Tyrosine-nitration of caspase 3 and cytochrome c does not suppress apoptosis induction in squamous cell carcinoma cells

The influence of tyrosine nitration of cytochrome c and caspase 3 on apoptosis induction was investigated in an established squamous carcinoma cell line, OSC-4. The intracellular NO and O2(-) levels were increased up to about 110-120% and 140-180% of the control levels, respectively, after the treatment of OSC-4 cells with 5-FU (100 microg/ml), PLM (10 microg/ml), CDDP (10 microg/ml), or gamma-rays (20 Gy). The treatment of OSC-4 cells with ONOO(-) (1 mM) and the above anticancer agents induced tyrosine nitration of 14, 32 kDa protein among others and nitration of tyrosine residues of cytochrome c and caspase 3 was identified by the Western blotting of immunoprecipitates obtained by antibodies to these proapoptotic proteins. When cytochrome c and procaspase 3 were treated with ONOO(-), tyrosine nitration was increased in a ONOO(-)-dose dependent manner. Tyrosine nitration of cleaved (17 kDa) caspase 3, however, was not induced by ONOO(-). Procaspase 3 in the cytosol of HeLa cells was activated by the addition of ONOO(-)-treated as well as ONOO(-)-untreated cytochrome c. In addition, cleavage of ICAD and PARP were not suppressed in OSC-4 cells by pretreatment with ONOO(-). Activity of cleaved caspase 3 was not suppressed at low concentrations or by treatment with ONOO(-) or NO donors, SIN-1 and SNP. Furthermore, apoptosis of OSC-4 cells by the anticancer agents was not suppressed by ONOO(-). In conclusion, these results suggest that nitration of tyrosine residues of cytochrome c and procaspase 3 is induced by chemoradiotherapy but their nitration does not suppress cancer cell apoptosis.     

Truncus arteriosus revisited: An angiographic demonstration

Summary The 1965 reclassification of truncus arteriosus by Van Praagh and Van Praagh greatly enhanced our understanding of this interesting anomaly. This brief review article attempts to illustrate the various types of truncus arteriosus identified in this classification by demonstrating their angiographic features. Reemphasis of the usefulness of this classification should help students of congenital heart disease recognize the advantages of a uniform diagnostic approach to this entity.     

Allodynia and hyperalgesia in adjuvant-induced arthritic rats: time course of progression and efficacy of analgesics

The complete Freund's adjuvant (CFA)-induced arthritic rat model has extensively served as a laboratory model in the study of arthritic pain. However, the time courses of allodynia and hyperalgesia and the efficacies of different analgesics have not fully been analyzed in this model. Mechanical allodynia, thermal and joint hyperalgesia, and other disease development parameters (body weight, mobility, paw volume, and joint stiffness) were measured on postinoculation days (PIDs) 0 to 28 in rats. Acute analgesic efficacies of drugs were evaluated on PID 9 when degrees of allodynia, hyperalgesia, and joint stiffness in the ipsilateral paw reached almost the maximum, although those in the contralateral paw changed only slightly. In the ipsilateral paw, thermal hyperalgesia reached the maximum on PID 1, whereas mechanical allodynia and joint hyperalgesia progressively developed during the first 7 or 8 days, being tuned in to arthritis development. In the contralateral paw, thermal hyperalgesia never occurred, whereas mechanical allodynia and joint hyperalgesia developed after PID 11. Morphine and tramadol had full efficacies for all the pain parameters tested at sedation-inducing doses. Indomethacin and diclofenac significantly but partially improved thermal and joint hyperalgesia. Amitriptyline significantly reduced thermal and joint hyperalgesia only at sedation-inducing dose. Acetaminophen, carbamazepine, and gabapentin had, at the most, very small efficacies. In conclusion, the present study provided integrated information about the time course of pain and other disease development parameters in the CFA-induced arthritic rats, and clarified acute efficacies of different categories of analgesics for the allodynia and hyperalgesia.     

Alteration of Platelet Serotonin in Patients with Chronic Tension-Type Headache During Cold Pressor Test

Change of 5-hydroxytryptamine (5-HT) concentrations in platelets from patients with chronic tension-type headache (TH) and controls were observed during cold pressor test (CPT). Before cold stimulation, 5-HT concentrations in platelets from patients with TH were significantly lower than those found in controls. One minute after the start of cold stimulation, 5-HT levels in platelets from patients with TH were significantly much lower than in the controls, as control levels rose and TH levels fell. The results show that, under stress, the absorbance of 5-HT into the platelets in patients with TH is reduced. It is suggested that, in patients with TH, there are abnormalities of 5-HT uptake into platelets and factors which cause release of 5-HT from platelets.     

