Involvement of two distinct signalling pathways in IGF-1-mediated central control of hypotensive effects in normotensive and hypertensive rats.

AIMS: Insulin-like growth factor-1 (IGF-1) is abundantly expressed in the nucleus tractus solitarii (NTS). In a previous study, we revealed that the induction of nitric oxide (NO) production in the NTS reduces blood pressure (BP). It is well known that both acute administration and chronic administration of IGF-I reduce BP. The aim of this study was to evaluate the short-term hypotensive effect of IGF-1 in the NTS and to delineate the underlying molecular mechanisms of IGF-1 in the NTS of normotensive WKY rats and spontaneously hypertensive rats (SHRs). METHOD: Microinjections of the phosphatidylinositol 3-kinase (PI3K) inhibitor LY294002 and the MAP kinase-ERK kinase (MEK) inhibitor PD98059 into the NTS in WKY rats and SHRs were used to study the involvement of IGF-1-induced depressor effects. RESULT: An IGF-1 (7.7 pmol) injection into the NTS resulted in a significant decrease in BP and HR in WKY rats and SHRs. Immunoblotting and immunohistochemical analysis showed that the microinjection of LY294002 (0.6 pmol) or PD98059 (3.0 pmol) into the NTS attenuated the IGF-1-induced depressor effects and Akt or ERK phosphorylation in WKY rats. An attenuation effect of LY294002, but not PD98059, was found in the SHRs. However, the mRNA and protein expression levels of the IGF-1R showed no significant differences in the NTS of the WKY rats and the SHRs. CONCLUSION: These results suggest that distinct Akt and ERK signalling pathways mediated the IGF-1 control of the central depressor effects in WKY rats and SHRs. ERK signalling defects may be associated with the development of hypertension.

Blood trace minerals concentrations and oxidative stress in patients with obstructive sleep apnea.

BACKGROUND: Obstructive sleep apnea (OSA) is associated with increased oxidative stress. Certain essential trace minerals have shown to play an important role in the maintenance of redox homeostasis. We determined the concentrations of trace minerals in OSA patients and assessed their relationships to OSA severity as indicated by the apnea/ hypopnea index (AHI). METHODS: We enrolled 44 patients with newly diagnosed mild to moderate OSA and 20 without OSA. The following parameters were measured: polysomnographic values of nocturnal sleep; plasma trace minerals zinc (Zn), copper (Cu), iron (Fe), and erythrocyte selenium (Se); oxidative stress status; and plasma high-sensitivity C-reactive protein (hs-CRP) and tumor necrosis factor-α (TNF-α). RESULTS: Compared to controls matched for age, gender, and body mass index, OSA patients had lower concentrations of plasma Zn and erythrocyte Se and higher plasma concentrations of Cu and Fe. OSA patients had significantly higher plasma concentrations of hs-CRP, TNF-α, and malondialdehyde (MDA), and lower erythrocyte antioxidant enzyme glutathione peroxidase (GPx) and superoxide dismutase activities. Significant differences in all the above parameters were also found in patients with moderate OSA compared to those with mild OSA. Furthermore, AHI values correlated significantly with neck circumference, GPx activity, and MDA, hs-CRP, and TNF-α concentrations in OSA patients. AHI values were also negatively associated with concentrations of plasma Zn and erythrocyte Se, but were positively linked to plasma concentrations of Fe and Cu. CONCLUSIONS: Abnormal concentrations of these trace minerals may reflect oxidative damage and inflammatory response, thus increasing the severity of OSA.

Post-angioplasty far infrared radiation therapy improves 1-year angioplasty-free hemodialysis access patency of recurrent obstructive lesions.

