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Scanning-electron microscopy studies of 'ground substance' in the cement lines, resting lines, hypercalcified rings and reversal lines of human cortical bone.

Adult human cortical bone was decalcified and stained with Harris's hematoxylin, which has a strong affinity for cement lines, resting lines, hypercalcified rings and reversal lines. Small pieces of bone tissue containing these morphological entities were then dissected and manipulated mechanically so as to disrupt them partially. Scanning electron microscopy of these regions of interest revealed the presence of relatively large concentrations of amorphous 'ground substance' which was morphologically distinguishable from the fibrous collagen fiber bundles located in the adjoining collagen-rich lamellae. The methodology used in this study confirms the existence of the above-mentioned regions as true morphological entities and also makes possible enzymatic studies designed to identify the major biochemical components constituting the observed 'ground substance'.

Bone and Bones↗

Effect of direct cell-to-cell interaction between the KM-102 clonal human marrow stromal cell line and the HL-60 myeloid leukemic cell line on the differentiation and proliferation of the HL-60 line.

Hematopoietic cellular interaction was investigated in a coculture of the human clonal marrow stromal line, KM-102, and the myeloid leukemia cell line, HL-60. In the coculture, a large number of HL-60 cells remained in the supernatant but some of them became firmly attached to KM-102 cells. The attached HL-60 cells showed little positive reaction in the NBT test when the culture was supplemented with 10(-9) to 10(-7) M 1 alpha, 25-dihydroxyvitamin D3. In contrast, differentiation in the supernatant HL-60 cells was strikingly responsive to the agent in a dose-dependent way. Furthermore, the complete inhibition was observed in the incorporation of [3H]thymidine into the attached HL-60 cells with autoradiography, but 23.6% of the supernatant cells moderately incorporated [3H] thymidine into their nuclei. There was no attachment between HL-60 cells and stromal cells from human thymus and lymph node, or between lymphocytic leukemia cells and marrow stromal cells. These results indicate that there is direct cellular interaction between myeloid leukemic cells and marrow stromal cells which modulates the proliferation and differentiation of the myeloid leukemic cells. This modulation by marrow stromal cells is more strongly affected by this interaction than by exogenously added differentiation-inducing agents. Apparently marrow stromal cells produce a definitive milieu for the proliferation and differentiation of myeloid leukemic cells.

Bone Marrow Cells↗

Effect of crossing a line selected for increased shank width with two commercial sire lines on performance and walking ability of turkeys.

Published research indicated that a line (FL) of turkeys selected for increased shank width and exhibiting good walking ability (WA) and improved leg structure was useful in improving WA of a cross with an unimproved commercial sire line while remaining competitive in BW and body conformation. The purpose of the present study was to determine whether the FL line was useful in improving WA of two improved commercial primary-breeding sire lines and to study the inheritance of BW traits in the pure lines and reciprocal crosses of the pure lines. Samples of a primary breeding sire line from each of two major commercial turkey breeders were obtained as unpedigreed eggs and designated as Lines A and B. Lines A and B were reciprocally crossed with the FL line that had been selected for increased shank width for 16 generations. Pure lines and reciprocal crosses were produced by artificially mating 10 males to 15 females using pooled semen. The same semen pools were used to produce both the pure lines and the crosses. Traits measured included BW at 8, 16, and 20 wk of age, and length, width, and depth of the shank, width of the breast, and WA at 16 wk of age. The A and B lines were larger and had wider breasts and narrower and shorter shanks than the FL line. Line B was larger than Line A. Shank measurements and WA score did not differ between Lines A and B in either sex. Breasts of Line B females were wider than those of Line A females, but there was no sire line difference in breast width of males. Heterosis in BW was greater in Line A crosses (average of 4.6% with a range of 2.5 to 7.2% for BW at different ages) than in Line B crosses (average of 2.2% with a range of 0.1 to 5.3%). Heterosis in WA score was negative (crosses had better WA than pure lines) for males [14% for the Line A and FL crosses (P < or = 0.001) and 4% for the Line B and FL crosses (P < or = 0.05)], but was positive and nonsignificant for females of the above crosses (average of 2.6 %). With one exception, heterosis was not an important source of variation for shank width or shank depth and there was a low level of heterosis for shank length. No heterosis was observed for breast width. Reciprocal effects were an important source of variation in BW and shank measurements for females but not for males in both sets of crosses. The results of the present study indicated that use of the FL line to improve WA of improved primary-breeding commercial sire lines was not a feasible alternative. The slight improvement noted in WA of the male crosses was offset by the reduction in BW and breast width of the crosses relative to the pure commercial sire lines.

