Deep Vein Thrombosis in Patients with Severe Motor and Intellectual Disabilities, Especially Diagnosis and Prevention of Recurrence for Chronic Thrombosis-Serial Changes of Sonography and D-Dimer.

Most patients with severe motor and intellectual disabilities (SMID) have restricted mobility capability and have been bedridden for long periods because of paralysis of the extremities caused by abnormal muscular tonicity due to cerebral palsy and developmental disabilities. Such patients are associated with a high risk of complications like deep vein thrombosis (DVT). Here, we report twelve patients (42.9%) with DVT among 28 patients with SMID during prolonged bed rest. However, we did not detect thrombosis in the soleal veins, finding it mostly in the femoral and common femoral veins. We applied anticoagulant therapy (warfarin), and carefully followed up the cases with DVT, regulating the warfarin dosage at prothrombin time-international normalized ratio (PT-INR) values around two to prevent recurrence of chronic thrombosis. Regarding laboratory data for the coagulation system, there were no cases above 5 µg/ml for the D-dimer and there were significant differences between the DVT and non-DVT groups in the D-dimer levels. The plasma levels of D-dimer in patients with DVT diminished to less than 1.0 µg/ml after warfarin treatment. Concerning sudden death (4.2%) in patients with SMID, we have to be very careful of the possibility of pulmonary thromboembolism due to DVT. Therefore, we should consider the particularity of the underdeveloped vascular system from underlying diseases for the evaluation of DVT. A detailed study of DVT as a vascular complication is very important for the smooth medical care of SMID, and serial assessment of compression Doppler ultrasonography of the lower extremities, as a noninvasive examination and measurement of D-dimer, is very helpful. (This article is a translation of Jpn J Phlebol 2014; 25: 34-42.).

Relationship between Specific Distributions of Isolated Soleal Vein Thrombosis and Risk Factors.

OBJECTIVE: The relationship between specific distributions of isolated soleal vein thrombosis (SVT) and risk factors was investigated. SUBJECTS AND METHODS: The subjects included 93 patients with SVT diagnosed with ultrasonography. RESULTS: In the acute thrombus distribution, the thrombi of central veins were significantly more frequent than the thrombi of medial veins in the unilateral SVT. The thrombi of central veins were not more significantly frequent than the thrombi of medial veins in the bilateral SVT. CONCLUSION: The risk factors of bilateral SVT are considered to be different from that of the unilateral SVT. (English translation of J Jpn Coll Angiol 2013; 53: 159-166).

Relation between Isolated Venous Thrombi in Soleal Muscle and Positive Anti-Nuclear Antibody.

OBJECTIVE: In patients with isolated soleal vein thrombosis (SVT), the relation between acute thrombi and positive anti-nuclear antibody (ANA) was investigated. SUBJECTS AND METHODS: The subjects were 116 lower extremities in 86 patients with SVT. They were diagnosed and examined by ultrasonography and blood serum analysis (D-dimer, ANA), and had been followed up every three months. RESULTS: They had acute SVT in 35 limbs (30%) and chronic SVT in 86 limbs (70%), and they had positive ANA in 63%. They had recurrent SVT in 26%, and all were positive for ANA. CONCLUSION: ANA-positivity might be a risk factor for acute thrombi in patients with SVT. (*English Translation of J Jpn Coll Angiol 2010; 50: 417-422.).

The effect of different frequencies of stretch on human dermal keratinocyte proliferation and survival.

The effects of cyclic stretch on mitogen-activated protein kinase (MAPK) and apoptosis in keratinocytes are not well understood. The aim of this study is to compare the effect of high frequency repetitive (HF) stretch to intermittent (I) stretch on human dermal keratinocytes proliferation and survival. Cultured human dermal keratinocytes were exposed to either repetitive HF or I stretch. Cell number was measured by coulter counter, DNA synthesis was assessed by BrdU staining, and apoptosis was assessed by terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL) staining. The activation of p38 MAPK, ERK 1/2, and AKT was assessed by immunoblotting. p38 MAPK, ERK 1/2, and AKT exhibited no change after HF stretch, while AKT and Homo sapiens BCL-2-antagonist of cell death (BAD) were significantly activated after I stretch. After experiencing I stretch for 2 d, keratinocyte proliferation rates were significantly decreased. This decrease was most likely not due to apoptosis as TUNEL-positive cells only increased for cells treated with an AKT inhibitor.

Inhibition of development of experimental aortic abdominal aneurysm in rat model by atorvastatin through inhibition of macrophage migration.

