Geriatric nutritional risk index as the prognostic factor in older patients with fragility hip fractures.

This study investigated the long-term survival and incidence of secondary fractures after fragility hip fractures. The 5-year survival rate was 62%, and the mortality risk was seen in patients with GNRI < 92. The 5-year incidence of secondary fracture was 22%, which was significantly higher in patients with a BMI < 20. BACKGROUND: Malnutrition negatively influences the postoperative survival of patients with fragility hip fractures (FHFs); however, little is known about their association over the long term. OBJECTIVE: This study evaluated the ability of the geriatric nutritional risk index (GNRI) as a risk factor for long-term mortality after FHFs. METHODS: This study included 623 Japanese patients with FHFs over the age of 60 years. We prospectively collected data on admission and during hospitalization and assessed the patients' conditions after discharge through a questionnaire. We examined the long-term mortality and the incidence of secondary FHFs and assessed the prognostic factors. RESULTS: The mean observation period was 4.0 years (range 0-7 years). The average age at the time of admission was 82 years (range 60-101 years). The overall survival after FHFs (1 year, 91%; 5 years, 62%) and the incidence of secondary FHFs were high (1 year, 4%; 5 years, 22%). The multivariate Cox proportional hazard analysis revealed the risk factors for mortality as older age (hazard ratio [HR] 1.04), male sex (HR 1.96), lower GNRI score (HR 0.96), comorbidities (malignancy, HR 2.51; ischemic heart disease, HR 2.24; revised Hasegawa dementia scale ≤ 20, HR 1.64), no use of active vitamin D3 on admission (HR 0.46), and a lower Barthel index (BI) (on admission, HR 1.00; at discharge, HR 0.99). The GNRI scores were divided into four risk categories: major risk (GNRI, < 82), moderate risk (82-91), low risk (92-98), and no risk (> 98). Patients at major and moderate risks of GNRI had a significantly lower overall survival rate (p < 0.001). Lower body mass index (BMI) was also identified as a prognostic factor for secondary FHFs (HR 0.88 [p = 0.004]). CONCLUSIONS: We showed that older age, male sex, a lower GNRI score, comorbidities, and a lower BI are risk factors for mortality following FHFs. GNRI is a novel and simple predictor of long-term survival after FHFs.

Giant Rashba-type spin splitting in bulk BiTeI.

There has been increasing interest in phenomena emerging from relativistic electrons in a solid, which have a potential impact on spintronics and magnetoelectrics. One example is the Rashba effect, which lifts the electron-spin degeneracy as a consequence of spin-orbit interaction under broken inversion symmetry. A high-energy-scale Rashba spin splitting is highly desirable for enhancing the coupling between electron spins and electricity relevant for spintronic functions. Here we describe the finding of a huge spin-orbit interaction effect in a polar semiconductor composed of heavy elements, BiTeI, where the bulk carriers are ruled by large Rashba-like spin splitting. The band splitting and its spin polarization obtained by spin- and angle-resolved photoemission spectroscopy are well in accord with relativistic first-principles calculations, confirming that the spin splitting is indeed derived from bulk atomic configurations. Together with the feasibility of carrier-doping control, the giant-Rashba semiconductor BiTeI possesses excellent potential for application to various spin-dependent electronic functions.

Topological surface states with persistent high spin polarization across the Dirac point in Bi2Te2Se and Bi2Se2Te.

Helical spin textures with marked spin polarizations of topological surface states have been unveiled for the first time by state-of-the-art spin- and angle-resolved photoemission spectroscopy for two promising topological insulators, Bi(2)Te(2)Se and Bi(2)Se(2)Te. Their highly spin-polarized natures are found to be persistent across the Dirac point in both compounds. This novel finding paves a pathway to extending the utilization of topological surface states of these compounds for future spintronic applications.

Experimental verification of PbBi2Te4 as a 3D topological insulator.

The experimental evidence is presented of the topological insulator state in PbBi2Te4. A single surface Dirac cone is observed by angle-resolved photoemission spectroscopy with synchrotron radiation. Topological invariants Z2 are calculated from the ab initio band structure to be 1;(111). The observed two-dimensional isoenergy contours in the bulk energy gap are found to be the largest among the known three-dimensional topological insulators. This opens a pathway to achieving a sufficiently large spin current density in future spintronic devices.