OBJECTIVE: To explore the role of far infrared (FIR) radiation therapy for hemodialysis (HD) access maintenance after percutaneous transluminal angioplasties (PTA). METHODS: This was a prospective observational study. Eligible patients were those who received repeated PTA with the last PTA successfully performed within 1 week before the study enrollments. Consecutively enrolled patients undergoing successful HD treatments after PTA were randomly assigned to the FIR-radiated group or control group without radiation. FIR-radiated therapy meaning 40-minute radiation at the major lesion site or anastomosed site three times a week was continued until an end-point defined as dysfunction-driven re-PTA or the study end was reached. RESULTS: Of 216 participants analyzed, including 97 with arteriovenous grafts (AVG) (49 FIR-radiated participants and 48 control participants) and 119 with arteriovenous fistulas (AVF) (69 FIR-radiated participants and 50 control participants), the FIR-radiated therapy compared with free-radiated usual therapy significantly enhanced PTA-unassisted patency at 1 year in the AVG subgroup (16.3% vs. 2.1%; p < .01), but not the AVF subgroup (25.0% vs. 18.4%; p = .50), and this accounted for the overall improved patency rates (21.4% vs. 10.3%; p = .02). CONCLUSIONS: This study suggests FIR-radiated therapy improves PTA-unassisted patency in patients with AVG who have undergone previous PTA.

NO donor KMUP-1 improves hepatic ischemia-reperfusion and hypoxic cell injury by inhibiting oxidative stress and pro-inflammatory signaling.

This study investigates whether KMUP-1 improves hepatic ischemia-reperfusion (I/R) and hypoxic cell injury via inhibiting Nox2- and reactive oxygen species (ROS)-mediated pro-inflammation. Rats underwent ischemia by occlusion of the portal vein and hepatic artery for 45 minutes. Reperfusion was allowed for 4 h. Serum was used for analysis of aspartate aminotransferase (AST) and alanine aminotransferase (ALT). DNA extracted from liver homogenate was analyzed by electrophoresis to observe the fragmentation. Lipid peroxidation (LPO) was evaluated by measuring thiobarbituric acid-reactive substances (TBARS). NO and ROS contents were measured using Griess reagent and 2'-7'-dichlorofluorescein, respectively. Proteins levels were visualized by Western blotting. Liver damage was observed under a microscope. Intravenous KMUP-1 (0.25, 0.5 and 1 mg/kg) reduced I/R-induced ALT and AST levels, DNA fragmentation, ROS and malondialdehyde (MDA) and restored the NO levels of I/R rats. KMUP-1 protected the liver architecture from worsening of damage and focal sinusoid congestion, increased endothelium NO synthase (eNOS), guanosine 3', 5'cyclic monophosphate (cGMP), protein kinase G (PKG) and the B-cell lymphoma 2/Bcl-2-associated X protein (Bcl-2/Bax) ratio, attenuated phosphodiesterase 5A (PDE-5A) and cleaved caspase-3 expression in I/R-liver. In hypoxic HepG2 cells, KMUP-1 increased cGMP/PKG, restored peroxisome proliferator-activated receptor-gamma (PPAR-gamma) and decreased matrix metalloproteinases-9 (MMP-9), Rho kinase II (ROCK II), hypoxia-inducible factor-1alpha (HIF-1alpha) and vascular endothelium growth factor (VEGF). KMUP-1 protects liver from I/R-injury and hypoxic hepatocytes from apoptosis-associated free radical generation and pro-inflammation by restoring/increasing NO/cGMP/PPAR-gamma, reducing ROS/Nox2 and inhibiting ROCKII/MMP-9.

Modulation of baroreflex function by rosiglitazone in prediabetic hyperglycemic rats.

Baroreflex sensitivity (BRS) is abnormal in the prediabetic state. This study was conducted to determine effects of chronic rosiglitazone (RSG), an insulin sensitizer, on BRS in prediabetic hyperglycemic (PDH) rats induced by nicotinamide and streptozotocin. The fasting and postprandial blood glucose levels were 5.6-6.9 and 7.8-11.0 mmol/l, respectively. Rats were treated with RSG or saline for 12 weeks. BRS response to phenylephrine (PE-BRS) or sodium nitroprusside (NP-BRS) was determined by linear regression method. Cardiac sympathetic and parasympathetic influences were determined by autonomic blockades. In the saline-treated PDH rats, PE-BRS was enhanced early at week 4 and became greater at week 12. Abnormalities in NP-BRS and cardiac autonomic influences were found only after week 12. Four weeks of RSG treatment normalized blood glucose levels but not PE-BRS. All altered cardiovascular variables were completely restored by 12 weeks of RSG treatment. The correlation between BRS and blood glucose levels in saline-treated PDH rats was significant at week 12, but no correlation was found in RSG-treated rats. In conclusion, hyperglycemia, even in the prediabetic state, may play a role in BRS abnormalities. RSG treatment early in the prediabetic state may normalize BRS via cardiac autonomic modulation, besides its anti-hyperglycemic action.