Recently, atherosclerosis has been considered to be the result of inflammation. Interestingly, hydroxymethylglutaryl-coenzyme (HMG-Co) A inhibitors (statins), which are clinically used as lipid-lowering agents, have been reported to have various anti-inflammatory effects. As abdominal aortic aneurysm (AAA) is a common degenerative condition associated with atherosclerosis, this study was designed to investigate the inhibitory effect of a statin, atorvastatin, on aneurysm formation apart from its lipid-lowering effect. We employed an elastase-induced rat AAA model, as statins do not lower cholesterol in rats. Mean aneurysm diameter was significantly smaller in the atorvastatin treatment group as compared to control at 4 weeks after surgery (P<0.05). Interestingly, atorvastatin inhibited the expression of ICAM and MCP-1, followed by the suppression of macrophage recruitment into the aortic wall at 1 week after operation. A significant reduction in MMP-12, but not MMP-2, -3 and -9, expression was also observed by treatment with atorvastatin at 1 week after surgery. In addition, synthesis of collagen and elastin in the vascular wall were significantly increased by atorvastatin. Here, the present study demonstrated a direct effect of atorvastatin to inhibit the progression of aortic aneurysm, independent of its lipid-lowering effect. This study suggests new therapeutic aspects of statins to inhibit the progression of aneurysms.

Hemodynamic and humoral effects of atrial natriuretic peptide on pulmonary circulation after cardiac surgery.

PURPOSE: Human atrial natriuretic peptide (h-ANP) elicits biological effects such as natriuresis, diuresis, and vasodilation, and plays a role in regulating pulmonary circulation. We conducted this clinical study to define its role and elucidate its mechanisms. METHODS: Twelve consecutive adult patients scheduled to undergo cardiac surgery with cardiopulmonary bypass (CPB) were prospectively selected for this study. After the completion of surgery, h-ANP was infused from the right atrium through a Swan-Ganz (S-G) catheter. Blood samples for measurement of ANP and cyclic guanosine monophosphate (cGMP), the second messenger of ANP, were drawn from the pulmonary artery (PA) through the S-G catheter and from the left atrium (LA) through the left atrial pressure line, before and after the infusion of h-ANP. Hemodynamic values were measured at the same time. RESULTS: After the h-ANP infusion, the plasma levels of ANP were significantly lower in the LA than in the PA, whereas the plasma levels of cGMP were significantly higher in the LA than in the PA. The infusion of h-ANP decreased the mean PA pressure significantly, and the systolic PA pressure remarkably. CONCLUSION: The infusion of h-ANP after cardiac surgery stimulates the secretion of cGMP from the pulmonary vascular bed and dilates the PA, thereby decreasing the PA pressure.

Effect of different frequencies of tensile strain on human dermal fibroblast proliferation and survival.

The aim of this study is to compare the effect of a high-frequency repetitive (HF) stretch or an intermittent (I) stretch on the cell proliferation and survival of human dermal fibroblasts and to determine the activation of any relevant signal pathways. Cultured human dermal fibroblasts were exposed to either HF or I stretch. Cell number was measured by counting, while DNA synthesis was assessed by 5-bromo-2'-deoxyuridine (BrdU) staining and apoptosis by terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling staining. To investigate the potential mechanisms of repetitive strain on the proliferation and survival of fibroblasts, the activation of relevant transduction pathways, such as p38 mitogen-activated protein kinase (MAPK), extracellular signal-regulated kinase (ERK)1/2, AKT, and BAD, was assessed by Western blot. In addition, the effect of inhibition of these pathways on the fibroblast response was also studied. After either HF or I stretch for 7 days, fibroblast number was significantly decreased and there were less BrdU-positive cells. The numbers of apoptotic and/or necrotic fibroblasts were not affected. p38 MAPK and ERK1/2 were significantly activated after HF stretch, but AKT and BAD were significantly activated after I stretch. The inhibitors of p38 MAPK and MAPK/ERK kinase as well as dominant-negative AKT reduced cell number after both HF and I stretch but these pathways were not critical for the stretch-induced decrease in cell number.

Hypertension accelerated experimental abdominal aortic aneurysm through upregulation of nuclear factor kappaB and Ets.

