bluuebunny/crossref_metadata_2025 huggingface dataset:
Characteristics of Mast Cells in Chediak-Higashi Mice: Light and Electron Microscopic Studies of Connective Tissue and Mucosal Mast Cells
Authors: Crowle Patricia K. , Phillips Dwight E.
Published: October 2008

The beige mouse, a homologue of the Chediak-Higashi syndrome in man, possesses abnormally large granules in many tissue cells. The granules in the mucosal mast cells (MMC) of the small intestine of beige and littermate C57BL/6J mice were examined after infecting the mice with the intestinal parasite, Nippostrongylus brasiliensis. MMC in both beige and littermate mice had irregular granules which contained paracrystalline substructures embedded in an amorphous matrix. Granules were not observed in fusion with the cell membrane. Instead, in late-stage mast cells, the granule membrane broke down, the granule contents were spread throughout the cytoplasm, and the cell organelles disintegrated. Unlike connective tissue mast cells, MMC were poorly demonstrated with formalin fixation and toluidine blue staining.
The Cell Surface Sulphydryl Content of Metastatic Variants of B16 Murine Melanoma
Authors: Sherbet G.V.
Published: October 2008

The cell surface sulphydryl content of three metastatic variants of the B16 murine melanoma has been determined using isoelectric equilibrium techniques. The F1 variant, which has no ability for natural metastasis, and the F10 variant with moderate metastatic ability appeared to have no detectable surface thiol groups. The variant BL6, which shows a high degree of natural metastasis, possessed surface thiol groups. The variants were found to be heterogeneous in isoelectric distribution. Three subpopulations were identified based on isoelectric criteria. The size of the pi 5.0 subpopulation appeared to increase with metastatic ability. A proportion of this subpopulation, approximately 4% in the F10 and 10% in the BL6, was found to possess surface thiol groups. In the BL6 line, 10–20% of the pi 4.6 subpopulation possessed surface thiol groups. The surface negative charge density of the cell types showed no correlation with their natural metastatic ability.
Effects of Prostanoid Precursors and Indomethacin on Chick Embryonic Cartilage Growth in Organ Culture
Authors: Kirkpatrick Charles James , Mohr Winfried , Haferkamp Otto
Published: October 2008

Using the Fell technique of organ culture of 8-day chick embryo femoral and tibial rudiments, the effects of indomethacin, dihomo-gamma-linolenic acid and arachidonic acid on limb rudiment linear growth and differentiation were investigated. Indomethacin (50 and 100 µmol/l) elicited a statistically significant decrease in rudiment linear growth without affecting differentiation or cell structure. Dihomo-gamma-linolenic acid and arachidonic acid, both at 100 µmol/l, exerted no effect on limb rudiment linear growth or differentiation. From previous work, it has been shown that PGAi and PGBi caused a marked inhibition of linear growth, PGAi being cytotoxic. The failure of the prostaglandin (PG) precursors to reproduce these effects suggests that PGA or PGB biosynthesis in embryonic chondrocytes plays no significant role in cartilage growth regulatory mechanisms. Moreover, the growth inhibitory effect of the PG cyclooxygenase inhibitor, indomethacin, suggests that a product of arachidonic acid metabolism via the cyclooxygenase pathway may promote cartilage growth.
Uncontrolled Growth of Tumour Stromal Fibroblasts in vitro
Authors: Delinassios John G. , Kottaridis Stavros D. , Garas John
Published: October 2008

The disturbance of the growth control mechanisms in tumour cells in vivo may be manifested as uncontrolled growth of the tumour stroma in vitro. Stromal fibroblast-like finite cell lines produced from benign or malignant human breast tissue specimens exhibited cell overlapping which ranged from multilayers to dense piling up colonies, while cells derived from normal tissues exhibited intense contact inhibition of growth and locomotion, under the same culture conditions. 6 out of 13 lines derived from malignant tissues and 2 out of 5 lines derived from benign lesions exhibited the phenomenon of ‘periodic appearance of piling up colonies’.
Characteristics of 85 Pediatric Tumors Heterotransplanted into Nude Mice
Authors: Neely John E. , Ballard Edgar T. , Britt Alfred L. , Workman Linda
Published: October 2008

81 primary pediatric tumors and 4 tumor lines were heterotransplanted into nude mice with an overall success rate of 38.3%. There was variability in success between tumor types. Bone sarcomas were highly successful while brain, lymphoid, and benign tumors in general did poorly. With increasing passage lag times decreased but actual growth rates in general did not change. Results suggested that tumors obtained prior to therapy which grew in nude mice were more likely to recur in the patient. During the observation period 7 spontaneous mouse tumors developed, distinguished from human tumors by histology, cytogenetics, and isoenzyme studies.
Studies on Thymocyte Subpopulations in Guinea Pigs
Authors: Sandberg G. , Söder O. , Kölare Susanne , Ernström U.
Published: October 2008

Thymocytes were separated according to increasing buoyant density into the three subpopulations la (25% of recovered cells), lb (20%) and II (55%), and according to binding to peanut agglutinin (PNA) into PNA+ (65 %) and PNA– cells (35 %). The frequency of PNA+ was 56% in la, 60% in lb and 66% in population II. Electronic cell volume determinations disclosed mean volumes of 160 fl for la, 130 fl for lb and 100 fl for population II. PNA+ and PNA– cells were very similar as regards cell volume. Thus, PNA+ and PNA– cells are remarkably uniformly distributed among cell categories of different density and cell volume. The rapidly cycling thymocytes, regarded as the most immature cells in the thymus, and the target cells for a thymocyte growth factor both belonged to the PNA+ cells of population la. The mitogen-responsive thymocytes also belonged to population la, but were PNA-. The largest subpopulation of thymocytes, apparently corresponding to the small, non-cycling cortical cells, were recovered as PNA+ cells of population II.
Bithorax Phenocopy and Pattern Formation
Authors: Ho Mae-Wan , Bolton Eleanor , Saunders Peter T.
Published: October 2008