Aging↗

Transfection of DNA from a chloroethylnitrosourea-resistant tumor cell line (MER+) to a sensitive tumor cell line (MER-) results in a tumor cell line resistant to MNNG and CNU that has increased O-6-methylguanine-DNA methyltransferase levels and reduced levels of DNA interstrand crosslinking.

A subclone of a human glioma tumor cell strain which was deficient at O-6-alkylguanine repair was transfected with DNA from an O-6-alkylguanine DNA alkyltransferase-positive human colon tumor cell line (HT29). The transfected subclone, which was selected by its resistance to N-methyl-N'-nitro-N-nitrosoguanidine (MNNG), acquired increased resistance to chloroethylnitrosourea (CNU), contained O-6-methylguanine-DNA methyltransferase activity, and failed to accumulate interstrand crosslinks in its DNA upon treatment with CNU. The subclone morphologically resembled its CNU-sensitive parental glioma cell strain. The close correlation between the increased cellular resistance, increased O-6-alkylguanine repair activity and the absence of crosslinking suggests that the formation of DNA crosslinks is one important mechanism of cytotoxicity produced by CNU, and that repair of DNA lesions by O-6-alkylguanine DNA alkyltransferase may partially prevent CNU-induced cytotoxicity.

Cell Line↗

Long-term egg production and heterosis in quail lines after within-line or reciprocal recurrent selection for high early egg production.

1. Four Japanese quail lines were developed using 13 generations of reciprocal recurrent (lines AA and BB) or within-line selection (lines DD and EE) for high egg number until 98 d of age. In these lines and their reciprocal crosses, egg production, egg weight and mortality were monitored for up to 21 months to evaluate the impact of selection method and line origin on long-term production and heterosis. 2. Both hen-housed total egg number (EN) and hen-day egg laying rate (ELR) were consistently lower in recurrent lines than in lines under within-line selection: after 13 months of test, the difference was -47.1 eggs for EN and -20% for ELR%, whereas mortality was similar at around 20%. 3. Line EE was the best pure line for early egg production (55.2 eggs at 98 d of age) and also gave eggs which were consistently about 2 g larger than those from line DD but it had the highest overall mortality (78.5%) of all lines. On average, line DD laid 399 eggs (EN) over the whole experiment, 84 more than Line EE. 4. Crossbreds from the 2 types of pure lines and line DD had similar patterns and rates of egg production during the whole experiment, as indicated by the similarity of the curves adjusted for those lines by using the monomolecular model for EN and the modified compartmental model for ELR. 5. However, heterosis was higher from recurrent selection lines and it increased more with time than did heterosis from within-line selection lines.

Animals↗

Properties of soleus muscle Z-lines and induced Z-line analogs revealed by dissection with Ca2+-activated neutral protease.

Rat soleus muscle Z-lines and Z-line anomalies induced by neostigmine methyl sulfate (NMS) and cat soleus muscle Z-lines and Z-line anomalies induced by tenotomy were examined by electron microscopy before and after dissection of muscle fibers with Ca2+-activated neutral protease (CAF) to elucidate structural properties of Z-lines and related Z-line-type structures. In both normal and treated muscles, interdigitation of thin (6-7 nm) filaments, which were continuous with I-filaments (actin) from adjacent sarcomeres, was observed at the Z-line in longitudinal section. Both neostigmine methyl sulfate and tenotomy treatments induced muscle atrophy associated with Z-line degradation, streaming, and irregular distribution and accumulation of Z-line material and Z-rod formation. Tenotomized muscle also was characterized by the presence of N-line-like bands and I-Z-I brushes. CAF digestion removed the electron-dense covering material from Z-rods and revealed a backbone of actin filaments. The origin of Z-rods, their structural similarity to Z-lines in longitudinal and cross section, and their susceptibility to CAF indicate that Z-rods are directly related to native Z-lines and are probably lateral polymers of a basic Z-line unit. The regular square net alignment (22 nm) of I-filaments (actin) in cross sections of I-Z-I brushes which contain no N-lines suggests that the I-square net arrangement near the Z-line is determined by Z-filament-actin filament interaction rather than by the N-line or other factors. The results suggest that I-filaments (actin) penetrate the mammalian Z-line and are Z-line constituents and that the width of Z-lines and the length of Z-rods are determined by the amount of overlap of actin filaments. The perpendicular periodicity of Z-rods and the zigzag-oblique arrowheadlike appearance seen in longitudinal sections of Z-lines are attributed to alpha-actinin.