In this study, we focused on the effect of hypertension on the transcription factors nuclear factor kappaB (NFkappaB) and ets in the mechanisms of abdominal aortic aneurysm (AAA), and we investigated how hypertension affects the progression of AAA. AAA was produced by elastase perfusion in hypertensive rats and normotensive rats. The size of AAA rapidly increased in hypertensive rats as compared with normotensive rats. Western blot analysis demonstrated that the expression of matrix metalloproteinase (MMP)-2, -3 , -9, and -12, as well as intercellular adhesion molecule, was increased in hypertensive AAA rats, accompanied by upregulation of NFkappaB and ets. Moreover, in situ zymography showed that the activity of MMPs was increased in the aorta of a hypertensive AAA model as compared with that in a normotensive AAA model. Interestingly, transfection of chimeric decoy oligodeoxynucleotide (ODN) resulted in significant inhibition of aortic dilatation both in normotensive and hypertensive rats at 4 weeks after transfection. Destruction of elastic fibers was also significantly inhibited by transfection of chimeric decoy ODN in both hypertensive rats and normotensive rats. The expression of MMP-2, -3, -9, and -12, as well as intercellular adhesion molecule, was significantly attenuated by the chimeric decoy ODN, accompanied by inhibition of the migration of macrophages. Also, the effect of chimeric decoy ODN was confirmed in an organ culture. The present study demonstrated that hypertension accelerated the progression of experimental AAA through upregulation of NFkappaB and ets. Inhibition of NFkappaB and ets could be a novel therapeutic strategy to treat AAA in hypertensive patients.

Resection of a cardiac tumor extending into the inferior vena cava presenting as Budd-Chiari syndrome.

This report describes the successful treatment of a case of cardiac adenocarcinoma with the clinical presentation as Budd-Chiari syndrome. Complete surgical excision of the atriocaval mass was successfully achieved under deep hypothermic circulatory arrest. Histopathological diagnosis of this tumor was tubular adenocarcinoma with positive immunostaining by carcinoembrionic antigen. Subsequent systemic search could not detect any evidence of extra-cardiac primary site and distant metastatic lesion. A 2-year follow-up without any adjuvant therapy revealed no sign of recurrence.

Identification of secernin 1 as a novel immunotherapy target for gastric cancer using the expression profiles of cDNA microarray.

Despite the discovery of multiple TAAs, only a limited number is available for clinical application, particularly against epithelial malignancies. In this study we searched for novel TAAs using expression profiles of gastric cancer examined with cDNA microarray, and identified the SCRN1 gene as a candidate. SCRN1 was confirmed to be expressed in five out of seven gastric cancers with semiquantitative RT-PCR. With Northern blot analysis, it was detected abundantly in the testis and ovary, but it was barely detectable in 14 other normal human adult organs. Colony formation assay revealed that its augmented expression is associated with promoted cell growth. As these expression profiles and functional features of SCRN1 appeared to be compatible with the characteristics of the hypothesized ideal TAAs, we examined whether SCRN1 protein contains antigenic epitope peptides restricted to HLA-A*0201. We synthesized the candidate peptides derived from SCRN1, and tried to induce CTLs with each peptide. The CTL clones were successfully induced with a peptide SCRN1-196 (KMDAEHPEL), and they lyzed not only the peptide-pulsed targets but also the tumor cells expressing both SCRN1 and HLA-A*0201 endogenously. These results strongly suggest that SCRN1-196 is an epitope peptide restricted to HLA-A*0201. Furthermore, we synthesized an anchor-modified peptide SCRN1-9 V (KMDAEHPEV), in which leucine at position 9 was substituted for valine to increase the binding affinity to the HLA-A*0201 molecules. The CTL clones induced by SCRN1-9 V also recognized tumor cells expressing its natural SCRN1 protein endogenously. These results strongly suggest that SCRN1 is a novel TAA and these peptides, both native and modified, may be applicable for cancer vaccines to treat gastric cancer.

Role of AKT in cyclic strain-induced endothelial cell proliferation and survival.

Endothelial cells (ECs) are exposed to repetitive cyclic strain (CS) in vivo by the beating heart. The aim of this study was to assess the influence of CS amplitude and/or frequency on EC proliferation and survival and to determine the role of AKT in CS-induced EC proliferation and survival. Cultured bovine aortic ECs were exposed to 10% strain at a frequency of 60 (60 cpm-10%) or 100 (100 cpm-10%) cycles/min or 15.6% strain at a frequency of 60 cycles/min (60 cpm-15.6%). AKT, glycogen synthase kinase (GSK)-3beta, BAD, and cleaved caspase-3 were activated by CS in ECs. Increasing the magnitude or frequency of strain resulted in an earlier phosphorylation of GSK-3beta, although the magnitude of phosphorylation was similar. After CS at 60 cpm-10% for 24 h, the number of nontransfected ECs was significantly increased by 8.5% (P < 0.05). We found that the number of apoptotic ECs was slightly decreased with exposure to CS. ECs transfected with kinase-dead AKT (KA179) as well as plasmids containing a point mutation in the pleckstrin homology domain of AKT (RC25) not only prevented AKT, GSK-3beta, and BAD phosphorylation but also inhibited the CS-induced increase in cell number as well as the CS-induced protection against apoptosis (both P < 0.05). The ratio of 5'-bromo-2'-deoxyuridine-positive cells was increased when ECs transfected with RC25 and KA179 as well as nontransfected ECs and ECs transfected with Lipofectamine 2000 were exposed to CS. We conclude that AKT is important in enhancing the survival of ECs exposed to CS but is not involved in EC proliferation.

