Hiromi Suzuki

Dok-1 and Dok-2 are negative regulators of lipopolysaccharide-induced signaling

Endotoxin, a bacterial lipopolysaccharide (LPS), causes fatal septic shock via Toll-like receptor (TLR)4 on effector cells of innate immunity like macrophages, where it activates nuclear factor κB (NF-κB) and mitogen-activated protein (MAP) kinases to induce proinflammatory cytokines such as tumor necrosis factor (TNF)-α. Dok-1 and Dok-2 are adaptor proteins that negatively regulate Ras–Erk signaling downstream of protein tyrosine kinases (PTKs). Here, we demonstrate that LPS rapidly induced the tyrosine phosphorylation and adaptor function of these proteins. The stimulation with LPS of macrophages from mice lacking Dok-1 or Dok-2 induced elevated Erk activation, but not the other MAP kinases or NF-κB, resulting in hyperproduction of TNF-α and nitric oxide. Furthermore, the mutant mice showed hyperproduction of TNF-α and hypersensitivity to LPS. However, macrophages from these mutant mice reacted normally to other pathogenic molecules, CpG oligodeoxynucleotides, poly(I:C) ribonucleotides, or Pam3CSK4 lipopeptide, which activated cognate TLRs but induced no tyrosine phosphorylation of Dok-1 or Dok-2. Forced expression of either adaptor, but not a mutant having a Tyr/Phe substitution, in macrophages inhibited LPS-induced Erk activation and TNF-α production. Thus, Dok-1 and Dok-2 are essential negative regulators downstream of TLR4, implying a novel PTK-dependent pathway in innate immunity.

Magnetic resonance imaging of osteomyelitis in the mandible: Comparative study with other radiologic modalities

Magnetic resonance imaging of 14 histopathologically confirmed cases of osteomyelitis of the mandible wasretrospectively reviewed. The findings of magnetic resonance imaging were compared with conventional radiography, computed tomography, bone scintigraphy, and histopathologic examinations. All lesions in bone marrow were shown as areas of low (64%) or low-to-intermediate (36%) signal intensity on T1-weighted images, and areas of high (29%), mixed (high and low, 21%; high and intermediate, 36%) or low (14%) signal intensity on T2-weighted images. Histopathologically, high T2-weighted signal intensity areas that showed enhancement after contrast injection corresponded to active infection. These were not collections of pus but were predominantly areas of granulation tissue. Magnetic resonance imaging showed larger areas of abnormality than plain radiography or computed tomography. Bone scintigraphy did not accurately reveal the locations of lesions but showed heterogeneous increased uptake in all patients. MRI was an extremely useful technique for assessing osteomyelitis of the mandible.

Comparison of cerebral activity during teeth clenching and fist clenching: a functional magnetic resonance imaging study

Using functional magnetic resonance imaging (fMRI), we compared the cerebral activity during bilateral light fist-clenching and light-teeth clenching to provide more information on the central processing mechanisms underlying awake bruxism. Fourteen subjects participated in our study. Statistical comparisons were used to identify brain regions with significant activation in the subtraction of light fist clenching and light teeth clenching activity minus baseline. Participants also evaluated the perceived effort of clenching for each task, using a visual analogue scale of 0-100, after fMRI was performed. Bilateral light fist-clenching significantly activated the bilateral sensorimotor cortex, while light teeth-clenching was significantly associated with activation of the bilateral sensorimotor cortex, supplementary motor area, dorsolateral prefrontal cortex, and posterior parietal cortex. The VAS scores for fist clenching and teeth clenching were not significantly different. As light teeth-clenching activates a more extensive cortical network compared with light fist-clenching, we suggest that the teeth clenching may induce a more complex cerebral activity compared with the performance of a hand motor task. The clinical significance of these findings remains unknown but could perhaps be related to the propensity to trigger awake bruxism.

Imaging tumors of the minor salivary glands

Magnetic resonance imaging evaluations of nine histopathologically confirmed minor salivary gland tumors were made retrospectively and compared with evaluations of images obtained by computed tomography. All tumors had low-to-intermediate T1 signal intensities and intermediate-to-high T2 signal intensities. Malignant tumors had an irregular margin in all but one case. Benign tumors invariably had well-defined margins. In terms of tumor margination, the magnetic resonance imaging findings correlated well with the histopathologic findings. Magnetic resonance imaging demonstrated the internal architecture of the minor salivary gland tumors multidirectionally and was superior to computed tomography in this respect and in the ability to locate the tumors.

Magnetic resonance appearance of bone marrow in the mandible at different ages

To differentiate pathologic states of bone marrow from normal age-related marrow conversion, magnetic resonance appearances of the mandible were analyzed cross-sectionally according to age in 73 normal volunteers. Low signal intensities of red marrow first changed to high signal intensities in the anterior region of the mandibular body. With increase in age, marrow conversion was observed in the premolar/molar region, angle, ramus, and condyle regions in that order. No red marrow was recognized in the body of the mandible of subjects more than 30 years old. After the age of 30, the majority of the subjects showed high signal intensities in the entire bone marrow of the mandible. However, one subject over 30 years of age showed some portions of red marrow in the ramus and condyle. This knowledge about age-related marrow conversion of the mandible on magnetic resonance imaging will be useful in the visual differentiation from abnormal bone marrow, such as anemias, inflammatory diseases, infiltrative neoplastic disorders, and metastatic diseases.

