Search PubMed⌕ Search

Biomedical subjects

K L White

Publications and source records attributed to K L White.

At least 109 records · Page 6Linked to original sources
PubMed1993

Mouse Hepa 1c1c7 hepatoma cells produce complement component C3; 2,3,7,8-tetrachlorodibenzo-p-dioxin fails to modulate this capacity.

Previous studies from this laboratory have shown that 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) decreased complement component C3 levels in female B6C3F1 mouse serum following in vivo acute or subchronic exposure (White et al., 1986). Since TCDD is a hepatotoxic compound and more than 90% of serum C3 is produced by the liver, studies were undertaken using mouse Hepa 1c1c7 (Hepa 1) hepatoma cell line to determine if TCDD acts directly on hepatocytes to inhibit C3 production. The C3-producing capacity of Hepa 1 cells was first examined. When confluent Hepa 1 cell monolayers were cultured in 24-well plates with serum-free medium, a detectable amount of C3 (14.1 +/- 0.8 ng/ml) was secreted as early as 1 h after culture and reached a plateau at 12 h (68.3 +/- 4.9 ng/ml). Furthermore, the sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) analysis demonstrated that the molecular weight of C3 in culture supernatant corresponded to that present in mouse serum. Human recombinant IL-1 beta (hrIL-1 beta), a known inducer of complement C3, at doses as low as 1 unit/ml increased the C3 production to 158% of control after 24 h of incubation. The effect of hrIL-1 beta was dose dependent, and the maximum tested dose of 10 units/ml increased C3 production to 256% of control. When cells were directly exposed to TCDD at concentrations from 10(-10) to 10(-6) M, there was no inhibitory effect on production of C3. TCDD also failed to block the stimulatory effect of 10 units/ml hrIL-1 beta added to the culture 1 h later. To verify that cultured Hepa 1 cells were able to respond to TCDD, 7-ethoxyresorufin O-deethylase (EROD) activity was measured under the same conditions. TCDD dose-dependently increased EROD activity of Hepa 1 cells at 24 h following exposure. The activity reached 56.7 +/- 3.0 pmol/min/mg protein with 10(-9) M TCDD, compared with 10.7 +/- 1.7 pmol/min/mg protein of vehicle-exposed cells. Our results indicate that the direct interaction of TCDD with Hepa 1 cells does not affect their C3-producing capacity, although EROD activity, a characteristic response mediated by the cellular TCDD/Ah receptor, was induced. The lack of effect of TCDD in vitro suggests that the decrease of serum C3 levels observed in vivo may result from an indirect effect of TCDD on hepatocytes.

Animals↗
PubMed1993

Modulation of liver intracellular C3 in mice by 2,3,7,8-tetrachlorodibenzo-p-dioxin.

Earlier studies from this laboratory have shown that the complement system, especially the component C3, in female B6C3F1 mice is suppressed following TCDD exposure in vivo. However, the direct exposure of TCDD in vitro does not affect the C3-producing capacity of two types of hepatoma cells, as well as mouse primary hepatocytes. To investigate the effect of TCDD on C3 production by the liver following in vivo exposure, liver intracellular C3 levels and pro-C3, the precursor of the secreted C3, were examined in the present study. The results demonstrated that there was a dose-dependent increase of liver intracellular C3 levels (from 138% to 175% of control) immediately following TCDD (from 10 to 40 micrograms/kg) exposure. This increase was rapid (4 h after exposure), but transient (less than 2 h), and was not accompanied by an alteration of serum C3 levels. Studies using sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) analysis showed that the increase in liver intracellular C3 levels resulted from, at least partially, an increase in intracellular pro-C3. Serum C3 levels did not decrease until d 3 after exposure, when both liver intracellular C3 levels and pro-C3 in TCDD-treated mice were not different from those of the control mice. These results indicated that the modulation of liver intracellular C3 by TCDD did not correlate with the suppression of serum C3 levels following exposure.

