Density gradient centrifugation of tumor cells from needle biopsies and their respective source tumors: a comparison of density distributions

A needle biopsy technique was applied to a murine fibrosarcoma (FSa) tumor system. FSa tumors of 8 mm diameter in size were biopsied and then made into single cell suspensions. Resulting density profiles of cells from both sources were compared following centrifugation in Renografin gradients. In all cases, there was excellent agreement between the density profiles of the material from each of the biopsies and the corresponding solid tumors.     

Density gradient centrifugation of cells separated from multicellular tumor spheroids.

Cells from a murine fibrosarcome (FSa) have been grown in vitro as multicell tumor spheroids (MTS). The growth rate of these MTS was determined. Following selected periods of growth, MTS were made into a single cell suspension and separated on linear density gradients of Renografin. While only 1 population of cells were separated from small spheriods (400 mum diameter), at least 3 subpopulations of tumor cells were separated and isolated from large spheroids (800 mum in diameter).     

Density gradient centrifugation of cells separated from multicellular tumor spheroids

Summary Cells from a murine fibrosarcoma (FSa) have been grown in vitro as multicell tumor spheroids (MTS). The growth rate of these MTS was determined. Follwing selected periods of growth, MTS were made into a single cell suspension and separated on linear density gradients of Renografin. While only 1 population of cells was separated from small spheroids (400 m diameter), at least 3 subpopulations of tumor cells were separated and isolated from large spheroids (800 m in diameter).This investigation was supported in part by grants numbered CA-18628 and CA-06294, awarded by the National Cancer Institute, DHEW.     

Neocortical transplants in the rat brain: an ultrastructural study

Summary Electron microscopic analysis of neocortical transplants in the cerebellum of the host animals showed that the nerve cells, glial cells, and neuropil of the transplants were normal. These transplants showed anatomical integration with the host brain through various regions of interface. Neuropil interfaces were found to have a high density of synaptic profiles, and medullary interfaces had a very small number of synaptic profiles.Acknowledgment. Research supported by Contract No. N00014-83 from the Office of Naval Research, Department of the Navy to E.N.A., and N.I.H. Research Grant No. NS-08817 to G.D. Das.     

Neocortical transplants in the rat brain: an ultrastructural study

Electron microscopic analysis of neocortical transplants in the cerebellum of the host animals showed that the nerve cells, glial cells, and neuropil of the transplants were normal. These transplants showed anatomical integration with the host brain through various regions of interface. Neuropil interfaces were found to have a high density of synaptic profiles, and medullary interfaces had a very small number of synaptic profiles.     

Establishment and characterization of 3 human cell lines derived from intracerebral metastatic tumors]

Twelve human tumors were cultivated in vitro by the trypsinization technique and 3 cell lines were established. Each of these 3 lines showed individual characteristics which were maintained during passages. After inoculation into nude mice, 2 lines induced tumors which possessed the histologic features of the original human tumors (melanoma and carcinoma of stomach), the other line derived from a breast carcinoma was rejected.     

Different types of synaptic vesicles in axons of the retractor penis muscle of the bull

Summary Three types of axon profiles were observed in the smooth muscle of the retractor penis and the penile artery of the bull: 1. profiles containing small granular vesicles, presumably representing adrenergic axons; 2. profiles containing small agranular vesicles, presumably representing cholinergic axons; 3. profiles containing numerous large and small granular vesicles. The third type of profile was not found in the vas deferens or the metatarsal artery. It is therefore possible that this type of profile represents the non-adrenergic, non-cholinergic inhibitory nerves, the presence of which has previously been pharmacologically indicated in these tissues.This study has been supported by grants from the Sigrid Jusélius Foundation (to O. E.), from the Medical Research Council of the Academy of Finland (to E. K.) and from the Magnus Bergvall Memorial Fund (to N. O. S.). Excellent technical assistance by Mrs.Paula Hasenson is gratefully acknowledged.     

