The FGFR1 inhibitor PD173074 induces mesenchymal-epithelial transition through the transcription factor AP-1.

BACKGROUND: Epithelial-mesenchymal transition (EMT) is a crucial process in cancer progression that provides cancer cells with the ability to escape from the primary focus, invade stromal tissues and migrate to distant regions. Cell lines that lack E-cadherin show increased tumorigenesis and metastasis, and the expression levels of E-cadherin and Snail correlate inversely with the prognosis of patients suffering from breast cancer or oral squamous cell carcinoma (OSCC). Moreover, recent studies have shown that most EMT cases are regulated by soluble growth factors or cytokines. Among these factors, fibroblast growth factors (FGFs) execute diverse functions by binding to and activating members of the FGF receptor (FGFR) family, including FGFR1-4. Fibroblast growth factor receptor 1 is an oncoprotein that is involved in tumorigenesis, and PD173074 is known to be a selective inhibitor of FGFR1. However, the roles of FGFR1 and FGFR1 inhibitors have not yet been examined in detail. METHODS: Here, we investigated the expression of FGFR1 in head and neck squamous cell carcinoma (HNSCC) and the role of the FGFR1 inhibitor PD173074 in carcinogenesis and the EMT process. RESULTS: Fibroblast growth factor receptor 1 was highly expressed in 54% of HNSCC cases and was significantly correlated with malignant behaviours. Nuclear FGFR1 expression was also observed and correlated well with histological differentiation, the pattern of invasion and abundant nuclear polymorphism. Fibroblast growth factor receptor 1 was also overexpressed in EMT cell lines compared with non-EMT cell lines. Furthermore, treatment of HOC313 cells with PD173074 suppressed cellular proliferation and invasion and reduced ERK1/2 and p38 activation. These cells also demonstrated morphological changes, transforming from spindle- to cobble stone-like in shape. In addition, the expression levels of certain matrix metalloproteinases (MMPs), whose genes contain activator protein-1 (AP-1) promoter sites, as well as Snail1 and Snail2 were reduced following PD173074 treatment. CONCLUSION: Taken together, these data suggest that PD173074 inhibits the MAPK pathway, which regulates the activity of AP-1 and induces MET. Furthermore, this induction of MET likely suppresses cancer cell growth and invasion.

Implications for the role of cognate interactions in in vitro human B cell activation by Staphylococcus aureus Cowan I and pokeweed mitogen.

Human B cell-triggering mechanisms were investigated using the polyclonal activators Staphylococcus aureus Cowan I (SAC) and pokeweed mitogen (PWM). When the cultures of B cells, T cells, and monocytes were stimulated for 5 d by SAC or PWM, B cells could be activated by both mitogens to proliferate and secrete Ig. Even when T cells were substituted by T cell-derived soluble factors, SAC-stimulated B cells could differentiate into Ig-secreting cells. In contrast, interactions of B and T cells for at least the first 6 h of culture were necessary for the B cell triggering by PWM. Experiments that allow a more precise delineation of the B cell-triggering mechanisms by PWM demonstrated that interactions of B cells with T4+ but not T8+ cells are required for the B cell triggering; anti-Ia or anti-T4 antibody can block this triggering; in contrast, anti-T3 or anti-T8 antibody do not exert any effects on the B cell triggering. However, all these monoclonal antibodies could not modulate the ability of B cells that had been already activated by PWM to respond to T cell-derived factors. These data suggest that SAC can directly activate B cells, while cognate interactions between Ia-like antigens on B cells and T4+ cells are essential for B cell triggering by PWM. Furthermore, once B cells are triggered, they will proliferate, differentiate, and secrete Ig in response to T cell-derived factors; Ia-like antigens or T cell differentiation antigens may not be involved in the processes in this cascade.

Studies of immune functions of patients with systemic lupus erythematosus: antibodies to desialized, rather than intact, T cells preferentially bind to and eliminate suppressor effector T cells.