Elevated umbilical cord serum TARC/CCL17 levels predict the development of atopic dermatitis in infancy.

BACKGROUND: Thymus-and-activation-regulated chemokine (TARC; CCL17) is related to both allergy and pregnancy, but the relationships of maternal and umbilical cord blood CCL17 to atopic dermatitis (AD) development have not yet been examined. Objective Seventy paired full-term and normal vaginal delivery newborns and their mothers were enrolled in this study. METHODS: To elucidate the pathogenesis and fetomaternal inheritance of AD in infancy, CCL17, IFN-γ-inducible protein 10 kDa (IP-10; CXCL10), soluble HLA-G (sHLA-G), IgE and eosinophil counts were examined using sera from 70 paired umbilical cord and maternal blood samples. RESULTS: Serum CCL17 (r(s) =0.340, P<0.001) and sHLA-G (r(s) =0.600, P<0.001) levels showed high correlations between umbilical cord and maternal blood. Umbilical cord serum levels of CCL17 from neonates destined to develop AD in infancy were higher than in those from neonates who showed no signs of AD during infancy (median 1586.9 vs. 819.6 pg/mL, P<0.001). Serum levels of CCL17 were higher in mothers with AD than in those without AD (median 909.6 vs. 214.1 pg/mL, P<0.001). High umbilical cord serum levels of CCL17 were associated with infantile AD development even in 62 neonates born to mothers without AD (median 1514.4 vs. 740.6 pg/mL, P<0.001) and 38 neonates born to mothers with no allergies (median 1624.2 vs. 740.6 pg/mL, P<0.001). The summary estimates for umbilical cord serum CCL17 in the diagnosis of infantile AD were: sensitivity 85.7% (95% confidence interval: 72.8-98.7), specificity 73.8% (60.5-87.1), positive predictive value 68.6% (53.2-84.0) and negative predictive value 88.6% (78.0-99.1). CONCLUSION AND CLINICAL RELEVANCE: These findings suggest that the umbilical cord blood CCL17 may be involved in the pathogenesis of infantile AD and in fetomaternal inheritance. Serum levels of CCL17 from umbilical cord blood may be a predictive marker for AD in infancy.

Comparative histopathological analysis between tenosynovitis and joint synovitis in rheumatoid arthritis.

AIMS: To clarify the histological and biological features of tenosynovitis accompanying rheumatoid arthritis (RA). METHODS AND RESULTS: Synovial tissue was obtained from the wrist joint and extensor tendon of the digits of six RA patients and the sections were examined by haematoxylin and eosin staining and immunohistochemical analysis. RA tenosynovitis exhibited the typical histological features of RA joint synovitis, including hyperplasia of the synovial lining and infiltration of leucocytes, largely CD4+ T cells and CD68+ macrophages. Notably, there was a significant correlation in the number of CD4+ T cells and CD68+ macrophages between the tenosynovium and joint synovium in each individual. Real-time reverse transcriptase-polymerase chain reaction analysis revealed similar mRNA expression patterns of various inflammatory mediators in tenosynovitis and joint synovitis. It was also observed that synovial fibroblasts isolated from the tenosynovium behaved in a manner similar to those isolated from the joint synovium with regard to proliferation and the production of inflammatory mediators. CONCLUSIONS: The histopathological features of RA tenosynovitis were indistinguishable from those of joint synovitis. Therefore, it is suggested that the ongoing inflammation is driven by similar mechanisms in the tenosynovium and joint synovium and that RA is probably a tissue-specific disease which targets systemic synovial tissues.

Development of the modified sum-peak method and its application.

As the sum-peak method requires the total count rate as well as the peak count rates and the sum peak count rate, this meets difficulties when a sample contains other radionuclides than the one to be measured. To solve the problem, a new method using solely the peak and the sum peak count rates was developed. The method was theoretically and experimentally confirmed using (60)Co, (22)Na and (134)Cs. We demonstrate that the modified sum-peak method is quite simple and practical and is useful to measure multiple nuclides.