In four different lines of Drosophila melanogaster the highest frequencies of bithorax phenocopies result from ether treatment during the precellular blastoderm stages. The spatial characteristics of the phenocopy spots are dependent on the precise timing of phenocopy induction, and are not in accord with those predicted from the model of sequential determination of whole compartments put forward by Garcia-Bellido et al. in 1976. Ether treatment for 13 successive previous generations gave rise to changes in the spatiotemporal characteristics of the phenocopy response. Our results suggest the involvement of continuous diffusion-like processes in the prepattering event which is affected by ether.
Bithorax Phenocopy and Pattern Formation
Authors: Ho Mae-Wan , Saunders Peter T. , Bolton Eleanor
Published: October 2008

Phenocopy maps are constructed from the spatial pattern of the phenocopy spots using a distance function defined on the frequencies with which two compartments transform separately and jointly. The temporal changes in distances between different compartments in turn enable us to make inferences concerning the diffusion-like processes involved in prepattern formation of the metathorax. A model is presented which accounts for the data in terms of the propagation of two wavefronts in sequence and in approximately orthogonal directions. Sensitivity to transformation by ether is limited in time to the interval between the passing of the two wavefronts. Changes caused by 13 successive generations of ether treatment can be inteφreted in terms of alterations in the timing and rates of wave propagation. A comparison of phenocopy and fate maps suggests that the prepatteraing processes affect primarily the blastoderm cytoplasm or surface membrane.
Immunological Competence in Osteopetrotic (Microphthalmic) Mice
Authors: Schneider Gary B. , Marks, Jr. Sandy C.
Published: October 2008

Osteopetrosis in microphthalmic (mi) mice is characterized by excessive skeletal mass and reduced bone resorption. The skeletal defects can be corrected by the transfer of mononuclear spleen cells from normal httermates. These studies suggest that osteopetrotic mutants might also demonstrate defective immune functions. Several laboratories have demonstrated significantly reduced responses to T and B cell mitogens by spleen cells from mi and op mice and op, osteopetrotic, rats. The problem with these latter studies is that different populations of cells have been compared in mutants and normal littermates because the spleen is a focus of extramedullary hemopoiesis in osteopetrotic animals. To circumvent this problem, cellular suspensions of mononuclear cells from the spleen and mesenteric lymph node were separated using Ficoll-Hypaque from mi and normal littermates 5 and 10 weeks of age. Under appropriate culture conditions these cells were exposed to different concentrations of concanavalin A, phytohemagglutinin and lipopolysaccharide for 3 days and 3H-thymidine for the last 24 h. In all cases, the response to optimal concentrations of the three mitogens was higher for the mi lymph node cells than controls. Contrary to previous studies, the spleen cell response to all three mitogens was only slightly lower in the mi cultures. The cellular composition of the lymph node and spleen samples tested in the mitogen assays was also evaluated by fluorescence microscopy using FITC-conjugated monoclonal antibodies directed against specific cell surface markers. The percentage of B cells, monocytes and macrophages, total T cells, and suppressor/cytotoxic T cells was found to be similar in mi and normal mice using Ficoll-Hypaque isolates of spleen and lymph nodes. These results indicate a normal or slightly enhanced immune function in the mi mutant when similar cell populations are compared.
Abnormal Eye Development in Drosophila after Treatment with Vinblastine
Authors: Wolsky Alexander
Published: October 2008

Drosophila cultures, kept on a water-added medium containing 0.16–2.5 µg/ml vinblastine (Velban, Lilly), produced offspring with a high percentage of eye abnormalities. These were eyes completely missing or reduced (moderately or severely) in size, of abnormal shape (‘kidney eyes’) and partially transdetermined into hairy outgrowths resembling antennae. Several of the abnormalities were phenocopies of known mutations, like ‘eyeless’ or ‘erupt’. The abnormally shaped and partially transdetermined eyes had characteristics, namely the appearance of abnormalities in the anterior region of the eye field, which permit conclusions about the probable mechanism of their production during development. These conclusions are in agreement with known facts concerning the mechanism of development of the insect eye and the mode of action of vinblastine.
The Localization and Secretion of Type IV Collagen in Synovial Capillaries by Immunohistochemistry Using a Monoclonal Antibody against Human Type IV Collagen
Authors: Matsubara Tsukasa , Trüeb Beat , Fehr Kurt , Rüttner Jacques R. , Odermatt Bernhard F.
Published: October 2008

The localization and the secretion of type IV collagen in synovial capillaries have been investigated by detecting the antigenic determinant of the major triple helix of human type IV collagen. Type IV collagen was indicated to be localized mainly in the lamina densa of basement membranes (BM) and to be secreted by both endothelial cells and pericytes. The pericytes secreted this collagen to both surfaces facing endothelial cells and the interstitial connective tissue. On the contrary, the direction of type IV collagen secretion by the endothelial cells was strictly confined to one side, namely towards the surface facing the BM. The absence of the antigenic determinant in rough endoplasmic reticulum and Golgi apparatus of the endothelial cells and pericytes indicated that the major triple helix of type IV collagen is mainly formed in the secretory vesicles after budding from the Golgi apparatus.
Polyoma Virus Transformation and Two-Dimensional Electrophoresis Patterns of Baby Hamster Kidney Cell Native Proteins
Authors: Latner A.L. , Marshall T.
Published: October 2008

