The BULT Melanoma: A Spontaneous Transplantable Tumor in Mice

The BULT melanoma originated at Brown University as a spontaneous, small black nodule on the tail of an adult female mouse of the LT/Ch strain. Histological examination of a portion of the tumor indicated that it was intradermal and consisted predominantly of heavily melanized, ovoid to fusiform cells with melanin-laden macrophages scattered among them. The BULT melanoma has been maintained in LT/Ch mice for approximately 5 years by periodic transplantation, at first subcutaneously on the flanks and, more recently, intramuscularly in the hind legs. The shift in transplantation site was made following a marked decline in the growth of subcutaneous grafts. The transplants have retained the uniform deep-black melanization and general histology of the primary melanoma. Numerous melanosomes at all stages of development are found within the melanoma cells. DOPA-positive cytoplasmic vesicles are abundant. Occasional autophagic vacuoles containing clusters of melanosomes are also present. A few metastases from the transplanted melanoma have been observed in lymph nodes and on one occasion in the lungs. When grown in vitro, BULT melanoma cells do not require special growth promoting agents (e.g., TPA; cAMP) in order to proliferate. The BULT melanoma differs in one or more respects from each of the other three transplantable spontaneous mouse melanomas widely used in cancer research. In addition, it arose in a strain of mice characterized by the spontaneous death of melanocytes while the latter are engaged in synthesizing eumelanin within hair follicles. Karyotypic analysis of cultured cells showed a modal chromosome number of 68 with a range of 58–72 chromosomes.     

Coupling of M3 Acetylcholine Receptor to Gq16 Activates a Natriuretic Peptide Receptor Guanylyl Cyclase

Muscarinic activation of tracheal smooth muscle (TSM) involves a M₃AChR/heterotrimeric-G protein/NPR-GC coupling mechanism. G protein activators Mastoparan (MAS) and Mastoparan-7 stimulated 4- and 10-fold the NPR-GC respectively, being insensitive to PTX and antibodies against Gα_i/o subfamily. Muscarinic and MAS stimulation of NPR-GC was blocked by antibodies against C-terminal of Gα_q16, whose expression was confirmed by RT-PCR. However, synthetic peptides from C-terminal of Gα_q15/16 stimulated the NPR-GC. Coupling of α_q16 to M₃AChR is supported by MAS decreased [³H]QNB binding, being abolished after M₃AChR-4-DAMP-alkylation. Anti-i₃M₃AChR antibodies blocked the muscarinic activation of NPR-GC, and synthetic peptide from i₃M₃AChR (M₃P) was more potent than MAS increasing GTPγ [³⁵S] and decreasing the [³H]QNB activities. Coupling between NPR-GC and Gα_q16 was evaluated by using trypsin-solubilized-fraction from TSM membranes, which displayed a MAS-sensitive-NPR-GC activity, being immunoprecipitated with anti-Gα_q16, also showing an immunoreactive heterotrimeric-G-β -subunit. These data support the existence of a novel transducing cascade, involving Gα_q16β γ coupling M₃AChR to NPR-GC.     

Seasonal dynamics of crustacean zooplankton, heterotrophic nanoflagellates and bacteria in a shallow, eutrophic lake

1. The seasonal development of crustacean zooplankton, heterotrophic nanoflagellates (HNF) and bacteria was examined in Grosser Binnensee, a shallow, eutrophic lake in northern Germany. The grazing impact of Daphnia on bacteria and nanoflagellates was estimated from field data on population abundances and from clearance rates obtained in laboratory experiments. 2. The seasonal succession of zooplankton showed distinct peaks of Daphnia magna, cyclopopid copepods, Bosmina longirostris and Daphnia galeata and D. hynlina. The population dynamics of Dapfinia had the strongest impact on all sestonic components. Daphnia maxima coincided with clearwater phases, and were negatively correlated with particulate organic carbon (POC), HNF and phytoplankton. Bacterial abundance was only slightly affected although daphnids were at times more important as bacterial consumers than HNF, as estimated from measured bacterial clearance rates. Other crustaceans (copepods, Bosmina) were probably of minor importance as grazers of bacteria and nanoplankton. 3. HNF abundance varied from 550 ml^?1 to more than 30000 ml^?1. HNF appeared to be suppressed by daphnids and reached highest densities when copepods dominated the metazooplankton. The variation in HNF abundance was not reflected in the concentration of heterotrophic bacteria, which fluctuated rather irregularly between 5 and 20 ± 10⁶ ml^?1. Long filamentous bacteria which were probably resistant to protozoan grazing, however, appeared parallel to the development of HNF. These bacterial cells, although small in number, could comprise more than 30% of the total bacterial biomass.     

