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PubMed2003

Gene expression of vitamin D hydroxylase and megalin in the remnant kidney of nephrectomized rats.

BACKGROUND: Regulation of vitamin D hydroxylase genes in the early stage of chronic renal failure is not fully understood. Using nephrectomized rats, we examined changes in mRNA levels of CYP27B1 (25-hydroxyvitamin D3-1 alpha-hydroxylase), CYP24 (25-hydroxyvitamin D3-24-hydroxylase), and vitamin D receptor in relation to megalin, recently found to participate in renal vitamin D metabolism. METHODS: A rat model of moderate renal failure was induced by 3/4 nephrectomy. Plasma parameters, including vitamin D metabolite concentrations, were measured at weeks 2, 4 and 8, and poly(A)+ RNA extracted from the remnant kidneys was subjected to Northern blot hybridization. RESULTS: Plasma creatinine concentration at week 2 was 0.40 +/- 0.02 mg/dL in the sham-operated and 0.93 +/- 0.15 mg/dL in the nephrectomized rats, and both values remained constant up to week 8. Plasma concentrations of 25(OH)D3, 1 alpha,25(OH)2D3, and 24,25(OH)2D3 were unchanged between nephrectomized and sham-operated rats at week 8. Intact parathyroid hormone (PTH) increased at week 8 in nephrectomized rats. CYP27B1 mRNA in nephrectomized rats did not vary at week 2, but increased approximately two- and four-fold at weeks 4 and 8, respectively, compared to the sham-operated rats. CYP24 and megalin mRNAs, on the other hand, began to decline as early as at week 2 in nephrectomized rats and kept decreasing throughout the experiment. The expression of vitamin D receptor was modestly but significantly decreased only at week 8. CONCLUSION: Coordinated and reciprocal alterations of the increase in CYP27B1 mRNA and the decrease in CYP24 mRNA may play a pivotal role in maintaining the plasma level of 1 alpha,25(OH)2D3 in the face of reduced nephron mass and/or megalin expression.

25-Hydroxyvitamin D3 1-alpha-Hydroxylase↗
PubMed1999

[Histological characteristics and clinical significance of Japanese prostate cancer with low levels of prostate specific antigen of 4.0 ng/ml or lower].

PURPOSE: They set a normal limit of prostate specific antigen (PSA) to 4.0 ng/ml in Tandem R assay at most institutions. We investigated clinical and histological characteristics of prostate cancer based on whole mount step-section histology of radical prostatectomy specimens, and taking notice of Japanese prostate cancer whose levels of PSA are less than 4.0 ng/ml in normal levels. MATERIALS AND METHODS: One hundred and twenty-two patients underwent radical prostatectomy for clinically resectable prostate cancer at University Hospital from February 1992 to April 1997. Clinicopathological findings were stratified according to the preoperative PSA levels in 111 patients without preoperative endocrine therapy. Immunohistochemical study for PSA was conducted in 7 randomly selected patients. RESULTS: Of the patients 22 (19.8%) had normal (4.0 ng/ml or lower) preoperative serum PSA. Mean tumor volume in this PSA range was 1.5 cm3 with one pT 0 case included. Pathologically organ confined, potentially curable disease (< pT 3) was found in 17 (77.3%) patients and extracapsular extension and seminal vesicle invasion in 5 (23.8%), respectively. No patients had positive pelvic lymph nodes. Well differentiated tumors of Gleason scores 2-4 were found in 9 (40.9%) of the patients, moderately differentiated tumors (Gleason scores 5, 6) in 5 (22.7%) and poorly differentiated histology (Gleason scores 7-10) in 7 (31.8%). Sixteen (72.7%) patients had clinically significant tumors (> 0.5 cm3, Gleason score > or = 7). All 7 patients had positive staining for PSA, but its intensity did not correlate with serum PSA levels. CONCLUSIONS: Many prostate cancers found in surgical specimens were clinically significant despite the low levels of PSA and potentially curable by definitive treatment. Age, co-morbidity and other clinicopathological variables as well as PSA levels should all be taken into account when treatment options are discussed.