The protein extracts of baby hamster kidney cells (BHK 21/C13) before and after malignant transformation by polyoma virus were resolved by isoelectric focusing in the first dimension and electrophoresis on concave 4-24% (w/v) polyacrylamide gradient gels in the second dimension. The resulting two-dimensional patterns were visualised by ultrasensitive silver staining. Major differences were observed in the protein distribution patterns of the two cell lines. Brief trypsinisation of the cells, prior to extraction of cell protein, had little effect upon the protein patterns of BHK cells. Similar treatment of the transformed cells markedly reduced the staining intensity of a string of spots which were present in far greater abundance than in the original BHK cells. The findings suggest that these proteins are located on the surface of the transformed cells and probably under the surface prior to transformation.
A Test for the Effects of Low-Temporal-Average-Intensity Pulsed Ultrasound on the Rat Fetus
Authors: Child Sally Z. , Carstensen Edwin L. , Davis Henry
Published: October 2008

In 1978, Pizzarello and co-workers reported that exposure of rat fetuses to diagnostic levels of ultrasound caused marked reduction in fetal weight. Replicates of these experiments have been conducted and, in addition, exposures at 10 times the peak and average intensity were used. No effect on fetal weight, numbers of living fetuses or resorptions could be attributed to ultrasound at either exposure level.
Monoclonal Antibodies to the Collagen Binding Domain of Human Plasma Fibronectin
Authors: Tarone Guido , Amedeo Maria Rosa , Di Renzo Maria Flavia , Comoglio Paolo
Published: October 2008

Five independent hybrids producing monoclonal antibodies to human plasma fibronectin have been obtained by fusing P3/X63-Ag8 myeloma cells with immune mouse splenocytes. The specificity of these monoclonal antibodies (MABs) for fibronectin was demonstrated by three independent tests: binding to the purified soluble molecule, immunofluorescence staining of insoluble extracellular matrices produced by endothelial cells in vitro, immunostaining of fibronectin tryptic peptides after separation on SDS-PAGE and transfer to nitrocellulose sheets. Two antibodies (MAB 29 and 52) recognized selectively human fibronectin while the others (MAB 5, 30 and 59) reacted also with plasma fibronectin from calf, hamster and chicken. Four distinct epitopes were recognized by the MABs studied. MAB 5, 30, 52 and 59 reacted with distinct antigenic sites, while MAB 29 and 52 bind to the same site. Antigenic fragments were identified by immunostaining of fibronectin tryptic peptides. MAB 5 reacted with a collagen binding fragment with a molecular weight of 120 K. In addition, each of the MAB 29, 30, 52 and 59 reacted with peptides with a molecular weight of 40 K that bind to gelatin. Since these antibodies do not inhibit fibronectin-collagen interaction, it is concluded that their corresponding epitopes are clustered in a region close, but not coincident, to the collagen binding site of fibronectin.
A Transparent Chamber for the Dorsal Skin Fold of Athymic Mice
Authors: Falkvoll Knut H. , Rofstad Einar K. , Brustad Tor , Marton Per
Published: October 2008

A transparent chamber designed for insertion in the dorsal skin fold of athymic mice is described. The chamber is based on ingrowth of a layer of granulation tissue from the surrounding subcutis of the animals. The thickness of the tissue layer in the chamber is accurately given by an exchangeable chamber component and the construction allows repetitive access to the tissue inside the chamber. A freely movable skin flap is prepared surgically on the back of the animals, and the chamber is inserted in this flap in a second operation about 2 weeks later. In the absence of air bubbles, infectious foci and excessive adipose tissue in the chamber, ingrowth of granulation tissue is completed 2–4 weeks after the insertion. When infections of the skin enclosing the chamber and atrophy of the skin flap are prevented, the functioning time of the transparent chamber is determined by the life span of the animals. In the present study the athymic mice remained healthy for 4–5 months after insertion of the chamber although they were kept under conventional conditions, but were given antimicrobial covering. Since athymic mice accept some human xenografts, both normal tissues and neoplasms, vital microscopical studies of such xenografts implanted into the chamber can be performed. Preliminary results of such studies on a human malignant melanoma are presented.
Effects of X-Radiation on the Retina of the Albino Rabbit as Viewed with the Transmission Electron Microscope
Authors: Newton James C. , Barsa-Newton Mary C. , Wisniewski Henry M. , Wen G.Y.
Published: October 2008

The eyes of albino rabbits were exposed in vivo to 7,000 rad of X-radiation. The retinae were dissected and examined with a transmission electron microscope 24 and 72 h following irradiation. Damage after 24 h was minimal, but after 72 h it was very pronounced. In both cases the damage was restricted to the photoreceptors.
Effects of Follicle-Stimulating Hormone, Luteinizing Hormone and Prolactin on Left and Right Gonads of Female Chick Embryos in Culture
Authors: Milano Elena Grassi
Published: October 2008

