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IEC-6 intestinal cell death induced by bovine milk alpha-lactalbumin.

Potent inhibition of cell proliferation was found for commercial preparations of bovine alpha-lactalbumin on cultured intestinal cell line IEC-6 albeit lot-dependent. The inhibition was irreversible and a single exposure to the culture medium containing alpha-lactalbumin of an active lot for a period as short as 30 min was enough to provoke cell death, possibly through apoptosis. The oligomer fraction from size exclusion chromatography was significantly robust, while the monomer fraction remained totally inert, in inducing cell death. Incubation at 37 degrees C for 5 d with 30% trifluoroethanol in acetate, pH 5.5, in a slowly rotating test tube rendered the monomer fraction cytotoxic. Again, the resulting inhibitory activity was found in the oligomer fraction from size exclusion chromatography, with emergence of subtle peaks at 22- and 30-kDa. Furthermore, the occurrence of SDS-stable 30-kDa as well as 20-kDa bands on electrophoresis was a common feature for alpha-lactalbumin with the activity inducing cell death. Thus, a certain dimeric state can be implicated in the cytotoxicity of bovine alpha-lactalbumin.

Animals↗

Studies of cytotoxicity and metabolic competence with the rat intestinal cell line IEC-17.

The characterization of metabolic activity of intestinal cells in culture may be of particular interest for the study of the effects of food additives and contaminants which, ingested through the diet, are absorbed primarily by the gut mucosa. IEC-17 cell line is derived from the intestine of newborn rat and has been shown to be competent for the metabolism of xenobiotics (Quaroni and Isselbacher, 1981), and to keep some degree of differentiation also in in vitro conditions. Our studies indicate that the xenobiotic metabolic activity of this cell line, as shown by measurement of 7-ethoxycoumarin (7-EC), can be induced by beta-naphthoflavone (NF) and not by phenobarbital (PB) and only at late subculturing stages, suggesting these cells undergo some kind of maturation in vitro. Induction of xenobiotic metabolism by beta-naphthoflavone seems also to elicit the toxicity of 9,10-dimethyl-1,2-benzathracene (DBA) but not that one of cyclophosphamide (CPA).

7-Alkoxycoumarin O-Dealkylase↗

Audiometric earphone calibration using the IEC 303 reference coupler: a cautionary note.

The diaphragm of a condenser microphone is exceedingly fragile and, in normal use, is covered by a protective grid. However, when the microphone is incorporated into the IEC 303 reference coupler for the calibration of audiometers or audiometric earphones, the Standard demands that the diaphragm be left exposed. Nevertheless, in order to avoid inadvertent mechanical damage, some users leave a protective grid on the microphone. The effects of two particular grids have been investigated and the magnitude of the resulting errors is indicated.

Audiometry↗

FDA Modernization Act of 1997: modifications to the list of recognized standards; availability; withdrawal of draft guidance "Use of IEC 60601 standards; medical electrical equipment"--FDA. Notice.

The Food and Drug Administration (FDA) is announcing the publication of the modifications to the list of standards that will be recognized for use in the premarket review process and withdrawing its draft guidance entitled "Use of IEC 60601 Standards; Medical Electrical Equipment." This will assist manufacturers who elect to declare conformity with consensus standards to meet all or part of medical device review requirements.

Device Approval↗

Induction of Salmonella enterotoxin (stn) gene expression by epithelial cells (IEC-6).

Prevalence of Salmonella enterotoxin (stn) gene among Salmonella enterica and S. bongori was investigated by polymerase chain reaction (PCR) and gene probe and its status of phenotypic expression was examined on chinese hamster ovary cells by cultivating the strains with conventional method for enterotoxin production and by cultivating the organisms in contact with intestinal epithelial cells of rats (IEC-6). All the 19 strains and serovars of S. enterica such as Typhimurium, Enteritidis, Newport, Weltevreden, Indiana, Gallinarum and Kentucky were found to carry stn gene as examined by PCR and gene probe but only a limited number of strains (13 out of 19) expressed phenotypically the enterotoxin when cultured by conventional method. Cultivation of organisms in contact with epithelial cells induced expression of stn gene phenotypically in all the 19 strains. In contrast to S. enterica, strains of S. bongori were found neither genotypically (stn) nor phenotypically (Stn) positive.