Aged↗
PubMedDate not supplied

Elevated levels of circulating cell-free DNA in the blood of patients with hepatitis C virus-associated hepatocellular carcinoma.

BACKGROUND: Circulating cell-free DNA is present in increased amounts in the blood of patients with one of several forms of cancer. MATERIALS AND METHODS: A real-time PCR assay with glutathione S-transferase pi (GSTP1) gene was used to measure cell-free DNA levels in the sera of 52 patients with hepatocellular carcinoma (HCC) associated with hepatitis C virus (HCV), which included 30 HCV carriers without known HCC and 16 HCV-negative non-cancer patients (controls). RESULTS: Cell-free DNA levels were significantly higher in the sera from HCC patients than in the sera from HCV carriers or the control subjects. Cell-free DNA levels were associated with the degree of tumor differentiation and size but not patient age, gender, TNM stage or levels of alpha-fetoprotein (AFP) or protein induced by vitamin K absence (PIVKA-II). The cell-free DNA assay had a sensitivity of 69.2% and a specificity of 93.3% in discriminating HCC and HCV carriers at the optimal cut-off value of 73.0 ng/ml, with an area of 0.90 (95% CI, 0.83-0.96) under the receiver operating characteristic curve. The discriminative power of cell-free DNA was superior to that of AFP or PIVKA-II. CONCLUSION: Our results showed that levels of circulating cell-free DNA are significantly increased in sera of patients with HCV-associated HCC, suggesting that circulating cell-free DNA may be a good biomarker specific for HCV-associated HCC.

Aged↗
PubMed2003

Functional recovery after simultaneous transplantation with neuro-epithelial stem cells and adjacent mesenchymal tissues into infarcted rat brain.

BACKGROUND: Cerebral infarction results in impairment of motor and cognitive functions. We performed intracranial transplantation of multipotential neuro-epithelial stem cells with mesenchyme into experimentally large ischemic lesions to study their potential to relieve deficits. METHODS: Wistar albino rats were subjected to transient middle cerebral artery occlusion for 60 minutes, producing an extensive ischemic lesion in the ipsilateral striatum and adjacent cerebral cortex. The rat mesencephalic neural plate at the early somite stage (embryonic day 10.5) together with adjacent ventral mesenchyme was used as donor material. We performed histological and immunohistochemical studies, with antibodies against tyrosine hydroxylase, and dopamine- and adenosine 3': 5'-monophosphate-regulated phosphoprotein 32 (DARPP-32; a striatal marker). Micro-angiograms were made by using Microfil silicone rubber. Morris Water-maze learning and treadmill task were employed to evaluate motor and cognitive functions. FINDINGS: A viable non-tumoral mass was recognized in the rat striatum, up to as long as 156 days after transplantation. There were many cells positive for tyrosine hydroxylase or DARPP-32 in the graft. Some of the DARPP-32 positive cells within the graft had extended their dendrites into host tissues. In the micro-angiograms, many fine vessels were observed within the graft and dilated vessels meandered around the graft. Transplanted animals recovered significantly better in motor and cognitive functions than animals injected with only culture medium. INTERPRETATION: Neuro-epithelial stem cells may follow several lines of differentiation; along the naturally genetically programmed line of differentiation, or along other cell lines depending on different environments. Grafting of neuro-epithelial stem cells with mesenchyme may merit a further study as a treatment for cerebral infarction.

Animals↗
PubMed2001

Morphometric characteristics of optic disk with disk hemorrhage in normal-tension glaucoma.