Organotypic cultures were made of right female gonads of chick embryos of 8 days, on medium containing follicle-stimulating hormone, luteinizing hormone or prolactin (PRL), and of left ovaries of the same age on the medium containing PRL. Results indicate that the explants cultured on PRL resemble most closely the normal ovaries in vivo. It is suggested that PRL may have a function in the morphogenesis of the female gonads of chick embryos.
Relationship between Karyotype of Tissue Culture Lines and Tumorigenicity in Nude Mice
Authors: Gershwin Eric , Lentz Daniel , Owens Robert B.
Published: October 2008

There are many factors, other than the absence of an immune response, that are responsible for the successful transplantation of xenografts to nude mice. We have defined, in this study, the relationship between karyotype and transplantability of xenografts into nude mice, using a library of 85 tumor and normal tissue culture lines. Each of these lines had been previously passaged in tissue culture for more than 10 generations. 53 of these 85 lines were classified as diploid, whereas 32 lines were classified as heteroploid. These tissue culture lines were transplanted to groups of 4-10 unmanipulated and antilymphocyte serum (ALS)-treated nude mice and serially monitored for 12 weeks. Only 19 % of the diploid lines were successfully transplanted into unmanipulated nude mice, and only 30% were successfully transplanted into ALS-treated nude mice. In comparison, 50% of the heteroploid lines were successfully grafted into unmanipulated nude mice, and a total of 60% were successfully transplanted into ALS-treated nude ice. 12 of the 53 diploid lines can be classified as being pseudodiploid on the basis of rearranged chromosomes or other minor karyotype anomalies. 8 of these 12 pseudodiploid lines were successfully transplanted into nude mice, further strengthening the relationship between abnormal karyotype and transplantability. Tumor karyotype is an additional parameter which should be added to the variables involved in the successful transplantation of tumors to nude mice.
Characterization of Human Bone Marrow Long-Term Cultures
Authors: Horikoshi Akira , Murphy, Jr. Martin J.
Published: October 2008

In an attempt to more closely simulate the native hematopoietic environment, human bone marrow bony matrix was cultivated in long-term human bone marrow cultures. Sternal bone marrow curettings (i.e., ‘bony matrix’) were cultured with and without autologous bone marrow single cell suspensions. Fresh media were provided at weekly intervals and the ‘harvested’ cells were assayed for CFU-gm (i.e., the granulocyte-macrophage precursor). Bone marrow bony matrix alone was not competent to maintain CFU-gm production, although early in the culture nucleated cells and CFU-gm were recruited from the bony matrix. A dose of 850 rad of X-radiation to the bone marrow matrix damaged the hematopoietic growth-promoting effects of the adherent cell layer, which was, however, rapidly reconstituted by an inoculation of intact autologous bone marrow cells. Additional investigations revealed that, whereas the addition of hydrocortisone to this culture system did not alter the maintenance of CFU-gm, recharging the cultures with fresh autologous marrow maintained CFU-gm for 10 weeks. These data indicate that (1) human bone marrow stroma is more sensitive to X-irradiation in vitro than in vivo, and (2) the bone marrow hematopoietic microenvironment provided by its bony matrix seemed to be less effective in vitro than in vivo with respect to the maintenance of hematopoietic precursor cells.
Aminoglutethimide – A New Endocrine Therapy in Breast Cancer
Authors: Harris Adrian L.
Published: October 2008

Aminoglutethimide is an effective treatment for advanced postmenopausal breast cancer, acting in a novel way. It inhibits peripheral and tumour aromatase, which converts androgens to oestrogens. Marked suppression of oestrone and oestradiol is produced. Aminoglutethimide was used as an inhibitor of adrenal steroid biosynthesis, but the concentrations required to inhibit aromatase in vitro are 10 times lower. This has led to its clinical evaluation in much lower doses. Doses of 125 mg twice daily produce oestrogen suppression equivalent to conventional doses of 1 g daily, and also similar tumour responses. Toxicity is much reduced by the lower dose.
Growth of Tumor Cells with Different Sensitivities for Murine Natural Killer Cells in Young and Adult Athymic Nude Mice
Authors: Romijn J.C.
Published: October 2008

Of four tumor cell lines, the murine YAC lymphoma, the human K562 lymphoma, and the human prostatic carcinomas PC3 and PC93, the susceptibility to murine natural killer (NK) cells as well as the tumorigenicity in young (3.5–4 weeks old) and in adult (8–10 weeks old) nude mice were studied. In young nude mice, which exhibited a lower level of NK cell activity than adult nude mice, the formation of solid tumors after inoculation of tumor cell suspensions occurred more frequently and with a shorter time lag than in adult animals. These effects were observed not only with the NK-sensitive YAC cells, but also with the relatively NK-insensitive PC3 and PC93 cells, indicating that also factors other than NK cell susceptibility may influence the growth of tumor cells in nude mice. Therefore, the use of young nude mice may enhance the rate of success of heterotransplantation of human tumors, regardless of the NK cell susceptibility of the tumor cells.
Differentiated Epidermal Outgrowths in the Planarian Dugesia gonocephala: A Model for Studying Cell Renewal and Patterning in Single-Layered Epithelial Tissue
Authors: Chandebois Rosine
Published: October 2008