Cesarean scar defect: correlation between Cesarean section number, defect size, clinical symptoms and uterine position.

OBJECTIVES: To determine the prevalence of clinical symptoms associated with Cesarean scar defects, and to determine the association between the size of these defects, clinical complaints, uterine position, and a history of multiple Cesarean sections. METHODS: In this cross-sectional study, Cesarean scar defects in women with a history of transverse lower-segment Cesarean section were measured by transvaginal ultrasound while being assessed for other gynecological conditions. The relationships between the size of the Cesarean scar defect and the clinical symptoms, uterine position and number of previous Cesarean sections were evaluated. Patients with other uterine pathologies, including endometrial hyperplasia, polyps, malignancy and submucosal myomas, were excluded from the study. RESULTS: During a 3-year period, 4250 women were assessed by transvaginal sonography, of whom 293 (6.9%) were diagnosed with Cesarean scar defects. Eighty-six patients were excluded due to other uterine pathologies. Altogether, 207 patients with Cesarean scar defects were included in this study. Women who had undergone multiple Cesarean sections tended to have larger scar defects (in width and depth) than did those who had undergone a single Cesarean section. Women with retroflexed uteri also tended to have wider defects than those with anteflexed uteri. Defect width was significantly greater in women with postmenstrual spotting, dysmenorrhea and chronic pelvic pain. CONCLUSIONS: Multiple Cesarean sections and retroflexed uteri are risk factors for larger Cesarean scar defects. The size of the Cesarean scar defect is associated with clinical symptoms such as postmenstrual spotting, dysmenorrhea and chronic pelvic pain.

Neuronal nitric oxide synthase activation is involved in insulin-mediated cardiovascular effects in the nucleus tractus solitarii of rats.

Neuronal nitric oxide synthases (nNOS) is distributed throughout the central nervous system (CNS) and has been proposed to modulate neuronal activity in the nucleus tractus solitarii (NTS). Here, we investigated whether the activation of nNOS is involved in insulin-induced cardiovascular responses in the NTS. Insulin (100 IU/ml) was unilaterally microinjected into the NTS, and the cardiovascular effects were evaluated before and after microinjection of the nNOS inhibitors 7-nitroindazole (7-NI) (5 pmol) and N(5)-(1-imino-3-butenyl)-l-ornithine (vinyl-L-NIO) (600 pmol). Western blot and immunohistochemical analyses were performed to determine nNOS phosphorylation levels after insulin or phosphoinositide 3-kinase (PI3K) inhibitor LY294002 microinjection into the NTS. Unilateral microinjection of insulin into the NTS produced prominent depressor and bradycardic effects in WKY rats. Pretreatment with the nNOS inhibitors 7-NI and Vinyl-L-NIO attenuated the cardiovascular response evoked by insulin in Wistar-Kyoto (WKY) rats. Moreover, Western blot analysis showed a significant increase in nNOS (16.5+/-0.4-fold; P<0.05; n=4) phosphorylation after insulin injection, whereas the PI3K inhibitor LY294002 abolished the insulin-induced effects. In situ nNOS phosphorylation was found to be increased in the NTS after insulin injection. Furthermore, co-immunoprecipitation assay showed Akt and nNOS can bind to each other as detected by phospho-Akt(S473) and phospho-nNOS(S1416) antibodies. In vitro kinase assay showed insulin activated Akt can directly phosphorylate nNOS(S1416). These results demonstrated that nNOS may couple with the activation of the insulin receptor, via the liberation of NO, in order to participate in central cardiovascular regulation of WKY rats.

Loop conization for the treatment of microinvasive carcinoma of the cervix.