Administration, Oral↗
PubMed1993

Production of complement component C3 in vivo following 2,3,7,8-tetrachlorodibenzo-p-dioxin exposure.

Exposure to 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) has been shown to decrease serum complement C3 levels in female B6C3F1 mice but failed to alter C3 production when added in vitro to either hepatoma cells (both human and mouse hepatoma cells) or mouse primary hepatocytes (Lin and White, 1993a). It has also been demonstrated that mouse liver intracellular C3 levels were not affected following TCDD exposure in vivo, while serum C3 levels were suppressed (Lin and White, 1993b). Therefore, further studies were undertaken to investigate the mechanism by which TCDD modulates newly synthesized serum C3 in vivo. Mouse serum C3 was depleted by an intravenous injection of 50 anti-complement units (ACU)/kg cobra venom factor (CVF). This dose of CVF depleted serum C3 levels to 9% of control at 24 h after treatment. Subsequently, serum C3 levels returned to 19% and 75% of the control level on d 3 and d 5. The recovery of serum C3 was then monitored following an acute oral exposure to 20 micrograms/kg TCDD. In mice exposed to both TCDD and CVF, serum C3 levels reached 15% and 69% of control on d 3 and d 5 after treatment; these results were not significantly different from those of mice treated with CVF alone. Furthermore, when the radiolabeled amino acid [3H]leucine was injected intravenously into either vehicle- or TCDD-treated mice, the incorporation of this labeled precursor into both C3 and other secreted plasma proteins was not inhibited by TCDD. These results demonstrated that TCDD did not decrease newly synthesized C3 in vivo. These studies provide additional support for the concept that TCDD does not act directly on hepatocytes to suppress C3 production. The lower serum C3 levels observed in vivo following TCDD exposure is not the result of a decrease in C3 production by hepatocytes.

Administration, Oral↗
PubMed1993

Comments on Navarro's review of Health services research: an anthology.

In preparing Health Services Research: An Anthology, the editors attempted to broaden the definition of health services research promulgated by the National Library of Medicine. Many articles in the Anthology focused on stemming the avalanche of medicine's unevaluated procedures and prescribing practices, and on broadening the horizons of the medical establishment. Another anthology is clearly needed to address Virchow's dictum that medicine is politics writ large.

Complementary Therapies↗
PubMed1992

Enhanced in vitro growth of murine fibroblast cells and preimplantation embryos cultured in medium supplemented with an amphipathic peptide.

Preliminary studies on the proliferative effects of lytic peptides were carried out using NIH 3T3 murine fibroblast cells and human lymphocytes. Cells were cultured in various concentrations of three different amphipathic peptides (SB-37, Shiva-1, and Vishnu), and enhanced proliferation was determined by uptake of 3H-thymidine with treated cells compared with control cultures. Enhanced proliferation of 3T3 cells was observed in cultures containing 50 microM or less SB-37. The primary study consisted of 263 four-cell- to eight-cell-stage mouse embryos from naturally bred mice and incubated in Whitten's medium containing 0.2, 1, or 10 microM of the amino terminus of an amphipathic cecropin B analog (Vishnu) or in Whitten's medium alone. Embryos were cultured to the hatched blastocyst stage, and effect of treatment was determined by the rate of growth to that stage of development. Statistical analysis revealed that culture in all three levels of Vishnu significantly accelerated in vitro growth of these stages of preimplantation embryos compared with controls. These results indicate that Vishnu promotes increased cleavage rates of embryos in vitro. A growth factor receptor clustering mechanism of action is proposed. This peptide may have some potential as an embryo culture medium additive to enhance in vitro growth rate.

3T3 Cells↗
PubMed1992

Electrofusion-induced intracellular Ca2+ flux and its effect on murine oocyte activation.

