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Biomedical subjects

M Spitz

Publications and source records attributed to M Spitz.

11 recordsLinked to original sources

Ovarian expression and function of neuropeptide systems in teleosts and anurans.

The hypothalamic-pituitary-gonadal axis regulates reproduction, sexual maturation, and spawning behaviours. Its evolutionary origins trace back to primitive jawless fish and has been well characterized in teleosts. Recent advances in multi-species genome sequencing, annotation, and experimental approaches for identifying and characterizing key regulators have advanced understanding of neuroendocrine regulation in teleost reproduction, reshaping existing models. Early studies in amphibians established that steroids are critical regulators of final oocyte maturation. Subsequent work in anurans revealed complex interactions among theca cells, follicular cells, and oocytes, supporting a three-cell model in which oocytes contribute to their own steroidogenic environment, challenging the traditional two-cell view of ovarian steroidogenesis. In teleosts, however, direct evidence that oocytes support steroid precursor delivery to theca and follicular cells is limited, and whether a comparable three-cell model applies remains an open hypothesis. Across both taxa, the roles of locally produced neuropeptides in coordinating interactions among theca cells, follicular cells, and oocytes remain largely uncharacterized. Here, we provide a short review of the localization and potential autocrine/paracrine functions of neuropeptides in teleost and amphibian ovaries and discuss existing knowledge gaps. We identify opportunities to leverage detailed localization studies that map neuropeptides to specific ovarian cell types and developmental stages, and discuss how integrating traditional and emerging experimental approaches can advance comparative studies in ovarian endocrinology. This work will improve our understanding of reproductive regulation in fishes and frogs, with applications in captive breeding, aquaculture, and endocrine disruption research.

Autocrine

Monoclonal antibodies to crosslinked fibrin degradation products (XL-FDP). I. Characterization and preliminary evaluation in plasma.

Monoclonal antibodies (mabs) were raised against X-oligomers, the earliest soluble fragments released from crosslinked fibrin (XL-FN), by the action of plasmin. Two of the mabs (NIBn 52 and NIBn 123) were monospecific for X-oligomers in that they showed no binding to fibrinogen, the plasmic fragments of fibrinogen (D and E) and non-crosslinked fibrin (X, Y, D and E), or the terminal digestion product of XL-FN, fragment DD-E. One other mab (NIBn 178) was panspecific for X-oligomers in that it exhibited a weak affinity for fibrinogen. The mabs were used to develop a two-site immunoradiometric assay (IRMA) and an enzyme-linked immunospecific assay (ELISA) which permitted the specific measurement of X-oligomers directly in plasma, rather than in serum. This immunoassay is a true assay of fibrinolysis as distinct from fibrinogenolysis and may be a potential aid in the diagnosis and evaluation of thrombosis. In preliminary studies, the assay detected low levels of X-oligomers in normal plasma and elevated levels in patients with disseminated intravascular coagulation.

Antibodies, Monoclonal

Studies on the human basophil IgE receptor. Generation and characterization of monoclonal antibodies against the receptor.

Anti-human basophil IgE receptor (Fc epsilon R) monoclonal antibodies (McAbs) were generated and characterized. The production of these antibodies was facilitated by both a single step procedure for the enrichment of basophils using Ficoll-Paque centrifugation and a double staining technique combining the APAAP with alcian blue. By the double staining technique we could recognize the specific anti-basophil antibodies because the cells binding the antibody--APAAP positive cells--also had alcian blue-stained basophil granules. Blocking experiments showed that the McAb was directed against the Fc epsilon R since preincubation with human IgE blocked the binding of the McAb and preincubation with the antibody blocked the binding of IgE to basophils and inhibited the formation of IgE rosettes.

Animals

Single step enrichment of human peripheral blood basophils by Ficoll-Paque centrifugation.

A simple, rapid and economical method for the enrichment of basophils from peripheral blood with high recovery is described. Peripheral blood was layered over a Ficoll-Paque cushion and centrifuged. The Ficoll-Paque layer was aspirated taking care not to disturb the interphase and the buffy coat. The Ficoll-Paque layer contained most of the basophils. A recovery of 82% (65-95%) and a 23.5-fold enrichment of basophils with their morphology and biological activity preserved was achieved.

