Heterogeneous phenotype of human glioblastoma: In vitro study

Glioblastomas (GBMs) are the most lethal primary brain tumours. Increasing evidence shows that brain tumours contain the population of stem cells, so‐called cancer stem cells (CSCs). Stem cell marker CD133 was reported to identify CSC population in GBM. Further studies have indicated that CD133 negative cells exhibiting similar properties and are able to initiate the tumour, self‐renew and undergo multilineage differentiation. GBM is a highly heterogeneous tumour and may contain different stem cell populations with different functional properties. We characterized five GBM cell lines, established from surgical samples, according to the marker expression, proliferation and differentiation potential. CD133 positive cell lines showed increased proliferation rate in neurosphere condition and marked differentiation potential towards neuronal lineages. Whereas two cell lines low‐expressing CD133 marker showed mesenchymal properties in vitro, that is high proliferation rate in serum condition and differentiation in mesenchymal cell types. Further, we compared therapy resistance capacity of GBM cell lines treated with hydroxyurea. Our results suggest that CSC concept is more complex than it was believed before, and CD133 could not define entire stem cell population within GBM. At least two different subtypes of GBM CSCs exist, which may have different biological characteristics and imply different therapeutic strategies. Copyright © 2013 John Wiley & Sons, Ltd.     

A comparative study of the soil zinc fractions determined by chemical methods and electro-ultrafiltration (EUF) and their relations to zinc nutrition in rice

Rice (IR 42) was grown on two soils differing in zinc status for 30 days with and without Zn under submerged conditions in pots. The fate of soil zinc was characterized by extraction of the soil successively with copper acetate and sodium hypochlorite and by EUF extraction. Most of the applied zinc was extracted by copper acetate and represented as complexed fraction. There exists a close and significnat relation between Cu(OAc)₂-extractable zinc and Zn extracted by EUF for 5 minutes at 50 volts (r=0.98). The EUF-extractable zinc and Cu(OAc)₂-extractable zinc were significantly correlated with the zinc content in the plant (r=0.82). The data from this investigation suggest the possibility of Zn fractionation with the EUF technique and the fractions obtained agree closely to those determined by chemical methods. The results obtained indicate that Zn in soil is held by weak organic bonding and that the extractions by Cu(OAc)₂ and/or EUF-5 minutes serve as a useful basis for extimating zinc availability in rice soils.     

Variation in Male Fertilities and Pairwise Mating Probabilities in Picea glauca

Frequencies of multilocus male gametes in seeds collected from clones in several blocks of a white spruce seed orchard were analyzed as part of a 2-yr study of mating system variation in this species. Observed frequencies of male gamete types departed significantly from those expected assuming equal male fertilities among clones. Male gamete frequencies in seed crops were significantly heterogeneous among clones within blocks, and among blocks within clones. Clonal male fertilities were estimated from male gamete frequency data. These estimates were highly skewed, with a small proportion of the clones contributing male gametes to the majority of the seed. The estimates were significantly heterogeneous among clones within blocks, and among blocks within clones. Between-year variation in clonal male fertilities was also pronounced, with male fertilities of some clones changing by as much as three orders of magnitude. Clonal male fertility was significantly correlated with clonal male cone production in both years. These results are important with regard to assumptions made for the estimation of general combining ability, average genetic correlation among progeny from single parents, and expected response to selection in open-pollinated plant populations.     

Light and electron microscopical immunocytochemistry of 5'-nucleotidase in rat cerebellum

5'-Nucleotidase in nervous tissue has so far not been localised at the ultrastructural level using immunocytochemical techniques. We have now applied monoclonal antibodies and a polyclonal antiserum raised against this ecto-enzyme and describe the distribution of 5'-nucleotidase antigenicity in rat cerebellum both at the light and electron microscopic levels. Within all cerebellar layers, 5'-nucleotidase immunoreactivity was found on plasma membranes of glial elements, i.e. Bergmann glial cell processes crossing the molecular layer, astrocytic end-feet around blood vessels and glial cell extensions surrounding single Purkinje cells. In the granular layer, 5'-nucleotidase immunoreactivity was present on glial membranes interposed between granule cells. Neuronal cells or processes were devoid of immunoreactivity. The immunocytochemical results were compared with conventional 5'-nucleotidase histochemistry. Both techniques showed the same ecto-localisation of the enzyme and favour the view of 5'-nucleotidase being predominantly situated at glial plasma membranes.     

