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Culture of blastomeres from in vitro-matured, fertilized, and cultured bovine embryos.

A culture system was devised to study the differentiation of bovine blastomeres. Blastomeres (2-13 per well) from embryos produced by in vitro maturation, fertilization, and culture of oocytes obtained from slaughterhouse ovaries were cultured for 10 days in 24-well culture plates on feeder layers in blastomere culture medium (BCM: equal parts tissue culture medium 199 and low-glucose Dulbecco's modified Eagle's medium with 10% fetal bovine serum). Ovine embryonic fibroblasts and STO cells were superior to bovine and mouse embryonic fibroblasts as mitotically inactivated feeder cells. Over five studies in which four blastomeres from an embryo were added to each culture well, an average of one colony per well formed from the blastomeres. The colonies continued to grow throughout the culture period, and most colonies resembled trophectoderm in their cellular characteristics, although some cultures contained a mixture of trophectoderm and endoderm. When the number of blastomeres cultured in each well was varied from 2-8, the number of colonies formed was proportional to the number of blastomeres added. Blastomeres from day 5 and day 6 embryos produced fewer colonies than did those from day 4 embryos, perhaps as a result of differentiation and tighter blastomere adhesion resulting in damage during their separation. The absence of serum did not alter the number of colonies formed. A number of growth factors, including LIF, OM, PGDF alpha, and FGF4, had no effect on the number of colonies, the size of colonies, or their alkaline phosphatase staining score beyond that provided by the feeder layer on serum when present. Blastomeres did not form colonies in the absence of feeder layers.

Animals↗

Development of a new culture system for human lymphokine-activated killer cells: comparison with a conventional static culture method.

We recently developed a new culture system based on dialysis perfusion (designated JCC-device) for the generation and expansion of human lymphokine-activated killer (LAK) cells (Murata et al., 1990). More recently we have scaled up the volume of the culture vessel of the JCC-device from 100 ml to 400 ml for clinical use. In the present study, using this new 400 ml JCC-device, we cultured human lymph node lymphocytes (LNL) obtained from 8 surgical patients with primary lung cancer, and investigated the cellular characteristics in comparison with a conventional batchwise culture system using tissue culture dishes. With the JCC-device, the cell density reached a maximum 2.7 x 10(7) cells/ml with greater than 90% viability by the appropriate exchange of perfusion medium and by making additions at the appropriate intervals for recombinant interleukin-2 (rIL-2). The expansion fold of LNL with the JCC-device, ranging 6.6- to 19.2-fold (mean 13.8-fold), was not significantly different from that in dish cultures. There was no marked difference in cell surface phenotypes between the two culture systems in 7 out of 8 cases. As for LAK activity of LNL, the JCC culture was either superior or equal in 4 out of 8 cases, but inferior in the other 4 cases to the conventional dish cultures. In the latter cases, the usage of serum for the JCC culture was limited, which might have resulted in the low LAK activity.(ABSTRACT TRUNCATED AT 250 WORDS)

Cell Count↗

Toward a culturally anchored ecological framework of research in ethnic-cultural communities.

Proposed a research framework (the "cube" model) in which community psychologists working in ethnic-cultural communities can make appropriate decisions on conceptual and methodological issues from a culturally anchored, ecological-contextualist perspective. The intent of the model is to articulate ethnic-cultural heterogeneity in community research by elucidating three metamethodological issues: (a) definition of an ethnic-cultural community, (b) applicability of cross-cultural theories and methods to ethnic-cultural community research, and (c) geographical or ecological stability of an ethnic-cultural community. The model posits that ethnic-cultural community research can be conceptualized as a three-dimensional structure that represents an interaction among research questions, methods, and cultural complexity (referring to the extent to which an ethnic-cultural group is defined in a larger ecological context or community both at the individual and collective levels). Future directions for research were discussed in terms of the utility and the limitations of the proposed research model.

Cross-Cultural Comparison↗

Isolated mouse pancreatic islets in culture: effects of serum and different culture media on the insulin production of the islets.

Various conditions for tissue culture of collangenase-isolated mouse pancreatic islets were studied in an attempt to optimize the maintenance of glucose stimulated insulin biosynthesis and release in the cultured specimens. Islets which had been cultured at a physiological glucose concentration (5.5 mmol/1) in the absence of serum had an impaired glucose-stimulated insulin biosynthesis and release as well as a reduced insulin content. Thus, insulin biosynthesis was three times higher after culture in a serum supplemented medium. Further, the insulin secretion of islets cultured in the presence of serum was markedly enhanced in acute incubations with high concentrations of glucose. This response was most pronounced in islets which had been cultured free-floating. A comparison between different culture media showed that islets cultured in RPMI 1640 had the highest insulin production. The present data suggest that the most favourable conditions for long-term storage of isolated islets in culture may be obtained when the islets are maintained as free-floating explants in a culture medium consisting of RPMI 1640 supplemented with serum.

