A technique for the culture of Barrett's oesophageal cells

Establishment of cells in tissue culture from Barrett's columnar epithelium has been difficult. The aim of this study was to develop a successful tissue culture method employing a serumfree medium for cultivation of Barrett's epithelial cells. Fragments of Barrett's mucosal tissue were explanted in a 3:1 mixture of Dulbecco's modification of Eagle's medium and Ham's F12, to initiate the outgrowth of epithelial cells. Subsequently, a commercial serum-free medium (formulated for the growth of keratinocytes) was used for the propagation of Barrett's oesophagus cells without fibroblast growth. Cells established in culture retained their epithelial morphology, stained positive for cytokeratin, and contained Alcian blue (pH 2.5) and periodic acid-Schiff reagent-positive/diastase-resistant vacuoles, confirming their origin from Barrett's epithelium. Electron microscopy showed tonofilaments, microvilli and desmosomes. Coating the surface of culture vessels was not required and four cell strains could be passaged up to 20 times with no fibroblast growth, in the keratinocyte serumfree medium.     

Systemic 5-fluorouracil producing an inflammatory response in porokeratosis

Topical 5-fluorouracil has been used as an effective treatment for porokeratosis. Upon its treatment, an inflammatory effect occurs with the topical 5-fluorouracil. We report a case of a patient with disseminated superficial actinic porokeratosis displaying a comparable inflammatory process following therapy with systemic 5-fluorouracil used to manage a metastatic breast cancer.     

Elevated expression of Bcl-2 and Bcl-x by intestinal intraepithelial lymphocytes: resistance to apoptosis by glucocorticoids and irradiation

Administration of glucocorticoids or exposure to ionizing radiation in vivo results in a rapid cell death of thymocytes. We report that murine small intestinal intraepithelial lymphocytes (IEL) are resistant to both steroid- and radiation-induced deletion. This is due to resistance to apoptosis, as evidenced by the absence of detectable apoptotic IEL nuclei in situ after in vivo glucocorticoid treatment. IEL express normal levels of glucocorticoid receptors and these receptors bind [3H]dexamethasone to equivalent levels as other lymphocyte populations. Thus, their survival is due to post-receptor signaling mechanisms. Many IEL express high levels of Bcl-2 and that of these Bcl-2high IEL are largely TCR gamma delta +. Those IEL that do express high levels of Bcl- 2 are CD8 alpha + beta - CD4-. In addition, IEL express Bcl-x, another protein shown to be involved in the protection of cells from apoptotic signals. IEL represent the first lymphocyte population in vivo shown to have high levels of expression of both molecules, that otherwise occur only in activated lymphocytes in vitro. These data suggest that the Bcl- 2+Bcl-x+ IEL are activated cells and not an effete population of cells necessarily destined to die. Also, the high levels of Bcl-2 and Bcl-x in this in vivo activated population supports the in vitro correlate of protection from activation-induced cell death.      

Central diabetes insipidus in a newborn with deletion of chromosome 7q

Abstract: We report an infant with midline craniofacial defects and holoprosencephaly due to chromosome 46, XY, del (7) (pter →q34) who presented at 1 week of age with central diabetes insipidus. The importance of hypothalamic-pituitary endocrine investigation in patients with this syndrome, and more generally, in patients with midline craniofacial malformation or holoprosencephaly is emphasized. As infants with chromosome 7q deletion bear close phenotypic resemblance to infants of Trisomy 13, chromosomal confirmation and karyotype banding is mandatory to establish an accurate diagnosis and for genetic counselling of their parents.     

Percutaneous retrieval of a chronic foreign body with both intravascular and extravascular components

Removal of unwanted intravascular foreign body is a useful but infrequent procedure carried out by interventional radiologists. We study a patient who had a long guidewire left in her body following central venous catheter placement by a surgeon. The guidewire was later found in situ, with both intravascular and extravascular components in continuity. We successfully removed the guidewire without causing any complications. Standard interventional techniques, Amplatz gooseneck snare (Microvena, White Bear Lake, MN, USA) and 6‐Fr Multipurpose catheter were used.     