Potential role of microvessel density in predicting radiosensitivity of T1 and T2 stage laryngeal squamous cell carcinoma treated with radiotherapy.

Curative radiotherapy is the first choice of therapy for T1 and T2 stage laryngeal squamous cell carcinoma (LSCC) patients to preserve their phonation. Patients with recurrent tumors who undergo salvage surgery require prolonged nasal feeding. Therefore, clinical interest has been focused on elucidating a predictive factor indicating which tumors are likely to be radiosensitive before radiotherapy. We analyzed the relations between radiosensitivity and clinicopathological factors (gender, tumor location, histological factors, and clinical tumor-node-metastasis stage), expression of apoptosis-related proteins (p53, bax, bcl-2), apoptotic index using the terminal deoxynucleotidyltransferase-mediated nick end labeling method, expression of cell proliferation-related proteins (Ki-67-labeling index and epidermal growth factor receptor overexpression) and microvessel density (MVD, vessels/field = 0.391 mm2) in biopsy specimens from 31 LSCC patients given radiotherapy (total radiotherapy dose of 52-70 Gy over 4-6.5 weeks). Univariate analysis revealed that tumors with a high MVD (> or =35 vessels/field) showed better radiosensitivity than those with a low MVD (<35 vessels/field, P = 0.008) and that a high Ki-67-labeling index (> or =40%) was weakly associated with radiosensitivity (P = 0.056). Multivariate analysis and Kaplan-Meier analysis showed that MVD alone had significant predictive power for radiosensitivity in T1 and T2 stage LSCCs after radiotherapy (P = 0.012, 0.0003, respectively). No significant association between clinicopathological factors, or of overexpression of p53, bax, bcl-2, epidermal growth factor receptor, or apoptotic index, with radiosensitivity was found. These results indicate that MVD is a potentially useful clinical factor predicting radiosensitivity for patients with early stage LSCCs before treatment.

Transfection with mutant p53 gene inhibits heat-induced apoptosis in a head and neck cell line of human squamous cell carcinoma.

PURPOSE: To confirm that human cancer cells show p53-dependent heat sensitivity through an apoptosis-related mechanism, we examined the heat sensitivity and Bax-mediated apoptosis after heating in a human squamous cell carcinoma cell line, SAS, with identical genetic backgrounds except for the p53 status. MATERIALS AND METHODS: We performed colony formation assay, Western blotting and analyses of apoptosis, using the SAS cells transfected with pC53-248 vector with mutant p53 gene (SAS/Trp248 cells) or the cells transfected with pCMV-Neo-Bam vector (SAS/neo cells) as a control. RESULTS: SAS/Trp248 cells showed heat resistance due to the dominant negative nature of mp53, compared with SAS/neo cells. The incidence of DNA ladders and apoptotic bodies increased markedly after heating in SAS/neo cells, but increased very little in SAS/Trp248 cells. CONCLUSION: These results suggest that heat resistance brought by mp53-transfection is p53-dependent and closely correlates with the induction of apoptosis in human squamous cell carcinomas.

Limb allografts in rats immunosuppressed with FK506. I. Reversal of rejection and indefinite survival.

We have tested the effects of FK506 (FK), a new immunosuppressive agent, on a rat limb allograft model. Histoincompatible BN limb allografts were rejected in untreated F344 hosts within 11 +/- 1 days (mean +/- SD) after operation. A single injection of 2 mg/kg, 10 mg/kg, or 50 mg/kg of FK on the day of limb transplantation (day 0) significantly prolonged graft survival in a dose-dependent manner--i.e., mean limb survival times (MST) based on gross signs of skin rejection were 16 +/- 3 days, 51 +/- 6 days, or 104 +/- 17 days, respectively (P less than 0.01). Delayed treatment with a single injection of 10 mg/kg of FK at when early signs of rejection were visible (day 7 or day 10) reversed the ongoing rejection. The MSTs in these groups were comparable to that of those treated with the same dosage of FK on day 0. The FK-induced unresponsiveness toward limb allografts was donor-specific because limb-allografted. FK-protected rats could not accept the skin grafts from a third-party donor. In the next set of experiments, rats were given a single administration of 10 mg/kg of FK on the day of limb allograft, followed by intermittent injections of 3 mg/kg of FK once a week. This regimen produced complete graft survival for more than 200 days, though Pneumocystis carinii pneumonia occurred in most of the recipients. These results represent the unique effects of FK in preventing or reversing the graft rejection and in inducing indefinite survival in this animal model of composite tissue allografts.