Animals↗

Importance of hepatocyte culture conditions in dimethylnitrosamine-induced suppression of antibody response in the mixed cultures of murine hepatocytes and splenocytes.

The role of the hepatocyte culture media in dimethylnitrosamine (DMN)-induced suppression of antibody responses by splenocytes against sheep erythrocytes (SRBCs) was investigated using the mixed culture system of murine hepatocytes and murine splenocytes. It was observed that hormone-supplemented complete media was required for hepatocyte cultures to optimally activate DMN to its immunosuppressive form(s). In the absence of the hormone supplement, the concentration of DMN required to produce a 50% suppression (IC50) was increased by over 10-fold (i.e., compare the IC50 in complete media of less than 0.5 mM to the IC50 in basal media of almost 10.0 mM). In contrast, the activation of cyclophosphamide (cytoxan, CTX), which was used in these studies as a comparative control, was not affected by the absence of the hormone supplement. These results indicate that the observed effect on the activation of DMN was not due to a generalized loss of metabolic capability of hepatocytes cultured without hormones. To examine the role of drug metabolizing capabilities of hepatocytes in the differential activation of DMN, we compared phase I and phase II enzyme activities of hepatocytes cultured for 24 h in either basal media or hormone-supplemented complete media. Our results indicated that there was a significant decrease of phase I monooxygenase activities of cultured hepatocytes when compared to freshly isolated hepatocytes. However, our results failed to show any difference in the activities of hepatocytes cultured in the two media. Most notably, there was no difference in the activity of either high- or low-affinity DMN demethylase, as measured by the generation of formaldehyde. We observed a similar profile with phase II conjugative capabilities, specifically sulfotransferase and glucuronyltransferase. These results indicate that the activation of DMN to its immunosuppressive form(s) can be modulated in the co-culture system by culturing hepatocytes under different conditions. Because we failed to show any differences in the metabolic capabilities of hepatocytes cultured under the two media conditions, the results suggest that the modulation of immunosuppressive activity may not be related to a change in the generation of the immunosuppressive metabolite(s).

Animals↗

Effect of holding technique and culture drop size in individual or group culture on blastocyst development after ICSI of equine oocytes with low meiotic competence.

The effect of medium-to-embryo ratio on blastocyst development of equine embryos from oocytes with compact cumuli was evaluated in the present experiment. In addition, two methods for holding oocytes before in vitro maturation were compared. In Experiment 1, oocytes cultured with roscovitine for 16-18h before maturation were fertilized by intracytoplasmic sperm injection and cultured individually in 2.5, 5, 10 or 50microl droplets. In Experiment 2, oocytes were either cultured with roscovitine or held in a modified M199 with 20% serum at room temperature (EH treatment) for 16-18h, then matured, fertilized and cultured in groups at 5microl medium per embryo. In Experiment 3, oocytes were held in the EH treatment, then were matured and fertilized. In Study 3.1, injected oocytes were cultured individually in drop sizes as for Experiment 1; in Study 3.2, groups of 2-7 oocytes were cultured in fixed drop sizes of 5 or 50microl. Blastocyst development rates of individually-cultured embryos were not significantly different among drop sizes in either Experiment 1 or 3 (15-29%). In Experiment 2, blastocyst rates were not significantly different between holding treatments (17-23%). In Experiment 3, for group-cultured oocytes, blastocyst development was not significantly different between 5 and 50microl drops (39 and 27%, respectively). In conclusion, compact-cumulus oocytes may be effectively held in the EH treatment before maturation, and single culture of equine embryos yields acceptable blastocyst development. The greatest blastocyst rate (39%) was obtained with group culture in a 5microl droplet.

Animals↗

The dispersed cell culture as model for functional studies of the subcommissural organ: preparation and characterization of the culture system.

