On-line capillary isoelectric focusing-mass spectrometry for quantitative analysis of peptides and proteins

On-line capillary isoelectric focusing-mass spectrometry (cIEF-MS) was applied to determine concentrations of peptides and proteins using angiotensin II and human tetrasialo-transferrin as the model samples. The concentration of the carrier ampholyte was optimized for both resolution and ion intensity. cIEF-MS employing 1% Pharmalyte 3-10 and a sheath liquid containing water/methanol/acetic acid (50/49/1) resolved angiotensin I and II (5 microM each, DeltapI=0.2) at an Rs value of 2.29. The determined concentration of angiotensin II (0.1-5 microM) well correlated (R=0.999) with that obtained by the conventional RP-HPLC method. The limit of detection was 0.22 microM, which was about 10 times lower than that by UV detection (2 microM). The repeatability and accuracy were <15 and <11%, respectively. cIEF-MS was also applied to determine human tetrasialo-transferrin concentration. The good linearity (R2=0.998) was also observed between the transferrin concentration (0.5-1.2 g/L) and peak area ratio (IS; beta-lactoglobulin B) with acceptable accuracy (<1.9%) and repeatability ( approximately 10% at 1g/L).     

Inhibitory effects of fungal bis(naphtho-gamma-pyrone) derivatives on nitric oxide production by a murine macrophage-like cell line, RAW 264.7, activated by lipopolysaccharide and interferon-gamma

Inhibitory effects of six fungal bis(naphtho-gamma-pyrone) derivatives on nitric oxide (NO) production by a murine macrophage-like cell line, RAW 264.7, which was activated by lipopolysaccharide and interferon-gamma were examined. Among these derivatives, chaetochromin (4) (IC(50): 0.8 microM), cephalochromin (1) (IC(50) 1.5 microM), and dihydroisoustilaginoidin A (6) (IC(50) 2.8 microM) exhibited strong inhibitory activity. The bis(naphtho-gamma-pyrone) derivatives did not affect the enzyme activity of inducible nitric oxide synthase (iNOS). However, these derivatives significantly reduced both the induction of iNOS protein and iNOS mRNA expression. These results suggest that the bis(naphtho-gamma-pyrone) derivatives have the pharmacologic ability to suppress NO production by activated macrophages.     

Successful topical application of a new antifungal agent, micafungin, in the treatment of refractory fungal corneal ulcers: report of three cases and literature review

PURPOSE: To report the efficacy of topical application of a new antifungal agent, micafungin (MCFG), in the treatment of yeast-related corneal ulcers. DESIGN: Noncomparative interventional case reports. PARTICIPANTS: Three patients with yeast-related corneal ulcer after keratoplasty recalcitrant to conventional antifungal treatment for 4 weeks were recruited in this study. METHODS: Topical 0.1% antifungal MCFG eye drops were applied in 3 patients with yeast-related corneal ulcer every hour while awake until epithelialization. After epithelialization, the frequency of eye drops was reduced to 5 times a day. MCFG eye drops were discontinued 1 month after the disappearance of stromal infiltration in each case. The patients underwent best corrected visual acuity (BCVA) measurements, slit-lamp examination, fluorescein-dye staining, and anterior segment photography. Corneal scrapings and cultures of surgical materials were also performed. MAIN OUTCOME MEASURES: Changes in ulcer size, stromal infiltration, fluorescein dye staining, and BCVA were looked for. RESULTS: All corneal ulcers epithelialized within 14 days after commencement of application of 0.1% MCFG eye drops. Yeasts were detected from corneal smears in all eyes. Two cases revealed positive culture isolates for Candida albicans and Candida parapsilosis. No recurrence of fungal keratitis was observed in any of the cases throughout the follow-up periods. CONCLUSION: Topical 0.1% MCFG eye drops seem to be an effective and a promising option in the treatment of refractory yeast-related corneal ulcers.     

