Sphingosine 1-phosphate protects rat liver sinusoidal endothelial cells from ethanol-induced apoptosis: Role of intracellular calcium and nitric oxide

In alcoholic liver disease, ethanol-induced damage to sinusoidal endothelial cells (SECs) appears to be important in the progression of liver damage. However, little is known about the mechanisms responsible for protection of SECs against ethanol-induced injury. To elucidate the role of sphingosine 1-phosphate (S1P), which is stored in platelets and may be released from them on their activation, we investigated the effect of S1P on rat liver SECs in primary culture. Pretreatment of cells with 1 mumol/L S1P attenuated ethanol-induced apoptosis. Electron microscopy confirmed this protective effect of S1P on damaged SECs in liver tissues after perfusion of ethanol. In the absence of ethanol, S1P increased DNA synthesis as determined via incorporation of bromodeoxyuridine. S1P also ameliorated the decreased DNA synthesis of cells induced by ethanol. Addition of S1P to cells induced an increase in intracellular calcium concentrations and NO production in cells. Western blotting revealed that S1P significantly induced the activation of endothelial NO synthase (eNOS), but not Akt, and that S1P-induced activation of eNOS was blocked by trifluoperazine, a calmodulin inhibitor. Furthermore, N(G)-nitro-L-arginine methyl ester, a NO synthase inhibitor, cancelled the effect of S1P on DNA synthesis, apoptosis, and NO production in vitro as well as the protective effect of S1P on cell damage in situ. In conclusion, the biological effect of S1P is at least partially mediated by Ca(2+)-sensitive eNOS activation and subsequent NO formation; extracellular S1P could contribute to sinusoidal protection and remodeling in alcoholic liver injury.     

Gene expression profiles in peripheral blood as a biomarker in cancer patients receiving peptide vaccination

BACKGROUND:

Because only a subset of patients show clinical responses to peptide‐based cancer vaccination, it is critical to identify biomarkers for selecting patients who would most likely benefit from this treatment.

METHODS:

The authors characterized the gene expression profiles in peripheral blood of vaccinated patients to identify biomarkers to predict patient prognosis. Peripheral blood was obtained from advanced castration‐resistant prostate cancer patients, who survived for >900 days (long‐term survivors, n = 20) or died within 300 days (short‐term survivors, n = 20) after treatment with personalized peptide vaccination. Gene expression profiles in prevaccination and postvaccination peripheral blood mononuclear cells (PBMCs) were assessed by DNA microarray.

RESULTS:

There were no statistically significant differences in the clinical or pathological features between the 2 groups. Microarray analysis of prevaccination PBMCs identified 19 genes that were differentially expressed between the short‐term and long‐term survivors. Among the 15 up‐regulated genes in the short‐term survivors, 13 genes, which were also differentially expressed in postvaccination PBMCs, were associated with gene signatures of granulocytes. When a set of 4 differentially expressed genes were selected as the best combination to determine patient survival, prognosis was correctly predicted in 12 of 13 patients in a validation set (accuracy, 92%).

CONCLUSIONS:

These results suggested that abnormal granulocytes present in the PBMC faction may contribute to poor prognosis in advanced prostate cancer patients receiving personalized peptide vaccination. Gene expression profiling in peripheral blood might thus be informative for devising better therapeutic strategies by predicting patient prognosis after cancer vaccines. Cancer 2012;118: 3208–21. © 2011 American Cancer Society.     

Orthostatic hypotension at the introductory phase of haemodialysis predicts all-cause mortality.

BACKGROUND: Since the predictive value of orthostatic hypotension (OH) at the introductory phase of haemodialysis (HD) is unknown, we examined the association between OH and all-cause death in patients who started HD between 1987 and 2001. METHODS: More than three consecutive blood pressure measurements before HD treatments (pre-HD BP) were made on each of 304 patients who had recently been started on HD and were in a stable condition. OH was defined as a drop in systolic BP of >20 mmHg or in diastolic BP of >10 mmHg after standing. RESULTS: Of 304 patients, 42% had OH. OH was significantly associated with pre-HD supine systolic BP; its severity was significantly associated with a past history of cerebrovascular disease and pre-HD supine systolic BP. During a mean follow-up of 4.0+/-3.0 years (range 0.1-13.2 years), 136 deaths were recorded. A multivariate Cox proportional hazards model analysis demonstrated that OH and a past history of cerebrovascular disease were independent predictors of all-cause death. The comparison by Kaplan-Meier analysis of the overall survival of patients with and without OH was significant. CONCLUSIONS: Our findings validate OH at the introductory phase of HD as a novel independent predictor of all-cause mortality among HD patients.     