The subcommissural organ (SCO) is an enigmatic secretory gland of the brain, which is believed to be derived from ependymal (glial) precursor cells. We here developed a dispersed cell culture system of the bovine SCO as an approach to functional analyses of this brain gland. Tissue of the bovine SCO obtained from the slaughterhouse was papain dissociated either directly after dissection or after preparation of SCO explants. The latter had been maintained for 4-6 weeks in organ culture. The dispersed cells were cultured for up to 14 days and continuously tested for their secretory state by immunostaining of their secretory product. With respect to the morphology of the SCO cells (shape, processes, nucleus), no difference was found between the culture of freshly dissociated SCOs and that of dissociated SCO explants. In all cases, the dissociation caused a dedifferentiation; typical elongated cells were formed increasingly after 1 day of culture. Thereafter, only the cellular size increased, whereas the shape and the viability of the cells remained unchanged. Proliferating SCO cells were never observed. The culture obtained from fresh SCO tissue contained more glia cells and fibrocytes than the culture prepared from SCO explants. The proliferation of glia cells and fibrocytes was suppressed by blocking the mitotic activity with cytosine-beta-D-arabino furanoside (CAF). The cytophysiological features of the cultured dispersed cells of both origins did not differ as demonstrated by classical histology, by immunocytochemistry for the secretory products of the SCO, by the characteristics of calcium influx into the cytoplasm ([Ca2+]i) and cyclic adenosine monophosphate (cAMP) after stimulation with adenosine-5-triphosphate, substance P or serotonin, and by the activation of the transcription factor cAMP-responsive element-binding protein. Because of the maintenance of their viability, their capacity to release the secretory product into the culture medium, their receptive capacity, and their signal transduction pathways, we conclude that the dispersed cell culture system, especially that obtained from SCO explants, represents an appropriate and useful model for functional studies of the mammalian SCO.

Adenosine Triphosphate↗

In search of cultural diversity: recent literature in cross-cultural and ethnic minority psychology.

The purpose of this study was to identify where the most work on cross-cultural and ethnic minority psychology is being published and who the most productive authors are. The journals that published the most articles on cross-cultural and ethnic minority issues from 1993 to 1999 and the most prolific authors on these issues were identified by PsycINFO. Cross-cultural research is cross-national, whereas ethnic minority research involves groups of color within the United States. The citation impact of these journals and authors was determined from the 1997 Social Sciences Citation Index. The results suggest that there is very limited overlap between the literatures in cross-cultural and ethnic minority psychology. Most of the research in these areas is published in specialty journals, and there is a paucity of this research in prestigious journals. Perceived or actual barriers to publication in prestigious journals may cause some to seek specialty journals as outlets for research on cultural diversity. The top scholars in cross-cultural psychology are primarily men of European ancestry, whereas most of the top scholars in ethnic minority psychology are ethnic minority men and women. Strategies to increase the prominence of cultural diversity in the psychology literature include combining cross-cultural and ethnic minority psychology, increasing the number of editorial board members of prestigious journals having expertise in cultural diversity, and increasing the quality of specialty journals. Psychology will remain ill-equipped to face the challenges of the new millennium without increased attention to cultural diversity.

Cultural Diversity↗

A tumor necrosis factor-responsive long-term-culture-initiating cell is associated with the stromal layer of mouse long-term bone marrow cultures.

Long-term bone marrow cultures provide a model for the study of hematopoiesis. Both an intact, adherent stromal layer and hematopoietic stem cells are necessary components in these cultures. Mycophenolic acid treatment of mouse long-term bone marrow cultures depletes them of all assayable hematopoietic precursors. The residual stromal cells are functional and support hematopoiesis if new progenitor cells are supplied. We now show that these mycophenolic acid-treated stromal cell cultures contain cells capable of hematopoietic differentiation without the addition of new progenitors. When treated with tumor necrosis factor alpha (20-200 units/ml), the apparently pure stromal cultures undergo an intense burst of hematopoietic activity. After 4 days such cultures contain approximately 2 x 10(6) hematopoietic cells and, by 1 week, they are indistinguishable from control long-term cultures that were not treated with mycophenolic acid. These results suggest that the stromal cultures either contain hematopoietic stem cells that are maintained quiescent and mycophenolic acid-resistant, perhaps by intimate contact with the stroma, or contain adherent cells that can be induced to differentiate into hematopoietic stem cells. These stem cells are primitive, in that they are capable of multilineage development in the long-term cultures, but are unable to form spleen colonies or myeloid colonies in semisolid medium. These data demonstrate that the adherent fraction of cultured bone marrow contains very primitive hematopoietic cells and that tumor necrosis factor alpha activates their proliferation and differentiation. They also suggest a strategy for obtaining the earliest progenitors free of other, more mature cell types.