Animals↗

Genetic variation in pure lines and crosses of large-bodied turkey lines. 3. Growth-related measurements on live birds.

An experimental line (F) selected over 34 generations for increased 16-wk BW was reciprocally crossed with a primary breeding sire line (C) from a large international turkey breeder to study the inheritance of growth-related traits measured on live birds. All genetic groups were grown intermingled in confinement with sexes reared in different houses. The traits measured included BW at 8, 16, and 20 wk of age and shank length, shank width, shank depth, breast width, and walking ability scores at 16 wk of age. Walking ability was rated from 1 to 5 with 1 representing birds with no leg defects and no difficulty walking and 5 indicating birds with extreme lateral deviations of the legs and great difficulty walking. Ratings of 2, 3, and 4 represented intermediate values. The F line had a different growth pattern than the C line with the F line being larger than the C line at 8 wk of age, but the reverse was true at 16 and 20 wk of age. The difference in BW between the C and F lines increased from 16 to 20 wk of age. The C line had wider breasts than the F line at 16 wk of age. The F line had longer shanks than the C line. Shank width was larger in the C line than the F line for females but not males. No line difference in shank depth was observed. Walking ability scores at 16 wk of age were lower (better) in the C line than in the F line for males but not females. Significant heterosis in BW of the crosses of the F and C lines was observed at all ages in males (range = 3.3 to 5.6%) and only at 8 wk of age in females (3.6%). These results were similar to an earlier study in which the F line was crossed with a primary-breeding sire line from 2 other international turkey breeders. No significant heterosis in the crosses of the C and F line was observed for breast width and shank measurements. Heterosis was significant for walking ability scores of females (-3.0%) but not males. Reciprocal effects, a measure of sex linkage and maternal influences, were noted only for shank length and the direction of the difference was not the same in the 2 sexes.

Animals↗

Characterization of lymphoma-derived cell lines: comparison of cell lines positive and negative for Epstein-Barr virus nuclear antigen. I. Physical, cytogenetic, and growth characteristics.

Sixteen lymphoid cell lines were derived from patients with undifferentiated lymphoma of Burkitt's or non-Burkitt's type. They were obtained directly from tumor biopsies, from serous effusions, or from bone marrow. In 10 of the cell lines, the Epstein-Barr virus (EBV) nuclear antigen (EBNA) was undetectable; the remaining 6 lines were EBNA-positive (EB-pos). Of the 16 lines, 15 were aneuploid, with detectable chromosome "14q+ markers (11 had +8;14 translocations). These 15 lines, which included the EBNA-negative (EB-neg) lines, were believed to be of tumor cell origin. The remaining line consisted predominantly of diploid cells derived from normal lymphocytes, but some cells of tumor origin were present. Four EB-pos cell lines derived from EB-neg tumors had an aneuploid karyotype consistent with an origin from tumor cells (including no.8;14 translocation in two), which suggested that either tumor cells were infected with EBV in vitro or a tiny fraction of EB-pos tumor cells (or potential tumor cells) present in vivo gave rise to the predominant cell of the line. EB-neg B-cell lines and EB-pos cell lines established from undifferentiated lymphomas differed greatly. EB-neg lines had consistently smaller electronic mean cell volumes and narrow-angle light scatter than did EB-pos lines. This finding correlated with a lower nuclear:cytoplasmic ratio in EB-pos lines. EB-neg lines also had higher saturation cell densities than did EB-pos lines under standard culture conditions. The data indicate either that EBV influences the morphologic and physiologic characteristics of lymphoid cell lines or that EB-neg B-cell lines and EB-pos cell lines are derived ultimately from different lymphocyte subpopulations or that both may apply.

Antigens, Viral↗

Response to second-line chemotherapy in patients with metastatic breast carcinoma previously responsive to first-line treatment: prognostic factors.