Neuroscience of cursive handwriting: a computational appraoch]

Based on the minimum torque change model (MTCM), Wada and Kawato (1995) proposed a computational model of cursive handwriting which includes the following assumptions. The brain represents via-points that the hand passes through in a trajectory. Cursive handwriting consists of consecutive reaching movements. The via-points for handwriting are retrieved from memory during actual handwriting. Mathematically extracted via-points based on MTCM proved to be practically identical to motor primitives measured by EMG suggesting that drastic directional changes in muscle movement may be determining the location of via-points. Based on computational model, we examined the via-points in the cursive handwriting of a patient with parietal lobe involvement accompaning agraphia. While the control subject showed basically the same numbers and locations in via-points regardless of the velocity of hand movement, the patient's number and locations of via-points differed depending on the velocity of hand movement. The results suggest that the control subject retrieved necessary via-points based on the velocity of handwriting. The results also may reflect the neuropsychological nature of agraphic hand writing in a patient with parietal lobe damage.     

A peptide derived from hepatitis C virus (HCV) core protein inducing cellular responses in patients with HCV with various HLA class IA alleles

C35‐44 peptide is a well known HLA‐A2‐restricted CTL epitope originating from hepatitis C virus (HCV) core protein. It was reported that the majority of HCV positive patients had significant levels of serum IgG specific to this peptide. This study addressed whether C35‐44 peptide could induce CTL activity restricted to various HLA class IA alleles or could not. This peptide demonstrated binding activity to HLA‐A*2402, ‐A*2601, ‐A*3101, and ‐A*3303 molecules, but not to HLA‐A*1101 by means of stabilization assay. This peptide also induced CTL activity restricted to each of them, except HLA‐A11⁺ peripheral blood mononuclear cells from HCV 1b⁺ patients by means of ⁵¹Cr‐release assay. With regard to HLA‐A2 subtypes, this peptide demonstrated binding activity to HLA‐A*0201 and ‐A*0206, but not to ‐A*0207 molecules. Furthermore, this peptide induced CTL activity from both the patients and healthy donors with all the HLA class IA molecules mentioned above by means of interferon‐γ production assay. These results may provide new insights for the development of a novel peptide vaccine against HCV compatible with various HLA class IA types. J. Med. Virol. 81:1232–1240, 2009. © 2009 Wiley‐Liss, Inc.     

Fulminating midbrain irradiation injury of pediatric brain tumor]

We report two children with post radiation midbrain damage causing severe neurological symptoms. A twelve-year-old boy with a four year history of hydrocephalus was diagnosed with tectal glioma, which endoscopic biopsy revealed to be low grade. He underwent gamma knife radiation surgery (central 24 Gy/peripheral 12 Gy). Two months later bilateral ptosis followed by total oculomotor palsy and drowsiness developed. Despite pulsed-steroid therapy the tumor size increased up to 4.6 times in volume. The tumor was totally removed and was diagnosed as an early delayed radiation reaction pathologically. His symptoms disappeared except for a slight upper gaze palsy. The second patient was a six-year-old girl with a medulloblastoma. Following total resection and a VP shunt she received conventional radiation therapy along with chemotherapy. After the final irradiation she became comatose (JCS II-2) and MRI revealed diffuse midbrain damage with acute aqueduct obstruction, which recovered in two weeks. Reports of irradiation injuries of the midbrain in childhood are rare but it should be considered as a possible cause of fulminant symptoms requiring emergency treatment. Because of midbrain anatomical complexity, midbrain radiation therapy requires great care, especially in children.     

High-level in vivo gene marking after gene-modified autologous hematopoietic stem cell transplantation without marrow conditioning in nonhuman primates.

The successful engraftment of genetically modified hematopoietic stem cells (HSCs) without toxic conditioning is a desired goal for HSC gene therapy. To this end, we have examined the combination of intrabone marrow transplantation (iBMT) and in vivo expansion by a selective amplifier gene (SAG) in a nonhuman primate model. The SAG is a chimeric gene consisting of the erythropoietin (EPO) receptor gene (as a molecular switch) and c-Mpl gene (as a signal generator). Cynomolgus CD34+ cells were retrovirally transduced with or without SAG and returned into the femur and humerus following irrigation with saline without prior conditioning. After iBMT without SAG, 2-30% of colony-forming cells were gene marked over 1 year. The marking levels in the peripheral blood, however, remained low (<0.1%). These results indicate that transplanted cells can engraft without conditioning after iBMT, but in vivo expansion is limited. On the other hand, after iBMT with SAG, the peripheral marking levels increased more than 20-fold (up to 8-9%) in response to EPO even at 1 year posttransplant. The increase was EPO-dependent, multilineage, polyclonal, and repeatable. Our results suggest that the combination of iBMT and SAG allows efficient in vivo gene transduction without marrow conditioning.     

A case of ovarian carcinoma with production of granulocyte colony-stimulating factor

A 60-year-old female with ovarian carcinoma showed marked leucocytosis. After exploratory laparotomy she had received carboplatin-based chemotherapy, but she died of respiratory failure. In this case the immunohistochemical examination of the tumour cells showed partial-positive staining for anti-G-CSF monoclonal antibody in addition to the elevation of the serum G-CSF concentration. This case is the first report of an ovarian carcinoma producing G-CSF.