The purpose of the study was to evaluate the specimen adequacy and diagnostic accuracy of loop conization in microinvasive carcinoma of the cervix. A retrospective study was conducted from 1997 to 2003 at the Colposcopic Clinic, Department of Gynecology and Obstetrics, Chang Gung Memorial Hospital, Taipei, Taiwan. Sixty-three consecutive patients with microinvasive carcinoma of the cervix receiving cold-knife conization (35 patients) or loop conization (28 patients) were included in the study. All patients underwent definitive hysterectomy. We reviewed the conization specimen together with the hysterectomied uterus to compare the two conization techniques with respect to the histopathologic interpretation and diagnostic accuracy. The mean depth of cone specimens was significantly less in the loop conization compared with cold-knife conization (1.65 versus 2.35 cm, P = 0.035). Regarding the application of conization, the loop conization was completed in a single slice in 27 patients (77.1%) and in multiple slices in 8 patients (22.9 %), in spite of encouragement to perform conization in a one-pass application when possible. However, the cold-knife specimens were invariably a single cone-shaped piece. As reviewed by microscopic examination, the rate of tissue transection was significantly higher in the loop group than in the cold-knife group (14.3% versus 0%, P = 0.04). Because of tissue transection and disorientation, pathologic evaluation of stromal status was inadequate in 11.4% (4/35) of the loop cones as opposed to none of the 28 cold-knife cones. After assessing the hysterectomy specimens, the clinical diagnoses in the loop group were downgraded in three patients compared with only one in the cold-knife group. Data from this investigation suggest that cervical cold-knife conization is superior to loop conization as a method to assess microinvasive cervical cancer.

Primary evacuation therapy for Cesarean scar pregnancy: three new cases and review.

We report three cases of first-trimester Cesarean scar pregnancy (CSP) treated with evacuation therapy. In one patient with a gestational age of 7 weeks and with deep sac implantation, there was massive uterine bleeding and uterine perforation following evacuation. The other two patients, both with a pregnancy < 7 weeks and with intrauterine gestational sacs, were successfully treated with surgical evacuation without complications. A MEDLINE search revealed 11 cases that fulfilled our criteria of a certain diagnosis of scar pregnancy and with evacuation as the primary treatment. The outcome of these 11 cases and that of our three new cases suggests that the combination of a gestational sac size corresponding to a gestational age >/= 7 weeks, coupled with a deep implantation, portend an unsatisfactory outcome of surgical evacuation as a means of treating CSP.

Prognostic value of age and histologic type in neoadjuvant chemotherapy plus radical surgery for bulky (>/=4 cm) stage IB and IIA cervical carcinoma.

The purpose of this study was to investigate pretreatment variables related to prognosis and to evaluate long-term outcome in patients with bulky early-stage cervical carcinoma who were enrolled into a protocol treatment of neoadjuvant chemotherapy (NAC) followed by radical surgery. Between 1988 and 1999, 162 women with previously untreated bulky (>/=4 cm) stage IB or IIA carcinoma of the uterine cervix registered to receive cisplatin, vincristine, and bleomycin for three cycles followed by radical hysterectomy and pelvic lymphadenectomy, were enrolled into this study. The 5-year relapse-free (RFS) and overall survival (OS) rates of the whole series were 65% and 69%, respectively. In multivariate analyses, age (< 35 vs. >/= 35-year-old: relative risk [RR], 2.06; 95% confidence interval [CI], 1.02-4.14; P = 0.043) and histologic type (adeno-adenosquamous vs. squamous carcinoma: RR, 2.60; 95% CI, 1.26-5.39; P = 0.010) were independent variables associated with RFS, and age (< 35 vs. >/= 35-year-old: RR, 2.53; 95% CI, 1.24-5.14; P = 0.010) and pre-NAC tumor size (> 5 vs. = 5 cm: RR, 2.06; 95% CI, 1.09-3.90; P = 0.027) were significantly related to OS. Narrowing down patient selection to squamous carcinoma and age older than 35 years with stratification of tumor size (break point of 5 cm) is recommended in future trials involving cisplatin-based NAC plus radical surgery.

Hyperbaric oxygen increases the lung's susceptibility to inhaled lipopolysaccharide in mice.