BACKGROUND: The aim of this retrospective study was to determine those prognostic factors associated with response to a second-line chemotherapy in patients with metastatic breast carcinoma (MBC) that was previously responsive to a first-line chemotherapy. METHODS: The 70 MBC patients studied had previously responded to a first-line chemotherapy, mainly anthracycline or anthracenedione-containing regimens. During first-line chemotherapy they had received treatment until the maximum response was obtained, at which time treatment was discontinued. Second-line chemotherapy regimens were of several types (48.5% with anthracycline). A study of prognostic factors associated with response to second-line chemotherapy was performed by univariate and multivariate analysis. RESULTS: Second-line chemotherapy achieved a 44% response rate, with a median response duration of 10 months. Survival was 13 months in the entire patient group, 22 months in responders, and 8 months in nonresponders. Univariate analysis identified seven factors related to patient response rate to second-line treatment. A better response rate to second-line chemotherapy was observed in patients with the following features: 1) chemotherapy free time (time between onset of metastatic disease and initiation of first-line) < 12 months (P = 0.03); 2) complete response to first-line chemotherapy (P = 0.013); 3) response duration to first-line chemotherapy > 14 months (P = 0.0001); 4) progression free interval (time between end of first-line treatment and initiation of second-line chemotherapy) > 11 months (P = 0.0001); 5) performance status at second-line treatment < 2 (P = 0.04); 6) tumor index at second-line chemotherapy < 4 (P = 0.05); and 7) treatment with an anthracycline-containing second-line regimen (P = 0.03). In multivariate analysis, only progression free interval was identified as being associated with response rate to second-line chemotherapy (P = 0.0001). CONCLUSIONS: Retained chemosensitivity appeared to be an important characteristic in patients responding to second-line chemotherapy.

Adult↗

Visually perceived vertical (VPV): induced changes in orientation by 1-line and 2-line roll-tilted and pitched visual fields.

We report a series of nine experiments which show that a single roll-tilted line in darkness induces changes of the orientation perceived as vertical (VPV) that are similar in magnitude and direction to those measured by Witkin and Asch (1948a) [Studies in space orientation. I. Perception of the upright with displaced visual fields. Journal of Experimental Psychology, 38, 762-782] with the classical square 4-sided frame, and we describe the configuration-independent mass-action rules by which the influences of the individual lines influences are combined. Clockwise (cw) and counterclockwise (ccw) orientations of a line produce cw and ccw displacements of the VPV setting, respectively, with effect magnitude increasing approximately linearly with line orientation (e.g., a 66.25 degrees - long line at 25 degrees horizontal eccentricity that varies in roll-tilt through +/-13.2 degrees around vertical generates a systematic variation in VPV over +/-7 degrees). The slope of the VPV-vs-roll-tilt function increases with line length along a negatively accelerated exponential function (length constant = 17.1 degrees). The influences of two bilaterally symmetric lines combine linearly and algebraically and the combined influence is linearly related to the sum of the VPVs for the 1-line components with a slope equal to 0.91 for short lines and 0.66 for long lines; thus, VPV for short lines manifests nearly complete additive summation, but for long lines, the 2-line VPV is nearer to the average of the VPV values for the two components measured separately. The effectiveness of the conjunction of two line segments within a visual scene does not depend on their separate orientations, only on their sum. Individual lines from pitched-only planes or from combinations of such planes generate identical influences to those generated from lines in frontoparallel planes with the same image orientations at the eye of the observer (their "retinal orientations"). Retinal orientation is the key to the induction of VPV change independently of the line's plane of origin.

Adult↗

Genetic relationship among lines and smooth muscle and ovarian follicular development within lines of Japanese quail in two long-term selection studies.

