Ron-mediated cytoplasmic signaling is dispensable for viability but is required to limit inflammatory responses.

Ron receptor activation induces numerous cellular responses in vitro, including proliferation, dissociation, and migration. Ron is thought to be involved in blood cell development in vivo, as well as in many aspects of the immune response including macrophage activation, antigen presentation, and nitric oxide regulation. In previous studies to determine the function of Ron in vivo, mice were generated with a targeted deletion of the extracellular and transmembrane regions of this gene. Mice homologous for this deletion appear to die early during embryonic development. To ascertain the in vivo function of Ron in more detail, we have generated mice with a germline ablation of the tyrosine kinase domain. Strikingly, our studies indicate that this domain of Ron, and therefore Ron cytoplasmic signaling, is not essential for embryonic development. While mice deficient in this domain are overtly normal, mice lacking Ron signaling have an altered ability to regulate nitric oxide levels and, in addition, have enhanced tissue damage following acute and cell-mediated inflammatory responses.

Inter-alpha-inhibitor binding to hyaluronan in the cumulus extracellular matrix is required for optimal ovulation and development of mouse oocytes.

This report characterizes the effects of excess hyaluronan (HA) upon the expansion of the cumulus oocyte complex (COC) within intact follicles and upon ovulation and oocyte viability in mice. Covalent linkage between heavy chains of the inter-alpha-inhibitor (IalphaI) family of serum glycoproteins and HA is necessary for optimal cumulus extracellular matrix (cECM) stabilization and cumulus expansion. Intravenous administration of HA oligosaccharides inhibited the binding of IalphaI to endogenous HA, disrupting the process of expansion and resulting in a reduction in the size of the cumulus mass. Western blot and immunocytochemical analyses of COCs from HA-treated animals demonstrated a reduction of IalphaI heavy chains within the cECM. Additionally, HA-treated immature animals ovulated 56.3% fewer COCs compared to control animals. The developmental potential of COCs in HA-treated animals was also tested. Extended periods of oviductal storage of COCs ovulated by HA-injected adult mice resulted in a reduction of normal embryos and a significant increase in the proportion of fragmented oocytes/embryos. These observations support the view that covalent binding of IalphaI heavy chains to HA is required for optimal cumulus expansion, extrusion of the COCs from the follicle at ovulation, and maintenance of oocyte viability within the oviduct.

The ovarian blood follicle barrier is both charge- and size-selective in mice.

This report characterizes the permeability and selectivity properties of the ovarian blood-follicle barrier. Proteins of similar size but opposite net charge possess strikingly different permeabilities with respect to this barrier. Inter-alpha-inhibitor (I alpha I, 220 kDa, pI approximately 6.2) is excluded from the follicle until an ovulatory stimulus, whereas immunoglobulin G (IgG, 155 kDa, pI approximately 6.5-7.0) passes into the follicle without an ovulatory stimulus. However, cationization of I alpha I results in its influx into the follicle in the absence of an ovulatory signal. Conversely, anionization of IgG results in its exclusion from the follicle unless an ovulatory stimulus (hCG administration) is provided. Molecular size also plays a role in blood-follicle barrier selectivity. For example, cationization of alpha 2-macroglobulin (pI approximately 8.5; 700 kDa) fails to facilitate its entry into unstimulated follicles. Conversely, negatively charged BSA (pl approximately 4.5; 66 kDa) passes freely into unstimulated follicles. These studies support the hypothesis that the blood-follicle barrier is size-selective but that charge sign and density play a role in the permeability of this barrier to proteins within an intermediate size range.

Spatial and temporal expression of 4f-rnp gene in Drosophila melanogaster.

We have recently discovered that a pre-messenger RNA (mRNA) encoded by the 4f-rnp gene in Drosophila melanogaster undergoes RNA editing by site-specific A-to-G conversions. In this report, we describe the spatial and temporal expression patterns of 4f-rnp mRNA transcripts during fly development. The 4f-rnp locus was mapped by polytene in situ hybridization experiments to the 4F1,2 bands at the distal tip of the X-chromosome, demonstrating that this gene is nuclear. We show that 4f-rnp is a single-copy gene by a combination of genomic Southern blot hybridization and restriction map analysis of clones isolated through chromosome walking techniques. Northern blot analysis indicates that two alternatively spliced messenger RNA transcripts, differing in length by 200-300 nucleotides, are synthesized in adult male and female flies, during embryogenesis and throughout the larval period. Using tissue in situ hybridization techniques that do not distinguish between the transcripts, we find that the distribution of 4f-rnp mRNAs is specific to germline tissue in the ovary, demonstrating that these transcripts are maternally produced. During embryogenesis, 4f-rnp transcripts are localized within cells of most tissues, but they have a very distinct localization pattern within the segmental ganglia of the central nervous system. We discuss the significance that RNA editing is expected to have for the predicted 4F-RNP protein products that may be expressed throughout fly development and in high abundance within the fly's central nervous system.

Chemosurgical reports: myxoid dermatofibrosarcoma protuberans.

Dermatofibrosarcoma protuberans (DFSP) is a fibroblastic tumor characterized by a high rate of recurrence following conventional surgical treatment. Several different histopathologic patterns exist, with the "cartwheel" pattern the most common. In this report, a patient with the unusual myxoid histopathologic pattern was successfully treated with Mohs surgery.

Lipoprotein secretion by rat liver Golgi apparatus. Lipoprotein particles and lipase activity.

Lipoprotein particles of the size range of very low density lipoproteins in smooth endoplasmic reticulum, peripheral elements of the Golgi apparatus, and secretory vesicles of the immature Golgi apparatus face are 55 to 80 nm in diameter. Particles in mature secretory vesicles are smaller (45 nm). Concomitant with the change in particle size, the lumina of mature vesicles increase in electron density. A technique to fractionate immature and mature secretory vesicles was based on precipitation of a cupric-ferrocyanide complex (Hatchett's brown) through the action of a NADH-ferricyanide oxido-reductase resistant to glutaraldehyde which is characteristic of the membranes of mature secretory vesicles and of the plasma membrane of liver. Mature secretory vesicle fractions so isolated were enriched in cholesterol and depleted in triglycerides relative to immature vesicles on a phospholipid basis. Lipase activity was present in secretory vesicle fractions of the Golgi apparatus as shown by biochemical analysis and by cytochemistry. Cytochemical studies showed lipase to be present in both mature and immature vesicles but most evident in immature vesicles. The findings suggest that some very low density lipoprotein particles are converted to particles of smaller diameter during transit through Golgi apparatus. A lipase-mediated hydrolysis of triglycerides may relate to the transformation.

Intellective characteristics of mothers of failure-to-thrive syndrome children.

The intelligence levels of three groups of mothers were ascertained by the vocabulary test of the Stanford-Binet Intelligence Scale. Eight mothers having children diagnosed as failure-to-thrive due to non-organic reasons (FTT-E), eight mothers with children diagnosed as failure-to-thrive due to physically traceable reasons (FTT-O), and eight mothers with children hospitalized due to reasons other than failure-to-thrive (C), were contrasted on intelligence, age, education and the presence or absence of the father in the home. Hypotheses regarding a disadvantage of FTT-E mothers on the four variables were supported. Implications of the results were discussed regarding programmes dealing with failure-to-thrive children and mothers. Further investigation in this area was proposed.