A tail length modifier gene discovered in the Japanese wild mice (Mus musculus molossinus)

The presence of the t haplotypes in strains derived from the Japanese wild mice (Mus musculus molossinus) was investigated. Crosses between the T/+ heterozygous short tailed mice and five normal tailed molossinus strains (MOL-ANJ, MOA, MOL-NEM, MOM and Mns) produced no tailless mice, indicating that these strains possess no t haplotype. In contrast, tailless mice were produced by a cross between the T/+ heterozygotes and a MOL-NIS strain. Mating experiments showed that the tailless character was due to an interaction between the T gene and an autosomal recessive gene carried by the MOL-NIS strain that expresses the short tail character under the homozygous condition. We have tentatively named this gene brachyury-interacting tail length modifier (btm). It remains to be investigated whether the btm gene is located in the t complex region or in the other locus.     

Formulation of a flowable liquid concentrate of Bacillus thuringiensis serotype H-14 spores and crystals as mosquito larvicide

Bacillus thuringiensis serotype H-14 spores and crystals, produced in 51 fermenters, were centrifuged and resuspended in emulsified palm olein to give 3.2 x 10(11) colony forming units (cfu)/ml. The suspension was mixed with a cassava-molasses-palm olein-charcoal (CMPC-2) mixture which served as the carrier, adhesive, dispersant and protectant. The final concentration of the formulation was 3.2 x 10(9) cfu/ml. The lethal concentrations capable of killing 50% of the test population (LC50) of CMPC-2 during 0, 1 and 2 years of storage at 32 +/- 4 degrees C were 0.056, 0.058 and 0.058 mg/ml respectively as against 0.054, 0.051 and 0.054 mg/ml for the Institut Pasteur Standard-1978 (IPS-78) during the corresponding period. The chi 2 tests showed that the results were homogeneous at P = 0.05. The relative potencies of the preparations were 964.3, 879.3 and 931 International toxic units (ITU) Aedes aegypti as compared with the 1000 ITU assigned to IPS-78. At 95% confidence limits there was no significant difference between the potencies of CMPC-2 and IPS-78. Field tests showed that CMPC-2 provided between 87.5 and 100% control of natural populations of Aedes spp. and Cutex spp. Sedimentation tests showed that CMPC-2 settled markedly during storage. This, therefore, required that the product be thoroughly shaken before use.     

Interstitial bone stress distributions accompanying ingrowth of a screen-like prosthesis anchorage layer

Recent development of screen-like bonded weaves of titanium wire for orthopaedic implant anchorage affords a unique opportunity for analytic studies of porous ingrowth micromechanics. The regular geometry of individual wires and the periodicity of the mesh weave are exploited in a series of two-dimensional finite element models, mapping interstitial bone stress fields as a function of ingrowth depth and wire size, shape, and spacing.

When the depth of bone ingrowth was less than one wire diameter, peak bone stresses always occurred at the leading (i.e. deepest) edge of bone ingrowth, immediately adjacent to the wire. As ingrowth depth approached a full wire diameter, peak local bone stresses were 2–9 times the nominal applied host bone stress, with greater stresses occurring for lower screen weave densities. Within multiple screen layers, the top layer consistently experienced the peak stress and transmitted most of the applied load, regardless of the number of underlying screen layers surrounded by bone. Neither wire size variations nor partial wire flattening substantially affected general trends in stress predictions.     

A comparison of three muscle pennation assumptions and their effect on isometric and isotonic force

Three different pennation angle assumptions are compared to experimental data from Huijing and Woittiez (Neth. J. Zool. 34, 21-32, 1984) that relate fibre length to angle of pennation changes. The assumptions tested are: (1) neglecting pennation; (2) assuming a fixed pennation; and (3) assuming a constant muscle volume and thickness resulting in pennation angle being dependent on fibre length. Each assumption is compared by transforming fibre force/length and force/velocity characteristics to muscle properties. In general, the fixed pennation assumption provides the worst estimate of muscle force output with a peak error of 0.31 Fo during isometric contractions at small muscle lengths. A better estimate of muscle force output was provided by neglecting pennation entirely. The assumption that the pennation angle changed with fibre length maintained an error of less than 0.05 Fo for most lengths and velocities tested and provided the best estimate of muscle force output.     

Sheep amniotic fluid has a protein factor which stimulates human fibroblast populated collagen lattice contraction.