Smooth muscle tumor and ovarian follicular development were studied in lines of Japanese quail selected for increased 4-wk BW (HW, P, and T) and their randombred controls (C and R1). The lines studied were from long-term selection studies at The Ohio State University (HW and R1) and The University of Georgia (P, T, and C). To study the genetic relationship among the lines in the two selection studies, the C, P, HW, and R1 lines were DNA-fingerprinted by digestion of the DNA with the HaeIII restriction enzyme and using Jeffreys' 33.6 probe. The BW of females at 4 wk of age and at the end of a 240-d egg production period were similar for the C and R1 lines. The BW of the selected lines was ranked P > T > HW for both measurements. Smooth muscle tumors were found in the oviducal ligaments adjacent to the magnum. A greater percentage of hens from the BW-selected lines had smooth muscle tumors of greater weight than the randombred control lines, which did not differ in tumor incidence or weight. The P and T lines had a greater incidence of multiple-lobed tumors than the HW line. Based on bandsharing (BS) of DNA fingerprints, the Georgia and Ohio lines did not appear to be closely related, suggesting that, perhaps, the smooth muscle tumors in the BW-selected lines in the two studies might have resulted from pleiotrophic effects of genes affecting growth or to genes closely linked to the growth genes. The BW-selected lines in both selection studies had more ovarian follicles in rapid development, which were of greater weight, than the randombred control lines. The HW line had a larger number of ovarian follicles in rapid development than the P and T lines. The percentage of hens with atretic follicles was greater in the BW-selected lines. The results of the present study suggest that the effect of BW selection on ovarian follicular development may occur early in selection (within the first 30 generations) and is not influenced by additional genetic changes in BW.

Animal Husbandry↗

Genetic variation in pure lines and crosses of large-bodied turkey lines. 1. Body weight, walking ability, and body measurements of live birds.

An experimental line of turkeys selected for increased BW (F) was reciprocally crossed with sire lines (designated A and B) from each of two major commercial breeders in order to study the inheritance of growth traits. All genetic groups were grown intermingled in confinement with the sexes reared in different houses. Traits measured included BW at 8, 16, and 20 wk of age; shank length, width, and depth; breast width; and walking ability scores at 16 wk of age. The F line had a different growth pattern than the two commercial sire lines. The F line had higher (males) or similar (females) BW in comparison to Line A at 8 wk of age, but by 16 wk of age, Line A was heavier than Line F in both sexes. The difference in BW between the F and A lines increased from 16 to 20 wk of age. Lines F and B did not differ in BW at 8 wk of age, but at 16 and 20 wk of age, Line B birds were heavier than those of F line. Reciprocal effects, a measure of sex linkage and maternal effects, were noted only for breast width of females in crosses of Lines A and F and for shank width and depth of males and 8-wk BW and shank depth of females in crosses of Lines B and F. Heterosis was an important source of variation in BW for males from both crosses. The percentage heterosis at the various ages ranged from 3.1 to 7.5. For females, heterosis (range = 2.6 to 4.9%) was only significant at younger ages (8 wk for the crosses of the A and F lines and 8 and 16 wk for the crosses of the B and F lines). No heterosis for breast width was observed in either cross. Heterosis in walking ability scores was significant only for males from the crosses of Lines B and F. The presence of heterosis was inconsistent for shank measurements.

Age Factors↗

Cell lines used in prostate cancer research: a compendium of old and new lines--part 1.

PURPOSE: This is part 1 of a 2-part review. Research into the molecular mechanisms underlying the various aspects of prostate cancer (PCa) requires the use of in vivo and in vitro model systems. In the last few years many new cell lines have been established by investigators from primary tissue sources and clonal derivatives of previously established lines. Therefore, the purpose of this 2-part review is to catalogue the current human cell lines developed for PCa research, as reported in the literature. Part 1 includes tissue culture cell lines derived from metastases, primary tumors and nonadenocarcinomas that were established without the use of transgenes. It also includes a section describing lines that have been contaminated with other lines, shown not to be of prostatic origin or whose identity is being challenged. MATERIALS AND METHODS: Prostate cell lines included in this review were identified by extensive searching of the literature using several strategies, including PubMed searches and book chapter reviews. RESULTS: In total we describe the derivation, phenotype, genotype and characterization of molecular markers expressed by approximately 200 lines and sublines used in PCa research, including those derived from primary tumors, metastases and normal prostate tissue. We paid particular attention to the expression of prostate specific antigen, androgen receptor, cytokeratins and other molecular markers used to indicate the status of PCa and the prostatic lineage of a given line. In an attempt to provide PCa researchers with a resource of information regarding new and established cell lines we have also created an online database of these PCa cell lines freely accessible via the World Wide Web at http://www.CaPCellLines.com. The web based interface allows researchers to peruse and print information regarding cell lines, add new cell lines and update or add new information regarding established cell lines. CONCLUSIONS: This compendium of cell lines currently used in PCa research combined with access to our on-line database provides researchers with a continually updated and valuable resource for investigating the molecular mechanisms of PCa.

Biomedical Research↗

The National Pork Producers Council Maternal Line National Genetic Evaluation Program: a comparison of six maternal genetic lines for female productivity measures over four parities.