These experiments were designed to monitor influx of extracellular Ca2+ into the murine ooplasm following a 1.56 kV.cm-1 direct current (DC) electrofusion pulse and subsequently to determine its effect on rate of activation. Pulse media consisted of non-electrolyte (0.3 M mannitol) and electrolyte (phosphate-buffered saline; PBS) media each containing 0.0, 0.05, or 0.9 mM Ca2+ (groups T1-T3 and T4-T6, respectively). Cumulus-free oocytes were incubated in 100 microliters drops of PBS containing 2 microM of the calcium indicator fluo-3/AM for 60 min at 37 degrees C. Fluo-3/AM-loaded oocytes were equilibrated for 7 min in assigned treatment media (T1-T6) prior to application of DC pulse. Change in fluorescent intensity was monitored for 6.5 min after DC pulse by photon counting spectrofluorometry. Fluorometric measurements demonstrate a dramatic rise in intracellular free Ca2+ (Ca2+i) following DC pulse is associated with Ca2+ ion concentration in the pulse medium. Significantly (P less than 0.01) higher Ca2+i levels were observed when 0.9 mM Ca2+ was added to the pulse medium (T3 and T6) compared with pulse medium containing lower Ca2+ ion concentrations (T1, T2, T4, and T5; P greater than 0.05). Differences (P less than 0.01) were observed in peak Ca2+i levels 18 sec after pulse with mean percent change in fluorescence of 5.1%a, 33.9%b, 112.7%c, 1.2%a, 9.3%a, and 99.9%c for T1-T6, respectively (values with different superscripts are significantly different at P less than 0.01). Increased oocyte membrane permeability to Ca2+ ion after DC pulse was observed for a minimum of 5 min after delivery of the 1.56 kV.cm-1 pulse.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1992

Effect of electrofusion pulse in either electrolyte or nonelectrolyte fusion medium on subsequent murine embryonic development.

This study was designed to determine what effect electropulse parameters would have on rate of fusion, lysis, and embryo viability when embryos were subjected to electrofusion treatment in nonelectrolyte or electrolyte pulse media. Previous experiments have shown electrolyte medium (i.e., phosphate-buffered saline; PBS) to have a positive effect on electric pulse-induced murine oocyte activation. In addition, these results also indicated that pulse media containing 0.9 mM Ca2+ induced a dramatic increase in the rate of murine oocyte activation compared with oocytes pulsed in media containing 0.0 or 0.05 mM Ca2+. Pronuclear or two-cell-stage embryos were obtained from superovulated prepubertal randomly bred Swiss (albino) female mice. Embryos were randomly assigned to three nonelectrolyte and three electrolyte treatment media. Nonelectrolyte media consisted of 0.3 M mannitol (T1), 0.3 M mannitol + 0.05 mM CaCl2 (T2), and 0.3 M mannitol + 0.9 mM CaCl2 (T3). Electrolyte media consisted of Ca(2+)-free PBS (T4), PBS containing 0.05 mM CaCl2 (T5), and PBS containing 0.9 mM CaCl2 (T6). Three experiments were carried out; the objective of the first was to determine the rate of fusion and rate of lysis in murine two-cell embryos placed in the two types of (0.3 M mannitol, T1-T3; and PBS, T4-T6) fusion media and subjected to a fusion procedure (3 V, 5 sec AC alignment pulse, followed by a 1.56 kV.cm-1, 99 microsec DC fusion pulse). Control two-cell embryos were placed in T1 for 2 min and did not receive a fusion pulse.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1992

Metabolism of benzo[a]pyrene by murine splenic cell types.