Antibodies, Anti-Idiotypic

Multiple proteins related to the soluble galactose-binding animal lectin revealed by a monoclonal anti-lectin antibody.

A monoclonal antibody (NIBy 142-36/8) raised against the soluble galactose-binding lectin of bovine heart muscle has been tested by solid-phase vinyl-plate radiobinding and nitrocellulose immunoblotting with homogenates of various bovine tissues, and the muscle tissues of pig, rabbit, chicken and rat. Muscle lectins of chicken, rabbit and rat differed from those of man and pig in their lack of reactivity with the 36/8 antibody. There was a good correlation of haemagglutinating activities and immunoreactivities of the bovine tissue homogenates, suggesting that the soluble galactose-binding protein is a major haemagglutinin in various tissues. Immunoblotting experiments revealed an array of antigenically active components in the homogenates in addition to the 13 and 26kDa proteins that were previously detected in preparations of purified lectin. These were in the range 36kDa to more than 200kDa, and a different spectrum of immunoreactive components was found in various cell types. Galactose-binding activity was demonstrable in 13, 26 and 36kDa components in certain bovine tissues, suggesting that the immunoreactive components of higher Mr may be inactive precursor forms of the lectin.

Animals

Use of 'single shot' intrasplenic immunization for production of monoclonal antibodies specific for human IgM.

We report here the use of 'single shot' intrasplenic injection of human IgM for immunization of mice to obtain splenocytes for use in the production of hybridomas secreting antibodies against human IgM. Fusion was performed 3 days after intrasplenic injection of 20 micrograms of myeloma IgM. IgM-specific antibodies were found in 12% of the fusion wells; only 1 well contained antibodies which cross-reacted with other immunoglobulin classes. Two monoclonal antibodies (McAbs) have been fully characterized as specific for different epitopes on Fc mu. These antibodies can be used to detect IgM on the surface of human B cells by immunofluorescence and in solution by solid-phase radiobinding assay or single radial immunodiffusion. Both McAbs can also detect IgM fragments by immunoblotting from non-reducing SDS-polyacrylamide gels.

Animals

Human B cell proliferation is stimulated by interleukin 2.

The proliferation of human B cells was studied for response to interleukin 2 (IL-2) produced in Escherichia coli using recombinant DNA technology. The IL-2 was found to be an homogenous preparation by sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) and immunoblotting using the anti-IL-2 monoclonal antibody DMS-1. IL-2 was found to stimulate B cell proliferation. Activation of the B cells using anti-IgM antibodies increased this response. Resting T cells from the same donors were found to be less reactive to IL-2. The results suggest that human B cell proliferation can be stimulated by IL-2 alone.

Antibodies, Anti-Idiotypic

Interleukin-2 in vivo: production of and response to interleukin-2 in lymphoid organs undergoing a primary immune response to heterologous erythrocytes.

In this report, we describe experiments which demonstrate that antigenic stimulation in vivo causes the appearance of cells in both spleen and lymph node which secrete interleukin-2 (IL-2). Cells also appear in these organs which proliferate in response to IL-2. For these experiments, sheep red cells (SRBC) were injected into the spleens or footpads of mice, and cell suspensions from spleens or popliteal lymph nodes prepared at various times after antigenic stimulation. These cells were assayed for their ability to respond to IL-2, and their cell culture supernatants for secreted IL-2. The proliferative response to IL-2 steadily increased following SRBC injection to reach a peak at Day 2 for spleen cells and at Day 3 for lymph node cells. Maximal production of IL-2 was displaced from the maximal response to the lymphokine by peaking one day later for both organs. Our results strongly implicate the participation of IL-2 in the in vivo immune response and suggest the existence of in vivo regulatory mechanisms, which can control the time of IL-2 production and also the appearance of cells with receptors for IL-2.

Animals

The human fetal auditory evoked potential.

The brain-stem auditory evoked potential (BAEP), a sensitive test of the functional status of the neonatal brain, has not been studied in utero since no practical technique for human fetal recording is available. We have developed a simple recording technique which allows continuous monitoring of the fetal AEP during labor. Waves I, III and V of the fetal brain-stem AEP have been consistently identified. Wave form morphology, interpeak latencies, and latency-intensity relations are similar to postnatal recordings. Middle latency potentials have also been recorded, with wave forms that correspond to the neonatal middle latency AEP.

Brain Stem