Patients with systemic lupus erythematosus (SLE) were found to have in their plasma antibodies specific for desialized T cells. Adsorption studies with intact or desialized T cells indicated that SLE anti-T cell antibodies consisted of two populations with different target cell specificities, one capable of recognizing unique determinants on desialized T cells and another able to bind to both intact and desialized T cells. Normal T cells did not remove the antibodies specific for desialized T cells. moreover, the antibodies to desialized T cells were not removed by adsorption with either desialized non-T cells or desialized erythrocytes. Thus, the antibodies to desialized T cells recognize a determinant that is unique to a T cell subset and also includes a sugar. Inhibition studies with various sugars indicated that lactose was the most potent inhibitor of antibody binding. The anti-desialized T cell antibody appears to recognize a T cell determinant which includes lactose, probably in the form of a beta-galactosyl residue, but which also includes additional T cell determinants. The antibodies to desialized T cells were found to bind preferentially to concanavalin A-induced autorosetting T cells, which had been already demonstrated to contain suppressor effector cells. Indeed, such antibodies were effective in eliminating suppressor effector function without interfering with T cells necessary for such activation (such as precursor or inducer cells). Finally, studies of patients with SLE yielded a highly significant correlation (r = 0.92) between impaired suppressor effector function of their cells and the presence of antibodies to desialized T cells in their plasma.

Interrelationships between blood pressure, sodium, potassium, serum cholesterol, and protein intake in Japanese.

Interrelationships among blood pressure (BP), sodium (Na), potassium (K), dietary protein, and serum cholesterol level (Chol) were examined in 62% (1120) of 1818 Japanese inhabitants of both sexes aged over 30 years who lived in a rural village in Japan. Fasting single-spot urine specimens were collected in the morning to measure Na, K, urea nitrogen (UN), inorganic sulfate (SO4), and creatinine (Cr). The Cr ratios of Na, K, UN, SO4, Na/K, and SO4/UN were analyzed by multiple regression analysis to determine independent associations with BP together with age, obesity index, hematocrit (Hct), Chol, triglyceride (TG), and fasting serum glucose level (Glu). Except for Na/Cr in men, Na/Cr and Na/K were found to be independently and positively related to BP, particularly to systolic BP (SBP). In contrast, K/Cr and SO4/UN (an index related to the dietary score of sulphur-containing amino acids derived mainly from animal protein) were both negatively associated with SBP, and UN/Cr (an index of total protein intake) was positively associated with SBP in men. Chol was linked to BP negatively in men but positively in women. Age, obesity index, TG, and Hct were generally positively and significantly related to BP in both sexes. The results confirmed on epidemiological grounds the positive link of Na and the negative link of K to BP within a single population in Japan. They further suggest, although only in men, that there is a negative relationship of Chol and dietary animal protein with BP.

Dysautoregulation of the cerebral circulation in primary systemic amyloidosis.

A case of primary systemic amyloidosis associated with marked dysautoregulation of the cerebral circulation in response to hypotension is reported. The major amyloid protein was immunoglobulin A lambda type. An autonomic nerve function test showed severe and extensive disturbances of the autonomic nervous system. The cerebrovascular reactivity to 5% CO2 inhalation was spared. To our knowledge, this is the first case to demonstrate that amyloid peripheral autonomic neuropathy can be accompanied by dysautoregulation of the cerebral circulation.

Antinuclear antibodies in healthy aging people: a prospective study.

In order to evaluate the expression of antinuclear antibodies (ANA) in normal elderly individuals over time and clinical significance, a cross-sectional ANA testing in healthy Japanese was performed, followed by annual evaluations of ANA positive aged (> or = 65 years) and a control group. ANA was more prevalent in the aged (11.4% vs. 3.8%) and most were persistent after 4 years. Anti-ssDNA and anti-histone antibodies were increased in aged ANA positive as compared to ANA negative controls. Except for a history of spontaneous abortion, there was no differences in clinical findings. HLA DRB1*0901 and the DQB1*0602 + 0302 + 0303 set of alleles were increased in ANA positive. Therefore, ANA in the aged were persistent, apparently directed toward chromatin elements, and shared MHC associations with autoimmune diseases. Longer follow-up may be necessary to improve the evaluation of clinical significance of ANA in the aged.

Familial occurrence of impaired interleukin-2 activity and increased peripheral blood B cells actively secreting immunoglobulins in systemic lupus erythematosus.