Association between intratumoral free and total VEGF, soluble VEGFR-1, VEGFR-2 and prognosis in breast cancer

Vascular endothelial growth factor (VEGF) receptors consist of three cell-membrane type receptors (VEGFR-1, VEGFR-2 and VEGFR-3), and soluble form of VEGFR-1 (sVEGFR-1), an intrinsic negative counterpart of the VEGF. In this study, we measured intratumoral protein levels of free and total VEGF, VEGFR-2 and sVEGFR-1 from 202 primary breast cancer tissues and examined their prognostic values. A significant inverse correlation was found between free or total VEGF and oestrogen receptor (ER) status (P=0.042 and 0.032, respectively). A univariate analysis showed that low sVEGFR-1 and high total VEGF were significantly associated with poor prognosis in disease-free survival (DFS) and overall survival (OS). The ratio of sVEGFR-1 to total VEGF was a strong prognostic indicator (DFS: P=0.008; OS: P=0.0002). A multivariate analysis confirmed the independent prognostic values of total VEGF and the ratio of sVEGFR-1 to total VEGF. In subgroup analysis, total VEGF was a significant prognostic indicator for ER-positive tumours but not for ER-negative tumours, whereas sVEGFR-1 was significant for ER-negative tumours but not for ER-positive tumours. In conclusion, the intratumoral sVEGFR-1 level, VEGF level and the ratio of sVEGFR-1 to total VEGF are potent prognostic indicators of primary breast cancer, and might be relevant to ER status.     

Cytologic factors associated with prognosis in patients with peripheral adenocarcinoma of the lung measuring 3 cm or less in greatest dimension

BACKGROUND: Recently, peripheral lung adenocarcinomas (PLA) measuring < or = 3 cm in greatest dimension often have been diagnosed using diagnostic radiology. The objective of the current study was to determine which cytologic factors are associated with a favorable outcome and an unfavorable outcome in patients with PLA. METHODS: Imprint smears from 134 patients with PLA were examined. Sixteen cytologic factors, including necrosis, cellular distribution, overlapping of cell clusters, cluster aggregation, cluster size, cluster thickness, nuclear irregularity, nuclear size, variation in nuclear size, multinucleated cells, intranuclear inclusions, type of intranuclear inclusions, appearance of nucleoli, eosinophilic nucleoli, multinucleoli, and mitosis, were evaluated using univariate and multivariate analyses. A counting method was used to determine the prognosis for individual patients. RESULTS: In the univariate analysis, a cluster size that measured > or = 831 microm in short dimension (P = 0.0011), moderate or severe nuclear irregularity (P = 0.0030), > or = 5 multinucleated cells per 100 tumor cells (P = 0.0047), moderate or severe variation in nuclear size (P = 0.0061), medium or large nuclear size (P = 0.0169), and > or = 1 mitotic cell per 100 tumor cells (P = 0.0412) were associated significantly with a poor outcome. In the multivariate analysis, cluster size in short dimension (P = 0.0018), multinucleated cells (P = 0.0066), and nuclear irregularity (P = 0.0310) were found to be independent prognostic factors. CONCLUSIONS: The combination of cytologic features using intraoperative imprint smears, namely, cluster sizes < or = 830 microm in short dimension, < or = 4 multinucleated cells per 100 tumor cells, and mild nuclear irregularity, may provide favorable predictive information in patients with PLA.     

Effect of saline flush using dual injector on abdominal dynamic contrast enhanced computed tomography

