Non-tumor mast cells cultured in vitro on a honeycomb-like structured film proliferate with multinucleated formation.

Mast cells are released from bone marrow into the circulatory system as immature precursors and differentiate upon their arrival at diverse organs and tissues. Because mast cell functions can be altered in these tissues, we propose that mast cells are sensitive to their surrounding microenvironment. To examine the morphological responses of mast cells, we cultured a proliferative mouse non-tumor cell line of mast cells (NCL-2 cells) on a honeycomb-like structured polystyrene film (HCF) representing a microenvironmental scaffold. In this study, the NCL-2 cells cultured on the HCF proliferated without apoptosis. Furthermore, NCL-2 cells cultured on 3- and 5-µm HCFs exhibited multinuclear formation. These observations of different NCL-2 cell morphologies and proliferation rates on HCF scaffolds with different hole sizes suggest that mast cells undertake specific proliferative shapes depending on the surrounding microenviroment. Moreover, HCFs may lead to the regulation of mast cell differentiation. FROM THE CLINICAL EDITOR: This team reports on the development of a honeycomb-like structured film to study mast cell differentiation of non-cancerous origin, demonstrating that different microenvironments provided by different honeycomb hole sizes determine the morphology of the differentiated cells.

Quantitative analysis of thyroid-stimulating hormone messenger RNA and heterogeneous nuclear RNA in hypothyroid rats.

Thyroid-stimulating hormone (TSH) stimulates the synthesis and release of thyroid hormones including triiodothyronine (T3) and thyroxine (T4). Semiquantitative analyses using northern blot and in situ hybridization suggested that TSH gene transcription is upregulated under conditions of hypothyroidism. However, no quantitative analysis of TSH gene expression using real-time polymerase chain reaction (PCR) has been reported. In this study, we quantitated the TSHbeta messenger ribonucleic acid (mRNA) level as well as the TSHbeta heterogeneous nuclear ribonucleic acid (hnRNA) level in the anterior pituitary of hypothyroid rats, by real-time PCR using the LightCycler system. The hnRNA is the primary deoxyribonucleic acid (DNA) transcript, which reflects the transcription rate more reliably than the mRNA because of its short half-life. In the anterior pituitary of rats with methimazol-induced chronic hypothyroidism, both mRNA and hnRNA expression of TSHbeta were upregulated fourfold relative to normal rats (n=4). Our method provides a rapid and accurate measure of gene transcription. In the present report, we described a technique for accurate measurement of TSHbeta hnRNA level.

Delayed effect of administration of COX-2 inhibitor in mice with acute cerebral ischemia.

This study was designed to investigate the time lag of effect of NS398, selective COX-2 inhibitor, on infarct volume and neurologic deficits in mice with experimentally-induced cerebral ischemia. Our results showed that the beneficial effect of NS398 on reduction of infarct volume and improvement of neurologic deficits were noted more than 24 h after the injection regardless of the administration timing.

Effects of aminoguanidine on cerebral ischemia in mice: comparison between mice with and without inducible nitric oxide synthase gene.

The expression of inducible nitric oxide synthase (iNOS) is induced in the late stage of cerebral ischemia, and NO produced by iNOS contributes to delays in recovery from brain neuronal damage. To examine the importance of iNOS in the above process, we compared the effects of aminoguanidine (AG), selective iNOS inhibitor, on infarct volume and neurological deficit in iNOS null mice and normal mice subjected to brain ischemia. Neurological deficits in iNOS null mice at 24 h post-ischemia were mild compared with those of normal mice, and did not improve by AG treatment at 96 h post-ischemia. The mean infarct volume of iNOS null mice was smaller than that of normal mice at 96 h post-ischemia, and was not influenced by AG treatment. In contrast, AG reduced the infarct volume in normal mice to levels similar to those of saline-treated iNOS null mice at 96 h post-ischemia. Our results indicated that AG suppresses iNOS activity in mice with brain ischemia to level equivalent to those seen in iNOS knockout mice, confirming that this enzyme is involved in ischemic brain injury.

Differential effects of forced swim-stress on the corticotropin-releasing hormone and vasopressin gene transcription in the parvocellular division of the paraventricular nucleus of rat hypothalamus.