Animals↗

Impact assessment of IEC campaign during anti-malaria month, June 1998 through KABP study.

To assess the IEC Campaign during anti malaria month (June, 98), a base line KABP study through pre-assessment and post-assessment was conducted in the State of Gujarat (four districts namely Ahmedabad, Dang, Panchmahal and Baroda). The study was carried out based on questionnaire (open and closed) developed by NAMP in the randomly selected population. A rapid assessment of the current level of KABP among a sample of population before and after the observance of anti-malaria month. The results of the KABP study revealed that there is definite impact (between 2.18 to 30%) and change in the KABP of the local people, where intensive and continuous I.E.C. activities are being undertaken. In order to achieve the desired change in knowledge, attitude, behaviour and practice of the respondents, it requires continuous I.E.C. campaign throughout the year. The attitude, behaviour and practice in the hard core areas need special efforts, where maximum efforts are required to bring a change in. It should be taken more intensively in the problematic and hardcore areas on priority basis.

Attitude to Health↗

The green tea polyphenol (-)-epigallocatechin-3-gallate blocks nuclear factor-kappa B activation by inhibiting I kappa B kinase activity in the intestinal epithelial cell line IEC-6.

The I kappa B kinase complex (IKK) mediates activation of the transcription factor nuclear factor-kappa B (NF-kappa B). We previously showed that green tea polyphenols inhibited endotoxin-mediated tumor necrosis factor-alpha (TNF alpha) production by blocking NF-kappa B activation. In this study, we evaluated whether green tea polyphenols inhibit NF-kappa B by blocking IKK activity. We assessed IKK activity by detecting changes in phosphorylation of an I kappa B alpha-glutathione S-transferase (GST) fusion protein. IEC-6 cells pretreated with an extract of green tea polyphenols (GrTPs; 0--0.4 mg/ml) had diminished TNF alpha-induced IKK and NF-kappa B activity. Of the various GrTPs, (-)-epigallocatechin-3-gallate (EGCG) was the most potent inhibitor. We next examined whether EGCG inhibited activated IKK. In cytosolic extracts of TNF alpha-stimulated cells, EGCG inhibited phosphorylation of I kappa B alpha-GST (IC(50) > 18 microM) consistent with inhibition of IKK activity. Using other polyphenols, we showed that the gallate group was essential for inhibition, and antioxidants were ineffective in blocking activated IKK. Importantly, EGCG decreased IKK activity in cytosolic extracts of NIK transiently transfected cells. This latter finding showed that our findings were not related to nonspecific kinase activity. In conclusion, EGCG is an effective inhibitor of IKK activity. This may explain, at least in part, some of the reported anti-inflammatory and anticancer effects of green tea.

Animals↗

The cytotoxicity of hydrophobic bile acids is ameliorated by more hydrophilic bile acids in intestinal cell lines IEC-6 and Caco-2.

Bile acids, especially those with hydrophobic properties, are known to possess cytotoxicity. However, the mechanisms responsible for the cytotoxicity of bile acids are still under investigation. On the other hand, the hydrophilic bile acid, ursodeoxycholic acid has been reported to exhibit therapeutic effects against cytotoxic hydrophobic bile acids. The aim of the present study was to investigate the cytotoxicity of individual bile acids and combinations of bile acids using the intestinal cell lines IEC-6 and Caco-2 cells. The cytotoxicities of individual bile acids and the effects of various bile acid combinations were evaluated using the MTS [3-(4, 5-dimethylthiazol-2-yl)-5-(3-carboxymethoxy-phenyl)-2-(4-sulfophenyl)-2H-tetrazolium] assay. The bile acids induced cytotoxic effects depending on their hydrophobicity except for hyodeoxycholic acid. In the study for the effects of combined bile acids, not only ursodeoxycholic acid but other hydrophilic bile salts such as cholic acid and hyocholic acid exhibited cytoprotection against deoxycholic acid-induced cytotoxicity. Moreover, even some hydrophobic bile acids, such as chenodeoxycholic acid also exhibited cytoprotection. It is possible that the cytotoxicity of hydrophobic bile acids is ameliorated by more hydrophilic bile acids under certain conditions. The understanding of the precise mechanism of this phenomenon remains to be determined.

