The assessment of in vitro modulation of milk fat globule membrane expression by human breast carcinomas

Although knowledge of functional differentiation and tumour-associated changes of breast carcinomas can be gained by the application of antibodies directed against the milk fat globule membrane, more significant information may be obtained by assessment of the potential of breast carcinomas to modulate their antigenic phenotype. In this study, the extend to which primary tumours can undergo modulation in vitro has been investigated, with consideration of the suitability of organ culture in combination with the immunohistochemical detection of two milk fat globule membrane epitopes, HMFG1 and HMFG2, as methods for detecting this. The preservation of three of the 30 carcinomas assessed, all poorly differentiated, was poor after 3 days of culture. The viability of the other 27 was variable, and was greater in the better differentiated tumours and with the addition of insulin. Expression of the milk fat globule membrane epitopes was generally well maintained. Six of the carcinomas showed a significant change in antigen expression, with this being more frequent in tissues incubated with insulin. Hence, a small group of carcinomas have been identified which appear to have a greater capacity to undergo functional differentiation. Organ culture is considered to be a suitable method for maintaining the tissues in vitro for such evaluation, but the problems encountered in quantifying the immunohistochemical staining, because of antigenic heterogeneity, were such that it is suggested that other approaches be employed.     

In search of the central respiratory neurons: II. Electrophysiologic studies of medullary fetal cells inherently sensitive to CO2 and low pH.

Although extensively pursued, the central respiratory neurons have remained elusive. We departed from the more conventional physiologic and morphologic methods of system and tissue examination and cultured dissociated fetal rat cells (Fitzgerald et al., J Neurosci Res 33:579-589, 1992) from the area of the nucleus ambiguus and the nucleus tractus solitarius located within the 2 mm rostral to the obex. Pacemaker-like cells, with a regular single or bursting activity, studied at 3-5 weeks of age, responded to very small pulses of CO2 (50 ms) and low pH with an increase in spike frequency and a decrease in spike amplitude. Other irregularly beating or silent cells did not respond or else required very large pulses (> 200 ms) to do so. The pacemaker cells also responded to hypoxia induced by administration of sodium hydrosulfite with an increase in spike frequency and amplitude; high oxygen (> 600 torr) and adenosine produced a decrease in electrical activity. Most of these cells were multipolar after staining with antibodies to neuron-specific enolase (NSE) and Fragment C of tetanus toxin. They did not stain for choline acetyltransferase (ChAT). The results suggest that these cultured cells, expressing a phenotype inherently responsive to CO2 and low pH, have the characteristics of central respiratory chemoreceptors, and may be involved in the generation of the respiratory rhythm.     

Midnight reckonings: on a question of knowledge and nursing

Abstract The paper contrasts understandings of knowledge grounded in Enlightenment norms with the departures from those norms taken by some strands of feminism and hermeneutics, as well as the contributions made by the writing of Michel Foucault. A reading of Foucault's writings on knowledge, power and the discursive constitution of self and world is offered as a potentially useful frame within which to raise questions about nursing, nurses and knowledge.     

Expression of human T-cell lymphotropic virus type-1 gene products in the short-term cultured skin tissues of an adult T-cell leukemia/lymphoma patient with cutaneous manifestations.

Adult T-cell leukemia/lymphoma (ATLL) is recognized as a disease etiologically associated with human T lymphotropic virus type-1 (HTLV-1) infection, but, neither viral replication nor specific virus antigen expression have been detected on ATLL cells distributed in organs, including skin. To examine the latent expression of HTLV-1 in the cutaneous lesions of ATLL patients, we cultured the lesional skin tissues in vitro and applied immunofluorescence staining with mouse monoclonal antibodies Lt-4, GIN-14, and F10, which react with p40tax, p19 and gp21, respectively. We recognized HTLV-1 specific antigens on clustered ATLL cells only in the deeper dermis of the skin after 24 hrs cultivation of the lesional skin tissue from an ATLL patient in RPMI-1640 medium supplemented with 20% fetal calf serum. In the electron microscope, we observed HTLV-1 like particles, 80-140 nm in diameter with envelope and core structures, in the same tissue specimen. These findings suggest that HTLV-1 gene products may be expressed in the skin lesions of ATLL patients and involved in the pathogenesis of skin eruptions in cutaneous type ATLLs. To our knowledge, this is the first report that envisages the potency of intracutaneous HTLV-1 expression in vivo.     

