Peritoneal cavity B cells are precursors of splenic IgM natural antibody-producing cells

Peritoneal cavity B-1 cells are believed to produce IgM natural Abs. We have used alpha1,3-galactosyltransferase-deficient (GalT(-/-)) mice, which, like humans, produce IgM natural Abs against the carbohydrate epitope Galalpha1,3Gal (Gal), to demonstrate that peritoneal cavity B-1b cells with anti-Gal receptors produce anti-Gal IgM Abs only after LPS stimulation. Likewise, peritoneal cavity cells of GalT(-/-) and wild-type mice do not produce IgM Abs of other specificities without LPS stimulation. Development of Ab-secreting capacity is associated with loss of CD11b/CD18 (Mac-1) expression. In contrast, there are large numbers of cells producing anti-Gal and other IgM Abs in fresh splenocyte preparations from GalT(-/-) and (for non-Gal specificities) wild-type mice. These cells are Mac-1(-) but otherwise B-1b-like in their phenotype. We therefore hypothesized a pathway wherein peritoneal cavity B cells migrate into the spleen after activation in vivo and lose Mac-1 expression to become IgM Ab-producing cells. Consistent with this possibility, splenectomy reduced anti-Gal Ab production after immunization of GalT(-/-) mice with Gal-positive rabbit RBC. Furthermore, splenectomized B6 GalT(-/-), Ig micro -chain mutant ( micro (-/-)) (both Gal- and B cell-deficient) mice produced less anti-Gal IgM than nonsplenectomized controls after adoptive transfer of peritoneal cavity cells from B6 GalT(-/-) mice. When sorted GalT(-/-) Mac-1(+) peritoneal cavity B cells were adoptively transferred to B6 GalT(-/-), micro (-/-) mice, IgM Abs including anti-Gal appeared, and IgM-producing and Mac1(-) B cells were present in the spleen 5 wk after transfer. These findings demonstrate that peritoneal cavity Mac-1(+) B-1 cells are precursors of Mac-1(-) splenic IgM Ab-secreting cells.     

Identification and characterization of human Golgi nucleotide sugar transporter SLC35D2, a novel member of the SLC35 nucleotide sugar transporter family

We report the molecular cloning of SLC35D2, a novel member of the SLC35 nucleotide sugar transporter family. The gene SLC35D2 maps to chromosome 9q22.33. SLC35D2 cDNA codes for a hydrophobic protein consisting of 337 amino acid residues with 10 putative transmembrane helices. Northern blot analysis revealed the SLC35D2 mRNA as a single major band corresponding to 2.0 kb in length. SLC35D2 was localized in the Golgi membrane and exhibited around 50% similarity with three nucleotide sugar transporters: human SLC35D1 (UDP-glucuronic acid/UDP-N-acetylgalactosamine transporter), fruitfly fringe connection (frc) transporter, and nematode SQV-7 transporter, the latter two being involved in developmental and organogenetic processes. Heterologous expression of SLC35D2 protein in yeast indicated that UDP-N-acetylglucosamine is a candidate for the substrate(s) of the transporter. The sequence similarity, subcellular localization, and transporting substrate suggest that SLC35D2 is a good candidate for the ortholog of frc transporter, which is involved in the Notch signaling system by providing the fringe N-acetylglucosaminyltransferase with the substrate. We also describe the identification and categorization of the human SLC35 gene family.     

An association between oxidative stress and radiation-induced lymphomagenesis

It is generally thought that reactive oxygen species (ROS) play an important role in carcinogenesis. However, direct evidence supporting this idea is still lacking. In the present study, we measured ROS in thymocytes at the thymic prelymphoma stage in C57BL/6 mice. Mice (n = 20) were irradiated at 1.6 Gy/week for 4 consecutive weeks and the levels of ROS were measured 8 to 11 weeks later by dehydrorhodamine 123, which accumulated in mitochondria and became fluorescent dye upon oxidation. Unirradiated littermates (n = 17) served as controls. Thymic prelymphoma cells were diagnosed by the aberrant CD4/CD8 staining profile and monoclonal or oligoclonal T-cell receptor gene rearrangement. A significant fraction of mice (11/13) bearing thymic prelymphoma cells exhibited elevated levels of ROS in thymocytes (P < 0.001). The result is consistent with the hypothesis that ROS may play an important role in radiation carcinogenesis.     

Histologic changes in thyroid glands from great cormorant (Phalacrocorax carbo) in Tokyo Bay, Japan: possible association with environmental contaminants

We compared morphologic changes in thyroid glands of great cormorants (Phalacrocorax carbo) from the Tokyo Bay and Lake Biwa areas in Japan with presence of residues of polychlorinated dibenzo-dioxins (PCDDs), polychlorinated dibenzo-furans (PCDFs), and coplanar polychlorinated biphenyls (Co-PCBs). Prominent morphologic changes in thyroid glands included increased density of small follicles and increased number of epithelial cells surrounding follicular lumens. The extent of morphologic changes in the thyroid gland was higher in cormorants captured from Tokyo Bay than in those captured from Lake Biwa. Increased thyroid change in cormorants from the Tokyo Bay area was associated with significantly higher levels of PCDFs and Co-PCBs. Thus, we suggest that morphologic changes in thyroid glands from the cormorants are associated with increased levels of dioxin contamination in Japan.     