PURPOSE: To evaluate the morphometric characteristics of the optic disk in eyes with and without disk hemorrhage in normal-tension glaucoma. METHODS: This was a prospective study conducted at Gifu University Hospital of 50 eyes of 50 patients with normal-tension glaucoma (12 men, 38 women; age, 56.5 +/- 14.1 years) who had developed new disk hemorrhage at the time of enrollment and 58 eyes of 58 patients with normal-tension glaucoma (20 men, 38 women; age, 56.7 +/- 12.4 years) with no history of disk hemorrhage during the follow-up period of more than 2 years. Age and global indexes of the visual field were matched. We morphometrically compared the optic disk with and without hemorrhage using a scanning laser tomograph. Global and sector analyses were made of the optic disk structural parameters. RESULTS: There was no significant difference in the global values of the disk parameters between the disk hemorrhage and the nonhemorrhage groups. However, the inferotemporal values for the rim area, rim volume, mean retinal nerve fiber layer thickness, and retinal nerve fiber layer cross-section area in the disk hemorrhage group were significantly smaller than those in the nonhemorrhage group (P <.05). In the disk hemorrhage group, moreover, the values for the rim area, rim volume, and retinal nerve fiber layer cross-section area in the inferotemporal sector with hemorrhage were significantly smaller than those in the same sector without hemorrhage (P <.05). CONCLUSION: Localized damage of the disk rim and retinal nerve fiber layer at the inferotemporal sector was prominent in eyes with disk hemorrhage.

Female↗
PubMed2002

Repair of the thoracic aorta by transaortic stent grafting (open stenting).

BACKGROUND: The late results of direct open stent grafting of the aortic arch for aortic arch repair have not been reported previously. METHODS: Between September 1997 and December 2000 19 patients underwent open stent grafting with carotid artery bypass for thoracic arteriosclerotic aneurysms (TAA) of the distal aortic arch. In addition, 21 patients underwent open stent grafting with total aortic arch replacement for Stanford type A acute aortic dissection and 7 patients underwent stenting with carotid bypass for Stanford type B chronic aortic dissection. RESULTS: The early mortality rate was 11% for TAA, 10% for type A dissection, and 0% for type B dissection. Whereas none of the TAA or type A dissection required a second operation on the thoracic aorta, 1 TAA patient died 6 months postoperatively after sudden aortic rupture and 1 type B patient required descending aortic replacement because of ulceration caused by the stent graft at 11 months postoperatively. On follow-up computed tomography scan, in TAA patients, true aneurysms excluded by the stent graft showed early thrombosis, but the absorption of thrombosed aneurysms started from 1 to 6 months postoperatively and gradually progressed. In patients with type B chronic dissection, the false lumen showed early thrombosis and the true lumen was dilated at the central portion of the graft, which might increase turbulent flow by interaction with the stent. In patients with type A acute dissection, the false lumen showed both early thrombosis and early absorption. CONCLUSIONS: Early and late results of open stenting are acceptable and follow-up computed tomography scan may be able to predict late results of open stenting.

Adult↗
PubMed2001

Diversity of the damage recognition step in the global genomic nucleotide excision repair in vitro.

The XPC-HR23B complex, a mammalian factor specifically involved in global genomic nucleotide excision repair (NER) has been shown to bind various forms of damaged DNA and initiate DNA repair in cell-free reactions. To characterize the binding specificity of this factor in more detail, a method based on immunoprecipitation was developed to assess the relative affinity of XPC-HR23B for defined lesions on DNA. Here we show that XPC-HR23B preferentially binds to UV-induced (6-4) photoproducts (6-4PPs) as well as to cholesterol, but not to the cyclobutane pyrimidine dimer (CPD), 8-oxoguanine (8-oxo-G), O6-methylguanine (O6-Me-G), or a single mismatch. Human whole cell extracts could efficiently excise 6-4PPs and cholesterol in an XPC-HR23B-dependent manner, but not 8-oxo-G, O6-Me-G or mismatches. Thus, there was good correlation between the binding specificity of XPC-HR23B for certain types of lesion and the ability of human cell extracts to excise these lesions, supporting the model that XPC-HR23B initiates global genomic NER. Although, XPC-HR23B does not preferentially bind to CPDs, the excision of CPDs in human whole cell extracts was found to be absolutely dependent on XPC-HR23B, in agreement with the in vivo observation that CPDs are not removed from the global genome in XP-C mutant cells. These results suggest that, in addition to the excision repair pathway initiated by XPC-HR23B, there exists another sub-pathway for the global genomic NER that still requires XPC-HR23B but is not initiated by XPC-HR23B. Possible mechanisms will be discussed.