PURPOSE: We tested the hypothesis that some abnormalities of immune functions are genetically controlled in patients with systemic lupus erythematosus (SLE). SUBJECTS AND METHODS: We used a phytohemagglutinin-induced interleukin-2 (IL-2) activity assay and a spontaneous plaque-forming cell assay to evaluate T-cell and B-cell function, respectively, in 34 clinically healthy family members of six SLE probands. RESULTS: Impaired IL-2 activity was found in 15 of the 29 consanguineous relatives. There was no relation between the household relatives and the nonhousehold relatives; none of the five nonconsanguineous household persons had abnormal results. Results for the B-cell assay were abnormal in 22 of the 29 consanguineous relatives. The B-cell abnormalities were more commonly observed in the consanguineous household relatives; four of the five nonconsanguineous household relatives also had abnormal assay results. CONCLUSION: The findings indicate that the impaired IL-2 activity in relatives appears to strongly correlate with a genetic relationship. Although the evidence favors a genetic basis for the B-cell abnormalities, environmental effects may also contribute to the familial occurrence of these abnormalities.

Effects of administration of dopamine and L-DOPA to dogs on their plasma level of dopamine sulfate.

The effect of intravenous administration of dopamine (DA) or L-3,4-dihydroxyphenylalanine (L-DOPA), its immediate precursor, on the level of DA sulfate in dog plasma was examined, to clarify the source and physiological significance of DA sulfate which is present at high level in the plasma. After DA administration, the plasma level of free DA increased markedly, but the level of DA sulfate did not change. However, after administration of L-DOPA, the levels of both free DA and DA sulfate increased greatly. After a single injection of L-DOPA, increase in the level of free DA was transient, but that of DA sulfate persisted for a long time. These results suggest that some of the DA sulfate in dog plasma is formed from circulating L-DOPA, not from circulating DA, and that formation of DA conjugate may play a role in regulating the plasma level of free DA.

Impairment of lymphocyte function in patients with myelodysplastic syndrome and its correction by addition of Ca2+ ionophore and phorbol myristate acetate.

T and B cell functions were evaluated in eight patients with myelodysplastic syndrome (MDS). We studied the response to stimulation with phytohemagglutinin, concanavalin A, and anti-CD3 monoclonal antibody (OKT3 mAb), and found impairment of response in T cells from MDS patients in comparison with age-matched controls. The decreased proliferative response was not restored by combining normal T cells with monocytes from MDS patients, or by using T cells from MDS patients together with normal monocytes. Furthermore, proliferative responses in the autologous and allogenic mixed lymphocyte reactions were significantly decreased in MDS patients. B cells from MDS patients incorporated significantly less tritiated thymidine than B cells from controls when stimulated with Staphylococcus aureus Cowan I (SAC), and SAC stimulation failed to cause efficient immunoglobulin production by MDS B cells. However, T cells from MDS patients reacted normally when stimulated by phorbol myristate acetate plus a Ca2+ ionophore (ionomycin). Abnormal lymphocyte function may contribute to the failure of hematopoietic regulation and may thus play an important role in the pathogenesis of MDS. The finding that the addition of specific signals allowed the stimulation pathway to function may have important therapeutic implications.

c-myc protein expression during cell cycle phases in differentiating HL-60 cells.

We examined c-myc protein expression in cell cycle phases during differentiation induction of HL-60 cells by flow cytometry using an indirect immunofluorescence method. In exponentially proliferating HL-60 cells, c-myc protein was expressed in a cell cycle dependent manner. During the differentiation induction of HL-60 cells with dimethylsulfoxide (DMSO), c-myc protein was rapidly down-regulated in the G1/0 specific phase prior to the appearance of differentiation associated markers. Our results indicate that c-myc protein functions in the G1/0 specific phase in cellular differentiation, and the rapid down-regulation of c-myc protein in G1/0 phase is closely associated with initial differentiation programs.

Effects of an antithrombotic agent (MD-805) on progressing cerebral thrombosis.