Effects of tyramine on noradrenaline outflow and electrical responses induced by field stimulation in the perfused rabbit ear artery.

In the perfused rabbit ear artery the basal outflows of noradrenaline (NA) and 3,4-dihydroxyphenylglycol (DOPEG) were less than 1 ng g-1 and 1-2 ng g-1 wet weight of tissue respectively. Field stimulation increased outflows of NA and DOPEG in a frequency-dependent manner, and they reached the maximum value at frequencies over 5 Hz. Tyramine (1 X 10(-6) -1 X 10(-4) M) increased basal outflow of NA and DOPEG, in a dose-dependent manner. This effect was not blocked by tetrodotoxin (TTX, 3 X 10(-7) M), but was prevented by pretreatment with 6-hydroxydopamine (6-OHDA). Tyramine increased the field stimulation-induced outflow of NA but not that of DOPEG in a dose-dependent manner. Cocaine (1 X 10(-5) M) reduced the increased outflow of NA induced by tyramine at rest and during field stimulation, without modifying DOPEG-outflow. Guanethidine (5 X 10(-6) M), increased outflows of NA and DOPEG at rest, and reduced the NA outflow induced by field stimulation. Pretreatment with guanethidine (5 X 10(-6) M) did not block the action of tyramine on NA and DOPEG basal outflows. Additional application of guanethidine during the presence of tyramine did reduce the outflow of NA induced by field stimulation, but did not modify the outflow of NA and DOPEG at rest. Tyramine at concentrations over 1 X 10(-5) M depolarized the smooth muscle membrane of the rabbit ear artery. After chemical denervation with 6-hydroxydopamine (6-OHDA) the depolarizing action of tyramine was reduced. Tyramine-induced depolarization was attenuated by prazosin (5 X 10(-6) M) or phentolamine (5 X 10(-6) M), but not by guanethidine (5 X 10(-6) M). In 6-OHDA-denervated tissues, tyramine-induced depolarization was attenuated by phentolamine but not by prazosin. Field stimulation evoked excitatory junction potential (e.j.p.), slow depolarization and spike potential in the rabbit ear artery. Tyramine reduced, while guanethidine blocked these electrical responses. Tyramine did not alter the facilitation process of e.j.ps. In tissues pretreated with guanethidine, tyramine evoked either no electrical response or a slow depolarization during field stimulation. The slow depolarization was blocked by prazosin. Tyramine reduced the NA content of tissues in a dose-dependent manner (by 31% at 10(-4) M). Guanethidine (5 X 10(-6) M) reduced the NA content by 20%. 10 We conclude that in the rabbit ear artery, tyramine depolarizes the smooth muscle membrane indirectly by releasing neuronal NA which acts on alpha-adrenoceptors, and directly by an action on the smooth muscle cells. Two NA compartments (guanethidine-sensitive and tyramine-sensitive NA) could be identified. Field stimulation releases the former with associated generation of ej.p. and slow depolarization whilst the release of the latter is not accompanied by ej.p. generation.

Transmitter release modulated by alpha-adrenoceptor antagonists in the rabbit mesenteric artery: a comparison between noradrenaline outflow and electrical activity.

