identified as a probable binding domain for the 67 kDa LBP. Previous studies have identified a region of the 67 kDa LBP which may be involved in laminin interaction, although not necessarily via the peptide 11 domain. We have identified a second site within the amino acid coding sequence of the 67 kDa LBP which also shows biological activity both in vitro and in vivo. A peptide with this sequence, LBP residues 205-229, binds laminin-1 in a peptide 11 inhibitable manner. The receptor-derived peptide modulates invasion of basement membrane matrix in vitro and inhibits experimental lung colony formation when injected along with B16BL6 mouse melanoma cells. However, pretreatment of the melanoma cells with the peptide enhances lung colony formation. Thus, the interaction of the 67 kDa LBP with basement membrane matrix appears to involve a complex series of events including multiple adhesive sites and tight regulation of cell surface expression.

Amino Acid Sequence↗

Characterization and modeling of P-type electrosensory afferent responses to amplitude modulations in a wave-type electric fish.

The first stage of information processing in the electrosensory system involves the encoding of local changes in transdermal potential into trains of action potentials in primary electrosensory afferent nerve fibers. To develop a quantitative model of this encoding process for P-type (probability-coding) afferent fibers in the weakly electric fish Apteronotus leptorhynchus, we recorded single unit activity from electrosensory afferent axons in the posterior branch of the anterior lateral line nerve and analyzed responses to electronically generated sinusoidal amplitude modulations of the local transdermal potential. Over a range of AM frequencies from 0.1 to 200 Hz, the modulation transfer function of P-type afferents is high-pass in character, with a gain that increases monotonically up to AM frequencies of 100 Hz where it begins to roll off, and a phase advance with a range of 15-60 degrees. Based on quantitative analysis of the observed gain and phase characteristics, we present a computationally efficient model of P-type afferent response dynamics which accurately characterizes changes in afferent firing rate in response to amplitude modulations of the fish's own electric organ discharge over a wide range of AM frequencies relevant to active electrolocation.

Action Potentials↗

Microcomputer-driven digital data processing of mouse actograph activity.

The system is designed to measure the ambulatory activity of each of a set of eight mice, using a group of actographs driven by an inexpensive microcomputer, thus avoiding unnecessary usage of a costly instrument over an extended period. A toric actograph was chosen since it is sensitive to ambulatory activity even at low amplitudes. In this computerized system, actograph data are recorded on audio cassettes during the experiment, and later reread by an Apple II microcomputer for storage on floppy disks. This makes it possible to perform all types of mathematical treatment at a later date. Since the experiments last for several days and the environment is kept constant, activity rhythms can be studied. The data processing takes place off-line and employs digital filtering, which is shown to be preferable for the presentation of temporal data to a simple smoothing, by a moving average. Spectral analysis is obtained through a discrete Fourier transform. Two methods of reconstitution of temporal curves from the frequency analyses are presented: an inverse Fourier transform, or summation of the Fourier series.

Animals↗

Regulation of the rat brain endothelin system by endogenous beta-endorphin.

Several lines of evidence indicate that the central endogenous opioid and endothelin (ET) system regulate each other. To explore this idea further, we determined the effect of intracerebroventricular (i.c.v.) administration of anti-beta-endorphin IgG (rabbit) on the expression level of the opioid, corticotropin-releasing hormone and endothelin receptors, and tissue concentration of ET-1. Three days after implanting cannula into the lateral ventricle, male Sprague-Dawley rats were administered 10 microl (i.c.v.) of either control rabbit IgG (2.5 microg/microl) or anti-beta-endorphin IgG (2.5 microg/microl) on days 1, 3 and 5. On day 6, animals were euthanized and caudate, cortex and hippocampus collected for Western blot analysis. Anti-beta-endorphin IgG down-regulated ET-A receptor protein expression in the caudate (51%), but had no effect on the expression of mu, delta, kappa opioid, ET-B, CRH-1 and CRH-2 receptors in any brain region. Anti-beta-endorphin IgG increased tissue ET-1 levels in the caudate by 30.3%. [35S]GTP-gamma-S binding assays demonstrated that anti-beta-endorphin IgG increased the efficacy of [D-Ala2-MePhe4, Gly-ol5]enkephalin without altering its potency in caudate. Control experiments showed that there was no detectable rabbit IgG in caudate, cortex and hippocampus samples. These results suggest that beta-endorphin in the CSF coordinately regulates ET-1 levels and the ET-A receptor in rat caudate. These findings support the hypothesis that CSF neuropeptides have regulatory effects and further demonstrate a link between opioid and ET system.