Sutured incisional wounds made in fetal sheep and rabbits heal without scarring. Fetal sheep excisional wounds can close by contraction, but those in fetal rabbits do not. In vivo and in vitro evidence suggests that rabbit amniotic fluid inhibits wound contraction. The question arises: does sheep amniotic fluid promote wound contraction because their fetal wounds close by contraction? Sheep amniotic fluid (SAF) from 100 and 125 days gestation was tested in fibroblast populated collagen lattice (FPCL) system, an in vitro model of wound contraction. SAF stimulated FPCL contraction in a dose responsive manner. SAF from a 100 day fetus was more stimulating than a 125 day SAF. SAF enhanced FPCL contraction in the presence or absence of serum. SAF was fractionated by size, using column chromatography. It yielded a fraction with an estimated molecular weigh near 40,000 daltons, which stimulated FPCL contraction. The factor was inactivated by proteolytic digestion and heat denaturation. This protein fraction which stimulates FPCL contraction is not related to (1) actin-myosin filaments enhanced contraction by ATP-induced cell contraction, (2) promotion of fibroblast elongation on glass surface or in collagen, or (3) increased cell number by enhanced fibroblast duplication in a collagen matrix. A mechanism for SAF promotion of FPCL contraction was investigated but not identified.     

Characteristics of taurine transport in rat liver lysosomes

Taurine (2-aminoethanesulfonic acid) is a unique sulfur amino acid derivative that has putative nutritional, osmoregulatory, and neuroregulatory roles and is highly concentrated within a variety of cells. The permeability of Percoll density gradient purified rat liver lysosomes to taurine was examined. Intralysosomal amino acid analysis showed trace levels of taurine compared to most other amino acids. Taurine uptake was Na(+)-independent, with an overshoot between 5-10 minutes. Trichloroacetic acid extraction studies and detergent lysis confirmed that free taurine accumulated in the lysosomal space. Kinetic studies revealed heterogeneous uptake with values for Km1 = 31 +/- 1.82 and Km2 greater than 198 +/- 10.2 mM. The uptake had a pH optimal of 6.5 and was stimulated by the potassium specific ionophore valinomycin. The exodus rate was fairly rapid, with a t1/2 of 5 minutes at 37 degrees C. Analog inhibition studies indicated substrate specificity similar to the plasma membrane beta-alanine carrier system, with inhibition by beta-alanine, hypotaurine, and taurine. alpha-Alanine, 2-methylaminoisobutyric acid (MeAIB), and threonine were poor inhibitors. No effects were observed with sucrose and the photoaffinity derivative of taurine NAP-taurine [N-(4-azido-2-nitrophenyl)-2-aminoethanesulfonate]. In summary, rat liver lysosomes possess a high Km system for taurine transport that is sensitive to changes in K+ gradient and perhaps valinomycin induced diffusional membrane potential. These features may enable lysosomes to adapt to changing intracellular concentrations of this osmotic regulatory substance.     

Prostaglandins do not mediate the actions of cholera toxin on pancreatic acini or gastric chief cells from the guinea pig

Recent reports suggest that prostaglandins, rather than cAMP, play a major role in mediating cholera toxin-induced water and electrolyte secretion from rabbit intestinal loops. We examined the role of prostaglandins in mediating toxin-induced pancreatic and gastric exocrine secretion. In these tissues, indomethacin, a potent inhibitor of prostaglandin synthesis, did not alter the stimulatory effects of cholera toxin on increases in cellular cAMP or enzyme secretion. Moreover, the addition of cholera toxin did not alter prostaglandin E2 release from either tissue. In contrast to their effects in rabbit intestinal loops, prostaglandins do not regulate cholera toxin-induced enzyme secretion from the guinea pig pancreas or stomach.     

Continuous activation of primitive hematopoietic cells in long-term human marrow cultures containing irradiated tumor cells.