Suppression of growth defects of α-amylase secreting Escherichia coli by signal sequence fusion

Abstract In a previous study, we have described unusual cross-reactions among monoclonal antibodies (Mabs) to bacteria and in particular to the Inaba and Ogawa serotypes of Vibrio cholerae . In this study, the extent to which the binding sites of both antibodies and antigens overlap has been investigated by competitive binding and idiotypic analysis. The competitive binding data indicate that the cross-reactive binding of the Inaba Mabs to the Ogawa vibrios can be abolished by incubation with higher affinity Ogawa Mabs. However, rabbit antiserum raised against the Inaba series does not react with the Ogawa series, indicating that anti-Inaba Mabs do not share idiotypic determinants with anti-Ogawa Mabs. The results therefore suggest that the two sets of antibodies recognise different determinants which are closely related in spatial terms, and which consequently do not permit simultaneous binding of the two types of monoclonal antibody.     

Acardiac fetus in a triplet pregnancy

The acardiac monster represents one of the most severe but rare congenital anomalies. It occurs only in multiple gestations associated with vascular anastomoses between the affected fetus and its co-twin. The prenatal diagnosis of an acardiac fetus must be suspected in any multiple gestation in which cardiac activity cannot be documented sonographically in a growing fetus. We report an acardiac fetus occurring in a spontaneously conceived triplet pregnancy. A review of the literature, including pathogenetic theories and sonographic reports, is discussed.     

A coordinate relationship between theGALK and theTK1 genes of the Chinese hamster

Chinese hamster cells in culture were treated with various concentrations of thymidine, 5-bromodeoxyuridine, trifluorothymidine, and 2-deoxy-D-galactose. Selection was made for deficiencies in the activities of galactokinase and thymidine kinase. Selection in the presence of thymidine, 5-bromodeoxyuridine, and trifluorothymidine was expected to produce clones deficient in thymidine kinase only, whereas those deficient in galactokinase were expected to be selected in the presence of 2-deoxy-D-galactose. However, it was found that clones growing in the presence of these inhibitors were frequently deficient in both enzymes. Or if a clone was deficient in only one, the deficiency frequently was not expected according to the selection procedure. This indicates some sort of coordinate relationship between the two gene loci, GALK and TK1, which specify galactokinase and thymidine kinase, respectively. GALK and TK1 are linked in all primates and rodents in which linkage determinations have been made. It is therefore probable that this linkage has been conserved for a long period of time. It is suggested that the apparent relationship between the two genes shown by the data presented here, as well as by others, supports the conclusion that linkage has been conserved by natural selection and is therefore not fortuitous.     

Current-voltage relations of the apical and basolateral membranes of the frog skin

We determined the current-voltage (I-V) relations of the apical and basolateral barriers of frog skins by impaling the cells with an intracellular microelectrode and assuming that the current across the cellular pathway was equal to the amiloride-inhibitable current. We found that: (a) The responses in transepithelial current and intracellular potential to square pulses of transepithelial potential (VT) varied markedly with time. (b) As a consequence of these transient responses, the basolateral I-V relation was markedly dependent on the time of sampling after the beginning of each pulse. The apical I-V plot was much less sensitive to the time of sampling within the pulse. (c) The I-V data for the apical barrier approximated the I-V relations calculated from the Goldman constant field equation over a relatively wide range of membrane potentials (+/- 100 mV). (d) A sudden reduction in apical bath [Na+] resulted in an increase in apical permeability and a shift in the apical barrier zero-current potential (Ea) toward less positive values. The shift in Ea was equivalent to a change of 45 mV for a 10-fold change in apical [Na+]. (e) The transient responses of the skin to square VT pulses were described by the sum of two exponentials with time constants of 114 and 1,563 ms, which are compatible with the time constants that would be produced by an RC circuit with capacitances of 65 and 1,718 microF. The larger capacitance is too large to identify it comfortably with a true dielectric membrane capacitance.     

The two-sex multiethnic stable population model

A two-sex multiethnic stable population is a model with fixed mortality and fertility parameters that explicitly recognizes the behavior of males and females in at least two distinct ethnic groups. The presence of intergroup fertility may allow the different ethnic groups to grow at a constant rate in a population with a fixed sex-ethnic composition. The present paper considers a variety of rules for determining the ethnicity of intergroup births based on the ethnicities of the mother and father, and examines the mathematical models implied by those rules. Numerical examples are presented for a two-ethnic-group population in cases where intergroup births are shared equally by the two groups, are all considered members of one particular group, and are all members of the father's group. A special case of a more general model, where sons become members of the father's group and daughters become members of the mother's group, is also considered. The results suggest that when intergroup fertility is not uncommon, how ethnicity is determined can substantially influence the ethnic composition of the population.     

Molecular weights of Cucurbita sieve tube proteins

Summary By means of SDS-polyacrylamide-gel electrophoresis, molecular weights of 15000, 28000, 59000, 116000 and 220000 were determined for the main sieve tube proteins from Cucurbita maxima.