Hyperbaric oxygen (HBO2) has been shown to inhibit the adhesion function of beta(2)-integrin, which is important in mediating cell-to-cell adhesion and extravasation of inflammatory cells. In the present study, we examined the effects of HBO2 exposure on neutrophil infiltration and tissue injury in a model of acute lung inflammation induced by lipopolysaccharide (LPS) inhalation. Male C57BL/6 mice of 8 weeks old were exposed to 3 atmosphere absolute (ATA) 100% HBO2, 3 ATA hyperbaric air (HBA), or room air for 90 min. After exposure, they were exposed to aerosolized LPS solution (1 mg/ml) or saline in a plexiglass chamber for 10 min. Four hours after inhalation, bronchoalveolar lavage (BAL) was performed to determine protein concentration, LDH activity, total cells, and differential cell counts in the lavage fluid (BALF). Myeloperoxidase (MPO) content, lung histopathology, and plasma nitric oxide (NO) metabolite concentrations were also determined in separate sets of animals. We observed that LPS inhalation increased neutrophil number in the BALF, which was significantly inhibited by HBO2 but not HBA pre-exposure. However, MPO content in the lung was prominently increased by HBO2 pre-exposure, which correlated with increased PMN infiltration in lung tissues. Further, HBO2 plus LPS, but not saline inhalation caused a significant increase in the BALF protein level and LDH activity compared with that of LPS inhalation alone. LPS exposure induced significant increase in plasma NO metabolites, which was not potentiated by HBO2 pre-exposure. The inducible nitric oxide synthase inhibitor, aminoguanidine, significantly attenuated the increases in plasma NO metabolites and tissue MPO content as well as lung injuries. In summary, our data suggest that HBO2 pre-exposure increases the lung's susceptibility to inhaled LPS, which may be related to increased tissue neutrophil infiltration and dependent on interaction(s) between HBO2 exposure with LPS-induced nitric oxide production.

Down-regulation of basal Fos expression at nucleus tractus solitarii underlies restoration of baroreflex response after antihypertensive treatment in spontaneously hypertensive rats.

Antihypertensive therapy not only normalizes the elevated blood pressure but also restores the reduced baroreceptor reflex response associated with hypertension, although the underlying mechanism is not fully understood. We assessed the hypothesis that a reversal of the enhanced basal Fos expression seen during hypertension in nucleus tractus solitarii, the terminal site of baroreceptor afferents, underlies the restoration of baroreceptor reflex sensitivity after antihypertensive treatment. Male adult spontaneously hypertensive or normotensive Wistar-Kyoto rats received for 3 weeks captopril (100 mg/kg/day) added to their drinking water. Evaluated subsequently under pentobarbital anesthesia, captopril-treated spontaneously hypertensive rats exhibited significantly lowered systolic blood pressure and restoration of the sensitivity in baroreceptor reflex control of heart rate to levels comparable with Wistar-Kyoto rats. Reverse transcription-polymerase chain reaction analysis and immunohistochemical evaluation revealed concomitant down-regulation of basal expression in nucleus tractus solitarii of c-fos gene at both mRNA and protein levels. Captopril treatment, on the other hand, elicited no discernible effect on systolic blood pressure, cardiac baroreceptor reflex sensitivity or basal expression of Fos protein at the nucleus tractus solitarii of normotensive Wistar-Kyoto rats. From these findings we suggest that a down-regulation of basal Fos expression in nucleus tractus solitarii may contribute to the restoration of baroreceptor reflex sensitivity in spontaneously hypertensive rats that received antihypertensive treatment such as captopril.

Transthoracic and transesophageal echocardiographic diagnosis of aneurysm of the sinus of Valsalva: a review of five year experience.