Animals↗

Tissue culture studies on human pituitary tumours: long term release of anterior pituitary hormones into the culture medium.

A study was undertaken to determine the length of time that human pituitary tumours are capable of releasing anterior pituitary polypeptide hormones in vitro under basal conditions and to study the spectrum of hormone release by functioning and "non-functioning" pituitary neoplasms. Fragments from the pituitary tumours of 10 patients in the following categories: 1 Cushing's disease, 2 with amenorrhoea-galactorrhoea, 3 with acromegaly, and 4 with "non-functioning" pituitary tumours and from 2 normal human anterior pituitary glands were placed in primary culture immediately after surgery. The in vitro release of human growth hormone (hGH), prolactin (Prl), thyrotrophin (TSH), adrenocorticotrophin (ACTH), luteinizing hormone (LH), and follicle stimulating hormone (FSH) was measured by specific radioimmunoassays at the end of each week in culture. Hormone release was surveyed from 6 weeks to 6 months depending upon the survival of the culture. Hormone release patterns were compared with clinical and pathological data. In the initial week of the study, all 6 anterior pituitary polypeptides were detected in the media from the 2 control pituitaries and from 4 of the tumours (1 amenorrhoea-galactorrhoea and 3 acromegaly) in concentrations up to 100 ng/ml of medium while 5 of the 6 hormones were readily detectable in the media from 2 additional tumour samples (Cushing's disease and 1 "non-functioning" pituitary tumour). The media of the remaining 4 tumours contained at least 3 of the 6 hormones (1 amenorrhoea-galactorrhoea and 3 "non-functioning" pituitary tumours). After 6 months in culture, the 6 hormones were readily detectable in at least 1 of the 5 surviving cultures and hGH (up to 800 ng/ml) and LH were each detectable in the media from 2 cultures. Although most of the hormone concentrations in the media decreased with length of time in culture, there were 2 exceptions. First in the media from 5 of the 12 cultures from both controls and tumours, Prl concentrations increased after 50 to 80 days culture. This increase usually lasted for several weeks before Prl levels again began to decline. The second unusual finding occurred in a tumour from a patient with acromegaly in the media of which hGH levels rose from 60 ng/ml to 800 ng/ml between days 125 and 174. These findings of prolonged hormone release in vitro give promise of future usefulness of tissue culture methods for study of polypeptide hormone releasing mechanisms and long-term production of human anterior pituitary hormones for use in research and possible therapy.

Acromegaly↗

[Primary culture of human functioning pituitary adenoma in monolayer and collagen gel culture].

Human functioning pituitary adenomas (3 acromegalies, 3 prolactinomas) obtained at transsphenoidal hypophysectomy were dispersively embedded inside collagen gels and subjected to the conventional monolayer culture in Eagle's MEM medium containing fetal bovine serum. Basal secretion of growth hormone (GH) and prolactin (PRL) in the media of monolayer and collagen gel cultures were measured by radioimmunoassay (RIA) for 5 weeks. GH secretion in the culture media was initially high in all cases with acromegaly in both monolayer and collagen gel cultures. GH secretion in monolayer culture declined rapidly almost as a straight line on a semilogarithmic scale until 5 weeks, when GH level decreased around 10 ng/ml. GH secretion in collagen gel culture was preserved more than in monolayer culture, but declined slowly up to about 500 ng/ml within 5 weeks. PRL secretion in monolayer culture initially declined rapidly until 1 week, but after then remained constant or decreased slowly. In the other hand, PRL secretion in collagen gel culture remained almost constant for up to 5 weeks. It is concluded that this new culture method may provide suitable conditions for maintenance of the adenomas cells in vitro.

Adenoma↗

Effects of homocysteine on smooth muscle cell proliferation in both cell culture and artery perfusion culture models.