Impaired vision in the elderly: a preventable condition

BACKGROUND: Vision is thought to deteriorate with age as a number of factors in later life endanger eyesight. Assessment of the visual acuity of the elderly and identification of endangering factors help in detecting those with impaired vision which in turn impairs daily activities. OBJECTIVE: This study measured the visual acuity of the senior citizens and identified those with impaired vision. The probable contributing factors for impaired vision were studied with the aim of preventing visual impairment. METHOD: The study was part of a screening campaign for elderly glaucoma in the community. A convenience sample of ambulatory senior citizens from stratified localities had their visual acuity measured with a standard Snellen's chart. The test was repeated with pinholes if the visual acuity was less than 0.5. Those without improvement after pinhole were considered as having impaired vision. People with elevated intraocular pressure by the Pulsair were selected for examination by an ophthalmologist for ocular pathology. RESULTS: For the ambulatory population aged > or = 65 the mean visual acuity of either eye before pinhole was 0.3. Nearly 72% had impaired vision (visual acuity not corrected above 0.5 with pinhole). There was a significant association between this impairment and female sex, history of diabetes mellitus or glaucoma, cataract, and infrequent eye examination. CONCLUSION: Impaired vision is highly prevalent in the elderly ambulatory population, a condition which is preventable by tight surveillance of predisposing factors and regular simple measurement of visual acuity. The primary care setting is most suitable for these activities.      

Analysis of cartilage oligomeric matrix protein (COMP) in synovial fibroblasts and synovial fluids

We investigated the expression of cartilage oligomeric matrix protein (COMP) in normal and rheumatoid arthritis (RA) synovial fibroblasts. In situ hybridization (ISH) was conducted on synovial specimens from five RA patients applying specific probes for COMP or fibroblast collagen type I. ISH was combined with immunohistochemistry, applying antibodies to the macrophage marker CD68. Ribonuclease protection assay (RPA) and rapid amplification of 3'-cDNA ends (3'-RACE) were performed on total RNA from normal and RA synovial fibroblast cultures. Protein extracts from fibroblasts and culture supernatants were compared with synovial fluids and protein extracts from isolated chondrocytes by Western blot utilizing polyclonal and monoclonal antibodies (18-G3 mAb) to COMP. COMP mRNA was detected in fibroblasts of RA synovium by ISH, and in normal and RA synovial fibroblast cultures by RPA. 3'-RACE demonstrated sequence homology of chondrocyte and synovial fibroblast COMP along the coding sequence. COMP protein was detected in synovial fibroblasts and culture supernatants by immunoblot. Using polyclonal antibodies, the major portion of COMP from fibroblasts and culture supernatants was present as low-molecular-weight (LMW) bands, corresponding to those found in synovial fluids. These LMW COMP bands, however, were not detected in any of the cells or tissues tested using 18-G3 mAb. In protein extracts from chondrocytes and in COMP purified from cartilage, these LMW bands could not be detected. In conclusion, the data suggest that certain forms of COMP detected in synovial fluid are secreted from synovial fibroblasts and could be distinguished by specific mAbs from COMP secreted by chondrocytes.      

Blood use in patients undergoing repeat coronary artery bypass graft procedures: multivariate analysis

BACKGROUND : The prevailing clinical opinion is that patients undergoing repeat coronary artery bypass graft (CABG) operation require more blood transfusions than do patients undergoing primary CABG operation. To determine the extent of this increased demand and the variables responsible for it, the cases of 196 patients who had undergone primary procedures and 65 patients who had had repeat procedures at the same institution were reviewed. STUDY DESIGN AND METHODS : To analyze the differences in transfusion requirements for these two groups, the following data were obtained: number of transfusions given between the time of surgery and the time of hospital discharge; preoperative hemoglobin (Hb), hematocrit (Hct), prothrombin time, and platelet count; Hb and Hct at hospital discharge; time the patient was on cardiopulmonary bypass; number and type of grafts; estimates of intraoperative blood loss; and chest-tube blood shed during the first 48 hours after surgery. RESULTS : The groups were comparable with respect to age, body weight, preoperative Hb and Hct, number of grafts, and aspirin exposure. Patients in the repeat group had 35-percent greater blood loss and required 75-percent more blood components than did the patients undergoing primary procedures. The mean number of blood components transfused per patient was as follows: red cells, 3.8 +/? 0.5 units in repeat patients and 2.2 +/? 0.2 units in primary patients (p = 0.002); platelets, 2.9 +/? 0.9 vs. 1.1 +/? 0.2 (p = 0.043); fresh-frozen plasma, 1.6 +/? 0.4 vs. 0.8 +/? 0.1 (p = 0.044). Analysis of variables by regression method for repeat patients showed a predictive effect of blood loss (p < 0.0001), prolonged time on cardiopulmonary bypass (p < 0.0001), preoperative Hb (p = 0.0003), and aspirin exposure (p = 0.0094) on red cell transfusion rate in repeat patients (R-square = 0.7778, Prob > f = 0.0001). CONCLUSION : Repeat CABG patients have higher transfusion rates. These findings may be attributed to the increased microvascular bleeding, prolonged time on cardiopulmonary bypass, lower preoperative Hb, and the use of preoperative antiplatelet medications.