Litter (n = 8,424) and female performance records were collected in two breed-to-wean production units in order to evaluate genetic line differences for sow longevity and maternal performance over four parities. Lines evaluated were American Diamond Genetics, Danbred North America, Dekalb-Monsanto DK44, Dekalb-Monsanto GPK347, Newsham Hybrids, and National Swine Registry. Females within a line were derived from a minimum of 65 sires, 197 dams (three dams per sire), and a maximum of three daughters per dam, except in the GPK347, which were produced using semen from 12 Nebraska Index boars mated with Dekalb-Monsanto Line 34 females. All lines expressed 100% maternal heterosis. Mixed model statistical procedures were used with fixed effects including genetic line, parity, production unit, and two-way interactions. Random effects included a contemporary week of production and female for repeated records. Lactation length (average 15 d) was included as a linear covariate where appropriate. In total, 3,599 females entered as early-weaned pigs, 3,283 entered the breeding herd, 2,592 farrowed at least a single litter, and 1,656 and completed four parities. Line (P < 0.001) and parity (P < 0.001) effects were observed for virtually all traits measured. Ranges of genetic line differences averaged across parities were 1.76 pigs for total born, 1.45 pigs born alive, and 0.31 stillborn pigs per litter. Ranges of line differences in total and live litter weight were 1.4 and 1.3 kg, respectively. Ranges among lines, within Parities 1 through 4, for litter size at weaning were 0.56, 1.08, 0.91, and 0.64 pigs per litter, respectively. Line differences for weight (33.8 kg) and backfat depth (6.4 mm) at farrowing, lactation feed intake (8.7 kg), weight loss (5.0 kg), and backfat loss (0.87 mm) were observed. Extended wean-to-estrus interval was related to variation in weight, feed intake, and backfat loss in all lines except the GPK347. The GPK347 females farrowed and weaned the largest number of pigs, ate less feed in lactation, and lost more backfat and weight during lactation, yet they had the largest litters and the shortest wean-to-estrus intervals. Line x parity interactions existed for many traits due to small rank changes, but in general, the high- and low-ranked lines did not change. Genetic line differences in reproductive efficiency through four parities exist and must be recognized when choosing a female line.

Adipose Tissue↗

Cell lines used in prostate cancer research: a compendium of old and new lines--part 2.

PURPOSE: This is part 2 of a 2-part review. Research into the molecular mechanisms underlying the various aspects of prostate cancer (PCa) requires the use of in vivo and in vitro model systems. In the last few years many new cell lines have been established by investigators from primary tissue sources and clonal derivatives of previously established lines. Therefore, the purpose of this 2-part review is to catalogue the current human cell lines developed for PCa research, as reported in the literature. Part 2 describes tissue culture cell lines derived by the insertion of transgenes, including human telomerase reverse transcriptase, SV40 T antigen and human papillomavirus genes. Part 2 also includes xenograft lines that require propagation and passage in vivo in mice. MATERIALS AND METHODS: Prostate cell lines included in this review were identified by extensive searching of the literature using several strategies, including PubMed searches and book chapter reviews. RESULTS: In total we describe the derivation, phenotype, genotype and characterization of molecular markers expressed by approximately 200 lines and sublines used in PCa research, including ones derived from primary tumors, metastases and normal prostate tissue. We paid particular attention to the expression of prostate specific antigen, androgen receptor, cytokeratins and other molecular markers used to indicate the status of PCa and the prostatic lineage of a given line. In an attempt to provide PCa researchers with a resource of information regarding new and established cell lines we have also created an online database of these PCa cell lines freely accessible via the World Wide Web at http://www.CaPCellLines.com. The web based interface allows researchers to peruse and print information regarding cell lines, add new cell lines and update or add new information regarding established cell lines. CONCLUSIONS: This compendium of cell lines currently used in PCa research combined with access to our on-line database provides researchers with a continually updated and valuable resource for investigating the molecular mechanisms of PCa.

Antigens, Polyomavirus Transforming↗

Tumorigenicity, invasion and metastasis of the small cell lung cancer cell line NCI-H69 and two derivative lines MOG-H69V and MOG-H69VZ.