Positive effects of native shrubs on three specially protected cacti species in Durango,México

The spatial association between the specially protected cacti Coryphanta durangensis, Echinocereus longisetus and Peniocereus greggii and potential nurse plants was evaluated, as was their relative position to the sun under the crown of the latter in the southern Chihuahuan Desert. The soil temperature under potential nurse plants was lower than under direct sunlight. There was more nitrogen and organic matter in the soil under Prosopis laevigata trees than in soil under direct sunlight. There were 68 plants of C. durangensis, 59 plants of E. longisetus and 157 of P. greggii. Only one individual of C. durangensis and one of E. longisetus grew outside the shade of the crown of other plants. Echinocereus longisetus was not associated with any particular species, but grew more often than expected by chance in the northern segment of the crown (i.e. the area most shaded in the afternoon). Coryphanta durangensis and P. greggii grew more often under P. laevigata than expected by chance. The effect of other plants, and P. laevigata in particular, on the facilitation of growth of protected cacti species should be considered in management plans of the Chihuahuan Desert, where mesquite (P. laevigata) is often harvested for charcoal production.     

Ediacaran microbial colonies

Enigmatic discoidal fossils are common in Neoproterozoic sedimentary sequences and in the stratigraphic record pre-date the first appearance of diverse Ediacaran fossil assemblages. Termed 'medusoids', these Neoproterozoic discoidal fossils have generally been interpreted as coelenterate-grade organisms implying a radially symmetrical body plan for ancestral eumetazoans. Analysis of exceptionally preserved discoidal fossils from the White Sea area, however, indicates that most of these discoidal forms represent colonial microbes. Localized pyritization, for example, reveals the presence of a conspicuous filamentous substructure in Ediacaria , whereas concentric rings, radial sectors and central structures in Cyclomedusa and Paliella compare directly with Recent microbial colonies growing in a nutritionally heterogeneous environment. At least some Ediacaran discoids can be compared with extant concentric ring-shaped microbial colonies that grow in hypersaline microbial mats. Insofar as most of the remaining record of Ediacaran discoids can be attributed to the holdfast structures of non-radiate modular organisms, there is no support from the fossil record for identifying a radiate ancestry for the Metazoa.     

<Emphasis Type="Italic">E. coli</Emphasis> DNA Polymerase I and other Host Functions participate in T4 DNA Replication and Recombination

The recognition of bacterial functions involved in DNA metabolism of bacteriophage T4 might reveal interactions between different enzymes during DNA replication and recombination. To detect such functions we have studied the replication of complete and incomplete T4 chromosomes in various mutant strains of Escherichia coli that are defective in their own DNA metabolism. We found that several E. coli functions can substitute for phage functions in T4 replication and recombination and will discuss here the role of the E. coli pol A gene which codes for DNA polymerase I^1–4 and of the dna B and E genes^3,5.     

Production via good manufacturing practice of exofucosylated human mesenchymal stromal cells for clinical applications

Background

The regenerative and immunomodulatory properties of human mesenchymal stromal cells (hMSCs) have raised great hope for their use in cell therapy. However, when intravenously infused, hMSCs fail to reach sites of tissue injury. Fucose addition in α(1,3)-linkage to terminal sialyllactosamines on CD44 creates the molecule known as hematopoietic cell E-/L-selectin ligand (HCELL), programming hMSC binding to E-selectin that is expressed on microvascular endothelial cells of bone marrow (BM), skin and at all sites of inflammation. Here we describe how this modification on BM-derived hMSCs (BM-hMSCs) can be adapted to good manufacturing practice (GMP) standards.

Are the trichomes in corollas of Mutisieae (Asteraceae) really twin hairs?

The corollas of three species of Onoseris , 13 species of Trichocline and one species of Uechtritzia (Asteraceae, Mutisieae) exhibit trichomes that are morphologically similar to the twin hairs which are traditionally considered exclusive of the cypselas in Asteraceae. Four types of trichomes were found: 1-celled, 2-celled, 3-celled, and 4-celled. The 3-celled and 4-celled corolla hairs are those which most resemble the typical cypsela twin hairs. The ontogeny of the corolla hairs was analysed and coincidences with the ontogeny of the cypsela twin hairs were found. (1) An anticlinal division of the epidermal mother cell originates two hair cells which, in turn, originate the basal cells (sometimes one of them is reduced or similar to the epidermal cells) by oblique or periclinal subdivision. (2) In some cases the basal cell(s) of the 3- or 4-celled corolla hair elongates and reaches the same length as the hair cells. The same kind of trichomes have been demonstrated in cypselas of Mutisieae. (3) The 1- and 2-celled corolla hairs have already been described as variants of the cypsela twin hairs. Based on this evidence we conclude that the corolla hairs of Onoseris , Trichocline and Uechtritzia are twin hairs. It is hypothesized that the 1–4 corolla hairs could be involved in water absorption, as occurs in the cypsela twin hairs.  © The Linnean Society of London, Botanical Journal of the Linnean Society , 2002, 140 , 427–433.