Large deep wounds on the ventral side of a flatworm (Planaria) will not heal. Instead, the damage to the parenchyma in the wound’s roof will result in a differentiated swelling in the dorsal epidermis, above the wound which will eventually disappear with the disintegration of the underlying damaged tissue and a ventrodorsal hole appears in place of the wound. The dorsal epidermal outgrowth is formed by a number of excrescences, the development of which involves four successive stages. Their analysis suggests that epidermal cells are continuously produced by their own stem cells which remain unnoticed because their nuclei are hardly stainable. The daughter cells differentiate without information from either the underlying tissues or the basal epithelial membrane. During the first stage of this differentiation the cells become ciliated and motile, with some embryonic features. They then produce rhabdites and take up a columnar shape as they may become attached to the basal membrane. After wound setting the production of epidermal cells increases and the overcrowding of the basal membrane results in (1) detachment of stem cells and motile ciliated cells from the basal tissues, i.e. outgrowths; (2) stretching of columnar cells at the base of the outgrowths. When in the process of tissue disintegration the basal membrane of the epithelium also disappears, the cells remain in a single-layered epithelial configuration and retain their original polarity. These results are at variance with the generally accepted hypothesis that, in planarians, epidermal cells originate from the parenchyma and the epidermis is not an autonomous tissue.
Establishment and Characterization of an Epstein-Barr Virus Nuclear Antigen-Negative Hairy Cell Line
Authors: Lang Alois B. , Odermatt Bernhard F. , Gut Dorothea R. , Rüttner Jacques R.
Published: October 2008

Continuous cell lines have been established from spleen cells of patients with confirmed hairy cell leukemia (HCL). One cell line, HCL-Z1, lacks Epstein-Barr virus nuclear antigen (EBNA), grows attached to the substratum and retains typical features of hairy cells as revealed by transmission and scanning electron microscopy. HCL-Z1 differs morphologically from the three other EBNA-positive lymphoblastoid cell lines obtained (HCL-Z2, HCL-Z3, HCL-Z4) as well as from normal spleen cells or lymphocytes. The three lymphoblastoid cell lines derived from HCL patients show similar surface features as a line from a myeloma patient. Therefore, not all cell lines derived from HCL patients may be considered as representative of the patients leukemia cells.
Mechanism and Regulation of Natural Cytotoxicity
Authors: Kimber I. , Moore M.
Published: October 2008

Natural killer (NK) cells comprise a heterogeneous population of effector cells functionally and phenotypically distinct from B cells and mature antigen-sensitive T cells, with the capacity to spontaneously lyse target cells of widely different tissue provenance in a genetically unrestricted fashion. As such they have been widely implicated in immunosurveillance against neoplastic and virus-infected cells, as well as in the homeostasis of haematopoietic differentiation and regulation of immune function. In common with cytotoxic T cells, the lytic mechanism may be resolved into several discrete stages. Target cell recognition appears to involve several chemical entities, while susceptibility is also influenced by a multiplicity of factors operative at post-recognition stages of the lytic process. NK activity is subject to both positive and negative regulation. The potentiating effects of interferons and interleukin-2, products of activated T cells, indicate a possible pathway by which adaptive immune responses may augment natural cytotoxicity under local physiological conditions. Negative regulation is mediated by certain prostaglandins and a variety of cell types including macrophages, granulocytes and thymocytes as well as subsets of peripheral blood lymphocytes.
Effects of Murine Tumor Necrosis Factor on Friend Erythroleukemic Cells
Authors: Suyama Kimita , Goldstein Jack , Green Saul
Published: October 2008

Tumor necrosis inducing factor (TNF), a 140,000 molecular weight glycoprotein present in the serum of Corynebacterium parvum endotoxin-treated mice, was cytotoxic toward Friend virus-transformed erythroleukemic cells (FELC). These cells grow in culture as undifferentiated pro-erythroblasts but can be induced to differentiate in a limited fashion along the erythroid pathway to orthochromatic normoblasts by various agents such as dimethylsulfoxide (DMSO). Partially and highly purified preparations of TNF were cytotoxic toward logarithmically growing FELC whereas a comparable serum protein fraction from C. parvum treated mice or endotoxin from E. coli had no effect upon FELC viability. DMSO-induced cells were more sensitive to the action of TNF requiring only about half the concentration needed to produce 50% kill in noninduced cells. Inhibition of hemoglobin formation was TNF dose-related and could be decreased by 94%. TNF was also cytotoxic toward DMSO-induced cells in stationary phase and mitomycin C treated noninduced FELC. Neuraminidase modification of the surface of FELC increased the cytotoxicity of TNF by 50%. These results demonstrate that TNF destroys FELC whether they are nondividing, dividing or partially differentiated and suggest that TNF may accomplish this by affecting cell metabolism after internalization.
Antitumor Activity of Normal Human Globulin: Effects on Murine and Human Erythroleukemic Cells
Authors: Suyama Kimita , Goldstein Jack , Green Saul
Published: October 2008

The effect of the antitumor fraction isolated from the α2-globulin region of normal human serum (NHG-I) upon murine (FELC) and human (K562) erythroleukemic cells in vitro was determined. NHG-I inhibited the growth of both actively growing FELC and K562 cells in a dose-dependent manner. However, it had no effect upon mitomycin C treated FELC which were unable to divide nor upon dimethylsulfoxide-induced FELC in stationary phase. These results indicate that NHG-I has a cytostatic effect upon cell growth and suggests that its action may be dependent upon DNA synthesis. This is in marked contrast to TNF, the α2-globulin factor obtained from murine serum which is also not species-specific but whose action upon these cell lines is cytotoxic.
Repair Mechanisms Involved in Muscle Regeneration following Partial Excision of the Rat Gastrocnemius Muscle
Authors: Roth D. , Oron U.
Published: October 2008