Effects of alpha-adrenoceptor blockers (prazosin, yohimbine, phentolamine and phenoxybenzamine) on the outflow of noradrenaline (NA) and 3,4-dihydroxyphenylglycol (DOPEG) during perivascular nerve stimulation were observed in relation to electrical events in the rabbit mesenteric artery. Cocaine or imipramine increased the NA outflow and reduced the DOPEG outflow induced by nerve stimulation. In the absence of stimulation, cocaine and imipramine did not significantly modify the NA and DOPEG outflows. The alpha-adrenoceptor blockers we used enhanced the NA and DOPEG outflow during nerve stimulation, in a dose-dependent manner; the potency of the enhancement was higher for phentolamine and phenoxybenzamine than for prazosin and yohimbine. Higher concentrations (10(-5) M) of yohimbine reduced the NA and DOPEG outflows induced by nerve stimulation. Prazosin increased the DOPEG outflow in the absence of stimulation, and this effect was not inhibited by pretreatment with cocaine. Guanethidine increased the NA and DOPEG outflow in the absence of stimulation, and the NA outflow was reduced during nerve stimulation. These effects of guanethidine were prevented by pretreatment with cocaine or imipramine. Perivascular nerve stimulation evoked excitatory junction potentials (e.j.ps) and with high frequency stimulation, slow depolarization and spike potentials. Application of phentolamine, phenoxybenzamine or yohimbine enhanced, and of prazosin had no effect, on the amplitude of the e.j.p. Spike potentials were not affected by these alpha-adrenoceptor blockers. Slow depolarization ceased in the presence of prazosin, phentolamine or phenoxybenzamine, and was slightly inhibited by yohimbine. Guanethidine blocked all of these electrical responses induced by perivascular nerve stimulation. Application of prazosin, phentolamine or phenoxybenzamine did not alter the resting membrane potential of the smooth muscle cells. Depolarizations of smooth muscle membrane produced by exogenously applied NA were inhibited by prazosin, phentolamine or phenoxybenzamine. Yohimbine itself depolarized the membrane and the inhibitory effects on the NA-induced depolarization were weaker. We conclude that the smooth muscle membrane of the rabbit mesenteric artery possesses alpha 1-adrenoceptors. Increase in NA outflow by alpha-adrenoceptor antagonists during nerve stimulation was not always consistent with increase in e.j.p. amplitude, presumably due to involvement of actions other than alpha-adrenoceptor blockade with each of these antagonists.

Comparison of the effects of caffeine and procaine on noradrenergic transmission in the guinea-pig mesenteric artery.

The effects of caffeine and procaine on noradrenergic transmission in the guinea-pig mesenteric artery were investigated by recording electrical responses of smooth muscle cells and by measuring the outflow of noradrenaline (NA) and 3,4-dihydroxyphenylglycol (DOPEG) induced by perivascular nerve stimulation. Caffeine possessed dual actions on the membrane, i.e., at low concentrations (2.5 X 10(-4)-5 X 10(-4)M), it hyperpolarized the membrane and decreased the membrane resistance and at high concentrations (over 2.5 X 10(-3)M) it depolarized the membrane and increased the membrane resistance. Procaine (over 10(-4)M) consistently depolarized the membrane and increased the membrane resistance. The amplitude of the excitatory junction potential (e.j.p.) produced by perivascular nerve stimulation was increased by low concentrations of procaine (2.5 X 10(-5)-10(-4)M) or high concentrations (10(-3)-5 X 10(-3)M) of caffeine and was decreased by low concentrations of caffeine (2.5 X 10(-5)-10(-4)M) or high concentrations of procaine (5 X 10(-4)-10(-3)M). Higher concentrations of caffeine (over 5 X 10(-3)M) induced a spike potential on the e.j.p., while higher concentrations of procaine (over 2.5 X 10(-3)M) inhibited the generation of e.j.ps. Facilitation of e.j.ps produced by repetitive stimulation of perivascular nerves remained unchanged by caffeine, while it was enhanced by procaine at any given concentration (caffeine 2.5 X 10(-4)-10(-3)M; procaine 10(-4)-10(-3)M). The membrane depolarization produced by exogenously applied NA (10(-5)M) was not blocked by pretreatment with procaine. Conduction velocity of perivascular nerve excitation remained unchanged by application of caffeine (up to 5 X 10(-3)M), and was reduced by application of procaine (over 2.5 X 10(-4)M). Outflow of NA during perivascular nerve stimulation remained unchanged by caffeine (10(-4)-3 X 10(-3)M), while it was enhanced by procaine (over 2.5 X 10(-4)M). The outflow of DOPEG was slightly reduced by caffeine (10(-3)-5 X 10(-3)M) and by lower concentrations of procaine (10(-4)-2.5 X 10(-4)M) but was not altered by higher concentrations of procaine (10(-3)-5 X 10(-3)M). It is concluded that in the guinea-pig mesenteric artery, high concentrations of caffeine (over 10(-3)M) increased the e.j.p. amplitude which might be due to an increase in membrane resistance of the smooth muscle cells. No marked effect of caffeine was observed on transmitter release from the nerve terminals. Procaine (over 2.5 X 10(-4)M) increased transmitter release from perivascular nerves and blocked the re-uptake mechanism of released NA. The mechanisms underlying the decrease in ej.p. amplitude by procaine remain to be determined.