Treatment of Tension-type Headache With Tizanidine Hydrochloride: Its Efficacy and Relationship to the Plasma MHPG Concentration

Seventy-eight patients with tension-type headache (TH) were treated with tizanidine hydrochloride (tizanidine). Plasma 3-methoxy-4-hydroxyphenylglycol (MHPG) and serum free fatty acids (FFA) levels in these patients were determined before the treatment with tizanidine. Eighteen healthy volunteers composed the control group. Four weeks after the treatment with tizanidine 24 (31%) of 78 patients showed excellent improvement (excellent group); 28 (35%) showed moderate improvement (moderate group); 18 (23%) showed mild improvement (mild group); 7 (10%) showed no improvement and one (1%) showed worsening of her headache (no change and worsened group). The plasma MHPG levels in the excellent group were significantly higher than in the other groups, including the control group. The serum FFA levels in the excellent group were significantly higher than in the controls. In this study, 66% of the patients reported improvement in their headaches. Some patients with TH had high plasma MHPG levels and these patients in particular showed excellent improvement after the administration of tizanidine. Though there may be a placebo response to some extent, the clinical usefulness of tizanidine for TH seems to be excellent. Further study is necessary concerning the pharmacological effect of tizanidine and plasma MHPG levels in patients with TH.     

Platelet activation in muscle contraction headache and migraine

In migraine, the role of platelets is regarded as an important factor. We investigated plasma beta-thromboglobulin (BTG), platelet factor 4 (PF4), and 5-hydroxytryptamine (5-HT) in migraine patients and muscle contraction headache (MCH) patients during headache-free periods. The mean values of the plasma BTG, PF4, and 5-HT concentrations in the migraine group and the MCH group were significantly higher than those in healthy controls. The mean value of the plasma BTG concentration was significantly higher in the migraine group than in the MCH group, but the differences in the mean plasma PF4 and 5-HT concentrations between the two groups were not significant. Continuous platelet activation exists in both MCH patients and migraine patients. From the biochemical point of view, we have provided evidence for a similarity between migraine and MCH.     

Contrast-enhanced ultrasonography in the diagnosis of solid renal tumors.

OBJECTIVE: The purpose of this study was to evaluate the usefulness of contrast-enhanced ultrasonography (CEUS) in the diagnosis of solid renal tumors. METHODS: Twenty-nine patients with solid tumors detected on gray scale ultrasonography underwent resection for suspected renal malignancy. Findings of arterial phase contrast computed tomography (CT) and CEUS were compared for each diagnosis. RESULTS: Histopathologic examination of resected lesions showed malignancy in 26 patients (clear cell carcinoma, n = 18; papillary renal cell carcinoma, n = 6; collecting duct carcinoma, n = 1; and infiltrative urothelial carcinoma, n = 1) and benign tumors in 3 patients (oncocytoma, n = 2; and angiomyolipoma, n = 1). Contrast CT failed to show tumor blood flow in 5 of 29 patients, whereas CEUS showed this in all patients. Positive predictive values of CEUS and contrast CT in the diagnosis of renal malignancy were 100% and 82.8%, respectively. Among clear cell carcinomas, hypervascularity was observed on contrast CT in 16 of 18 patients and on CEUS in 17 of 18 patients. On the basis of hypervascularity, diagnostic sensitivity values for clear cell carcinoma were 94.4% for CEUS and 88.9% for contrast CT, whereas specificity values were 45.5% for CEUS and 72.7% for contrast CT. Among papillary cell carcinomas, contrast CT showed avascular lesions in 4 of 6 patients. However, CEUS showed blood flow in these lesions, leading to diagnosis of hypovascular renal tumors. CONCLUSIONS: Contrast-enhanced ultrasonography was more sensitive for detecting slight tumor blood flow than contrast CT and was useful in preoperatively diagnosing malignant hypovascular renal tumors but was less so for hypervascular renal tumors.     

Ventral approach for resecting hepatocellular carcinoma in the caval portion of the caudate lobe

Background

Resection of hepatocellular carcinoma located in the caudate lobe is challenging because this anatomical location is difficult to approach, especially the caval portion.

Haplotype of thrombomodulin gene associated with plasma thrombomodulin level and deep vein thrombosis in the Japanese population

INTRODUCTION: Thrombomodulin (TM) is an essential cofactor in protein C activation by thrombin. Here, we evaluated the contribution of genetic variations in the TM gene to soluble TM (sTM) level and deep vein thrombosis (DVT) in Japanese. PATIENTS AND METHODS: We sequenced the TM putative promoter, exon, and 3'-untranslated region in DVT patients (n=118). Among 17 genetic variations we identified, two missense mutations (R385K, D468Y) and three common single nucleotide polymorphisms (-202G>A, 2487A>T, 2729A>C) were genotyped in a general population of 2247 subjects (1032 men and 1215 women) whose sTM levels were measured. We then compared the frequency of these mutations in DVT patients with that in the age, body mass index-adjusted population-based controls. RESULTS: We identified one neutral mutation (H381) and three missense mutations (R385K; n=2, A455V; n=53 heterozygous, n=14 homozygous, D468Y; n=2) of TM in the DVT patients. Age-adjusted mean values of sTM were lower in C-allele carriers of 2729A>C than in noncarriers in the Japanese general population (women: 16.7+/-0.3 U/ml vs. 17.9+/-0.2 U/ml, p<0.01, men: 19.4+/-0.3 U/ml vs. 20.4+/-0.3 U/ml, p=0.03). Additionally, the CC genotype of this mutation was more common in the male DVT patients than in the male individuals of the general population (odds ratio=2.76, 95% confidence interval=1.14-6.67; p=0.02). This mutation was in linkage disequilibrium (r-square>0.9) with A455V mutation. CONCLUSIONS: TM mutations, especially those with a haplotype consisting of 2729A>C and A455V missense mutation, affect sTM levels, and may be associated with DVT in Japanese.