Corticotropin-releasing hormone (CRH) and vasopressin (AVP) colocalize in the parvocellular division of the paraventricular nucleus of the hypothalamus (PVN). We examined the effect of forced swim-stress on the CRH and AVP primary transcript (hnRNA) levels in the rat PVN by semi-quantitative in situ hybridization. CRH hnRNA increased markedly following 10-min swim-stress and returned to the basal level by 2 h. AVP hnRNA in the parvocellular division of the PVN, where AVP colocalizes with CRH, did not change significantly immediately after the swim-stress, but it did increase significantly 2 h after the stress. Pretreatment with dexamethasone abolished the increases in CRH and AVP hnRNA levels after the swim-stress. The present results demonstrate the differential effects of forced swim-stress on the CRH and AVP gene transcription in the parvocellular PVN, confirming the diverse response of the dual peptide-containing system in the face of acute stressful events.

The dubious effect of milnacipran for the treatment of burning mouth syndrome.

OBJECTIVE: Burning mouth syndrome (BMS) is a condition accompanied by oral burning symptoms, including glossal pain (glossodynia) without a detectable cause. Although BMS is a chronic-pain syndrome, only one self-controlled pilot study and some case reports have reported that milnacipran is effective for the treatment of chronic pain, including that caused by BMS. However, these papers assessed only pain, and the dosage of prescribed milnacipran varied from 30 to 150 mg/d in each patient. In this study, the dosage of prescribed milnacipran was set at 60 mg/d for 12 weeks for all patients, and depression and quality of life (QOL) were assessed in addition to pain. METHODS: Twelve patients with glossodynia participated in this study. Milnacipran was initiated at a dosage of 15 mg/d and then raised gradually to 60 mg/d after 4 weeks of treatment; this dose was continued until the end of the study (total of 12 weeks). The evaluation included the Hamilton Rating Scale for Depression, the Visual Analog Scale score for pain evaluation, the General Oral Health Assessment Index for oral-related QOL evaluation, and the Medical Outcomes Study's 36-Item Short-Form Health Survey (SF-36) for whole QOL evaluation. RESULTS: The Hamilton Rating Scale for Depression score decreased significantly after treatment with a 60-mg/d dosage of milnacipran for 12 weeks. However, the Visual Analog Scale pain, General Oral Health Assessment Index, and SF-36 scores did not change. CONCLUSIONS: A randomized, double-blind, placebo-controlled multi-institution trial of milnacipran will be essential to determine its effectiveness for the treatment of BMS.

The effectiveness of the Uchida-Kraepelin test for psychological stress: an analysis of plasma and salivary stress substances.

BACKGROUND: The hypothalamic-pituitary-adrenocortical (HPA) axis and sympathetic adrenomedullary (SAM) system are the major stress-response pathways. Plasma adrenocorticotropic hormone (ACTH) represents HPA axis activity, while plasma catecholamines are used as markers of the SAM system. Salivary alpha amylase (AA), chromogranin A (CgA), and immunoglobulin A (IgA) are candidate markers of stress activation, although their role has not been established. The Uchida-Kraepelin (U-K) test is a questionnaire that requires intense concentration and effort, and has been used as a tool to induce mental stress. However, it is not clear whether or not the test is effective as a psychological/mental stressor. METHODS: In this study, normal young women took the U-K test and serial measurements of plasma ACTH and catecholamines (dopamine, noradrenaline, and adrenaline) (n = 10), as well as salivary AA, CgA, and IgA (n = 16) before, during and after the test. RESULTS: We found no changes in any of these parameters at any time point during or after the U-K test. CONCLUSION: Our findings indicate that the U-K test is not a suitable for measuring the psychological/mental stress of young women because the plasma data showed that it did not affect the HPA axis and SAM system. The U-K test should be employed carefully as a psychological/mental stressor due to insufficient scientific evidence of its effectiveness. In addition, salivary AA, CgA, and IgA should not simply be compared with previous reports, because the mechanism of secretion and normal range of each salivary parameter remain unknown. Salivary AA, CgA, and IgA may not be suitable candidate markers of psychological/mental stress.

Response of spinal myoclonus to a combination therapy of autogenic training and biofeedback.