The objective of the present study was to determine which splenic cell type(s) of B6C3F1 mice was capable of metabolizing B(a)P. Separation of splenocytes based on density by centrifugation through discontinuous Percoll gradients along with immunomagnetic negative selection or antibody-mediated complement lysis was utilized to obtain highly enriched populations of splenocytes for B(a)P metabolism studies. Immunofluorescent cell staining in conjunction with flow cytometry and examination of Giemsa-stained cytospin cell preparations indicated that B- or T-cell populations of greater than 95% purity and an 80-90% pure population of splenic macrophages were routinely attained. Splenic cell populations were incubated with [3H]B(a)P for 24 hr. High-pressure liquid chromatography was used to separate and quantitate the B(a)P metabolites generated by the enriched splenic cell populations. The results of these studies demonstrate that the macrophage is the cell type responsible for the metabolism of B(a)P within the spleen. The major metabolites of B(a)P produced were as follows: an unidentified peak of polar metabolites containing polyhydroxylated metabolites, B(a)P-9,10-dihydroxy-9,10-dihydrodiol, and B(a)P-7,8-dihydroxy-7,8- dihydrodiol. Other splenic cell types examined, including B and T cells, polymorphonuclear cells, or the spleen capsule did not produce amounts of B(a)P metabolites significantly above background levels. Based on these findings, macrophages are the splenic cell types which metabolize B(a)P. As a result, macrophages may be the cell type targeted by B(a)P resulting in suppression of splenic humoral immune responses.

Animals↗
PubMed1992

Evaluation of murine splenic cell type metabolism of benzo[a]pyrene and functionality in vitro following repeated in vivo exposure to benzo[a]pyrene.

Recent studies have demonstrated that macrophages are the cell types capable of metabolizing benzo[a]pyrene (B(a)P) within the spleens of untreated mice. Since repeated exposure to B(a)P results in immunosuppression and B(a)P is known to induce cytochrome P450 levels, the first objective of this study was to investigate whether exposure of mice to B(a)P could increase the amounts of immunosuppressive B(a)P metabolites generated and/or alter the pattern of B(a)P metabolites formed by several different splenic cell types. Mice were dosed with a daily sc dose of 200 mg/kg B(a)P or vehicle for 4 days. Separation of splenocytes based on density by centrifugation through discontinuous Percoll gradients along with immunomagnetic negative selection or antibody-mediated complement lysis was used to obtain different splenic cell populations. Cells were incubated with [3H]B(a)P for 24 hr. High-pressure liquid chromatography was used to separate and quantitate B(a)P metabolites. Results indicate that splenic macrophages of B(a)P-treated mice produced significantly greater amounts of some metabolites compared to those of vehicle-treated mice. The three major metabolites produced were an unidentified peak of polar metabolites containing polyhydroxylated metabolites, B(a)P-9,10- and B(a)P-7,8-dihydrodiols. Other splenic cell types examined did not produce metabolite amounts significantly above (T-cells, PMNs, or the capsule) or just above (B-cells) background. The second objective was to investigate the splenic cell type(s) targeted by B(a)P resulting in suppression of humoral immunity. Separation-reconstitution studies along with in vitro sensitization techniques with several different antigens (sheep red blood cells (SRBC), dinitrophenyl-Ficoll (DNP-Ficoll), lipopolysaccharide (LPS)) were used to identify splenic target cells following exposure of mice to B(a)P (200 mg/kg/day, sc for 4 days). Findings indicate that in vitro plaque-forming cell (PFC) suppression was due to alterations in the adherent (macrophage) cell population. Exposure also suppressed the PFC response to the T-dependent antigen SRBC and the T-independent antigen DNP-Ficoll, but did not suppress the PFC response to the polyclonal antigen, LPS. These data suggest that B(a)P is targeting macrophages.

Animals↗
PubMed1992

Generation of 7,8-dihydroxy-9,10-epoxy-7,8,9,10-tetrahydrobenzo[a]pyrene by murine splenic macrophages.