The effects of the newly developed synthetic selective antithrombin agent MD-805 were examined in 11 patients with progressing cerebral thrombosis, as an open trial. Nine (82%) of 11 patients improved after 1 week of intravenous drip infusion. No hemorrhagic complications were observed during the therapy. These results suggest that MD-805 is a safe and effective agent for treatment of progressing cerebral thrombosis.

Pulmonary hypertension in connective tissue disease. Clinical analysis of sixty patients in multi-institutional study.

Clinical features and prognosis of sixty patients with connective tissue disease accompanied by pulmonary hypertension (PH) (26 MCTD, 20 SLE, and 14 PSS) reported retrospectively by multi-institutions were compared. Though the obtained data were incomplete and lacking in uniformity, no significant difference in the clinical features among the three diseases were observed except high incidence of pulmonary fibrosis and low % VC in PSS and PH patients. Statistically significant difference, however, was observed between live and dead patients of three diseases gathered in post sternal pain, pulmonary diastolic murmur, right ventricular hypertrophy on ECG and mean pressure of pulmonary artery. Higher incidence of anti-nRNP antibody was observed in SLE with PH and PSS with PH patients than with the general population. A quicker occurrence of PH and shorter survival time were observed in MCTD patients with PH than in SLE and PSS patients with PH.

Double-blind study with liposteroid in rheumatoid arthritis.

A multicentre double-blind comparative trial was performed in 138 patients with rheumatoid arthritis (RA) after biweekly intravenous or intramuscular injections of liposteroid (containing 2.5 mg of dexamethasone), which had been developed as a drug for targeting therapy of RA, and Decadron (containing 3.3 mg of dexamethasone) as a reference drug. The results showed a tendency to a significantly higher rate of improvement with lower frequency of side-effects in the liposteroid group than in the Decadron group. This study indicates that liposteroid is more useful for RA and that the separation of the efficacy and side-effects of steroids could be clinically confirmed to some extent.

Effect of long-term administration of buflomedil on regional cerebral blood flow in chronic cerebral infarction.

The influence of long-term oral administration of buflomedil on regional cerebral blood flow was investigated in 10 patients with chronic cerebral infarction and mild to moderate mental deterioration. Patients were given 150 mg buflomedil three times daily for a period of eight weeks. Regional cerebral blood flow (rCBF) was measured in patients before they began buflomedil treatment and during the last week of treatment. The interval of the rCBF measurements in the control subjects (n = 12) ranged from seven to forty-eight days. Mental function in the patients studied was evaluated by use of Hasegawa's simple intelligence scale for the aged and Zung's self-rating depression scale, before and during the last week of buflomedil treatment. The results showed a mean increase in rCBF of 10.5% +/- 12.1% in the patient group. The control group demonstrated virtually no change in rCBF measurements. Improvements in the mental function scores of the buflomedil-treated group were noted, and 5 of the 10 patients reported improvement in subjective symptoms.

Effect of single oral administration of nilvadipine on cerebral blood flow in chronic cerebral infarction.

UNLABELLED: The effect of nilvadipine, a newly developed calcium antagonist, on regional cerebral blood flow (rCBF) was investigated in 7 patients with chronic cerebral infarction. rCBF was measured by the 133Xenon inhalation method. Patients were given a single dose of 4 mg of nilvadipine after the first measurement of rCBF, and the second measurement was done one hour after the administration. All patients had hemiparesis and 2 of them had mild to moderate mental deterioration, but all patients could walk to the outpatient clinic by themselves. RESULTS: (1) rCBF of the affected side significantly increased by 22.7% after single oral administration of nilvadipine (p < 0.05). The increase of rCBF was significantly marked in frontal regions of the affected hemispheres. (2) No significant changes in blood pressure or end tidal partial pressure of carbon dioxide were observed during the examination. These results indicate that nilvadipine has a potent selective vasodilatory action on the cerebral arteries in patients with cerebral infarction.

Age-related changes in cerebral white matter measured by computed cranial tomography.

