Endocan as a prognostic biomarker of triple-negative breast cancer.

PURPOSE: Triple-negative breast cancer (TNBC) has aggressive characteristics and fewer treatment options than other subtypes. The purpose of this study was to explore prognostic biomarkers for TNBC that can be easily detected from the blood samples. METHODS: MDA-MB-231 and MDA-MB-231BR, a brain metastatic variant of the human TNBC cell line MDA-MB-231, were used as less and more aggressive models of TNBC, respectively. The extent to which the candidate gene/protein identified by RNA sequencing correlated well with aggressiveness of TNBC and how much protein was detected from the blood of tumor-bearing mice were evaluated. RESULTS: Both the in vitro proliferation and in vivo tumor growth of MDA-MB-231BR were more rapid than those of MDA-MB-231. RNA sequencing identified ESM1 as a gene that was expressed significantly more in MDA-MB-231BR than in MDA-MB-231, and qRT-PCR confirmed a significantly higher expression of ESM1 in MDA-MB-231BR xenograft in vivo. Furthermore, Kaplan-Meier analysis of relapse-free survival demonstrated that TNBC patients with high ESM1 expression had clearly worse relapse-free survival than those with low ESM1 expression, which was consistent with our preclinical findings. Endocan, a protein of ESM1 gene product, was successfully detected in both conditioned medium from MDA-MB-231BR and plasma samples from mice bearing MDA-MB-231BR xenograft, which showed a significantly distinct pattern from less aggressive MDA-MB-231. Moreover, bisulfite sequence analysis revealed that overexpression of ESM1 in MDA-MB-231BR might be attributed to DNA demethylation in an upstream region of the ESM1 gene. CONCLUSION: This study indicates that endocan could be used as a blood-based prognostic biomarker in TNBC patients.

Controlled Secretion of the Anticancer Protein MDA-7 from Engineered Mesenchymal Stem Cells.

Mesenchymal stem cells (MSCs) have been explored as a "live" carrier of cytokines for targeted cancer therapy, but, in earlier reports in the literature, the secretion process of therapeutic cytokines was not regulated. The purpose of this study was to generate MSCs to conditionally secrete the melanoma differentiation-associated gene-7 (MDA-7) tumor-suppressor protein. To control the secretion of MDA-7 from MSCs, a well-established tetracycline-controlled transcriptional activation system was incorporated into MDA-7 plasmid. MDA-7 gene expression was induced in the engineered MSCs only in the presence of doxycycline, as characterized by quantitative reverse transcription (qRT)-PCR. Enzyme-linked immunosorbent assay (ELISA) also revealed that the MDA-7 protein was secreted from the engineered MSCs only after the cells had been exposed to doxycycline. Both recombinant human MDA-7 protein and the conditioned medium from the engineered MSCs in the presence of doxycycline significantly inhibited tube formation of human umbilical vascular endothelial cells (HUVECs), indicating that our system could be used for targeted, antiangiogenic therapy. Overall, this study provides useful information on the potential use of engineered MSCs for the controlled secretion of therapeutic proteins, in this case MDA-7, for targeted cancer therapy.

Bilateral isolated coronary ostial stenosis in a middle-aged premenopausal woman with vasospastic angina: a case report.

Background: Vasospastic angina (VSA) is uncommon in premenopausal women who have less chronic endothelial injury causing vascular remodelling, considered to play a primary role in the pathogenesis for coronary vasospasms. Furthermore, vasospasms rarely occur in the bilateral coronary ostia. Isolated coronary ostial stenosis (ICOS), which often causes severe effort angina and requires surgical intervention, is more commonly reported in middle-aged women, with causes including fibromuscular dysplasia (FMD) and large-vessel vasculitis. However, ICOS associated with VSA is extremely rare. Case summary: A 50-year-old premenopausal Japanese woman presented with a complaint of typical chest pain due to angina during light exertion daily in the early morning hours since 3 years. Coronary angiography (CAG) revealed bilateral mild-to-moderate ICOS in addition to multi-vessel spasms involving the bilateral coronary ostia confirmed by the vasospasm provocation test using intracoronary acetylcholine injection. Tests to determine the cause of ICOS did not identify FMD or any other disease. The angina attacks alleviated after calcium channel blocker (CCB) administration without intervention for bilateral ICOS for 24 years since the first presentation. Moreover, coronary computed tomography angiography (CTA) performed 24 years after the first presentation showed no ICOS. Discussion: In our patient with typical and frequent VSA symptoms, CAG revealed both mild-to-moderate ICOS and the vasospasms in the bilateral coronary ostia. Fibromuscular dysplasia or large-vessel vasculitis was ruled out as the causes of ICOS. Vasospastic angina rarely occurred after the prescription of CCB, and coronary CTA 24 years after the first presentation showed no ICOS. Bilateral ICOS in our patient might be VSA related.