Different ras gene mutational frequencies in thyroid papillary carcinomas in Japan and Thailand.

The incidence and pattern of ras oncogene mutations in human malignancies demonstrate geographic and racial differences. For example, specificity of alterations is found in cholangiocellular carcinomas in Thai patients with a different etiology from those in Japanese patients. In the present study, a comparison of ras gene mutations in thyroid papillary carcinomas from Japanese and Thai patients was performed using single-strand conformation polymorphism and direct sequencing analyses. The incidence of ras mutation differed markedly in Japanese (two of 24 carcinomas, 8.3%) and Thai (five of 10 carcinomas, 50%) patients. In addition, all but one ras mutation occurred at codon 12 of the K-ras gene in the Thai cases. These results suggest that thyroid cancers in Thailand may be due to specific genetic and/or environmental factors.

Lack of p16/CDKN2 alterations in thyroid carcinomas.

Exons 1-3 of the p16/CDKN2 gene and exons 4-9 of the p53 gene were screened for mutations by single-strand conformation polymorphism (SSCP) analysis and direct sequencing of PCR-amplified DNA from human primary thyroid carcinomas and thyroid carcinoma cell lines. The samples included 21 papillary carcinomas, 2 undifferentiated carcinomas, 1 follicular carcinoma, 1 medullary carcinoma and 2 cell lines originating from thyroid undifferentiated carcinomas. No homozygous deletions and mutations in the p16/CDKN2 were observed in any of the primary tumors or cell lined. In contrast, one of the two undifferentiated carcinomas an both cell lines demonstrated point mutations in the p53 gene. These results that p16/CDKN2 gene alteration is not required for malignant transformation in the thyroid, while p53 gene mutations may play a role in the progression from differentiated to undifferentiated carcinoma.

Expression of the estrogen receptor in human thyroid neoplasms.

The expression and quantitation of the estrogen receptor (ER) in human thyroid tumors were examined by biochemical, immunohistochemical, and reverse transcriptase-polymerase chain reaction (RT-PCR) techniques. For this study, neoplasms, adenomatous goiters and adjacent normal thyroid tissues were obtained from 35 patients which included 10 cases of papillary carcinomas, 17 cases of adenomas and 8 cases of adenomatous goiters. Regardless of the histopathological subtype, ER was detected in 19% (5/27) of the neoplastic tissues with the mean value of ER content of 5.0 +/- 1.3 fmol/mg protein and the mean Kd value of 0.38 +/- 0.28 nM. ER was also detected, but at a lower concentration (2.8 +/- 1.6 fmol/mg protein), in the surrounding normal tissues. There was no significant difference between the neoplasms and adenomatous goiters with respect to the incidence of ER positivity and ER content. Furthermore, ER-positive specimens, as determined by both biochemical and immunohistochemical techniques, also showed the expression of ER mRNA detected by RT-PCR method. These results demonstrate that both ER mRNA as well as ER protein are expressed in thyroid neoplasms. This suggests the possibility that estrogen may affect the tumorigenesis or the progression of some thyroid neoplasms.

p53 tumor suppressor gene and ras oncogene mutations in hypopharyngeal squamous cell carcinomas.