BACKGROUND: Aneurysms of the sinus of Valsalva (SVA) are uncommon congenital lesions. The clinical presentations vary from asymptomatic to progressive heart failure following rupture of the aneurysm into an adjacent cardiac chamber. Retrograde aortogram is the diagnostic tool of choice preoperatively. Recent studies have demonstrated that the SVA can be accurately diagnosed using transthoracic two-dimensional, and color Doppler flow mapping, even for surgical preparation without cardiac catheterization. We report our 5-year experience of transthoracic echocardiography (TTE) and transesophageal echocardiography (TEE) in the evaluation of SVA. METHODS: Eleven adult patients with SVA with or without rupture were studied using both TTE and TEE. All of the diagnoses were subsequently comfirmed by either cardiac catheterization or surgical findings. RESULTS: Aneurysms originated in the right coronary sinus (n = 9) and noncoronary sinus (n = 2); they ruptured into the right ventricle in 5 patients and the right atrium in 5 patients. An unruptured right SVA was noted in 1 patient. Both TTE and TEE could identify the site of the aneurysm, rupture sites, and the receiving chamber equally well. Co-existent cardiac lesions included 11 cases of valvular aortic regurgitation (mild in 7, moderate in 2 and severe in 2). Two cases of perimembranous type ventricular septal defect (VSD) and 6 cases of supracristal type VSD (including 1 case of tetraology of Fallot, 3 supracristal, 1 muscular and 1 subaortic) were noted. Three cases were complicated with valvular vegetations (1 aortic valve, 1 aortic and tricuspid valve and 1 aortic and pulmonic valve). One patient had patent ductus arteriosus and 2 patients had pulmonic valvular stenosis. CONCLUSIONS: TEE provides clearer definition for the detailed anatomy of the ruptured sac and co-existent cardiac lesions than TTE through high resolution and closer approach. We conclude that TEE is a powerful complementary diagnostic tool in the evaluation of patients with SVA. TEE also provides additionally useful information for guiding the surgical approach and for assessing the operative results even without cardiac catheterization.

Significance of cardiac troponin I and creatine kinase release after coronary intervention.

BACKGROUND: Cardiac troponin I is a highly sensitive and specific marker for early detection of myocardial injury. Whether it can be used to monitor myocardial injury after coronary intervention is uncertain. This study was designed to measure the cardiac troponin I and creatine kinase (CK) after coronary intervention and investigate their clinical significance. METHODS: We measured cardiac troponin I and CK levels before intervention and 4 hours, 8 hours, 12 hours and 24 hours after apparently successful coronary intervention in 106 eligible patients. Nine patients were excluded due to missing data. We also followed up the clinical outcome to record major cardiac events (MACE). RESULTS: The frequency of cardiac troponin I increase after coronary intervention was higher than that of CK increase (40.2% vs 8.2%). The frequency of cardiac troponin I increase in the stent group was significantly higher than that in the PTCA group (49.2% vs 21.9%, p < 0.001). The frequency of cardiac troponin I increase was also higher than that of CK increase in patients with in-hospital events (58.8% vs 14.7%). CONCLUSIONS: Cardiac troponin I is more sensitive than creatine kinase in detecting myocardial injury after coronary intervention. The incidence of cardiac troponin I increase is significantly higher in patients undergoing stenting than in patients treated with balloon angioplasty only. The cardiac troponin I increase is more highly correlated with in-hospital events than is creatine kinase.

Modulation of catecholamine release by endogenous adenosine in the rat adrenal medulla.

Adenosine was shown to inhibit norepinephrine (NE) release from sympathetic nerve endings. The purpose of this study was to examine whether endogenous adenosine restrains NE and epinephrine release from the adrenal medulla. The effects of an adenosine receptor antagonist, 1,3-dipropyl-8-(p-sulfophenyl) xanthine (DPSPX), on epinephrine and NE release induced by intravenous administration of insulin in conscious rats were examined. Plasma catecholamines were measured by HPLC with an electrochemical detector. DPSPX significantly increased plasma catecholamine in both control rats and rats treated with insulin. The effect of DPSPX on plasma catecholamine was significantly greater in rats treated with insulin. Additional experiments were performed in adrenalectomized rats to investigate the contribution of the adrenal medulla to the effect of DPSPX on plasma catecholamine. The effect of DPSPX and insulin on epinephrine in adrenalectomized rats was significantly reduced compared with that of the controls. Finally, we tested whether endogenous adenosine restrains catecholamine secretion partially through inhibiting the renin-angiotensin system. The effect of DPSPX on plasma catecholamine in rats pretreated with captopril (an angiotensin-converting enzyme inhibitor) was reduced. These results demonstrate that under basal physiological conditions, endogenous adenosine tonically inhibits catecholamine secretion from the adrenal medulla, and this effect is augmented when the sympathetic system is stimulated. The effect of endogenous adenosine on catecholamine secretion from the adrenal medulla is achieved partially through the inhibitory effect of adenosine on the renin-angiotensin system.