Non-HDL cholesterol as a predictor of carotid atherosclerosis in the elderly.

The aim of the present study was to evaluate sclerotic lesions of the common carotid artery by ultrasonography in 921 in-patients aged 65 years and older (77 +/- 7 years) and investigate whether lipid levels were associated with carotid atherosclerosis. In men, an increased risk for carotid atherosclerosis was associated with increased levels of low-density lipoprotein cholesterol (LDL-C) and non-high-density lipoprotein cholesterol (non-HDL-C). Compared to men with the lowest tertile of LDL-C levels (< 83.4 mg/dl), the adjusted odds ratio was 2.502 (95% confidence interval: 1.426-4.390) in those with the middle tertile (83.4-115.2 mg/dl), and 2.688 (1.509-4.790) in those with the highest tertile ( > 115.2 mg/dl). Like the LDL-C level, the non-HDL-C level showed a positive and linear relationship with carotid atherosclerosis. Compared to men with the lowest tertile of non-HDL-C levels ( < 101 mg/dl), the adjusted odds ratio was 2.881 (1.633-5.081) for those with the middle tertile (101-135 mg/dl), and 2.990 (1.651-5.415) for those with the highest tertile ( > 135 mg/dl). Similarly, in women, an increased risk for carotid atherosclerosis was also positively and linearly associated with LDL-C and non-HDL-C. The Non-HDL-C level is a potential predictor of risk for carotid atherosclerosis in the elderly.

Association between abdominal wall fat index on ultrasonography and carotid atherosclerosis in non-obese men.

We tried to investigate whether accumulation of visceral fat, assessed by a simple but widely used ultrasonographic method, was associated with common carotid atherosclerosis in non-obese men ranging from 16 to 79 years old. The subjects were consecutive 297 male in-patients whose body mass index ranged from 18.5 kg/m(2) to 25 kg/m(2). An ultrasonographic evaluation using a 7.5 MHz linear type B-mode probe was performed by a specialist to determine the intima-media thickness (IMT) of the common carotid artery and the maximal thickness of peritoneal fat (Pmax) at the anterior surface of the liver and the minimal thickness of subcutaneous fat (Smin) of the abdomen. The Pmax/Smin ratio, which was termed the abdominal wall fat index (AFI), was then calculated. The mean age +/- standard deviation in this series was 65 +/- 13 (range, 15-79) years. Multiple regression analysis using IMT as an objective variable, adjusted by various risk factors as explanatory variables, showed that AFI [beta, 0.0538; 95% confidence interval (CI), 0.0116-0.0960] was a significant independent contributing factor along with known risk factors such as age, smoking status, systolic blood pressure, HDL-cholesterol and LDL-cholesterol. We found that AFI was useful in evaluating disorders of metabolism and atherosclerosis in non-obese men.

Metabolic syndrome as a predictor of ischemic stroke in elderly persons.