Laparoscopic bilateral partial adrenalectomy for adrenocortical adenomas causing Cushing's syndrome: report of a case.

Laparoscopic total adrenalectomy has become a standard technique for small adrenal tumors; however, bilateral adrenalectomy results in postoperative adrenal insufficiency, necessitating lifelong steroid replacement. To preserve adrenocortical function in a 41-year-old woman with bilateral adrenocortical adenoma (BAA) causing Cushing's syndrome, we performed laparoscopic bilateral partial adrenalectomy. We based our preoperative diagnosis of bilateral adrenocortical tumors causing Cushing's syndrome on the results of endocrinological investigations and imaging findings. Thus, we performed lateral transperitoneal laparoscopic bilateral partial adrenalectomy, preserving the adrenal glands, which were normal. Pathological examination of both tumors confirmed the diagnosis of adrenocortical adenoma. The patient had no postoperative complications, and her adrenocortical function was normal without steroid replacement at her 10-month follow-up. This report shows that Cushing's syndrome resulting from bilateral adenomas can be effectively treated by laparoscopic bilateral partial adrenalectomy as a minimally invasive, adrenocortical-preserving operation.

Off-pump axillo-coronary artery bypass grafting with porcelain aorta.

Coronary artery bypass grafting (CABG) in patients with severely diseased ascending aorta has been associated with high risk for cerebral vascular accidents due to atheromatous embolism. In this situation, aortic no-touch techniques are widely employed as most important surgical strategy to prevent these complications. A case of 75-year-old man with effort angina associated with porcelain ascending aorta was reported here. He successfully underwent off-pump axillo-coronary bypass grafting with saphenous vein graft and has remained uneventful during his follow-up period. The off-pump axillo-coronary artery bypass grafting seemed to be an appropriate procedure for coronary revascularization with severely diseased ascending aorta.

Prognostic significance of T helper 1 and 2 and T cytotoxic 1 and 2 cells in patients with non-small cell lung cancer.

BACKGROUND: We report the prognostic significance of peripheral and tumor-infiltrating Th1, Th2, Tc1 and Tc2 cells in lung cancer patients. MATERIALS AND METHODS: We evaluated the rates of interferon (IFN)-gamma+/CD4+ cells (Th1), interleukin (IL)-4+ /CD4+ cells (Th2), IFN-gamma+/CD8+ cells (Tc1), IL-4+ /CD8+ cells (Tc2), and the ratio of Th1 to Th2 and that of Tc1 to Tc2 among peripheral blood lymphocytes (PBL) and tumor-infiltrating lymphocytes (TIL), in 51 consecutive patients with non-small cell lung cancer, by detecting the intracellular cytokine production using three-color flow cytometry. RESULTS: Patients with a low Th1/Th2 ratio in peripheral blood lymphocytes had a significantly better prognosis than those with a high Th1/Th2 ratio (5-year survival rate: low: 74.7% vs. high: 50.3%; p=0.038). Patients with a low Th1/Th2 ratio in peripheral blood had a significantly better prognosis than those with a high Th1/Th2 ratio in pathological Stage II or III (5-year survival rate: low: 66.6% vs. high: 18.2%; p=0.018). CONCLUSION: A high Th1/Th2 ratio in peripheral blood is a negative prognostic factor, especially in pathological Stage II or III non-small cell lung cancer patients.

Identification of immunoglobulin superfamily 11 (IGSF11) as a novel target for cancer immunotherapy of gastrointestinal and hepatocellular carcinomas.

