Transfected MCF-7 cells as a model for breast cancer progression

Summary The MCF-7 human breast carcinoma cell line has been used as a recipient for eukaryotic plasmid expression vectors to determine the effects of growth factor and growth factor receptor overexpression on the estrogen-dependent, antiestrogen sensitive and poorly metastatic phenotypes exhibited by this line. Overexpression of some members of the erbB family of ligands and receptors were found to have some effects on these pheno-types. However, only when two members of the fibroblast growth factor family, FGF-1 and FGF-4, were overexpressed was progressivein vivo growth observed is either ovariectomized nude mice without estrogen supplementation or in mice that received tamoxifen treatment. FGF transfected cells also exhibited an increased ability to form micrometastases. The implications of these results with regard to the possible role of the paracrine and autocrine effects of angiogenic growth factor production in breast cancer progression are discussed.     

Inhibitory effects of retinoids on development of squamous metaplasia in rat mammary epithelial organoids cultured in Matrigel

Squamous metaplasia (SQM) developed in cultures of rat mammary organoids in reconstituted basement membrane, Matrigel, under either a complete hormone medium (CHM) or a serum-free mammary epithelium growth medium (MEGM). Organoids cultured in CHM gave rise to fewer such SQM (˜5%) than those in MEGM (˜16%). Formation of SQM was completely suppressed when retinoids were added to CHM. However, a few SQM were still observed in cultures in MEGM with added retinoids. Addition of 5% fetal bovine serum suppressed development of SQM cultured in MEGM. Delayed addition of retinoids also inhibited further development of SQM. Development of SQM from mammary epithelial cells is not common, and regulatory molecules other than retinoids apparently are involved in their formation and prevention.     

Development of a sampling and analytical method for 2,2-dichloro-1,1,1-trifluoroethane in workplace air

The use of 2,2-dichloro-1,1,1-trifluoroethane (compound number: HCFC-123) is growing in industry as a substitute for ozone-depleting chlorofluorocarbons (CFCs). Recently, liver-related illnesses have been reported from industries handling HCFC-123. However, information on worker exposure to the material is limited, and an acceptable sampling/analytical method is not available. The aim of this study was to develop a widely applicable sampling and analytical method to determine worker exposures to airborne HCFC-123 and to evaluate the performance of the method. A solid sorbent tube, containing two sections (400 mg in the front and 200 mg in the back) of activated coconut-shell charcoal was chosen for sampling airborne HCFC-123 vapor. The breakthrough volumes were 13.6 L at 3597 +/- 210.1 ppm (with a sampling airflow rate of 0.046 L/min) and 17.0 L at 1841 +/- 4.5 ppm (with sampling airflow rate of 0.046-0.050 L/min). Samples of HCFC-123 in the charcoal tube were stable for 7 days either at room temperature or in a refrigerator and a migration occurred within 14 days at room temperature. It is recommended that the HCFC-123 sample in activated charcoal tubes be stored either at room temperature or in a refrigerator and be analyzed within 7 days. The HCFC-123 in the charcoal tubes was desorbed into dichloromethane and analyzed using gas chromatography/ flame ionization detection. The limit of detection was 0.23 mg/sample, and the average desorption efficiency was 99.0%. The total coefficient of variation was 0.060, and the method accuracy was 16.6%. In conclusion, the performance of the sampling and analytical method developed for the determination of airborne HCFC-123 concentrations was acceptable to the NIOSH sampling and analytical criteria.     

Ultrasound therapy facilitates the recovery of acute pressure-induced conduction block of the median nerve in rabbits

Though the use of ultrasound for the treatment of carpal tunnel syndrome (CTS) or compression neuropathy has been described, its effect remains controversial. A test model of acute CTS was developed using rabbits. Acute median nerve compression was induced by the infusion of saline into the carpal tunnel under general anesthesia to elevate the intracarpal pressure. A reduction in the compound muscle action potential (CMAP) amplitude of the abductor pollicis was noted after intracarpal pressure increased. To investigate the efficacy of ultrasound in acute CTS, rabbits with acute median nerve compression were divided into 3 groups (10 each) and ultrasound was applied at different intensities to each group as follows: 1.5 W/cm(2) to group 1; 0.2 W/cm(2) to group 2; 0.0 W/cm(2) (sham) to group 3. A total of 10 treatment sessions were given over a period of 2 weeks. Following ultrasound application, the CMAP amplitudes showed significant improvement in group 1 compared to the other two groups (P < 0.05), indicating facilitated recovery from acute CTS in this pressure-induced median nerve compression rabbit model. The benefits of ultrasound application in a clinical setting must be verified by further clinical trials.     