INTRODUCTION: Clinical evidence indicates that certain types of movement disorders are due to psychosomatic factors. Patients with myoclonic movements are usually treated by a variety of therapeutic agents. Autogenic training (AT), a recognized form of psychosomatic therapies, is suitable for certain types of neurological diseases. We describe a patient with myoclonus who failed to respond to conventional medical therapy. His symptoms were exaggerated by psychogenic factors, especially anger. CASE PRESENTATION: A 42-year-old man was admitted to our hospital, Preventive Welfare Clinic, for severe paroxysmal axial myoclonus of the left shoulder and abdominal muscles. The initial diagnosis was "combination of spinal segmental myoclonus and propriospinal myoclonus". The myoclonic movements did not occur during sleep but were aggravated by bathing, alcohol drinking, and anger. Psychological examination indicated hostile attribution. Although considered not to be a case of psychogenic myoclonus, a "psychogenic factor" was definitely involved in the induction of the organic myoclonus. The final diagnosis was "combination of spinal segmental myoclonus and propriospinal myoclonus accompanied by features of psychosomatic disorders". The patient underwent psychosomatic therapy including AT and surface electromyography (EMG)-biofeedback therapy and treatment with clonazepam and carbamazepine. RESULTS: AT and EMG-biofeedback resulted in shortening the duration and reducing the amplitude and frequency of the myoclonic discharges. CONCLUSION: Psychosomatic therapy with AT and surface EMG-biofeedback produced excellent improvement of myoclonic movements and allowed the reduction of the dosage of conventional medications.

Human mast cells express corticotropin-releasing hormone (CRH) receptors and CRH leads to selective secretion of vascular endothelial growth factor.

Mast cells are critical for allergic reactions, but also for innate or acquired immunity and inflammatory conditions that worsen by stress. Corticotropin-releasing hormone (CRH), which activates the hypothalamic-pituitary-adrenal axis under stress, also has proinflammatory peripheral effects possibly through mast cells. We investigated the expression of CRH receptors and the effects of CRH in the human leukemic mast cell (HMC-1) line and human umbilical cord blood-derived mast cells. We detected mRNA for CRH-R1alpha, 1beta, 1c, 1e, 1f isoforms, as well as CRH-R1 protein in both cell types. CRH-R2alpha (but not R2beta or R2gamma) mRNA and protein were present only in human cord blood-derived mast cells. CRH increased cAMP and induced secretion of vascular endothelial growth factor (VEGF) without tryptase, histamine, IL-6, IL-8, or TNF-alpha release. The effects were blocked by the CRH-R1 antagonist antalarmin, but not the CRH-R2 antagonist astressin 2B. CRH-stimulated VEGF production was mediated through activation of adenylate cyclase and increased cAMP, as evidenced by the fact that the effect of CRH was mimicked by the direct adenylate cyclase activator forskolin and the cell-permeable cAMP analog 8-bromo-cAMP, whereas it was abolished by the adenylate cyclase inhibitor SQ22536. This is the first evidence that mast cells express functional CRH receptors and that CRH can induce VEGF secretion selectively. CRH-induced mast cell-derived VEGF could, therefore, be involved in chronic inflammatory conditions associated with increased VEGF, such as arthritis or psoriasis, both of which worsen by stress.

Interferon regulatory factor-1 immunoreactivity in neurons and inflammatory cells following ischemic stroke in rodents and humans.

Interferon regulatory factor-1 (IRF-1), a transcription factor that controls the expression of genes related to inflammation and injury, may be involved in the mechanisms of cerebral ischemia. In this study, we used immunohistochemistry to determine whether IRF-1 protein is up-regulated after cerebral ischemia, and to define the identity of the cells that express IRF-1 in the postischemic brain. In mice, IRF-1 immunoreactivity was present in intravascular neutrophils 24 h after middle cerebral artery occlusion. At 96 h, immunoreactivity was observed in neutrophils infiltrating the ischemic tissue and in neurons at the outer border of the ischemic territory. IRF-1 immunoreactivity was also found in neurons and inflammatory cells in the brain of patients who died 1-2 days after ischemic stroke. The neuronal expression of IRF-1, in conjunction with the finding that IRF-1 deletion is beneficial to the post-ischemic brain, suggests that expression of IRF-1-dependent genes in neurons plays a role in ischemic neuronal death.