Animals↗

Marked induction of IL-6, haptoglobin and IFNgamma following experimental BRSV infection in young calves.

Bovine respiratory syncytial virus (BRSV) has been identified worldwide as an important pathogen associated with acute respiratory disease in calves. An infection model has been developed reflecting accurately the clinical course and the development of pathological signs during a natural BRSV-infection. In the experiments described in the present study, calves were infected at 13-21 weeks of age and reinfected 14 weeks later. Blood samples from the entire infection period were analysed for acute phase protein (haptoglobin) by ELISA and for expression (mRNA level in peripheral blood mononuclear cells) of the cytokines interleukin-2 (IL-2), interleukin-4 (IL-4), interleukin-6 (IL-6) and interferon-gamma (IFNgamma) by quantitative real-time reverse transcribed polymerase chain reaction (RT-PCR). IFNgamma, interleukin-6 and haptoglobin were markedly induced together with development of clinical signs in response to the first infection with BRSV. The IFNgamma response was biphasic, with an early peak at day 1-3 post infection (p.i.) and a later increase between day 5 and 8 p.i. Reinfection also resulted in an induction of IFNgamma, but without induction of clinical signs, IL-6 and haptoglobin. These results indicate that early mediators connected with the innate responses are induced on a first encounter with the pathogen, but not on a second encounter (reinfection) where the adaptive immune system may act as the first line defence.

Animals↗

Quantitative postural load assessment for whole body manual tasks based on perceived discomfort.

Many Korean workers are exposed to repetitive manual tasks or prolonged poor working postures that are closely related to back pain or symptoms of musculoskeletal disorders. Workers engage in tasks that require not only handling of heavy materials, but also assuming prolonged or repetitive non-neutral work postures. Poor work postures that have been frequently observed in the workplaces of shipbuilding shops, manufacturing plants, automobile assembly lines and farms often require prolonged squatting, repetitive arm raising and wrist flexion and simultaneous trunk flexion and lateral bending. In most manufacturing industries, workers have to assume improper work postures repetitively, several hundreds of times per day depending on daily production rate. A series of psychophysical laboratory experiments were conducted to evaluate the postural load at various joints. A postural load assessment system was then developed based on a macro-postural classification scheme. The classification scheme was constructed based on perceived discomfort for various joint motions as well as previous research outcomes. On the basis of the perceived discomfort, postural stress levels for the postures at individual joints were also defined by a ratio scale to the standing neutral posture. Laboratory experiments simulating automobile assembly tasks were carried out to investigate the relationship between body-joint and whole-body discomfort. Results showed a linear relationship between the two types of discomfort, with the shoulder and low back postures being the dominant factor in determining the whole body postural stresses. The proposed method was implemented into a computer software program in order to automate the procedure of analysing postural load and to enhance usability and practical applicability.

Humans↗

Gait analysis in laying hens and broilers with and without leg disorders.

Selection of broiler strains for high body weight has changed the anatomical characteristics and in connection to that, the pattern of locomotion. In addition, rapid growing broilers show a high incidence of leg disorders which compromise the walking ability of the birds. Differences in the patterns of locomotion between laying hens and broilers and between broilers with and without leg disorders can be demonstrated and quantified by gait analysis. The gait pattern was recorded by videotracking. Three points of the body - the cloacal region and both feet joints - were marked by small patches of reflecting foils. The vertical and horizontal movements of the marked points were recorded by a camera in posterior position, while the bird was walking on a treadmill The camera was connected with a PC-operated videotracking system. The vertical and horizontal movements of the 3 marked points were recorded simultaneously and plotted against the time axis. Kinetograms show clearly the differences in the walking pattern between broilers and layers. Layers place the legs directly under the centre of gravity and, therefore, the body moves in a straight line. Broilers, in contrast, move the centre of gravity step by step laterally towards the position of the supporting leg. This pattern may be caused by the anatomical characteristics of broilers. Limping in broilers with leg problems can be measured by differences in the lateral and vertical movements of the right and left leg.

Animals↗

IL-12/IL-2 combination cytokine therapy for solid tumours: translation from bench to bedside.