The sequential cytological events of the regeneration process, after partial excision of the gastrocnemius muscle in the rat, were followed by light and electron microscopy. During the first 2 days after injury leukocytes and macrophages infiltrate into the traumatized area. Myogenic regeneration is then characterized by mainly two repair mechanisms. Mononucleated cells, that populate the excised area, most probably fuse together to give rise to newly formed multinucleated myotubes that further develop to striated myofibers. Another mechanism involves the repair of injured muscle fibers by the possible fusion of mononucleated cells with their necrotic cut ends. Consequently, by addition of nuclei and new muscular material, sarcoplasmic outgrowths from the injured fibers are formed. It is concluded that mainly two repair mechanisms are involved in the regeneration process following partial excision of a muscle: addition of new muscle fibers in a process similar to that of embryonic myogenesis and also meristic growth from the injured fibers.
Particles and Macrophages in Murine Peyer’s Patches
Authors: LeFevre M.E. , Warren J.B. , Joel D.D.
Published: October 2008

Mice were given 1 % suspensions of 5 insoluble particles (chrysotile asbestos, quartz, carmine, carbon, and iron oxide) in drinking water for 3 months. The particles were subsequently sought in intestinal Peyer’s patches by light microscopy. Carbon and iron oxide particles were visible in Peyer’s patch macrophages, particularly in the subepithelial region, but the other particles could not be detected. The findings suggest that particle surface properties as well as particle size govern accumulation in Peyer’s patches. The cytochemistry of sub-epithelial, mid-dome, tingible-body, and serosal macrophages of control mice indicated diversity of macrophages within the patch. Macrophages of asbestos-fed mice contained more lysosomes than macrophages of controls. Macrophage abundance in the dome apex was not significantly altered by asbestos ingestion. The other particles did not produce detectable alterations in macrophage morphology.
Early Recognition of Transplantation Antigens by T Lymphocytes
Authors: Ramseier Hansruedy
Published: October 2008

Within 2–4 h of interaction of parental spleen cells from naive mice or of their supernates with alloantigen-bearing F1hybrid spleen cells, a factor called soluble early product of immune recognition (SEPIR) is secreted. SEPIR could be revealed by its ability to enhance mixed leukocyte cultures (MLC) set up in suboptimal conditions. The factor appears to be generated by parental strain T but not B lymphocytes, is active at low concentration and acts in a pulse-like fashion. Its formation is triggered by unstimulated T cells reacting with H-2 antigens; no cytokine activity of IL 1, IL 2 or interferon character could be detected. It is suggested that the formation of SEPIR within the first few hours of MLC interaction is critically related to the further development of alloantigen-driven T cell proliferation. SEPIR might thus be the earliest discernible product of alloimmune recognition.
A Cell Line, ROSE 199, Derived from Normal Rat Ovarian Surface Epithelium
Authors: Adams Anne T. , Auersperg Nelly
Published: October 2008

A cell line, ROSE 199, derived from rat ovarian surface epithelium (ROSE) formed papillary structures which resembled, histologically, serous papillary cystadenomas of borderline malignancy seen in the human ovary. Crowded cultures produced two layers of cells separated by a thick layer of collagen fibers. Such cultures shed viable cells into the growth medium, while no cells were shed by short-term ROSE cultures. The resemblance to ovarian tumors exhibited by ROSE 199 cells in culture, reinforces the hypothesis that the common epithelial tumors of the ovary are derived from the ovarian surface epithelium. ROSE 199 cells, while retaining their epithelial morphology and ultrastructural characteristics, express stromal activity such as abundant collagen production. Perhaps this ability to express epithelial and stromal behavior is a contributing factor to the ready neoplastic transformation of the ovarian surface epithelium.
Suppression of Lymphocyte Activation by Plasma Lipoproteins: Modulation by Cell Number and Type
Authors: Okano Yukio , Macy Martha , Cardin Alan D. , Harmony Judith A.K.
Published: October 2008

Plasma lipoproteins containing apolipoproteins B and E, as well as delipidated water-soluble apoE, suppress lymphocyte activation by polyclonal T cell mitogens in vitro. This report establishes that apoBioo, isolated from human plasma LDL, also suppresses lymphocyte activation. Prereplicative mitogen-induced events as well as DNA synthesis and cell division are suppressed. A number of experimental variables influence the extent to which lipoproteins suppress lymphocyte activation. Lipoproteins isolated from different donors vary widely in suppressive potency. In addition, the extent of suppression depends on the cultured cell density: suppression at fixed concentration of lipoprotein or apolipoprotein decreases as the number of cells increases. When the total number of cells per culture and the suppressor concentration are both fixed, the extent of suppression decreases as the percent T cells or monocytes increases. In the lymphatic tissue where lymphocytes and accessory cells are concentrated, plasma lipoproteins may play a less important immunoregulatory role in normolipidemic subjects compared to that in subjects with hyperiipoproteinemia, particularly hypercholesterolemia, since the tissue concentration of lipoproteins in hyperlipidemic subjects is likely to be elevated.
Dose-Dependent Effect of 17β-Estradiol Determined by Growth Curves and Flow Cytometric DNA Analysis of a Human Breast Carcinoma (T61) Grown in Nude Mice
Authors: Brünner N. , Spang-Thomsen M. , Vindeløv L. , Nielsen A. , Engelholm S.A. , Svenstrup B.
Published: October 2008