DNA Damage↗
PubMed1995

Clinical aerosol inhalation cine-scintigraphy to evaluate mucociliary transport system in diffuse panbronchiolitis.

UNLABELLED: This study evaluates the mucociliary transport system in patients with diffuse panbronchiolitis using aerosol inhalation cine-scintigraphy (AICS). METHODS: Forty-one subjects, 10 healthy controls and 31 patients with diffuse panbronchiolitis, were studied. In addition, the mucociliary transport system was evaluated in 11 patients who had received erythromycin therapy for 3-8.3 yr. Following inhalation of 99mTc-human serum albumin aerosol for 3-5 min in a sitting position, the subjects were placed on the imaging table in the supine position and posterior images were obtained dynamically for 20 sec/frame over 2 hr with a gamma camera linked to a digital computer. The 360 20-sec serial frames were edited into a cinematographic presentation at 200-msec intervals. Clinical evaluation of the mucociliary transport system was based on the bolus movement of radioactive aerosol from the main bronchi to the trachea and the movement patterns, which were divided into four types using the movement in the controls as a standard (type I): type I, rapid and smooth movement; type II, slow movement; type III, stagnation at the carina; and type IV, complete stasis. RESULTS: All patients with diffuse panbronchiolitis had types III and IV, indicating that mucociliary transport system was severely impaired. Of the 11 patients on erythromycin therapy, 8 had movement pattern type IV and 3 had movement pattern type III before erythromycin therapy. In eight patients (72.7%), movement pattern was improved to type I or II after therapy. CONCLUSION: Aerosol inhalation cine-scintigraphy helps evaluate the clinical usefulness of erythromycin therapy in diffuse panbronchiolitis.

Aerosols↗
PubMed2000

Localization of initial lymph node metastasis from carcinoma of the thoracic esophagus.

BACKGROUND: Most surgeons consider esophageal carcinoma with lymph node involvement a systemic disease. However, it is possible that the disease may be localized in the earlier phases of lymphatic metastasis. The distribution of involved lesions in the initial phase of lymph node metastasis has not been thoroughly investigated yet. METHODS: Among 329 patients that underwent curative (R0 International Union Against Cancer [UICC]) esophagectomy with systematic mesoesophageal dissection, 51 cases of patients with only 1 involved lymph node (solitary involvement) were retrospectively investigated and compared with patients with multiple involved lymph nodes. The regional lymph nodes were divided into the thoracocervical junction group (lower deep cervical and recurrent nerve lymph nodes), perigastric group, and intrathoracic group. RESULTS: Lymph node involvement was limited to a solitary lymph node in 46% of lymph node positive patients with esophageal carcinoma confined to the wall (T1 and T2, UICC) and in 17% of lymph node positive patients with cancer that invaded the extramural layer (T3 and T4, UICC). Of patients with solitary involvement, 82% had a positive thoracocervical junction or perigastric lymph node. The 5-year survival rate in solitary involvement cases was 61%, and 65% when solitary involvement was not intrathoracic. Most of the 5-year survivors had involvement of a thoracocervical junction or perigastric lymph node and had not received systemic chemotherapy. CONCLUSIONS: Solitary involvement was not rare and not directly associated with a disseminated disease. Solitary involvement was commonly located in the thoracocervical junction or abdomen that are accessible without thoracotomy. Systematic dissection of the regional lymph nodes including thoracocervical junction and perigastric groups is recommended for resectable esophageal carcinoma at this time. However, less extensive dissection may be performed in selected cases if the sentinel lymph node concept proves valid.