Studies have demonstrated that macrophages are the cell types which metabolize benzo[a]pyrene (B[a]P) within the murine spleen. B and T cells, polymorphonuclear cells, or the splenic capsule did not produce amounts of B[a]P metabolites above those of background. Exposure of mice to B[a]P, a known inducer of isozymes of cytochrome P450, resulted in an increase in the amounts of some B[a]P metabolites generated by macrophages. Evaluation of the in vitro plaque-forming-cell response to several T-cell and macrophage-dependent antigens following in vivo exposure to B[a]P indicated that the macrophage is the cell type responsible for B[a]P-induced immunosuppression. While suggestive, the reported data have not definitively established that an enriched splenic macrophage population can generate 7,8-dihydroxy-9,10-epoxy-7,8,9,10-tetrahydro-B[a]P (BPDE) from B[a]P-7,8,-dihydroxy-7,8-dihydrodiol (B[a]P-7,8-diol). These data are critical to our hypothesis that the splenic cell type(s) which form BPDE will be the primary target cell responsible for B[a]P-induced immunosuppression. The first objective was to determine if splenic macrophages could generate BPDE. Enriched (80-90% purity) populations of macrophages were incubated with [3H]B[a]P for 24 hr. BPDE generated was quantitated by analysis of the cis- and trans-tetrol hydrolysis products of BPDE via HPLC procedures. Splenic macrophages generated BPDE from B[a]P. The cis syn was the predominate tetrol detected. Exposure of mice to B[a]P increased the amounts of the trans-anti-tetrols 2.2-fold, the trans-syn-tetrols 2.0-fold, the cis-anti-tetrols 1.8-fold, and the cis-syn-tetrols 2.6-fold above those formed by macrophages of vehicle-exposed mice. Both cytochrome P450- and peroxyl radical-dependent pathways are known to oxidize B[a]P-7,8-diol to BPDE. Since macrophages were found to generate BPDE, the second objective was to investigate which enzymatic pathway was responsible for its formation. The B[a]P-(+)-7,8-diol isomer has been shown to produce different specific BPDE isomers via the cytochrome P450 and peroxyl radical pathways. Macrophage populations were incubated with [3H]B[a]P-(+)-7,8-diol for 24 hr and the contribution of the cytochrome P450 and peroxyl radical pathways to BPDE formation determined by detection of syn-BPDE-hydrolysis and anti-BPDE-hydrolysis products, respectively. Based on the ratio of anti/syn BPDE-derived tetrol products, the results demonstrate that splenic macrophages can generate the BPDE by both a cytochrome P450-dependent and -independent (peroxyl radical) pathway. Macrophages are the cells which metabolize B[a]P within the murine spleen and may be the cell type responsible for B[a]P-induced suppression of humoral immune responses.

7,8-Dihydro-7,8-dihydroxybenzo(a)pyrene 9,10-oxide↗
PubMed1992

In vitro maturation and fertilization of swamp buffalo oocytes and their subsequent development.

The present experiment was carried out to evaluate the maturation, fertilization and subsequent embryo culture of swamp buffalo oocytes in vitro. The oocytes (n=273) were collected and morphologically graded based on the structure of cumulus-oocyte complexes as Grade 1 (compact, n=81), Grade 2 (expanded, n=70), Grade 3 (partially denuded, n=65) or Grade 4 (completely denuded, n=57). More than 60% of the in vitro matured oocytes co-cultured with capacitated spermatozoa demonstrated evidence of fertilization or cleavage to the 2-cell stage when either Grade 1 or 2 oocytes were used. The percentage of fertilized oocytes undergoing 2-cell stage cleavages from Grade 3 (53%) and Grade 4 (46%) groups was significantly lower (P<0.01) than that observed in the Grade 1 (64%) and Grade 2 (68%) groups. Development to the 6 to 8 cell stage substantiated fertilization of Grade 1 and 2 oocytes. These results demonstrated that swamp buffalo oocytes are capable of maturing in vitro, forming embryos, and developing at least to the 8-cell stage in culture medium alone.

Journal Article↗
PubMed1992

PGE2-independent immunosuppressive activity of horse trophoblast tissue.