Relationship between cellular ATP content and cellular functions of primary cultured rat hepatocytes in hypoxia

The importance of oxygen in maintaining the functional integrity of hepatocytes has been well established in a variety of experimental models, such as in vivo , perfused liver and isolated hepatocytes. However, one of the shortcomings of these systems is their short life span. Therefore, we have examined the effects of long-term hypoxia on cellular adenine nucleotide content and cellular functions, such as albumin production, urea production and DNA synthesis, in adult rat hepatocytes in primary culture. Hepatocytes were cultured at a density of 11 × 10⁴ and 5 × 10⁴ cells/0.18 mL per cm² for the study of albumin and urea production and DNA synthesis, respectively, at various oxygen tensions (20, 12, 8 and 5%) for 24 h. Cellular ATP content in cultured hepatocytes in hypoxia gradually declined, corresponding to the decrease in oxygen tension, and the cellular ATP level at 5% oxygen was approximately 20% of that at 20% oxygen. Albumin production also decreased in parallel with the decrease in cellular ATP content in cultured hepatocytes in hypoxia. However, even when cellular ATP content gradually declined corresponding with the decrease in oxygen tension in cultured hepatocytes in hypoxia, such as at 8 or 5% oxygen, urea production remained at a high level; in contrast, DNA synthesis was completely suppressed. These results suggest that the cellular ATP content decreases in cultured hepatocytes during long-term hypoxia in relation to oxygen tension and that the relationship between decreased ATP levels and liver function in cultured hepatocytes during hypoxia differs for albumin production, urea production and DNA synthesis.     

The effects of an HIV/AIDS educational programme on the anxiety level of nursing students

Acquired immunodeficiency syndrome (AIDS) is a serious disease that has special concern for the health care provider. AIDS has continued to grow despite control efforts. As the disease infectivity period remains lengthy, and the heterosexual population is affected to a greater degree, the level of anxiety has also risen despite educational endeavours. Many fears and anxieties have been associated with AIDS patients by health care workers. The reduction of stress, perceived risk and discomfort following educational efforts have been supported in past research. Educational programmes will need to be given for current health care workers at all levels as well as nursing students. Future nurses must be prepared to meet this challenge. This study was conducted using a convenience sample of nursing students at a university in western United States. Its purpose was to assess any changes that occurred in state anxiety following an educational presentation. Spielberge's State-Trait Anxiety Inventory was used as the measurement instrument. Some anxiety levels were significantly reduced.     

High level production of soluble single chain antibodies in small-scale Escherichia coli cultures

We have investigated the effect of growth and induction conditions on the production of soluble single-chain Fv antibody fragments in Escherichia coli under the control of wt lac promoter. The scFv was directed into the periplasmic space by a pelB leader sequence. Addition of sucrose to the medium gave a 15–25-fold increase in the yield of soluble scFv-phOx (3.0 mg/l) for bacterial shake-tube cultures and an increase of 80–150-fold (16.5 mg/l) for shake-flask cultures. Using flask culture in the presence of 0.4 M sucrose, a significant amount of scFv was released into the medium. We found that the scFv could be made to accumulate in the periplasm or be secreted into the medium by simply changing the incubation conditions and the concentration of the inducer. The ratio between soluble antibody fragments and insoluble scFv aggregates proved to be dependent on the strength of the promoter. Lowering the incubation temperature below 20°C had no effect on the yield of soluble antibody fragments in the periplasm, but they were no longer secreted into the medium. An example of high level production in shake-flask cultures and one-step purification by immobilized metal affinity chromatography (IMAC) is described for a soluble scFv specific for the T cell surface antigen CD3. The biological activity of the purified anti-CD3 scFv was demonstrated by flow cytometry. This method should be especially useful for the functional screening of a large number of clones in small-scale cultures.     

Efficacy of ivermectin for chronic strongyloidiasis: two single doses given 2 weeks apart

We assessed the efficacy and adverse effects of ivermectin for the treatment of chronic strongyloidiasis. Fifty patients were treated with a single dose of ivermectin at approximately 200 μg/kg, and the dose was repeated 2 weeks later. The eradication rate was 96% (48 of 50 patients) at 2 weeks after the first dose and 98% at 2 weeks after the second dose (49 of 50 patients). In the latter cases, no recurrence was noted at the end of the 4-month post-treatment follow-up period. The gastrointestinal symptoms observed most frequently before the treatment were borborygmus, constipation, and diarrhea. These symptoms improved in the majority of patients after the treatment. Side effects, including nausea and vomiting, occurred in 1 patient (2%), but these were mild and transient. Although abnormal values in laboratory tests were observed in 4 patients (8%), they were mild, and no particular clinical care was required. Thus, ivermectin showed an excellent antihelminthic effect, with little toxicity. Our results indicate that a regimen of two single doses of 200 μg/kg ivermectin, given 2 weeks apart, is clinically suitable for the treatment of chronic strongyloidiasis. Received: April 25, 2001 / Accepted: November 16, 2001     

大鼠小肠上皮细胞的体外原代培养

目的：探讨大鼠小肠上皮细胞原代培养的原理与方法以及培养过程中常见的问题和克服办法。方法：取生后1周SD大鼠的小肠上皮细胞，经分离纯化后进行原代培养。结果与结论：胶原酶Ⅺ型和中性蛋白酶Ⅰ型配合使用，可以分离到形态完整、容易贴壁生长的小肠绒毛细胞团。进一步利用离心技术纯化得到95％以上的小肠上皮细胞。结果显示，常规体外培养的肠上皮细胞是否增殖分化最终决定于用酶消化后的结构完整性，如果用0．25％胰酶消化，则细胞受损程度较大，不易贴壁生长。