Esophageal Neoplasms↗
PubMed2002

Prediction of the bitterness of single, binary- and multiple-component amino acid solutions using a taste sensor.

The purpose of this study was to develop a quick, quantitative, prediction method for the determination of the bitterness of solutions containing one or more of five amino acids (L-isoleucine, L-leucine, L-valine, L-phenylalanine, and L-tryptophan), using an artificial taste sensor. The bitterness of various solutions containing different concentrations (1, 3, 10, 30, and 100 mM) of five amino acids, singly and in combination, was estimated using a multichannel taste sensor and compared with the results of human gustatory sensation tests with nine volunteers. The relative response electric potential patterns were similar for all five amino acids. Large sensor outputs were observed in channels 1-4 (which are negatively charged) while there were no responses in channels 5-8 (positively charged). The sensor output for channel 1, which was the largest output value, was used for prediction of bitterness. The change of membrane potential caused by adsorption (CPA), which corresponds to aftertaste, could not be used as an explanatory variable since the adsorption of the amino acids to the sensor membrane was weak and CPA values were small. The bitterness intensity scores for single, binary, and multi-component amino acid solutions, could be easily predicted on the basis of the sensor output value of channel 1 using regression analysis. Principal component analysis of the sensor output data suggested that the sourness, astringency and/or smell of the solutions also played a role in the perception of bitterness.

Amino Acids↗
PubMed2000

Enhanced hepatic lipid peroxidation in patients with primary biliary cirrhosis.

OBJECTIVE: The mechanisms responsible for hepatic injury have not been fully clarified in primary biliary cirrhosis (PBC). It has recently been suggested that hepatic lipid peroxidation may be involved in the pathogenesis of PBC. The aims of the current study were to determine whether patients with PBC have evidence of enhanced hepatic lipid peroxidation and to evaluate its relationship to clinicopathological features. METHODS: Immunohistochemical detection of 4-hydroxynonenal (HNE) protein adducts was performed in the liver biopsies of 20 patients with PBC. Histological stages of PBC were evaluated. Orcein or Victoria blue staining was performed for detection of copper-associated proteins. The size of bile ducts was defined as the smallest diameter between the subepithelial basement membranes. RESULTS: All 20 patients had immunodetectable HNE protein adducts in the cytoplasm of damaged, but also intact, biliary cells. The mean diameter of bile ducts with HNE protein adducts was smaller than those without the adducts (61.0 +/- 1.9 vs 122.5 +/- 24.4 microm, respectively, p < 0.01). Out of 20 patients, 6 (30%) also had immunodetectable HNE protein adducts in hepatocytes preferentially located around the portal tracts. Most of the patients with hepatocytic HNE protein adducts had copper-associated protein granules in hepatocytes around the portal tracts and were classified as histological stage 3, whereas all of the patients without the adducts lacked copper-associated protein granules and were classified as histological stage 1 or 2. The patients with hepatocytic HNE protein adducts had higher levels of serum total bilirubin than did those without the adducts (2.9 +/- 0.9 vs 0.7 +/- 0.1 mg/dl, respectively, p < 0.01). CONCLUSIONS: Hepatic lipid peroxidation can occur in PBC and may be an early event in bile duct destruction. At advanced stages of PBC, hepatocellular lipid peroxidation may play a role in hepatocyte injury during cholestasis.

Aldehydes↗
PubMed2006

Tight regulation of transgene expression by tetracycline-dependent activator and repressor in brain.