BACKGROUND: Hyperhomocysteinemia is associated with increased risk for vascular disease. However, the pathogenic mechanisms of homocysteine are largely unknown. We evaluated the effects of homocysteine on smooth muscle cell (SMC) and endothelial cell proliferation in cell culture and on SMC proliferation of balloon angioplasty-injured arteries in a perfusion culture model. METHODS: Human and pig SMCs and endothelial cells were cultured with variable amounts of homocysteine for 72 h and the total cells were counted using a hemocytometer. Fresh pig carotid arteries were harvested from a local slaughterhouse and cultured in a newly designed artery perfusion culture system. Five groups of arteries (six per group) were cultured for 48 h under different conditions: normal control, balloon angioplasty injury alone, and injury with three different doses of homocysteine. Vessel viability was evaluated. SMC proliferation was assayed by bromodeoxyuridine (BrdU) DNA labeling. RESULTS: At concentrations equivalent to those in human hyperhomocysteinemia, homocysteine significantly stimulated both cultured human and pig SMC proliferation with a dose-dependent effect, while it inhibited cultured endothelial cell growth. Perfusion-cultured pig carotid arteries remained contractile in response to norepinephrine and relaxant to nitroglycerine, and viable cells were also isolated from the cultured arteries. SMC proliferation (BrdU index) showed significant differences among the groups. SMC proliferation was stimulated by vascular injury and further enhanced by homocysteine in a dose-dependent manner. The proliferative response occurred strongly on the luminal side of the vessel wall, with the effects tapering toward the adventitia. CONCLUSIONS: Homocysteine had a mitogenic effect on vascular SMCs and a cytotoxic effect on endothelial cells. This differential effect of homocysteine on vascular cells may represent a pathogenic mechanism of vascular lesion formation in patients with hyperhomocysteinemia.

Angioplasty, Balloon↗

Adult rat hepatocyte microcarrier culture. Comparison to the conventional dish culture system.

A technique has been devised to attach adult rat hepatocytes to collagen-coated dextran microcarriers. Cells were cultured serum-free for 2 d and their viability, enzyme activities, glucose metabolism, and hormone responsiveness were compared to data obtained from conventional dish cell culture. The two different culture methods showed no difference in cell viability and morphology. Microcarrier-cultured cells exhibited hormone responsiveness comparable to dish cultures; glycolysis could be activated three-fold by the sole addition of insulin, and gluconeogenesis was increased by 40 to 50% by glucagon. During the 48-h culture glucokinase and phosphoenolpyruvate carboxykinase activities declined at a similar rate in both culture systems. Long-term culture with 0.1 microM insulin prevented the decrease of glucokinase activity. Insulin responsiveness (activation of glycolysis) was still pronounced after 48 h in culture. The microcarrier technique establishes a new in vitro liver system in which acute and long-term hormonal actions can be investigated using the technical advantages of a suspension culture.

Animals↗

Enhancement of entomopathogenic nematode production in in-vitro liquid culture of Heterorhabditis bacteriophoraby fed-batch culture with glucose supplementation.

Nematode yield is a decisive factor for successful large-scale commercial production of entomopathogenic nematode. Various carbon sources were tested in in-vitro liquid culture to improve the yield of the entomopathogenic nematode Heterorhabditis bacteriophora. Canola oil was the optimal carbon source for nematode culture compared to carbohydrates when applied as a sole carbon source. However, when some of carbohydrates were applied together with canola oil, significant increases in nematode yield were observed. When 25 mg glucose ml(-1) was supplemented to 25 mg oil-based liquid culture medium ml(-1), the highest nematode yield, 3.62 x 10(5) infective juveniles, was achieved at 12 days, but nematode growth was suppressed at higher than 75 mg glucose ml(-1). A fed-batch culture process was introduced in nematode liquid culture consisting of two growth phases: bacteria and nematode. In the oil fed-batch culture, in which only glucose was initially added and oil was fed to the culture after the bacterial growth phase concurrent with nematode inoculation, nematode yield increased up to 4.25 x 10(5) infective juveniles ml(-1), while the batch culture resulted in 3.60 x 10(5) infective juveniles ml(-1). These results indicate that glucose is a superior carbon source for the bacteria, whereas canola oil is optimal for the nematode. The application of fed-batch culture provides significant enhancement of nematode yield in in-vitro liquid culture.

Animals↗

Fibronectin production by cultured human lung fibroblasts in three-dimensional collagen gel culture.

In vivo, fibroblasts are distributed in a three-dimensional (3-D) connective tissue matrix. Fibronectin is a major product of fibroblasts in routine cell culture and is thought to regulate many aspects of fibroblast biology. In this context, we sought to determine if the interaction of fibroblasts with a 3-D matrix might affect fibronectin production. To examine this hypothesis, fibronectin production by fibroblasts cultured in a 3-D collagen gel or on plastic dishes was measured by ELISA. Fibroblasts in 3-D gel culture produced more fibronectin than those in monolayer culture. Fibroblasts in 3-D culture produced increasing amounts of fibronectin when the collagen concentration of the gel was increased. The 3-D nature of the matrix appeared to be crucial because plating the fibroblasts on the surface of a plastic dish underneath a collagen gel was not different from plating them on a plastic dish in the absence of collagen. In addition to increased fibronectin production, the distribution of the fibronectin produced in 3-D culture was different from that of monolayer culture. In monolayer culture, more than half of the fibronectin was released into the culture medium. In 3-D culture, however, approximately two-thirds remained in the collagen gel. In summary, the presence of a 3-D collagen matrix increases fibroblast fibronectin production and results in greater retention of fibronectin in the vicinity of the producing cells.