Changes in the cerebral white matter in relation to aging were studied quantitatively by computed cranial tomography (CT) in 70 healthy subjects aged 30 to 94 years. There were no age-related changes in the CT number of the white matter (WMCT) in 41 younger subjects aged 30 to 65 years. But, there was a significant negative correlation between age and the WMCT in 29 elderly subjects aged 66 to 94 years. Brain atrophy was significantly correlated with the WMCT. The WMCT decreased with aging even in neurologically healthy elderly persons.

[An association of Shy-Drager syndrome with moyamoya disease--a case report].

A 58-year-old female was admitted to our hospital because of orthostatic syncope, and difficulty in urination and walking for 2 years. At age 35, she suffered from cerebral hemorrhage and was diagnosed as Moyamoya disease (MD). Neurological examination on admission revealed severe orthostatic syncope so that she could not stand. At that time, her blood pressure and heart rate were 104/74 mmHg and 78/min in supine position and 52/48 mmHg and 52/min in 60 degrees head-up position. She also had severe pollakisuria, cerebellar ataxia and dysarthria. Cranial nerves, motor strength and deep reflexes were normal. ECG, chest X-ray, and EEG were normal. Aschner, Czermak, and cold pressor tests revealed no response. Urodynamic study revealed autonomic bladder. MRI showed only enlarged fourth ventricle because of atrophy of the pons. Bilateral CAG revealed "Moyamoya" vessels in the cerebral basal regions. Cerebral blood flow (CBF) was measured by 133Xe inhalation method. Mean arterial blood pressure changed from 134 mmHg to 126 mmHg in 45 degrees head-up tilting and CBF decreased from 47.5 mg/100 g/min to 37 ml/100 g/min position. Though there was no relationship between SDS and MD, each one shows dysautoregulation of CBF, it is supposed that a severe orthostatic syncope attack was resulted from synergism of both effects.

[Diagnosis of cerebral amyloid angiopathy with cerebral hemorrhage by enzyme linked immunosorbent assay (ELISA) of cystatin C in the cerebrospinal fluid].

The lower level of cystatin C in cerebrospinal fluid (CSF) is one of the useful diagnostic markers of hereditary cerebral hemorrhage with amyloidosis in Iceland. We attempted to establish an assay to determine the level of cystatin C in CSF for diagnosis of CAA due to the deposition of cystatin C in CSF for diagnosis of CAA due to the deposition of cystatin C. We carried out the sandwich enzyme immunosorbent assay with the use of monoclonal mouse anti-cystatin C and polyclonal rabbit anti-cystatin C antibodies. CSF from nine cases of cerebral hemorrhage and fifty reference cases with other neurological diseases were examined. Four patients with cerebral hemorrhage showed a low level of cystatin C and clinical manifestations suggestive of CAA. Our study showed the feasibility of using ELISA for the diagnosis of cerebral amyloid angiopathy that causes cerebral hemorrhage with the deposition of cystatin C.

[The relationship between P300 latency and regional cerebral blood flow in patients with cerebral infarction in the territory of the deep perforators].

We studied the relationship between P300 latency and regional cerebral blood flow (rCBF) in nondemented patients with cerebral infarction. Subjects were 24 nondemented patients (mean age 64.1 years) who had a CT-proven infarct in the territory of the deep perforators of the internal carotid artery system and 53 controls (mean age 64.1 years). Prolongation of P300 latency with advancing age was observed in the both groups. There was no significant difference in P300 latency and rCBF between the two groups. There was a negative correlation between P300 latency and rCBF, especially in the bilateral fronto-parietal regions in the patient group. These results indicate that cognitive function assessed by P300 latency may be related to rCBF in the fronto-parietal region in the nondemented patients with lacunar infarctions.

[Immunoglobulin levels and blood parameters in inhabitants of a remote island].

To clarify the alteration of serum immunoglobulin levels with age, serum immunoglobulin and other parameters (age, symptoms, blood biochemical parameters, etc.) of inhabitants (213 persons; 63 men, 150 women) of a remote island in Shimane Prefecture were studied. Their subjective symptoms were collected by means of a questionnaire and various blood parameters were measured. These data were analyzed by t-test and multivariate analysis. The IgM level was decreased with age and each immunoglobulin level correlated with the total-protein level in serum. The IgG level of the subjects feeling fatigue was higher than that of the subjects not feeling fatigue.