An estrogen and progesterone receptor-positive human breast carcinoma (T61) grown in nude mice was exposed to 1.0, 0.1, 0.01, and 0.001 mg 17β-estradiol. These doses resulted in serum peak concentrations (day 1) of estradiol ranging from 3.5 × 10-8 to 6.9 × 10-10M. The effect of the treatment was evaluated using growth curves and flow cytometric DNA analysis. The treatment induced a dose-dependent growth delay and dose-dependent changes in the cell cycle distribution. The cell cycle changes comprised a decrease in the G1 phase, an accumulation of cells in the S phase, and an increasing fraction of polyploid cells. The results suggest that estradiol induces a dose-dependent cell killing effect in the T61 human breast carcinoma. The correlation between the treatment-induced growth delay and the effect on the cell cycle distribution indicates that the changes in the cell cycle are a reflection of the estradiol-induced cell destruction. Since no tumor growth stimulation could be observed even at very low serum estradiol concentrations, the T61 human breast carcinoma may represent a new aspect in the study of human breast cancer.
Differentiation of Chick Blood Island Erythroblasts into Erythrocytes and Stability in Long-Term Culture
Authors: Zagris Nikolas
Published: October 2008

Clusters of 20–70 erythroblasts from blood islands of early chick blastoderm were cultured in serum-free chemically defined medium for a 3-month period. The erythroblast cluster produces erythroid cells and hemoglobins characteristic of the primitive and definitive erythroid cell lines. It seems there is a progenitor erythroid cell(s) in the erythroblast cluster which starts and/or continues maturing along various pathways of hemopoietic differentiation under simple culture conditions. The erythroid character of these cells is stable during the 3-month culture period.
Multiple Labeling of Cellular Constituents by Combining Surface Reflection Interference and Fluorescence Microscopy
Authors: Opas Michał , Kalnins Vitauts I.
Published: October 2008

In this paper the technique for visualizing cytoskeleton in detergent-extracted cultured cells by surface reflection interference (SRI) microscopy after staining with the protein dye Coomassie Brilliant Blue (SRI-CooB technique) is used in conjunction with fluorescently-labeled antibodies or with other fluorescent probes to detect a number of constituents in the same cultured cell. Because SRI-CooB technique preferentially visualizes microfilament bundles along the ventral aspect of cells adhering to a glass substratum, we feel that this simple and rapid technique has great potential in studies of cell-substratum adhesiveness and of adhesion-related cytoskeletal organization.
Modulations in Mouse Hemopoiesis after Engraftment with Lewis Lung (3LL) Carcinoma Cells
Authors: Weinberg Sheila R. , Gruber Dale F. , Dubois Exum E. , Ledney David
Published: October 2008

Hemopoietic changes in male C57BL/6Cum BR mice engrafted with Lewis lung carcinoma (3LL) were evaluated between day 7, when palpable tumors were present, to day 30 postengraftment. All experimental animals demonstrated decreasing hematocrits (down 40% by day 30) with concurrent leukocytosis which by day 30 postengraftment had reached levels 13.4 times normal. The myelocytic/erythrocytic ratio for normal animals was 1:3 (bone marrow: spleen). The ratio for engrafted animals ranged between 10:1 and 40:1. This apparent shift in production priorities is even more significant in light of the fact that femoral bone marrow cellularity had decreased by 33% on day 17. Splenomegaly, evident by day 7, was seven times control by day 17. Clonogenic analysis of erythroprogenitor cell concentrations revealed an inverse relationship between bone marrow and spleen. 27 days after engraftment, splenic populations demonstrated significant increases in colony forming unit-erythroid (115-fold), burst forming unit-erythroid (7.4-fold), whereas bone marrow concentrations had decreased (6-fold). This report suggests that initiation of 3LL tumor in mice results in a change in the degree of hematopoietic priorities and participation of erythroid organs.
Biological Activity of Synthetic Subunits of Streptococcus Peptidoglycan
Authors: Pekárek J. , Rotta J. , Zaoral M. , Krchňák V. , Straka R. , Ježek J. , Rýc M.
Published: October 2008

Each of a series of synthetic peptidoglycan subunits and subunit analogues was injected in combination with streptococcus type M24 antigen extract. The substances tested were: (8a) N-acetylmuramyldipeptide (MDP) and the following derivatives thereof: MDP modified in positions C3 and C4, or with L-alanine substituted by L-2-aminobutyric acid or with the peptidic chain prolonged (by three lysines or a polylysine); (b) some synthetically prepared peptides: a hexapeptide, a tridecapeptide and an octadecapeptide. Configurations in positions C3 and C4 were found essential for the adjuvant effect. Adjuvant activity, though somewhat lower than in MDP, was pronounced in the analogue containing the L-2-aminobutyryl residue. Surprisingly, potent adjuvant effect was displayed by the hexapeptide; prolongation of the peptidic chain was not effective. The use of a polymeric carrier for MDP increased the adjuvant effect. Contrary to expectation, streptococcal antigens used with immunoadjuvant materials showed that induced delayed hypersensitivity was type related.
Suppression of B Cell Development and Antibody Responses in Mice with Polyclonal Rabbit and Monoclonal Rat Anti-IgM Antibodies
Authors: Cemy Andreas , Hügin Ambros W. , Sutter Stefan , Heusser Christoph H. , Bos Nico , Izui Shozo , Hengartner Hans , Zinkemagel Rolf M.
Published: October 2008