Methods to temporally and spatially regulate gene mutations will provide a powerful strategy to investigate gene function in the brain. To develop these methods, we have established a tightly regulated system for transgene expression in the forebrain using both a tetracycline (Tc)-dependent transcription activator (rtTA) and a repressor (TetR-Kruppel-associated box). In this system, the repressor binds to the Tc-responsive element (TRE) in the absence of doxycycline (Dox), leading to the repression of leaky activation of TRE-mediated transcription caused by weak binding of rtTA to TRE. Upon Dox administration, only the activator binds to TRE and activates transcription. We tested this system in cultured cells by bicistronically expressing both the regulators using an internal ribosome entry site (IRES). In COS-1, HeLa and SHSY5Y cells, leaky transcription activation led by rtTA in the absence of Dox was repressed without decreasing the level of activated transcription in the presence of Dox. Using this system, transgenic mice were produced that express both the regulators using IRES in the forebrain under the control of the alphaCaMKII promoter and were bred with transgenic mice carrying the TRE-dependent reporter transgene. In reverse transcription-polymerase chain reaction and in situ hybridization analyses of the forebrain in adult double transgenic mice, the treatment of Dox induces reporter mRNA expression, which was not detected before the treatment and after the withdraw of Dox following the treatment. These results indicate that this system allows the tight regulation of transgene expression in a Dox-dependent fashion in the forebrain and will be useful in investigating gene function in the brain.

Animals↗
PubMed1999

Antigen-specific, IgE-selective unresponsiveness induced by antigen-liposome conjugates. Comparison of four different conjugation methods for the coupling of antigen to liposome.

BACKGROUND: We have previously reported that ovalbumin (OVA) coupled with liposome via glutaraldehyde (GA) induced OVA-specific- and IgE-selective unresponsiveness in mice. METHODS: In this study, OVA-liposome conjugates were made using four different coupling protocols: via GA, N-(6-maleimidocaproyloxy) succinimide (EMCS), disuccinimidyl suberate (DSS) and N-succimidyl-3(2-pyridyldithio)propionate (SPDP) and the induction of antigen-specific IgG and IgE antibody production was investigated for each. In addition, antigen-specific cytokine production by spleen cells of mice immunized either with OVA-liposome or with OVA adsorbed with aluminum hydroxide was investigated. RESULTS: OVA-liposome conjugates coupled via GA or DSS did not induce anti-OVA IgE antibody production but induced substantial anti-OVA IgG antibody production. On the other hand, the induction of anti-OVA IgE unresponsiveness by OVA-liposome conjugates coupled via EMCS or SPDP was incomplete. The amount of interleukin 4 (IL-4) produced by spleen cells stimulated in vitro with OVA correlated well with anti-OVA IgE antibody production in donor mice. However, the production of no other cytokine, i.e., IL-2, IL-5, IL-10 or interferon-gamma, was correlated with in vivo IgE antibody production. CONCLUSION: OVA-liposome coupled via GA or DSS induced complete suppression of anti-OVA IgE production. The results in this study further suggest that the regulation of IgE antibody production does not necessarily correlate with so-called Th1 cytokine production.

Animals↗
PubMedDate not supplied

Prognostic significance of CD8+ T cell and macrophage peritumoral infiltration in colorectal cancer.

The purpose of this study was to evaluate the prognostic significance of CD8+ T cell and macrophage peritumoral infiltration in patients with colorectal cancer. A total of 97 adenocarcinomas of the colon and rectum were examined. Immunohistochemical staining was performed by the standard avidin-biotin-peroxidase complex method using antibodies to CD8 and CD68. Peritumoral infiltration by CD8+ T cells or macrophages was evaluated along the invasive margin of the cancer in each specimen. The area with the most abundant infiltration was selected, and the number of immunoreactive positive cells counted at x400 magnification. Patients were divided into two groups based on the degree of infiltration by each cell type: namely those with a high level of infiltration (more than the mean number of positive cells) and those with a low level of infiltration (less than the mean number of positive cells). Patients with a low level of macrophage infiltration had a significantly deeper depth of invasion than patients with a high level of macrophage infiltration (P=0.027). The percentage of patients with a high level of macrophage infiltration was significantly higher in vascular invasion-negative cases (46.7%) than in vascular invasion-positive cases (22.7%; P=0.045), and in lymph node metastasis-negative cases (52.9%) than in lymph node metastasis-positive cases (28.3%; P=0.014). Overall survival was significantly shorter for patients with a low level of CD8+ T cell infiltration than those with a high level of CD8+ T cell infiltration (P=0.01). The survival rate for patients with a high level of both CD8+ T cell and macrophage infiltration was 100%. In conclusion, both CD8+ T cell and macrophage peritumoral infiltration indicates anti-tumoral action in patients with colorectal cancer.