It has been proposed that PGE2 is an important immunosuppressant acting at the fetal-maternal interface during pregnancy. We have previously shown that horse conceptus-conditioned medium suppresses lymphocyte proliferation. This experiment was designed to determine if horse conceptus-derived immunosuppressive activity could be attributed to PGE2 production by the trophoblast tissue. Trophoblast tissue from 21-day-old conceptuses was cut into equal sections and cultured in the presence or absence of the prostaglandin inhibitor, indomethacin. Following culture, immunosuppressive activity and the concentration of PGE2 were determined for each sample of both horse-trophoblast conditioned medium (HTCM) and indomethacin-treated HTCM (I-HTCM). Suppressive activity was identified in lymphocyte proliferation assays via reduced [3H]thymidine uptake by pokeweed mitogen stimulated horse lymphocytes. A radioimmunoassay was used to quantify PGE2. While PGE2 production was greatly reduced in cultures containing indomethacin, trophoblast-derived immunosuppressive activity was not affected. These data indicate that PGE2 is not the primary immunosuppressant produced by horse trophoblast tissue.

Animals↗
PubMed1992

Risk assessment in immunotoxicology. I. Sensitivity and predictability of immune tests.

We have previously reported on the design and content of a screening battery involving a "tier" approach for detecting potential immunotoxic compounds in mice (Luster et al., 1988, Fundam. Appl. Toxicol. 10, 2-19). This battery has now been utilized to examine a variety of compounds by the NIEHS Immunotoxicology Laboratory, the National Toxicology Program-sponsored laboratories, and by the Cell Biology Department at the Chemical Industry Institute of Toxicology. The database generated from these studies, which consists of over 50 selected compounds, has been collected and analyzed in an attempt to improve future testing strategies and provide information to aid in quantitative risk assessment for immunotoxicity. Studies presented here have established the ability of each of the tests or test combinations in the screening battery to detect immunotoxic compounds. Efforts are currently underway using this database to determine the relationships between these immune tests and susceptibility to challenge with infectious agents or transplantable tumor cells. The present analyses indicated that the performance of only two or three immune tests are sufficient to predict immunotoxic compounds in rodents (greater than 90% concordance). The tests that showed the highest association with immunotoxicity were the splenic antibody plaque forming cell response (78%) and cell surface marker analysis (83%). The relationship between immunotoxicity and carcinogenicity, as well as genotoxicity, was also determined. These analyses suggested that potential immunotoxic compounds are likely to be rodent carcinogens (p = 0.019) although for compounds that are not immunotoxic the carcinogenic status is unclear. There was no relationship observed between immunotoxicity and mutagenicity as determined using in vitro genotoxicity tests. The significance of these observations is discussed in terms of the relationship between immunotoxicity tests and biological/toxicological processes concerned with human health (e.g., infectious disease).

Animals↗
PubMed1992

Involvement of interleukin 2 receptors in conceptus-derived suppression of T and B cell proliferation in horses.