Isolated paraaortic lymph node recurrence after definitive irradiation for cervical carcinoma.

PURPOSE: To evaluate the clinical features of isolated paraaortic lymph node (PALN) recurrence after definitive radiotherapy, and analyze the prognostic factors and effect of salvage treatment. METHODS AND MATERIALS: Of a total 876 patients who received pelvic radiotherapy after the diagnosis of primary cervical carcinoma, 26 were found to have isolated PALN recurrence as the first recurrent site, and these patients enrolled in this study. Only those with primary-site carcinoma controlled and who were free of other distant metastases were eligible. Nineteen of the 26 patients accepted salvage therapy. Fourteen patients accepted concurrent chemoradiation (CCRT), 1 accepted radiation to the paraaortic region, and 4 accepted chemotherapy alone. Clinical parameters evaluated included tumor markers (SCC and CEA) and image studies. RESULTS: Seven of the 26 patients were alive and disease-free. All 7 survivors had salvage treatment with radiation to the paraaortic region and concurrent cisplatin-based chemotherapy. None of the patients receiving chemotherapy or radiation alone enjoyed long-term, disease-free survival. The 5-year survival rate for isolated PALN recurrence of the 14 patients who accepted salvage concurrent chemoradiation (CCRT) was 51.2%. The presence of a clinical symptom at the time of PALN recurrence was analyzed. Seven of the 12 asymptomatic patients and none of the 14 symptomatic patients survived without disease after salvage treatment. The SCC levels at recurrence showed a statistically significant relationship to disease-free survival. CONCLUSIONS: An SCC level of < or = 4 ng/ml and a lack of symptoms at the time of recurrence were good prognostic factors in isolated PALN recurrence after primary radiation therapy. In addition to concurrent CCRT, periodical surveillance with tumor markers and imaging studies allowed early detection and salvage of those patients.

Negative predictive value of human papillomavirus test following conization of the cervix uteri.

OBJECTIVE: The goal of this study was to determine/evaluate the negative predictive value of human papillomavirus (HPV) testing following conization of cervix uteri. METHODS: A prospective analysis was undertaken on 79 cone biopsies of women with high-grade lesions (cervical intraepithelial neoplasia (CIN) III). HPV testing was performed on cervical smears before and after conization. We correlated the margin status (defined as positive cone margin or endocervical curettage status) and positive conization HPV status with the residual disease in a hysterectomy specimen. A Digene II kit was used to perform HPV testing. HPV detection was done by Hybrid Capture assay. RESULTS: Of the 79 patients, 47(59.5%) had positive margins after conization. HPV testing was positive in 37 cases (78.7%) and negative in 10 cases (21.3%). Residual disease was found in 31 of 47 (66%) postconization hysterectomy specimens. No residual lesions were found in HPV-negative cases. Of the 32 cases with negative margins following conization, HPV testing was negative in 25 cases (78%) and was positive in 7 cases (22%). Among these 25 cases with negative HPV tests, no residual lesion was detected, and in 7 HPV-positive cases, only one residual lesion was found. CONCLUSION: HPV testing is potentially an effective tool in predicting residual dysplasia after conization and could potentially assist in the decision between hysterectomy and conservative follow-up in women with CIN III.

Applications of the telomerase assay in peritoneal washing fluids.