To examine the potential role of p53 and ras gene mutations in hypopharyngeal tumorigenesis, twenty-eight primary hypopharyngeal carcinomas, obtained at biopsy or total pharyngolaryngectomy, were investigated. Exons 5 through 9 of the p53 gene and exons 1 and 2 of the H-, K-, N-ras gene were screened using a combination of immunohistochemistry and single-strand conformation polymorphism analysis of polymerase chain reaction products (PCR-SSCP). The targeted DNA sequences coding for p53 and ras were confirmed by direct DNA sequencing. Point mutations of p53 were found in 9 (32.1%) of the 28 cases, including one with a double mutation, 3 in exon 5, 1 in exon 6, 2 in exon 7 and 4 in exon 8. Positive nuclear immunostaining for p53 was evident in 14 (50.0%) lesions. Seven (25.0%) of the 28 demonstrated point mutations in the H-rns gene, and 11 (39.3%) showed positive cytoplasmic staining for I as. The 5-year survival rate was worse with than without p53 overexpression (p <0.05). The present results suggest that gene mutations, although they occur at a relatively low incidence, are involved in hypopharyngeal tumorigenesis with p53 expression being a prognostic factor.

Genetic alterations in thyroid carcinomas and adenomas detected by restriction landmark genomic scanning.

Alterations of genomic DNAs in 9 papillary thyroid carcinomas and 3 follicular thyroid adenomas were examined by restriction landmark genomic scanning, a 2-dimensional gel analysis that allows detection of deletions, amplifications and other rearrangements of genomic DNA. DNAs from both thyroid tumors and associated non-tumorous glandular tissues were cleaved with the restriction enzyme NotI end-labeled with P-32 and size-fractionated by 2-dimensional electrophoresis using HinfI in a second digestion. The altered spots in carcinomas and adenomas were compared with those in nontumorous samples. Five and 4 spots were commonly amplified in carcinomas and adenomas, respectively. One amplified spot was apparently specific only for carcinoma and was not detected in any of adenomas examined. In contrast, 12 spots reduced in intensity were frequently observed in tumors, although a subset of 5 were more sporadically affected in adenomas. The results indicate both common and distinct genetic abnormalities occurring in thyroid tumors, which may relate to the different biological behaviors of malignant and benign neoplasms.

CDDP induces p53-dependent apoptosis in tongue cancer cells.

We have investigated the CDDP sensitivities of two tongue cancer cell lines with differing p53 genetic status, one with wild-type p53 (SAS) and the other with mutant-type p53 (HSC-4). SAS was about 2 times more sensitive at the D10 dose and demonstrated increased p53 and Bax protein levels at 10 h after CDDP treatment on Western blot analysis. On the other hand, overexpression of p53 in HSC-4 was observed without CDDP treatment and no elevation of Bax could be detected. Apoptosis was observed after CDDP treatment in SAS but not in HSC-4 by Hoechst 33342-staining and electrophoresis methods. These findings indicate that p53 plays an important role in apoptosis as a positive regulator of Bax expression. It is suggested that p53 status may have predictive potential with regard to response to CDDP therapy.

Thrombin activates NF-kappa B through thrombin receptor and results in proliferation of vascular smooth muscle cells: role of thrombin in atherosclerosis and restenosis.

We investigated the role of thrombin in the pathogenesis in atherosclerosis and restenosis. First we examined the effect of thrombin on cultured human vascular smooth muscle cells (VSMC). We showed that thrombin acts as a mitogen on VSMC through thrombin receptor. The expression of thrombin receptor was increased in the cell lines of VSMC established from directional coronary atherectomy (DCA). This is more pronounced in the cells from patients with restenosis after PTCA. Next we investigated the signaling pathway from thrombin/thrombin receptor. Thrombin activates thrombin receptor resulting in the exposing of the agonist peptide domain (thrombin receptor agonist peptide, TRAP). The signal from thrombin/thrombin receptor activated protein C kinase, tyrosine kinase, and MAP kinase and resulted in NF-kappa B activation. Furthermore, treatment of the cells with antisense p65 oligodeoxynucleotides of NF-kappa B inhibited the thrombin-stimulated growth of VSMC in vitro. These results suggest that thrombin may have a role in the pathogenesis of atherosclerosis and restenosis after PTCA through the thrombin receptor.