Human hematopoietic cells can be maintained in vitro for many weeks in the absence of exogenously provided hematopoietic growth factors if an adequate stromal cell containing adherent layer is present. We have now extended the use of this type of long-term culture (LTC) system to create a model of perturbed hematopoiesis in which human tumor cells that constitutively produce a variety of factors are co-cultured together with normal human marrow cells. In the present study, we used the human bladder carcinoma cell line (5637) because these cells were known to produce not only a variety of factors active directly on hematopoietic cells but also factors that can stimulate hematopoietic growth factor production by human marrow stromal cells. Analysis of mRNA extracted from the adherent layer and measurement of growth factor bioactivity in the medium of established LTC of human marrow containing irradiated 5637 cells, showed increased levels of interleukin-1 and -6, as well as granulocyte and granulocyte-macrophage colony-stimulating factor production by comparison to control cultures. As in normal cultures, high proliferative potential clonogenic hematopoietic cells were found almost exclusively in the adherent layer of these co-cultures, but these primitive cells were maintained in a state of continuous turnover, in contrast to control cultures where the same cell types showed the expected oscillation between a quiescent and a proliferating state following each weekly change of the medium. A similar perturbation of primitive progenitor cycling was achieved by adding medium conditioned by 5637 cells twice a week to otherwise normal LTC. The presence of irradiated 5637 cells in the LTC or the addition of 5637 conditioned medium also resulted in modest (2- to 3-fold) but sustained increases in the total hematopoietic progenitor population, as well as in the final output of terminally differentiated granulocytes and macrophages. These findings indicate that primitive hematopoietic cells in LTC can be kept in a state of continuous activation for many weeks by appropriate endogenous or exogenous hematopoietic growth factor provision and that this does not necessarily lead either to their rapid exhaustion or to a large amplification in output of mature progeny.     

The signaling potential of the receptors for insulin and insulin-like growth factor I (IGF-I) in 3T3-L1 adipocytes: comparison of glucose transport activity, induction of oncogene c-fos, glucose transporter mRNA, and DNA-synthesis.

The receptors for insulin and insulin-like growth factor I (IGF-I) have in common a high sequence homology and diverse overlapping functions, (e.g., the stimulation of acute metabolic events and the induction of cell growth.). In the present study, we have compared the potential of insulin and IGF-I receptors in stimulating glucose transport activity, glucose transporter gene expression, DNA-synthesis, and expression of proto-oncogene c-fos in 3T3-L1 adipocytes which express high levels of both receptors. Binding of both hormones to their own receptors was highly specific as compared with binding to the respective other receptor (insulin receptor: KD = 3.6 nM, KI of IGF-I greater than 500 nM; IGF-I receptor, KD = 1.1 nM, KI of insulin = 191 nM). Induction of proto-oncogene c-fos mRNA by insulin and IGF-I paralleled their respective receptor occupancy and was thus induced by both hormones via their own receptor (EC50 of insulin, 3.7; IGF-I, 3.9 nM). Similarly, both insulin and IGF-I increased DNA synthesis (EC50 of insulin, 5.8 nM; IGF-I, 4.0 nM), glucose transport activity (EC50 of insulin, 1.7 nM; IGF-I, 1.4 nM), and glucose transporter (GLUT4) mRNA levels in concentrations corresponding with their respective receptor occupancy. These data indicate that in 3T3-L1 cells the alpha-subunits of insulin and IGF-I receptors have an equal potential to stimulate a metabolic and a mitogenic response.     

Macrophage membrane glycoproteins that bind Griffonia simplicifolia I-B4: effect on cytotoxicity and protein secretion.

Thioglycollate elicited peritoneal (TG-M?s) but not resident peritoneal M?s (R-M?s) were found to bind the lectin Griffonia simplicifolia isotype I-B₄ (GSI-B₄). This was demonstrated by ultrastructural studies and FACS analyses. Membranes from TG-M?s were isolated, separated on SDS-PAGE, electrotransferred onto nitrocellulose, and exposed to peroxidase-labeled GSI-B₄. These procedures revealed two major membrane glycoproteins of molecular weights 180,000 and 94,000 daltons that bound the lectin GSI-B₄ which has a specificity for recognizing terminal α-galactosyl residues. The presence of these epitopes on the two membrane glycoproteins was further substantiated by the fact that treatment of the membranes with α-galactosidase destroyed their capacity to bind GSI-B₄ and that α-D-galactopyranoside but not N-acetyl-D-glucosamine competitively inhibited GSI-B₄ from binding to the glycoproteins. Treatment of TG-M?s with GSI-B₄ reduced the capacity of interferon (IFN) and lipopolysaccharide (LPS), or IFN alone, to induce M? mediated cytotoxicity towards tumor cells by as much as 40%. GSI-B₄ also caused alterations in the pattern of biosynthetically ³⁵S-methionine labeled secreted proteins as early as 2 hours after contact with TG-M?s. Out of 35 discernible proteins on fluorograms of SDS-PAGE separated proteins, 5 were down-regulated and 9 were enhanced. It is suggested that the two novel M? membrane proteins may play a role in regulating the response of M? subpopulations to their humoral and cellular environments.