OBJECTIVE: To estimate the prevalence and risk of ischemic stroke associated with metabolic syndrome. METHODS AND PATIENTS: In 197 stroke patients (80 cases of atherothrombotic infarction, 97 lacunar infarction, 16 cardioembolic infarction, 4 others) and 356 age- and sex-matched control subjects aged 65 years and older in Seiyo Municipal Nomura Hospital, we investigated the association between metabolic syndrome and risk factor-dependent augmentation of ischemic stroke in subjects with several risk factors for atherosclerosis. The diagnosis of cerebral infarction in each patient was confirmed by CT findings of the brain and metabolic syndrome was defined as at least 3 of the 5 following conditions: visceral obesity, hypertension (HT), hypertriglyceridemia, low HDL-cholesterol and diabetes mellitus (DM). RESULTS: After adjustment for sex, age, and smoking, metabolic syndrome was significantly related to atherothrombotic infarction (odds ratio, 3.08; 95% confidence interval, 1.69-5.61). Of the individual components, visceral obesity, HT and DM were independent risk factors for atherothrombotic infarction. Increased risk for atherothrombotic infarction was also associated with increases in the 5 component conditions of the metabolic syndrome. CONCLUSION: The clustering of cardiovascular risk factors called metabolic syndrome increases the risk of cardiovascular morbidity, and its identification may thus be important in risk assessment and treatment of patients.

Metabolic syndrome amplifies the LDL-cholesterol associated increases in carotid atherosclerosis.

OBJECTIVE: Carotid intima-media thickness (IMT) is a useful surrogate marker of cardiovascular disease. In addition to low-density lipoprotein cholesterol (LDL-C), metabolic syndrome has been linked to the pathogenesis of atherosclerosis. The present study investigated whether the clustering of multiple components of metabolic syndrome has a greater impact on vascular parameters than individual components of metabolic syndrome, and assessed the association between carotid IMT and LDL-C and metabolic syndrome. METHODS: Carotid IMT was evaluated on B-mode ultrasonography in 760 patients (340 men aged 64+/-16 years and 420 women aged 69+/-13 years) in the Medical Department of Seiyo Municipal Nomura Hospital. The subjects did not demonstrate any clinical signs of cardiovascular disease. We investigated the association between carotid IMT and confounding risk factors including LDL-C and metabolic syndrome using the 2005 Japanese criteria. RESULTS: Carotid IMT increased with increasing numbers of metabolic syndrome components (p for trend<0.001). Multiple regression models, including age, sex, body mass index, smoking status, LDL-C, diabetes mellitus as well as each individual component of metabolic syndrome as continuous variables, showed that both metabolic syndrome (beta=0.100; p=0.029) and LDL-C (beta=0.210, p<0.001) were independent determinants of carotid IMT. Metabolic syndrome amplified the LDL-C associated increases in carotid atherosclerosis. CONCLUSIONS: Even after taking into account each individual component of MS, the clustering of visceral obesity with at least 2 of the 3 components, and LDL-C are independently associated with increased carotid IMT. This suggests that the components of metabolic syndrome interact to synergistically impact vascular thickness.

Association between uric acid and carotid atherosclerosis in elderly persons.

OBJECTIVE: Several cohort studies have shown a link between serum uric acid (SUA) and subsequent cardiovascular disease. However, such an association did not remain significant after adjusting for concomitant risk factors for atherosclerosis in some studies. Thus, the role of SUA as an independent risk factor remains controversial. We therefore investigated the association between SUA and sclerotic lesions of common carotid atherosclerosis. PATIENTS AND METHODS: We evaluated sclerotic lesions of the common carotid arterial intima-media thickness (IMT) by ultrasonography in 398 men aged 74+/-8 (range, 60-97) years and 521 women aged 75+/-8 (range, 60-104) years. To investigate the relation between SUA and various factors, all subjects were divided into three groups based on the tertile of SUA. RESULTS: Stepwise multiple linear regression analysis using IMT as an objective variable, adjusted by various risk factors as explanatory variables showed that SUA [B, 0.0099; 95% confidence interval (CI), 0.0022-0.0175] was a significant independent contributing factor along with known risk factors such as age, sex, smoking status, systolic blood pressure, diastolic blood pressure, antihypertensive drug use, HDL-cholesterol, and LDL-cholesterol. Multiple logistic regression analysis for carotid IMT as a tertile of SUA and dependent variables showed that the adjusted odds ratio was 1.25 (95% CI, 0.87-1.78) for those in the middle tertile (4.2-5.5 mg/dl), and 1.66 (95% CI, 1.16-2.39) for those in the highest tertile (5.6-14.1 mg/dl) compared to that for subjects in the lowest tertile of SUA levels (0.51-4.1 mg/dl). CONCLUSION: We suggest that SUA is a risk factor or marker for ultrasonographically determined IMT.