OBJECTIVE: The aim of this study was to detect telomerase activity in peritoneal ascites and to assess whether it can be used as an assistant tool for the early detection of ovarian cancer. METHODS: Telomerase activity was measured by TRAP assay in 47 patients with ovarian malignancies and 50 patients with benign uterine leiomyomas (control group). RESULTS: All 26 peritoneal washing cytology positive cases were telomerase positive. Of the 21 peritoneal washing cytology negative cases, 3 were telomerase positive. When these 3 were reevaluated for peritoneal cytology, malignant ascitis was identified in 1. All telomerase negative cases were negative for peritoneal washing cytology. The sensitivity and specificity of peritoneal cytology and telomerase testing in correlation with true malignant cells were 96 (26/27) and 100% (20/20) versus 100 (27/27) and 90% (18/20), respectively. The false negative rate of peritoneal cytology was 4.7% (1/21). The false positive rate of the telomerase test in relation to malignant ascites was 6.9% (2/29). CONCLUSION: Our preliminary results reveal a high sensitivity and specificity of both telomerase testing and conventional cytology in peritoneal fluids. Our data suggest that the telomerase test in peritoneal fluids can be used as an adjuvant to cytopathological methods in the diagnosis of malignant peritoneal ascites, particularly in cases of negative cytology. In these cases, a review of peritoneal histocytology is advised.

Value of human papillomavirus deoxyribonucleic acid testing after conization in the prediction of residual disease in the subsequent hysterectomy specimen.

OBJECTIVE: Our aim was to evaluate human papillomavirus deoxyribonucleic acid testing after conization in predicting residual disease in the subsequent hysterectomy specimen. STUDY DESIGN: A prospective study was conducted on 75 patients with grade 3 cervical intraepithelial neoplasia who had cone margins or endocervical curettage specimens showing disease and who elected to undergo hysterectomy after conization. All patients underwent high-risk human papillomavirus deoxyribonucleic acid testing by the Hybrid Capture II (Digene Corporation, Gaithersburg, MD) system before conization and at the time of hysterectomy (within 2-7 weeks after conization). The presence of human papillomavirus deoxyribonucleic acid in cells obtained by endocervical brush before hysterectomy was correlated with residual disease in the hysterectomy specimens. RESULTS: Of the 92 patients enrolled, 75 were eligible. Of these 75 patients, 52 (69.3%) had persistent human papillomavirus deoxyribonucleic acid after conization, and 27 (36.0%) of the 75 patients had residual cervical neoplasia in the hysterectomy specimens. Those with negative results for human papillomavirus deoxyribonucleic acid after conization were all (23/23) without residual disease in the uterus (100% negative predictive value). All those who had residual disease (27/27) had positive results for human papillomavirus deoxyribonucleic acid at the time of hysterectomy (100% sensitivity). Postconization human papillomavirus deoxyribonucleic acid status (odds ratio, 4.000; 95% confidence interval, 1.531-10.449; P =.005) and grade of dysplasia after endocervical curettage (classified as grade 2 cervical intraepithelial neoplasia or less severe disease vs grade 3 cervical intraepithelial neoplasia: odds ratio, 6.612; 95% confidence interval, 2.837-15.409; P =.0002) were significantly associated with residual tumor in the uterus. CONCLUSIONS: This prospective study confirms an excellent sensitivity and negative predictive value of human papillomavirus deoxyribonucleic acid testing after conization in predicting residual cervical neoplasia. A strategy of managing patients with grade 3 cervical intraepithelial neoplasia, based on postconization human papillomavirus deoxyribonucleic acid findings and endocervical curettage results, is proposed.

Cochlear gene delivery through an intact round window membrane in mouse.

Cochlear gene transfer studies in animal models have utilized mainly two delivery methods: direct injection through the round window membrane (RWM) or intracochlear infusion through a cochleostomy. However, the surgical trauma, inflammation, and hearing loss associated with these methods lead us to investigate a less invasive delivery method. Herein, we studied the feasibility of a vector transgene-soaked gelatin sponge, Gelfoam, for transgene delivery into the mouse cochlea through an intact RWM. The Gelfoam absorbed with liposomes and adenovirus, but not with adeno-associated virus (AAV), was successful in mediating transgene expression across an intact RWM in a variety of cochlear tissues. The Gelfoam technique proved to be an easy, atraumatic, and effective, but vector-dependent, method of delivering transgenes through an intact RWM. Compared with the more invasive gene delivery methods, this technique represents a safer and a more clinically viable route of cochlear gene delivery in humans.