Adenocarcinoma↗
PubMed2006

One-step palliative treatment method for obstructive jaundice caused by unresectable malignancies by percutaneous transhepatic insertion of an expandable metallic stent.

AIM: To describe a simple one-step method involving percutaneous transhepatic insertion of an expandable metal stent (EMS) used in the treatment of obstructive jaundice caused by unresectable malignancies. METHODS: Fourteen patients diagnosed with obstructive jaundice due to unresectable malignancies were included in the study. The malignancies in these patients were a result of very advanced carcinoma or old age. Percutaneous transhepatic cholangiography was performed under ultrasonographic guidance. After a catheter with an inner metallic guide was advanced into the duodenum, an EMS was placed in the common bile duct, between a point 1 cm beyond the papilla of Vater and the entrance to the hepatic hilum. In cases where it was difficult to span the distance using just a single EMS, an additional stent was positioned. A drainage catheter was left in place to act as a hemostat. The catheter was removed after resolution of cholestasis and stent patency was confirmed 2 or 3 d post-procedure. RESULTS: One-step insertion of the EMS was achieved in all patients with a procedure mean time of 24.4 min. Out of the patients who required 2 EMS, 4 needed a procedure time exceeding 30 min. The mean time for removal of the catheter post-procedure was 2.3 d. All patients died of malignancy with a mean follow-up time of 7.8 mo. No stent-related complication or stent obstruction was encountered. CONCLUSIONS: One-step percutaneous transhepatic insertion of EMS is a simple procedure for resolving biliary obstruction and can effectively improve the patient's quality of life.

Aged↗
PubMedDate not supplied

Increased basal levels of plasma nitric oxide in Type 2 diabetic subjects. Relationship to microvascular complications.

To assess the underlying mechanisms of decreased endothelial function and advanced vascular complications in patients with Type 2 diabetes, we determined basal levels of plasma nitric oxide (NO(x): NO(2)(-) and NO(3)(-)) using a newly developed high-performance liquid chromatography (HPLC)-Griess method in hospitalized 129 diabetic and 76 nondiabetic subjects, and examined their clinical characteristics. Serum lipid peroxide and advanced glycation end products (AGEs) as markers of oxidative stress were also measured, and intima-media thickness (IMT) of the carotid artery was evaluated as a marker of atherosclerosis. In diabetic subjects, microvascular complications were newly evaluated during their admission. There were no differences in age or sex between the diabetic and nondiabetic subjects. Although there was no difference in basal plasma NO(2)(-) levels between the two groups, the basal levels of plasma NO(3)(-) in diabetic subjects were significantly higher than those in nondiabetic subjects. Plasma NO(x) levels in neither diabetic nor nondiabetic subjects correlated with serum lipids, HbA1c, or IMT. In diabetic subjects, plasma NO(3)(-) levels were related not only to the presence of hypertension but also to advanced microvascular complications. Moreover, plasma NO(3)(-) levels were positively correlated with both serum lipid peroxide and AGEs. Multiple regression analysis revealed that serum AGEs level was strongly associated with plasma NO(3)(-) level. Thus, the findings are consistent with the hypothesis that decreased endothelium-dependent vasodilation in diabetic subjects is associated with the impaired action of NO secondary to its inactivation resulting from increased oxidative stress, rather than decreased NO production from vascular endothelium, and that abnormal NO metabolism is related to advanced diabetic microvascular complications.