Discrimination between adaptive and neutral amino acid substitutions in vertebrate hemoglobins

SummaryA discriminant analysis on the basis of the physicochemical properties of amino acid residues is developed to investigate the accumulation pattern of amino acid substitutions in a family of proteins. The application of this analysis to vertebrate hemoglobins reveals the following new results. (1) The major components of teleost fish and amphibian hemoglobins showing the Root effect are sharply discriminated from mammalian hemoglobins in several regions of the α and β chains, whereas shark, minor components of teleost fish and amphibian, reptile, and bird hemoglobins showing no Root effect exhibit a gradual change to mammalian hemoglobin in a straightforward way. This result suggests at least two lines of molecular evolution in vertebrate hemoglobins. (2) The nonadult hemoglobin chains are allocated to the latter line, i.e., tadpole, ξ, and π chains are similar to shark and trout I chains, and ∈ and γ chains are similar to some of the reptile chains. (3) In any case, most of the amino acid residues causing the discrimination are located near the sites that carry the amino acid residues conserved well throughout all classes of vertebrates, suggesting that modifications adapting to the respective living conditions or respiratory organs have taken place effectively near the amino acid residues essential for the manifestation of cooperative oxygen binding. (4) The amino acid residues at other sites are changed from one to another species even within the same class, showing a constant substitution rate as a whole. These amino acid substitutions may be nearly neutral, being under a weak functional constraint. The number of sites allowing such neutral substitutions is rather small, less than one-half of all the sites in the adult hemoglobins of bird and mammal, whereas it amounts to two-thirds in teleost fish hemoglobins.

Theoretical evaluation of transcriptional pausing effect on the attenuation in trp leader sequence

The effect of transcriptional pausing on attenuation is investigated theoretically on the basis of the attenuation control mechanism presented by Oxender et al. (Oxender, D. L., G. Zurawski, and C. Yanofsky, 1979, Proc. Natl. Acad. Sci. USA. 76:5524-5528). An extended stochastic model including the RNA polymerase pausing in the leader region is developed to calculate the probability of relative position between the RNA polymerase transcribing the trp leader sequence and the ribosome translating the transcript. The present study results in a new rationale that the transcriptional pausing site in the leader sequence makes the attenuation control both more sensitive as an on/off switch and less sensitive to variations in the concentration of cellular metabolites not connected with the need for expressing, or not expressing, the particular operon. It is also proposed that the transcriptional pausing diminishes the dependence of attenuation control characteristics on the number of nucleotides in the leader sequence. This result may be useful for understanding the attenuation control efficiencies of other amino acid leader sequences with different lengths of nucleotides.

Inhibition of human gingival carcinoma cell growth by prostaglandins

The inhibitory effects of various prostaglandins (PGA2, PGD2 and PGE2) on the growth of human gingival carcinoma cell line Ca 9-22 and human cervical carcinoma cell line HeLa were studied in vitro. 1. PGA2 and PGD2 significantly depressed Ca 9-22 cell proliferation in dose and time related fashion. 2. The inhibitory rate by PGs for Ca 9-22 cells exceeded than HeLa-cells. 3. IC50 value of PGD2 on growth of Ca 9-22 was calculated to be 4 micrograms/ml.

Central neurofibroma of the mandible —report of a case with an immunohistochemical and electron microscopical studies

Introduction Benign neurogenic tumors of the oral cavity are classified into several typings, such as, Schwannoma, neurofibroma, nerve sheath myxoma, multiple mucosal neuroma, amputation neuroma and others. Schwannoma is probably the most common among these tumors, while the neurofibroma is a rare neoplasm in the oral cavity, especially in the intraosseous region. Electron and immunohistochemical studies of this tumor are also uncommon and inconclusive. We recently encountered a case of central neurofibroma of the right mandible and examined it by light and electron microscopic studies and by immunohistochemistry. The reported cases of central neurofibroma of the jaw are also reviewed and discussed.

Theoretical analysis of single-round transcription experiments on trp leader region

A kinetic model is proposed to reproduce the time courses of the concentration change in paused leader RNA, terminated leader RNA, and readthrough RNA in the single-round transcription experiments on trp leader region of Escherichia coli and its mutants, L132, L75, and L75L135 (Winkler, M. E., and C. Yanofsky, 1981, Biochemistry, 20:3738-3744; Fisher, R., and C. Yanofsky, 1983, J. Biol. Chem., 258:9208-9212). This model fits the experimental results well and also captures the essential aspects of the processes of transcriptional pausing and termination. In the wild type template, under optimal conditions, it is found that the transcription rate at the pause and attenuation sites is of the same order of magnitude, 10,000-fold lower than the transcription rate at the other sites, and the high termination level at the attenuation site is attributable to a higher dissociation rate. This analysis also provides a clue as to how the template base change, various concentrations of ribonucleoside triphosphates, and the presence or absence of L-factor affect the transcription and dissociation rates to yield different termination levels at the pause or attenuation site. It also discusses the molecular mechanism of the transcriptional pausing and termination.