PURPOSE: To evaluate the efficacy of the saline flush technique for abdominal multidetector-row computed tomography (CT). METHODS: As a clinical study, 147 patients with chronic hepatic disease were divided into two groups: group A, given 100 cc of contrast material flushed with 50 cc of saline; and group B, given 100 cc of contrast material only. The difference in attenuation values between groups A and B, and cirrhotic liver and non-cirrhotic liver were evaluated. As an in-vitro experimental study, time vs. peak pressure curve was obtained changing injection devices. RESULTS: Group B showed a greater contrast enhancement effect in the early arterial equilibrium phase of the aorta and late arterial phase of the liver. In cirrhotic liver, the aorta showed greater contrast enhancement in the early arterial phase in group A. Attenuation values of hypervascular hepatic nodules showed no statistically significant difference between groups A and B. Time to reach peak pressure was prolonged when using saline flush devices in vitro. Flow of contrast material from the contrast material cylinder to the saline cylinder was also discovered. CONCLUSION: Saline flush causes peak arterial contrast to be strong for a short duration. Abdominal organ attenuation values are not necessarily increased in the saline flush method. It is important to know that contrast enhancement is dependent on the injection device.     

Study of automated segmentation of the cerebellum and brainstem on brain MR images

MR imaging is an important method for diagnosing abnormalities of the brain. This paper presents an automated method to segment the cerebellum and brainstem for brain MR images. MR images were obtained from 10 normal subjects (male 4, female 6; 22-75 years old, average 31.0 years) and 15 patients with brain atrophy (male 3, female 12; 62-85 years of age, average 76.0 years). The automated method consisted of the following four steps: (1) segmentation of the brain on original images, (2) detection of an upper plane of the cerebellum using the Hough transform, (3) correction of the plane using three-dimensional (3D) information, and (4) segmentation of the cerebellum and brainstem using the plane. The results indicated that the regions obtained by the automated method were visually similar to those obtained by a manual method. The average rates of coincidence between the automated method and manual method were 83.0+/-9.0% in normal subjects and 86.4+/-3.6% in patients.     

Internal mammary artery injury following subclavian vein catheterization

Inappropriate puncture of the internal mammary artery caused by subclavian vein puncture is a rare but potentially lethal complication. We report a case of 45-year-old woman with internal mammary artery injury following subclavian vein catheterization successfully treated by transcatheter arterial embolization using coils.     

Sequential evaluation of prostate edema after permanent seed prostate brachytherapy using CT-MRI fusion

PURPOSE: To analyze the extent and time course of prostate edema and its effect on dosimetry after permanent seed prostate brachytherapy. METHODS AND MATERIALS: Twenty patients scheduled for permanent seed (125)I prostate brachytherapy agreed to a prospective study on postimplant edema. Implants were preplanned using transrectal ultrasonography. Postimplant dosimetry was calculated using computed tomography-magnetic resonance imaging (CT-MRI) fusion on the day of the implant (Day 1) and Days 8 and 30. The prostate was contoured on MRI, and the seeds were located on CT. Factors investigated for an influence on edema were the number of seeds and needles, preimplant prostate volume, transitional zone index (transition zone volume divided by prostate volume), age, and prostate-specific antigen level. Prostate dosimetry was evaluated by the percentage of the prostate volume receiving 100% of the prescribed dose (V(100)) and percentage of prescribed dose received by 90% of the prostate volume (D(90)). RESULTS: Prostate edema was maximal on Day 1, with the median prostate volume 31% greater than preimplant transrectal ultrasound volume (range, 0.93-1.72; p < 0.001) and decreased with time. It was 21% greater than baseline at Day 8 (p = 0.013) and 5% greater on Day 30 (p < 0.001). Three patients still had a prostate volume greater than baseline by Day 30. The extent of edema depended on the transition zone volume (p = 0.016) and the preplan prostate volume (p = 0.003). The median V(100) on Day 1 was 93.6% (range, 86.0-98.2%) and was 96.3% (range, 85.7-99.5%) on Day 30 (p = 0.079). Patients with a Day 1 V(100) >93% were less affected by edema resolution, showing a median increase in V(100) of 0.67% on Day 30 compared with 2.77% for patients with a V(100) <93 % on Day 1. CONCLUSION: Despite the extreme range of postimplant edema, the effect on dosimetry was less than expected. Dose coverage of the prostate was good for all patients during Days 1-30. Our data indicate that postimplant dosimetry on the day of implant is sufficient for patients with good dose coverage (Day 1 V(100) >93%).