Molecular characterization of common fragile sites as a strategy to discover cancer susceptibility genes

The cytogenetic hypothesis that common fragile sites (cFSs) are hotspots of cancer breakpoints is increasingly supported by recent data from whole-genome profiles of different cancers. cFSs are components of the normal chromosome structure that are particularly prone to breakage under conditions of replication stress. In recent years, cFSs have become of increasing interest in cancer research, as they not only appear to be frequent targets of genomic alterations in progressive tumors, but also already in precancerous lesions. Despite growing evidence of their importance in disease development, most cFSs have not been investigated at the molecular level and most cFS genes have not been identified. In this review, we summarize the current data on molecularly characterized cFSs, their genetic and epigenetic characteristics, and put emphasis on less-studied cFS genes as potential contributors to cancer development.     

Transcriptome profiling of lung schistosomula,in vitro cultured schistosomula and adult Schistosoma japonicum

The schistosomulum is the main target of vaccine-induced protective immunity; however, most studies have utilized schistosomula produced by mechanical transformation of infective larvae followed by in vitro culture rather than larvae isolated directly from the lungs of infected mammals. Using transmission electron microscopy, we demonstrated that there was little difference in the ultrastructure of Schistosoma japonicum schistosomula obtained by the two methods. However, significant differences in gene expression profiles were apparent when we used an oligonucleotide microarray to compare the gene expression profiles of schistosomula obtained in vivo from lung tissue with those maintained in vitro, and with adult worms of S. japonicum. It is likely that host environmental factors, which cannot be reliably reproduced in vitro, do influence the growth, development and overall biology of schistosomes. Thus caution is urged when using in vitro-cultured schistosomes and mechanically transformed/cultured schistosomula in molecular, biochemical and immunological studies.     

Inhibition of transplanted mouse tumors by heterologous transfer RNA

Summary Injections of yeast tRNA to C57BL mice decreased takes and inhibited growth of syngeneic transplanted tumors. Mice remaining free of tumors as result of this treatment failed to develop tumors after challenge with 5×10⁴ cells of the same tumor.This work was supported in part by a grant from the Stanley Johnson Foundation, Berne, Switzerland.     

Tumorigenic activity of cloned polyoma virus DNA in newborn rats

Summary The proximal portion of the polyoma virus early region as well as the complete viral genome were cloned in pBR322. Recombinant plasmids induced tumors in newborn rats but only after linearization of the DNA by various restriction endonucleases.This work was financed by grant MA-6731 from the Medical Research Council and by a grant from the National Cancer Institute of Canada.     

Different types of synaptic vesicles in axons of the retractor penis muscle of the bull.

Three types of axon profiles were observed in the smooth muscle of the retractor penis and the penile artery of the bull: 1. profiles containing small granular vesicles, presumably representing adrenergic axons; 2. profiles containing small agranular vesicles, presumably representing cholinergic axons; 3. profiles containing numerous large and small granular vesicles. The third type of profile was not found in the vas deferens or the metatarsal artery. It is therefore possible that this type of profile represents the non-adrenergic, non-cholinergic inhibitory nerves, the presence of which has previously been pharmacologically indicated in these tissues.     

Interstrain variation of murine arylsulfatase C

Summary A method has been developed for the assay of arylsulfatase C in tissue extracts containing arylsulfatases A and B. Significant variation of enzyme activity was observed among 26 inbred murine strains. Activity differences were apparent at all stages evaluated between 1 and 70 days postnatal age. Arylsulfatase C from representative high- and low-activity strains exhibited similar Michaelis constants, temperature optima, pH optima, thermostabilities and inhibitor profiles.This work was supported in part by Biomedical Sciences Support Grant: HEW PHS RR 07030.     

Tumorigenic effect of topical mechlorethamine,BCNU and CCNU in mice

Summary Mice were painted with mechlorethamine (HN2), carmustine (BCNU) and lomustine (CCNU) for up to 33 weeks. HN2 was a potent carcinogen, producing squamous cell carcinomas in 9 of 33 mice in 1 series. BCNU was a weak carcinogen. CCNU produced no tumors in 1 series.Supported in part by PHS grant CA-14825 from the National Cancer Institute.     