Two adherent cell lines MOG-H69V and MOG-H69VZ have been isolated from a continuous cell line, NCI-H69, derived from human small cell lung cancer by Carney et al, [1987]. They have been established and characterised morphologically, biochemically, and for growth characteristics in vitro Khan et al (19). In the present study both the parental and the derivative lines have been investigated for invasiveness in vitro and in vivo. The parental line showed an early invasiveness compared with both the derivative cell lines. All cell lines formed tumours in nude mice with 100% take rate. Xenograft histology of all the cell lines revealed pleomorphic tumours, however the derivative lines showed areas of focal, large, spindle cells containing both acidic and neutral mucin, and spaces between the cells were found filled with alcianophilic, amorphous material. The parental line was invasive and metastatic. Tumours of both the derivative lines were non-metastatic under similar conditions. They were also investigated for neuroendocrine-cell marker expression. These data show that while the behaviour of the parental line was compatible with small cell lung cancer, that of the derivative lines was more indicative of non-small cell lung cancer, both in vitro and in vivo. As previous data suggested a common origin of the parental and the derivative lines, probably from a stem cell subpopulation present in the parental line, these lines represent a useful model for the study of phenotypic changes in lung cancer.

Animals↗

Cellular and karyotypic characterization of two doxorubicin resistant cell lines isolated from the same parental human leukemia cell line.

Separate mechanisms underlying the multidrug resistant (MDR) phenotype were identified in 2 independent approaches to select tumour cells resistant to low concentrations of doxorubicin (Dox) from the sensitive T cell leukemia cell line CCRF-CEM. The CEM/A7 cell line was selected at an initial concentration of 0.005 microgram/ml of Dox and maintained at 0.07 microgram/ml. In contrast, the CEM/A5 line was selected using an initial concentration of 0.01 microgram/ml and maintained in Dox at a concentration of 0.05 microgram/ml. P-glycoprotein expression was demonstrated in the CEM/A7 line but not the CEM/A5 line. Amplification of the mdrI gene was not observed in the CEM/A7 cell line. Both cell lines showed cross-resistance to a number of structurally unrelated cytotoxic drugs including anthracyclines and etoposide (VP-16), although only the CEM/A7 line was cross resistant to Vinca alkaloids. Immunoblots of total cell lysates of the CEM/A5 line have revealed almost undetectable levels of topoisomerase II alpha and beta in this line. Cytogenetic analyses of both lines revealed numerous karyotypic abnormalities which were present in the parental cell line as well as both resistant cell lines. The CEM/A7 line also demonstrated a duplication of part of the long arm of chromosome 7 which included the region containing the mdrI gene, a finding not seen in the parental or CEM/A5 line. CEM/A5, however, demonstrated an abnormality of chromosome 7, outside the region of the mdrI gene, and it also contained a deletion of the short arm of chromosome 2. Abnormalities in this latter region of genome have been associated with non-P-glycoprotein-mediated MDR.

ATP Binding Cassette Transporter, Subfamily B, Mem↗

Analysis of surface wave direction by the lateral line system of Xenopus: source localization before and after inactivation of different parts of the lateral line.

The turning responses of clawed toads (Xenopus laevis) to surface waves were examined in animals with an intact lateral line or with different combinations of lateral lines reversibly inactivated by CoCl2. The responses were characterized with respect to response frequency, turning accuracy, turning side, response time, and swim distance. After the inactivation most animals still responded to surface waves but the responses were different from those of animals with an intact lateral line. They also differed according to the combination of inactivated lines. In all experiments the responses for stimuli in some sectors of the surface did not differ from controls. The location of these sectors co-varied with the position of the intact lines, i.e., normal responses were found for frontal stimulus directions when head lines were intact and for caudolateral stimulus directions when trunk lines were intact. Their size was larger when lines on both sides of the body were intact and smaller when only lines on one side were intact. When the number of functional lines was reduced to one or two on one side of the body the turning angles shown within the sector of normal responses were maintained for stimulus directions outside these sectors. These results can be interpreted as indicating that head and trunk lines represent different "position values". When only a single line was functional the toads still turned towards the stimulus source more often than by chance. It is hypothesized that Xenopus uses two mechanisms to determine the direction of surface waves. One uses the position values of head and trunk lines; this mechanism is comparable to the "place value" postulated for individual head neuromasts of surface feeding fish. The other uses the information encoded in the activity pattern that is elicited in one line when the surface wave travels over the line. This second mechanism yields information about stimulus side but not about stimulus angle.

Animals↗