A broad range of approaches are under active investigation for the biological therapy of cancer, in particular, strategies directed at host immune response potentiation. These efforts have been fuelled by studies demonstrating the presence of an endogenous, but ineffective, host antitumour immune response and a greater understanding of the key factors which regulate this response. These mechanisms involve complex interactions between various effector cell populations, soluble factors and the tumour itself and are determined by the timing and relative intensity of positive and negative autoregulatory pathways, as well as a variety of immunosuppressive effects capable of mediating tumour self-defence. Based on these observations, immunotherapeutic regimens have been developed to potentiate antigen-specific sensitisation of effector cells with tumour vaccines/adjuvants, expand and amplify the number and function of effector cells, and to counteract suppressive pathways engaged by tumour cells themselves. Significant effort has focused on evaluating the use of exogenous cytokines, administered either systemically or locally into the tumour site via gene therapy. Several cytokines have demonstrated unique activity in the preclinical setting, including IL-2 and IFN-alpha -inducing cytokines such as IL12 and IL18. Most notably, later studies have now attempted to build on the clinical efficacy of IL-2 alone, to define combinations of agents with synergistic immunoregulatory and/or antitumour efficacy. Several lines of evidence suggest that IL-12 and IL-2 provide complementary immunoregulatory signals and have now shown that in combination, these two cytokines mediate synergistic antitumour activity in preclinical tumour models. This paper will review existing data regarding mechanisms of interaction between IL-2 and IL-12 in vitro and in preclinial models and describe future opportunities for the investigation of these potentially promising cytokines in the treatment of cancer.

Angiogenesis Inhibitors↗

Determination of transgene repeat formation and promoter methylation in transgenic plants.

The integration of transgenes into a plant host genome following Agrobacterium tumefaciens-mediated or direct transformation may occur as a single copy or in the form of tandem repeats. The latter has been associated with promoter methylation and silencing of transgenes. Thus, the early screening of such transgenic plants is desirable for ruling out future repeat-dependent transgene instability. We developed a simple PCR-based method in which primer pairs were specifically designed so that amplifications could only be obtained if the transgene was present in the form of multiple inserts in a transgenic line. The method was established using 35S-rolC transgenic aspen lines showing morphologically visible transgenic silencing. Later, it was possible to screen independent transgenic lines showing no visible marker gene expression. Furthermore, a method was developed in which positive PCR amplification was indicative of promoter methylation. The results were consistent and reproducible across different independent transgenic lines. The methods were quick, reliable, consistent and reproducible, and can be useful for routine screening of transgene silencing in lines derived from many different systems.

DNA Methylation↗

Involvement of the membrane-cytoskeleton in development of epithelial cell polarity.

The generation of cell surface polarity in transporting epithelial cells occurs in three distinct stages that involve cell-cell recognition and adhesion, cell surface remodelling to form biochemically and functionally distinct cell surface domains, and development of vectorial function. A widely used model system to study mechanisms involved in these stages is the Madin-Darby canine kidney (MDCK) cell line. Under appropriate growth conditions, MDCK cells develop in similar stages into polarized, multicellular epithelial structures. Analysis of membrane-cytoskeletal proteins ankyrin and fodrin during development of MDCK cell surface polarity shows that they gradually assemble into an insoluble protein complex on the basal-lateral membrane domain upon cell-cell adhesion, concomitantly with the redistribution of Na+,K(+)-ATPase, a marker protein of the basal-lateral membrane. Biochemical analysis shows that ankyrin, fodrin occur in a complex with Na+,K(+)-ATPase and the cell adhesion molecule uvomorulin in MDCK cells. A model is presented in which assembly of membrane-cytoskeletal complexes at sites of uvomorulin-induced cell-cell contact causes a remodelling of the cell surface distribution of specific membrane proteins which, in turn, contributes to the generation of epithelial cell surface polarity.

Animals↗

The heart of the newborn child: an anatomical study based upon transverse serial sections.

This study of the newborn child heart is based mainly on observations made on thin (0.2 mm) serial transverse sections of an entire thorax. Several features of the cardiovascular system associated with the fetal circulation are discussed. Despite considerable differences between the cardiac form in the newborn child and that in later postnatal life, the orientation of the heart in the newborn child has already attained many of its postnatal features. For example, it lies more nearly in the horizontal than the vertical plane and the atrial and ventricular septa are in line with each other at an angle of about 45 degrees to the median plane. This angulation of the septa determines that the right atrium and ventricle lie as much in front of the corresponding chambers of the left heart as they do to their right side. The cardiothoracic ratio and thoracic index are derived from a section through the thorax of the newborn child and these values are compared with those from a section through an adult man. A major purpose of this communication is to make a plea that the nomenclature given to all cardiac structures, even in the newborn child, should be consistent with the heart in situ in the anatomical position.

Adult↗