Bile Acids and Salts↗

Quality assurance procedures in radiotherapy--IEC specifications for equipment.

The International Electrotechnical Commission (IEC) worked out international standards for requirements and tests of electrical, mechanical and radiation safety as well as for definition and tests of functional performance characteristics of radiotherapy equipments (medical electron accelerators, gamma beam teletherapy and afterloading equipments, simulators and accessories) and for clinical dosimeters and terminology for medical radiology. A survey is given on the actual state of standardization projects. The problems of such standards are shown for the standard for functional performance characteristics of medical electron accelerators as example.

Humans↗

Structural requirements of analogues of polyamines for migration and growth of IEC-6 cells.

Healing of gastrointestinal mucosal lesions occurs through two processes: an early one involving cell migration and a later one in which cell division replaces lost cells. Both processes require the presence of polyamines, but the mechanism of action of these compounds is unknown. In the present study, we examined the ability of analogues of spermidine and spermine to support migration and growth of IEC-6 cells that have been grown in alpha-difluoromethylornithine to inhibit polyamines. All analogues of spermidine with the general formula x-3 (referring to the numbers of carbon atoms on either side of the central nitrogen), where x = 2-12, competed with spermidine for entry into the cells. However, in addition to spermidine (x = 4), only compounds for which x = 2, 3, or 6 supported migration and only those for which x = 2 or 7 supported growth. Spermine analogues 3-x-3, for which x = 3, 6, 9, or 12, competed for entry into the cells, but only compounds for which x = 3 or 6 supported migration and only the compound for which x = 3, in addition to spermine (x = 4), supported growth. In addition, analogues 2-3-2, 3-2-3, and 2-(3)2, a branched compound, supported both migration and growth but entered the cell via a mechanism different than that for spermidine and spermine. These data define some of the specific structural requirements for polyamines to produce their physiological effects.

Animals↗

[Participation in the IEC Workshop on Adolescent Sexual Health--a challenge to protect reproductive health/right in Latin America and Caribbean Region].

In order to explore the possibility of new global activity in nursing/midwifery to be performed by the WHO Collaborating Center for Nursing Development in Primary Health Care at St. Luke's College of Nursing, the center sent two faculty members to attend the IEC Workshop on Adolescent Sexual Health held in November 1996 in Mexico. The two representatives took part as resource persons and reported to the workshop on the activities of nurses and midwives for adolescent sexual health in Japan. During the session, they obtained information on the actual situation of adolescent sexual health in Latin America and Caribbean region and the novel strategies which were carried out in cooperation between governmental and non governmental organizations. The workshop also provided a good opportunity for international communication and information exchange with health workers in the region about adolescent sexual health. By participating in the workshop, we obtained first-hand information on various aspects of cooperation in international health.

Adolescent↗

Absorption enhancement, structural changes in tight junctions and cytotoxicity caused by palmitoyl carnitine in Caco-2 and IEC-18 cells.

Palmitoyl carnitine chloride (PCC) has been shown to be an effective enhancer of intestinal transport of hydrophilic molecules. The exact mechanism by which the epithelial barrier function is decreased is not clear. In an attempt to elucidate the mechanism of action of PCC, we studied the relationship among absorption enhancement, cell viability and tight junction protein localization in the human colonic Caco-2 cell line and the rat small intestinal cell line IEC-18. Filter-grown cells were exposed to 0 to 1 mM PCC for 30 min, and the efficacy of PCC treatment was determined by assessing the transepithelial electrical resistance and the apparent permeability for mannitol and PEG-4000. Membrane lysis and cytotoxicity were assessed by measurement of lactate dehydrogenase leakage and uptake of propidium iodide and neutral red. The immunolocalization of the tight junctional protein ZO-1 was quantified using CSLM and image-processing software. In both cell lines, PCC caused a dose-dependent decrease in transepithelial electrical resistance and a concomitant increase in the permeability for mannitol and PEG-4000. The transport enhancement was accompanied by an increase in apical membrane permeability and a reduction in cell viability. At higher PCC concentrations (>/=0.4 mM), the distribution of the tight junctional protein ZO-1 was changed and cells were unable to recover viability. PCC is effective as an absorption enhancer for hydrophilic macromolecules. However, lytic effects on the cell membrane and reduced cell viability were concomitant with transport enhancement.

Animals↗