We previously performed gene expression profile analyses of 20 intestinal-type gastric cancers, and identified a set of genes whose expression levels were elevated in cancer tissues compared to their corresponding non-cancerous tissues. In the present study we focused on the immunoglobulin superfamily 11 gene (IGSF11). Its expression was also elevated in colorectal cancers and hepatocellular carcinomas as well as intestinal-type gastric cancers. Northern blot analysis showed that it was expressed abundantly in testis and ovary. These data suggest that IGSF11 is a good candidate of cancer-testis antigen. Furthermore, suppression of IGSF11 by siRNA retarded the growth of gastric cancer cells. To investigate the possibility of clinical application of peptide vaccine to IGSF11, we synthesized candidate epitope peptides for IGSF11 and tested whether the peptides elicit IGSF11-specific CTL. As a result, we successfully established oligo-clonal CTL by stimulation with IGSF11-9-207 (ALSSGLYQC). In addition, we also established additional CTL using IGSF11-9V (ALSSGLYQV), anchor-modified peptides of IGSF11-9-207. These peptides showed IGSF11-specific cytotoxic activity in an HLA-A*0201-restricted fashion, suggesting that these peptides may be applicable for cancer immunotherapy. These findings have provided a novel insight into carcinogenesis of the stomach, colon and liver, and will be helpful for the development of novel therapeutic strategies to a wide range of human cancers.

Expression of hypoxia-inducible factor (HIF-1alpha), VEGF-C and VEGF-D in non-invasive and invasive breast ductal carcinomas.

BACKGROUND: Hypoxia-inducible factor 1 alpha (HIF-1alpha) is a transcription factor that may play an important role in tumour growth and metastasis by its regulation of angiogenesis and lymphangiogenesis to survive cellular hypoxia. Lymphangiogenesis is promoted by vascular endothelial growth factors (VEGF)-C and VEGF-D, but the correlation between the expression of HIF-1alpha and VEGF-C or VEGF-D in human breast carcinoma is not well elucidated. This study examined the pathobiological role of these molecules in human breast ductal carcinoma. MATERIALS AND METHODS: The expressions of HIF-1alpha, VEGF-C and VEGF-D were analyzed in 10 normal mammary epithelia, 12 fibroadenomas, 20 ductal carcinomas in situ (DCISs and 36 invasive ductal carcinomas (IDCs) by immunohistochemistry, comparing clinicopathological parameters. RESULTS: HIF-1alpha expression in nuclei was found in DCIS and IDC, but not in normal or fibroadenoma cells. The HIF-1alpha labelling index was significantly correlated with the degree of VEGF-C expression in IDC (p < 0.001), but not in DCIS. HIF-1alpha expression was significantly correlated with tumour necrosis and with the Van Nuys prognostic index (VNPI) (p < 0.05, p < 0.05, respectively) in the 20 DCISs. On the other hand, VEGF-D levels, but not those of HIF-1alpha and VEGF-C, were significantly higher in cases with lymph node metastasis and estrogen receptor expression in carcinoma cells (p < 0.01, p < 0.05, respectively) in the 36 IDCs. CONCLUSION: These findings suggest that HIF-1alpha is expressed in the early stage of mammary carcinogenesis, in which the expression correlates with necrosis in the DCISs and with VEGF-C expression in the IDCs, the latter resulting in a higher frequency of metastasis to regional lymph nodes.

Trapidil enhances the slowly activating delayed rectifier potassium current and suppresses the transient inward current induced by catecholamine in Guinea pig ventricular myocytes.

We examined the electrophysiological effects of trapidil on the ionic currents influencing the repolarization and on the transient inward current (ITi) that can cause triggered arrhythmia using the whole-cell patch-clamp technique in guinea pig ventricular myocytes. Trapidil shortened the action potential duration (APD) and increased the delayed rectifier potassium current (IK) in a concentration-dependent manner. The effect of trapidil on the rapidly and slowly activating components of IK (IKr and IKs, respectively) was studied by the envelope of tails test. Trapidil failed to affect IKr and selectively enhanced IKs. Trapidil increased the amplitude of the L-type Ca2+ current (ICa,L), with an acceleration of its inactivation, whereas isoproterenol, a beta-adrenoceptor agonist, increased the amplitude of the ICa,L in a different manner. Isoproterenol activated ITi; however, trapidil not only failed to facilitate ITi but also suppressed isoproterenol-induced ITi. The inhibitory effect of trapidil on isoproterenol-induced ITi is at least partly via a reduction of Ca2+ overload through an acceleration of ICa,L inactivation and/or a sarcoplasmic reticulum (SR) Ca channel modulation. These results suggest that trapidil does not prolong the QT interval and has an antiarrhythmic effect on arrhythmias elicited by triggered activity secondary to Ca2+ overload at much higher concentrations than clinical concentration.