Mice treated from birth with polyclonal, crude or affinity purified rabbit or monoclonal rat anti-mouse IgM antibodies [b-7-6 and C-2-23: Eur. J. Immunol. 14:753–757, 1984] were found to be heavily suppressed with respect to B-cell activities. Crude or affinity purified rabbit or monoclonal rat anti-mouse IgM gave comparable results as follows: (1) serum IgM was below detectable levels; (2) serum IgG was reduced to about 1–3% of normal levels; (3) free anti-IgM was always detectable; (4) IgM and/or Ò¡-light-chain positive cells as well as IgM-secreting cells were absent in various lymphoid organs; (5) the B-cell mitogen lipopolysaccharide was unable to induce proliferative responses; (6) primary antibody responses could not be induced against sheep red blood cells and phosphorylcholine; (7) lymphoid organs were reduced in size and B-cell areas were not populated with lymphocytes; (8) besides a 40% reduction in absolute lymphocyte numbers in the blood, we found increased platelet counts and a 10% eosinophilia in anti-IgM-treated mice.
Characterization of the Giant Myofiber in Bovine Skeletal Muscle
Authors: Sink J.D. , Mann O.M. , Turgut H.
Published: October 2008

Semitendinosus (ST) muscle samples were excised pre- and postrigor from 32 experimental animals (Bos taurus). Four intact and 4 castrate males were exsanguinated when they reached age endpoints of 8, 12, 16, and 20 months. The samples were sectioned, histochemically fiber-typed for SDH and ATPase activity, and examined for giant myofibers. Of the 32 muscles analyzed, only one (8-month intact male) postrigor ST contained a giant myofiber that was hematoxylin and eosin positive, SDH negative, ATPase positive, and 2 × the size of the surrounding fibers. Subsequent studies demonstrated these giant type II white myofibers are an anomaly of muscle contraction and not a distinct fiber type as has been previously reported. This is a first report of giant white or type II myofibers in postrigor bovine skeletal muscle.
Hydrocortisone Promotes the Neodifferentiation of Kirsten Murine Sarcoma Virus Transformed Human Skin Fibroblasts to Adipose Cells: Relevance to Oncogenic Mechanisms
Authors: Kopelovich Levy , Rich Robert F. , Wallace Addajane L.
Published: October 2008

Here we have demonstrated that transformation of human skin fibroblasts (SF) by the Kirsten murine sarcoma virus (KiMSV) is associated with their neodifferentiation into preadipose cells. Hydrocortisone (HC) promotes the transformation/neodifferentiation of such preadipocytes into mature fat cells. The effects of HC on the expression of adipocyte-containing foci and on the total number of transformed foci present in KiMSV-treated cultures appeared to be dose-dependent and was optimal at a concentration of about 500 ng/ml, or 1.25 × 10–6M. Although increasing serum concentrations (2–15%) increased the total number of transformed foci, it had no effect on the expression of adipocyte-containing foci in the presence of HC. The virus-induced preadipocytes undergoing spartial conversion in the presence of HC were capable of clonal expansion and extensive proliferative activity. In contrast, mature adipocytes were terminally differentiated and as such have lost their ability to proliferate. The results suggest a role for a ras oncogene and HC in the transformation/neodifferentiation of human cells that might ultimately lead to cancer in some fraction of such cells.
Low Frequencies of Apparently Fragile X Chromosomes in Normal Control Cultures: A Possible Explanation
Authors: Jenkins Edmund C. , Brown Ted , Brooks Judith , Duncan Charlotte J. , Sanz Maureen M. , Silverman Wayne P. , Lele Kusum P. , Masia Annette , Katz Edward , Lubin Robert A. , Nolin Sarah L.
Published: October 2008

Low frequencies of apparently fragile X [fra(X)] chromosomes have been reported in normal control, short-term, whole blood cultures, and they have been noted in both amniocyte and fetal blood cultures. However, there is currently no universal agreement on the lowest frequency for fra(X)(q27) that is diagnostic for the fragile X syndrome. Here, we present our observations on low levels of apparently fra(X) chromosomes in normal samples. We observed frequencies of 0.5% in short-term whole blood cultures and 0.9% in amniotic fluid cell cultures. In 1982, Steinbach et al. described nonspecific telomeric structural changes (TSC) and suggested that such low frequencies of apparently fra(X) chromosomes in normal material may be occurring by the same mechanism that is responsible for TSC formation. To determine if TSC formation can explain the significant baseline frequencies of fra(X) in normal controls, 10,457 cells were screened from 178 individuals referred for fra(X) analysis. Our findings indicated that TSC are not randomly distributed across chromosomes but tend to occur at specific sites. Based on our observations, we offer the hypothesis that the low frequency of apparent fra(X) in normal individuals may be due to nonrandom TSC distribution.
Colony Formation of Granulocyte (CFU-g) and Macrophage (CFU-m) Precursors in Serum- and Albumin-Free Culture: Effect of Transferrin on Clonal Growth
Authors: Iizuka Yoshikazu , Murphy, Jr. Martin J.
Published: October 2008

Clonal growth of mouse granulocyte and macrophage precursors were assayed in serum-free cultures without albumin. The number of granulocyte/macrophage colonies and clusters increased as transferrin (Trf) concentrations were increased in cultures containing serum-free L-cell-conditioned medium (LCM). On the other hand, cultures with LCM but without Trf produced relatively fewer colonies and clusters. These results indicate that Trf is one of the factors promoting the clonal growth of granulocyte and macrophage precursors in vitro. Although the presence of linoleic acid, α-thioglycerol, and dextran in the culture medium increased the number of granulocyte/macrophage colonies and clusters, these factors were not essential. Serum-free culture of mouse granulocyte and macrophage precursors provides a very useful system with which the activity and function of biological regulators of hematopoietic progenitors may be studied.