An estimation of the first positive Lyapunov exponent of the EEG in patients with schizophrenia

We studied the complexity of the electroencephalogram (EEG) in schizophrenic patients by estimating the first Lyapunov exponent (L1), which might serve as an indicator of the specific brain function in schizophrenia. We recorded the EEG from 25 schizophrenic patients (12 male, 13 female; age=25.1+/-7.0 years) fulfilling DSM-IV criteria and 15 healthy controls (9 male, 6 female; age=27. 8+/-4.2 years) at 16 electrodes, different from previous studies which recorded the EEGs at limited electrodes. We employed a method with an optimal embedding dimension to calculate the L1s. For limited noisy data, this algorithm was strikingly faster and more accurate than previous ones. Our results showed that the schizophrenic patients had lower values of the L1 at the left inferior frontal and anterior temporal regions compared with normal controls. These results for L1 in non-linear analysis have some differences from those for power ratios in linear analysis. These suggest that the non-linear analysis of the EEGs such as the estimation of the L1 might be a useful tool in analyzing EEG data to explore the neurodynamics of the brains of schizophrenic patients.     

Stair-step artifacts with single versus multiple detector-row helical CT

PURPOSE: To compare the effects of acquisition parameters on the magnitude and appearance of artifacts between single and multiple detector-row helical computed tomography (CT). MATERIALS AND METHODS: A cylindric (12.7 x 305.0-mm) acrylic rod inclined 45 degrees relative to the z axis was scanned at the isocenter and 100 mm from the isocenter with single detector-row (single-channel) helical CT (beam width, 1-10 mm; pitch, 1.0, 2.0, or 3.0) and multiple detector-row (four-channel) helical CT (detector width, 1. 25, 2.5, 3.75, and 5 mm; pitch, 0.75 or 1.5). The SD of radius measurements along the rod (SD(r)) was used to quantify artifacts in all 72 data sets and to analyze their frequency patterns. Volume-rendered images of the data sets were ranked by six independent and blinded readers; findings were correlated with acquisition parameters and SD(r) measurements. RESULTS: SD(r) was smaller in four- than in single-channel helical CT for any given table increment (TI). In single-channel helical CT, SD(r) increased linearly with beam width and geometrically with pitch. In four-channel helical CT, SD(r) measurements were directly proportional to the TI, regardless of the detector width and pitch combination used. Off-center object position on average increased SD(r) by a factor of 1.6 for single-channel helical CT and by a factor of 2.0 for four-channel helical CT. Subjective rankings of image quality correlated excellently with SD(r) (Spearman r = 0.94, P <.001). CONCLUSION: Artifacts are quantitatively and subjectively smaller with four- compared with single-channel helical CT for any given TI.     

Proteomic alterations of the variants of human aldehyde dehydrogenase isozymes correlate with hepatocellular carcinoma.

To develop novel markers for hepatocellular carcinomas (HCC), 10 cases were analyzed by 2-dimensional electrophoresis (2DE) and matrix-assisted laser desorption ionization mass spectrometry. Results were compared to those of paired adjacent nontumorous liver tissues. Comparative analysis of the respective spot patterns in 2DE revealed that 3 variants of class 3 aldehyde dehydrogenase (ALDH-3) newly appeared while 2 variants of ALDH-2 diminished to undetectable levels in HCC. However, 4 ALDH-1 variants with different pIs remained unaffected. This pattern of concomitant appearance and disappearance of ALDH-3 and ALDH-2 variants was consistently observed in all HCC tissues. Our results suggest that alterations of ALDH isozyme variants may be closely correlated to HCC and that proteomic analysis of these proteins might be a novel approach to identify the molecular events in detail during hepatocarcinogenesis.     

RET oligonucleotide microarray for the detection of RET mutations in multiple endocrine neoplasia type 2 syndromes.

Multiple endocrine neoplasia type 2 (MEN2) syndromes are inherited in an autosomal dominant fashion with high penetrance. There are three subtypes, namely, MEN2A (multiple endocrine neoplasia type 2A), MEN2B (multiple endocrine neoplasia type 2B), and familial medullary thyroid carcinoma. The variations in the RET gene play an important role in the MEN2 syndromes. In this work, we have developed a RET oligonucleotide microarray of 67 oligonucleotides to quickly detect RET mutations in MEN2 syndromes. The predominant RET mutations are missense mutations and are restricted to nine codons (codons 609, 611, 618, 620, 630, 634, 768, 804, and 918) in MEN2 syndromes. Missense mutations at codons 609, 611, 618, 620, and 634 have been identified in 98% of MEN2A families and in 85% of familial medullary thyroid carcinoma families. More than 95% of MEN2B patients also had a predominant mutation type at codon 918 (Met-->Thr). RET oligonucleotide microarray can detect RET missense mutations at these nine codons. Theoretically, a total of 55 missense mutation types can occur at eight codons (codons 609, 611, 618, 620, 630, 634, 768, and 804). RET oligonucleotide microarray is designed to detect all of these 55 missense mutation types at these eight codons and one predominant type at codon 918. Fifty-six oligonucleotides were designed for the 56 mutation types at nine codons, and 11 oligonucleotides were designed for the wild types and positive controls. We found RET mutations in all eight of the Korean MEN2A families (a total of 75 members; 27 affected members, 19 gene carriers, and 29 unaffected members) using the developed RET oligonucleotide microarray and an automatic sequencing. Because we found only five mutation types from eight MEN2A families, the international collaborations are required to see whether the RET oligonucleotide microarray may be used as a genetic diagnostic tool. Taken together, the RET oligonucleotide microarray can function as a fast and reliable genetic diagnostic device, which simplifies the process of detecting RET mutations.