Biomarkers↗
PubMed2000

Successful interferon therapy reverses enhanced hepatic iron accumulation and lipid peroxidation in chronic hepatitis C.

OBJECTIVES: Hepatic iron deposition has been reported in chronic hepatitis C (CH-C), and iron-induced lipid peroxidation may be involved in the pathogenesis of CH-C. The aims of the present study were: 1) to determine whether patients with CH-C have evidence of enhanced hepatic lipid peroxidation and to evaluate its relation to iron status, compared with that in patients with chronic hepatitis B (CH-B); and 2) to assess the effect of interferon (IFN) therapy on hepatic iron and lipid peroxidation. METHODS: In the liver biopsies of 40 patients with CH-C and 26 patients with CH-B, immunohistochemical detection of 4-hydroxy-2-nonenal (HNE)-protein adducts for evaluation of lipid peroxidation was performed, and hepatic iron status was biochemically and histologically assessed. In 16 CH-C patients with normal serum transaminases and undetectable serum HCV-RNA >6 months after the end of IFN treatment (responders) and in 11 nonresponders, hepatic HNE-protein adducts and siderosis were evaluated in pre- and posttreatment liver biopsies. RESULTS: Hepatocytic HNE-protein adducts and iron deposits were more abundant in the patients with CH-C than in those with CH-B. No correlation was found between the levels of hepatocytic HNE-protein adducts and hepatic iron status in either of the two groups. In the responders to IFN treatment for CH-C, hepatocytic HNE-protein adducts disappeared or attenuated with improvement of hepatic siderosis after the treatment, whereas IFN treatment did not improve hepatocytic expression of HNE-protein adducts and hepatic siderosis in the nonresponders. CONCLUSIONS: Patients with CH-C have evidence of enhanced hepatic iron accumulation and lipid peroxidation compared to those with CH-B. In CH-C, hepatic siderosis and lipid peroxidation are improved with successful IFN treatment. These results suggest that hepatic lipid peroxidation and iron may potentially play contributory roles in the pathogenesis of CH-C.

Adult↗
PubMed2004

Possible oncogenicity of subventricular zone neural stem cells: case report.

OBJECTIVE: The origin of brain tumors has attracted much controversy. Recent advances in neural stem cell biology coupled with the new concepts of central nervous system development have raised interesting possibilities regarding the oncogenic properties of neural stem/progenitor cells. To elucidate these putative properties further, the clinical materials from an infant brain tumor were analyzed, focusing on the relation with the neural stem/progenitor cells. METHODS: The expression of neural stem/progenitor cell markers in the tumor cells and the cellular components of the infant brain tumor were examined using immunohistochemistry. The tumor cell biology was analyzed both in culture and in the grafted brain environment. RESULTS: Three main bodies of evidence were demonstrated indicating that the tumor was of possible subventricular zone postnatal or adult normal neural stem cell origin. First, in the tumor specimen we demonstrated the strong positive expression of the neural stem/progenitor cell markers, nestin and Musashi-1. Second, immunohistochemistry revealed the presence of neuronal, astrocytic, and immature precursor cells in the tumor tissue, similar to the cellular components of the subventricular zone, thereby pointing to the subventricular zone as the possible origin of the tumor. The subventricular zone also is one of the strong candidates for the location of postnatal/adult neural stem cells. This cellular evidence was strengthened further by the clinicoradiological findings that demonstrated the involvement of the subventricular zone of the lateral ventricle by the tumor. Third, in the in vitro and in vivo experiments, a dynamic shift in expression patterns between neural stem cells (nestin, Musashi-1) and differentiated cells (glial fibrillary acidic protein, neuron-specific Class III beta-tubulin) markers was seen, similar to the proposed behavior of postnatal/adult neural stem cells in situ. CONCLUSION: These findings suggest that this brain tumor originated from neural stem cells located in the subventricular zone, and the further possibility of the general oncogenic potential of neural stem cells.

Brain Neoplasms↗