Severe hypoglycemia associated with insulin-like growth factor II-producing liver metastasis from gastric carcinoma treated with overnight total parenteral nutrition via a central vein catheter reserve port.

Hypoglycemia caused by insulin-like growth factor II is difficult to control. A 77-year-old woman was diagnosed with gastric cancer and multiple liver metastases in September 2006 and underwent chemotherapy; however, at that time there were no symptoms of hypoglycemia. From January 2007 onwards, hypoglycemic comas and symptoms of hypoglycemia began to appear frequently. Her serum level of insulin was normal; thus, we suspected the presence of big insulin-like growth factor II was causing the hypoglycemia. This was proven by Western immunoblotting and we diagnosed non-islet cell tumor hypoglycemia associated with gastric cancer. Overnight nutrition provided via a central venous catheter port to prevent hypoglycemia allowed the patient to become ambulant and to remain free of hypoglycemic coma at follow-up until her death 7 months later.

Evaluation of contrast-enhanced ultrasonography using perfluorobutane (Sonazoid(®)) in patients with small hepatocellular carcinoma: comparison with dynamic computed tomography.

This study aimed to elucidate the efficacy of contrast-enhanced ultrasonography (CEUS) with perfluorobutane (Sonazoid(®)) in the diagnosis of hepatocellular carcinomas (HCCs), particularly small HCCs, by comparing the results with dynamic computed tomography (Dy-CT). Seventy-nine nodules in 69 patients with chronic liver disease, suspected as HCCs were studied. The nodules were selected based on the results of B-mode ultrasonography and/or Dy-CT conducted between January and August 2007. The nodules were divided into two groups: the S-group with tumors ≤2 cm (49 nodules), and the L-group with tumors >2 cm (30 nodules). Typical HCCs were defined, and the nodules were enhanced and shown as defects in the arterial and late phase of Dy-CT, respectively. Target lesions were scanned using CEUS, and the results were compared with those of Dy-CT. The L-group nodules diagnosed as HCCs using Dy-CT were also diagnosed as HCCs using CEUS. In the S-group, the diagnostic sensitivity of CEUS was 94.7% and the specificity was 81.8%. We diagnosed two liver tumors that were detected by CEUS but not by Dy-CT; biopsies revealed one tumor to be a well-differentiated HCC and the other to be an atypical adenomatous hyperplasia. The sensitivity and specificity of CEUS against HCC were high even in the small-size HCCs. Thus, Sonazoid is useful in the screening for small HCCs.

Primitive neuroectodermal tumor of the pancreas.

The primitive neuroectodermal tumor (PNET) of the pancreas, a member of Ewing's sarcoma family of tumors, is extremely rare. We treated a 37-year-old Japanese man who had a solitary pancreatic tumor 40 mm in diameter and multiple hepatic tumors with surgical resection. The PNET was positive for CD99 on immunohistochemical staining. Fluorescence in situ hybridization (FISH) was also performed, which revealed a Ewing sarcoma breakpoint region 1 (EWSR1) 22q12 rearrangement. According to the Japan-Ewing protocol, chemotherapy with Ifomide (ifosfamide), etoposide, vincristine, and cyclophosphamide was given after surgery. To the best of our knowledge, to date 13 PNET cases have been reported with a mean age for all patients of 19.3 years old. Surgical resection was performed in most cases and some patients received postoperative chemotherapy. The clinicopathologic characteristics and management of this extremely rare disease are also discussed.

Brain metastasis from hepatocellular carcinoma treated with a cyber-knife.

A 70-year-old right-handed Japanese man who had undergone surgical resection for hepatocellular carcinoma (HCC) 2 years earlier was diagnosed with lung metastasis 3 months before consulting our hospital with a headache and visual field disturbance. Head computed tomography revealed a brain tumor with an intracerebral hemorrhage. Using (99m)Tc-PMT (pyridoxal-5-methly-triptophan) scintigraphy, we determined that the brain tumor was metastasis from the HCC and utilized the cyber-knife for treatment. The prognosis of patients with brain metastasis from HCC has been reported to be poor. Use of the cyber-knife was non-invasive, and proved to be effective for improving prognosis and quality of life.