Blotting, Northern↗

Cultural attitudes, knowledge, and skills of nursing faculty toward patients from four diverse cultures.

This study used a convenience sample of 94 nursing faculty recruited among attendees of two professional conferences to describe faculty attitudes toward and perceived level of confidence in cultural knowledge of patients representing four ethnic groups. The study also explored relationships between these variables and the respondents' demographic characteristics. Respondents completed the Cultural Attitudes Scale, the Cultural Self-efficacy Scale, and a demographic survey and wrote a response to an open-ended question. Mean scores indicated moderately positive attitudes toward and moderate-to-high confidence levels in cultural knowledge related to caring for four ethnic groups. Three variables accounted for 33% of the variability in level of confidence in skill in transcultural care: cultural content was included in the curriculum of the employing nursing school, education courses in culture, and the predominant ethnicity of the patients for whom care was provided. Repeated exposure to persons from other cultures was the most frequently cited means of increasing personal comfort in caring for ethnically diverse patients. Only 53% of the respondents remembered cultural content in their own academic preparation. Findings suggest that cultural exposure opportunities enhance faculty-perceived confidence in knowledge and attitudes toward people from other cultures.

Adult↗

Comparison of BACTEC 460 and MGIT 960 systems for the culture of Mycobacterium avium subsp. paratuberculosis S strain and observations on the effect of inclusion of ampicillin in culture media to reduce contamination.

To compare the utility and diagnostic accuracy of BACTEC and MGIT culture systems, a total of 41 pooled faecal samples, each containing faeces from one sheep infected with the S strain of Mycobacterium paratuberculosis and four uninfected sheep was cultured. The MGIT culture system did not support the growth of the S strain of M. paratuberculosis from faeces within the time frame of the experiments, although a laboratory adapted S strain grew slowly in MGIT provided that sufficient bacteria were inoculated. In contrast, C strain grew rapidly in MGIT. The sensitivity of culture was calculated relative to the infection status of the animals, none of which had clinical signs of ovine Johne's disease. The overall sensitivity of pooled faecal culture in the BACTEC culture system was 21.9% (95% confidence limits, 10.5-37.6), a figure dependant on the proportion of multibacillary cases. The sensitivities of the BACTEC culture system for pools containing animals with multibacillary and paucibacillary lesions were 100.0% (95% confidence limits, 47.2-100.0) and 17.8% (95% confidence limits 6.06-36.8), respectively. The contamination rate of BACTEC cultures was 9.7% compared to 14.3% for MGIT. The effect of 100 microg/ml ampicillin on the S strain of the M. paratuberculosis was examined and in both BACTEC and MGIT media it delayed growth by about 1 week. The composition of MGIT medium, particularly presence of vancomycin hydrochloride, slowed the growth of the S strain. The low content of egg yolk was considered to be another possible factor. The radiometric BACTEC culture system remains the best alternative for the culture of S strain and is recommended in circumstances where the genotype (s) of the strains present in a region/farm is either unknown or S strain.

Ampicillin↗

The cultural formulation: a method for assessing cultural factors affecting the clinical encounter.

The growing cultural pluralism of US society requires clinicians to examine the impact of cultural factors on psychiatric illness, including on symptom presentation and help-seeking behavior. In order to render an accurate diagnosis across cultural boundaries and formulate treatment plans acceptable to the patient, clinicians need a systematic method for eliciting and evaluating cultural information in the clinical encounter. This article describes one such method, the Cultural Formulation model, expanding on the guidelines published in DSM-IV. It consists of five components, assessing cultural identity, cultural explanations of the illness, cultural factors related to the psychosocial environment and levels of functioning, cultural elements of the clinician-patient relationship, and the overall impact of culture on diagnosis and care. We present a brief historical overview of the model and use a case scenario to illustrate each of its components and the substantial effect on illness course and treatment outcome of implementing the model in clinical practice.

Attitude to Health↗