The mechanism by which a horse conceptus-derived immunosuppressive factor (HCS) of M(r) > 100,000 inhibits lymphocyte proliferation was investigated. The factor was obtained from the culture supernatants of 20-day-old horse conceptuses; activity, identified by reduced uptake of [3H]thymidine by mitogen-stimulated lymphocytes, was greatest (P < 0.01) in cultures stimulated by mitogen from pokeweed. HCS also suppressed cell proliferation stimulated by phytohaemagglutinin (P < 0.01), but had no effect on lipopolysaccharide-stimulated cells (P > 0.05). Data from a fluorescence-activated cell sorter indicated that supplementation with HCS reduced the number of T cells in phytohaemagglutinin-stimulated cultures and suppressed proliferation of T and B cells in pokeweed-mitogen-stimulated cultures compared with controls. Cell proliferation was greater (P < 0.01) in cultures supplemented with HCS 24 h after stimulation than in those treated at the start of stimulation, and was even greater (P < 0.01) when cells were treated 48 h after stimulation. The removal of HCS from treated lymphocyte cultures resulted in complete recovery of cell responsiveness, and stimulated proliferation of treated cells did not differ (P > 0.05) from that of control cells. The addition of stimulated equine lymphocyte supernatant to cultures supplemented with HCS did not significantly increase (P > 0.05) cell proliferation in response to pokeweed mitogen. Addition of recombinant human interleukin 2 (rIL-2) to HCS-treated cultures did not alter the suppressive activity of HCS, although cell proliferation was greater in cultures supplemented with rIL-2 than in controls (P < 0.01). HCS inhibition of IL-2 receptor (IL-2R) function was investigated using an IL-2-dependent murine cytolytic T lymphocyte cell line; the fraction of HCS of M(r) > 100,000 had no effect (P > 0.05) on proliferation of IL-2-dependent murine cytolytic T lymphocyte cells induced by rIL-2. Together, these data suggest that HCS suppresses proliferation of T lymphocytes during the early stages of cell activation by inhibiting IL-2R interaction and that this suppression interferes with interactions between T cells and B cells, thereby also indirectly inhibiting proliferation of B cells. The potent immunosuppressive capacity of HCS may be one factor responsible for inhibiting cell-mediated fetal allograft rejection during pregnancy.

Animals↗
PubMed1992

All-trans retinoic acid in the treatment of acute promyelocytic leukaemia.

All-trans-retinoic acid (ATRA) is known to induce differentiation of promyelocytes in vitro and also to induce remission of acute promyelocytic leukaemia in vivo. We treated 11 patients with poor prognosis acute promyelocytic leukaemia (APL) with ATRA and obtained seven complete and one partial remission. Remissions took one to three months to achieve and were associated with adverse effects including dry skin and bone pain. In eight patients the white cell count rose above 20 x 10(9)/L within the first ten days of retinoic acid treatment and this was associated with the development of pulmonary leukostasis in three patients which was fatal in one. Another two patients died of intracranial haemorrhage also within the first ten days. ATRA is a promising new agent in the induction therapy of this particular category of acute leukaemia.

Adult↗
PubMed1992

The use of image analysis to evaluate the development of uterine and oviduct epithelial cells during in vitro culture. A potential quality assurance procedure for in vitro laboratories.

The objective of this study was to establish selection criteria for morphologic assessment of cell quality using a computer image-analysis system. Uterine and oviduct epithelial cells were isolated from the reproductive tracts of cyclic cows using a trypsin solution. Harvested cells were cultured in Tissue Culture Medium-199 with 10% fetal bovine serum and maintained at 37 degrees C with 5% carbon dioxide in air. Subcultures of different confluent monolayers were established using a weak trypsin solution. Morphologic assessment of cell integrity and viability were made visually, with electron microscopy and image analysis. After morphologic assessment, cells were assigned quality scores. Evaluations were conducted on cells during primary culture and following the first and third subpassages. Image-analysis evaluation was conducted on cells using a silicon-intensified target camera and computer-based software. Gray-level density was determined on cells from each flask. There was no difference (P greater than .05) between sample replicates, indicating repeatable measurements were obtained with the image-analysis system. In addition, there was no significant difference in parameters measured between uterine and oviduct cells; therefore, data from both cell types were combined for further statistical comparisons. Gray-level density values for combined uterine and oviduct cells during primary culture and following the first and third subpassages were as follows: 139.7, 154.5, and 173.3, respectively. There was an increase (P less than .05) in gray-level density values with each subpassage of the cell populations. Furthermore, gray level was influenced (P less than .01) by cell quality for combined uterine and oviduct cell populations. These results indicate that epithelial cell quality during in vitro culture may be effectively determined using image analysis. This approach should not be overlooked in establishing quality control measures for uterine and oviduct cells before mammalian embryos are cocultured in vitro.

Animals↗