Understanding the stoichiometric limitation of herbivore growth: the importance of feeding and assimilation flexibilities

Ecological stoichiometry suggests that herbivore growth is limited by phosphorus when this element in the diet is < 8.6 μg P mg C^?1 (C : P atomic ratio > 300). However, in nature, it is not necessarily related to the relative phosphorus content in diets. This may be the result of complex feeding and assimilation responses to diets. We examined these possibilities using herbivorous plankton fed mono‐specific and mixed algae varying in phosphorus content of 1.6 to 8.1 μg P mg C^?1. The herbivores showed a 10‐fold growth rate difference among the diets. Growth rates related poorly with phosphorus content in the diets (r² = 0.07), better with P ingestion rate (r² = 0.41) and best with phosphorus assimilation rate (r² = 0.69). Inclusion of assimilation rates for carbon and fatty acids increased 7% of the explained growth variance. These results indicate that the feeding and assimilation flexibilities play pivotal roles in acquiring a deficient element and in regulating growth rate.     

Ophidian l-amino acid oxidase. The nature of the enzyme–substrate complexes

1. To investigate the kinetics of ophidian l-amino acid oxidase, V and K_m were determined for phenylalanines that were substituted in every ring position with groups of various size and reactivity, and for a few ring-substituted tryptophans and histidines. The venom of one representative from each of three major classes of poisonous snakes, Naja melanoleuca, Vipera russelli and Crotalus adamanteus, served as a source of the ophidian l-amino acid oxidase. Both crude and crystalline enzyme from the venom of C. adamanteus were tested. 2. The introduction of a benzene ring into glycine and alanine caused some increase of V and a very marked depression of K_m. 3. With the exception of fluorine, residues in the ortho position of phenylalanine led to a decrease of V. The rates induced by various substitutions follow the pattern: meta ≥ para ≥ ortho. Within the halogen series, the effects become more pronounced with increasing atomic number. 4. Ring substitution in heterocyclic amino acids also affected the V values markedly. For methyl-substituted tryptophans the pattern was: 5-methyl ≥ 6-methyl ≥ 4-methyl. In a few instances ring substitution accounts for a considerable elevation of V, as shown for β-quinol-4-ylalanine and its 6-methoxy derivative. 5. The kinetic constants appear to be unaffected by relatively high concentrations of the corresponding d-amino acids. 6. A general principle that permits a uniform interpretation of a vast body of information is suggested. It is based on the assumption that most substrates form not only eutopic but also dystopic complexes with the enzyme. The latter, in contrast with the former, do not permit the formation of reaction products. K values for eutopic and dystopic complexes are computed. Similar concepts have been presented to elucidate the action of α-chymotrypsin (Hein & Niemann, 1962) and of monoamine oxidase.     

Ceramide accelerates dephosphorylation of extracellular signal-regulated kinase 1/2 to decrease prostaglandin D(2) production in RBL-2H3 cells.

In the present study, the effect of ceramide on antigen-stimulated phosphorylation of extracellular signal-regulated kinase (ERK) in the mechanism responsible for regulating production of prostaglandin (PG) D(2) was investigated in the mast cell line, RBL-2H3 cells. Cell-permeable C(6)-ceramide (N-hexanoylsphingosine) suppressed antigen-stimulated phosphorylation of ERK1/2 and p38 mitogen-activated protein kinase. Ceramide also inhibited production of PGD(2) and an increase in the activity of cytosolic phospholipase A(2) (cPLA(2)), whereas it did not influence the tyrosine phosphorylation of major cellular proteins in response to antigen. The ceramide-induced inhibition of ERK1/2 phosphorylation and of cPLA(2) activation was suppressed by orthovanadate, a tyrosine phosphatase inhibitor, but not by okadaic acid, a serine/threonine phosphatase inhibitor. Addition of ceramide to the lysate prepared from antigen-stimulated cells reduced the phosphorylated ERK1/2, and orthovanadate effectively prevented the reduction. These results suggest that ceramide accelerates the dephosphorylation of phosphorylated ERK1/2 via activation of a protein tyrosine phosphatase, thus preventing activation of cPLA(2) and production of PGD(2).     

Pregnancy-induced sclerosis in the myometrial vessels of cynomolgus monkeys (Macaca fascicularis)

Sclerotic changes were found histologically in the myometrial vessels of 27 out of 33 cynomolgus monkeys (Macaca fascicularis). The sclerosis was composed of fibrous proliferations and intimal thickening. These changes were not observed in nulliparous cases, but were found only in multiparous ones. The findings suggest that the sclerotic changes in the myometrial vessels of cynomolgus monkeys are a pregnancy-induced phenomenon.     

Potassium and Chloride Effluxes during Excitation of Characeae Cells

Action potentials of intact and tonoplast-free Characeae cellswere studied in relation to the effluxes of K^$ and Cl^-. K^$ effluxwas monitored by atomic absorption spectroscopy or direct measurementof ⁴²K efflux. Cl^- efflux was monitored by the Ag-AgCl electrodemethod. A large K^$ efflux was always observed during excitations irrespectiveof whether the cell was intact or tonoplast-free. In tonoplast-freecells, the permeability of the plasma membrane for K^$ at theplateau of the prolonged action potential was almost the sameas that in the resting state. In many cases, Cl^- efflux wasalso observed during excitations of intact cells although somecells showed action potentials without a detectable increasein Cl^- efflux. Almost all tonoplast-free cells generated actionpotentials with no associated increase in Cl^- efflux. Theseresults indicate that some ion other than Cl^- carries inwardcurrent which balances the outward current carried by K^$. Calciumion is discussed as the most probable candidate. (Received November 18, 1983; Accepted June 12, 1984)