Proliferation and cell loss of human leukemic cell subpopulations in liquid culture

A kinetic study was performed on leukemic blasts from patients with acute myeloid leukemia, separated into 2 subpopulations by a specific density gradient. The growth curve and the [3H]-thymidine uptake were simultaneously analyzed. While cumulative nucleotide uptake fitted with the growth kinetics in the low-density fraction, such a concordance was not found in the high-density subpopulation. That indicated the occurrence of simultaneous growth and loss in the high density fraction, which could not be evaluated by a simple numerical determination.     

Decreased monosaccharide transport in renal brush-border membrane vesicles of spontaneously hypertensive rats

Na(+)-dependent D-glucose and D-galactose transport were studied in brush-border membrane vesicles (BBMVs) from kidney cortex isolated from both spontaneously hypertensive rats (SHR) and their normotensive genetic control Wistar-Kyoto (WKY) rats. Initial rates and accumulation ratios of Na(+)-dependent D-glucose and D-galactose transport were significantly lower in SHR compared with WKY, the observed decreases being similar for both substrates. To explain the reduction in sugar transport by renal BBMVs, the density of Na(+)-dependent sugar cotransporters was studied in BBMVs from kidney cortex isolated from SHR and WKY rats. Phlorizin-specific binding and Western blot analysis indicated a reduction in the density of the cotransporters in SHR relative to WKY rats. This reduction was similar to those found for the initial rates and accumulation ratios for D-glucose and D-galactose in SHR. Na+ uptake, studied using 22Na+, was significantly increased in SHR, so the observed reduction in sugar transport could be due to disruption of the Na+ gradient between renal BBMVs in SHR. Furthermore, a significant decrease in the activity of Na(+)-K(+)-ATPase was observed in SHR. In conclusion, changes in the density of the Na(+)-dependent sugar cotransporter and in the Na+ gradient across the brush-border membranes might be involved in the observed reduction in sugar transport by renal BBMVs from SHR.     

Shoot regeneration fromAgrobacterium tumefaciens-induced tumors of a tropical timber tree,Fagraea fragrans

Summary Agrobacterium tumefaciens A208 with nopaline plasmid pTiT37 was used to obtain stem tumors on plantlets ofFagraea fragrans grown in vitro. Bacterial elimination and tissue proliferation were simultaneously achieved by growing tumors on cefatoxime medium. After some tissue growth the shoots regenerated. An examination of these showed the presence of nopaline, indicating genetic transformation by T-DNA.     

Correlation of histochemical and physiological properties of muscle units in the striped skunk

Summary Single motor units in striped skunk medial gastrocnemias were isolated and physiologically characterized. Individual muscle unit fibres tagged by glycogen depletion were analyzed histochemically and found to have histochemical profiles consistently correlated with the physiological findings.This work was supported by the Organized Research Committee, Northern Arizona University.     

Tumor incidence in Visna virus inoculated mice

Summary Mice (female Swiss albino) inoculated when newborn with Visna virus had tumors in 77% of cases when examined 8–12 months later. The tumors were mainly of the mammary carcinoma type. The tumor incidence in noninfected control animals was only 20%. In contrast, no increased incidence of tumors was observed among Visna virus-inoculated inbred mice (BALB/c, CBA and DBA) with low incidence of spontaneous mammary carcinoma.This study was supported by grant No. B75-16X-4511-01 of the Swedish Medical Research Council.     

Spatial distribution of signal and adaptive sensitivity in the receptive field surrounds of cat retinal ganglion cells

Summary A comparison was made between adaptive and signal sensitivity profiles of the surround response mechanism of cat retinal ganglion cells. The 2 profiles were found to be similar for X cells but the surrounds of Y cells appear to pool adaptation over a smaller retinal region than they pool signals. Acknowledgments. We wish to thank Mr O. Navarro for his valuable technical assistance. This research is supported by Public Health Service grant No. EY 00701.