Topical progesterone as support for the luteal phase in induced cycles

e efficacy and tolerability of 2.5% natural vaginal progesterone as luteal phase support in pharmacologically induced cycles has been evaluated. MATERIALS AND METHODS: On the basis of the administered therapy, the population studied was subdivided at random into four different groups. Sixty patients came to the Sterility Autonomous Department of the Niguarda Ca' Granda Hospital in Milan from November 1994 to January 1996. Both ultrasonographic and plasmatic hormonal parameters have been evaluated on a monthly basis. RESULTS: The plasma progesterone levels and ultrasonographic endometrial thickness average values resulted more important in groups treated with topical progesterone; in these same groups a greater percentage of pregnancies was observed. CONCLUSIONS: The results obtained demonstrated that the drug studied can be recommended as a valid luteal phase support in pharmacologically induced cycles.     

An interactive activation approach to object processing: effects of structural similarity, name frequency, and task in normality and pathology

We present a computational model of the processes involved in retrieving stored semantic and name information from objects, using a simple interactive activation and competition architecture. We simulate evidence showing a cross-over in normal reaction times to make semantic classification and identification responses to objects from categories with either structurally similar or structurally dissimilar exemplars, and that identification times to objects from these two different classes correlate differentially with measures of the structural similarity of objects within the category and the frequency of the object's name. Structural similarity exerts a negative effect on object decision as well as naming, though this effect is larger on naming. Also, on naming, structural similarity interacts with the effects of name frequency, captured in the model by varying the weight on connections from semantic to name units; frequency effects are larger with structurally dissimilar items. In addition, (1) the range of potential errors for objects from these two classes, when responses are elicited before activation reached a stable state, differ--a wider range of errors occur to objects from categories with structurally similar exemplars; and (2) simulated lesions to different locations within the model produce selective impairments to identification but not to semantic classification responses to objects from categories with structurally similar exemplars. We discuss the results in relation to data on visual object processing in both normality and pathology.     

Odor, drug and toxin analysis with neuronal networks in vitro: extracellular array recording of network responses

Neurons, by virtue of intrinsic electrophysiological mechanisms, represent transducers that report the dynamics of cell death, receptor-ligand interactions, alterations in metabolism, and generic membrane perforation processes. In cell culture, mammalian neurons form fault-tolerant, spontaneously active systems with great sensitivity to their chemical environment and generate response profiles that are often concentration- and substance-specific. Changes in action potential patterns are usually detected before morphological changes and cell damage occur, which provides sensitivity and reversibility. Such biological systems can be used to screen rapidly for novel pharmacological substances, toxic agents, and for the detection of certain odorants. Existing simple culture preparations can already be employed effectively for the detection of chemical compounds. So far, three strategies have been investigated in pilot experiments: (1) Substance-dependent major changes in spontaneous native activity patterns. All synaptically active agents (e.g. glutamate, strychnine, N-methyl D-aspartic acid) as well as metabolic poisons generate such changes. (2) Substance-dependent changes in network oscillations via disinhibition. The regularized, oscillatory activity is altered by synaptically and metabolically active substances, ion channel blockers, and toxins. (3) Detection of paroxysmal responses indicating major, pathological membrane currents in large subpopulation of cells. We have explored these three strategies via 64 channel array recordings using spontaneously active murine spinal cord cultures. The glycine receptor blocker strychnine reliably generated increased multichannel bursting at 5-20 nM and regular, coordinated bursting above 5 microM. During biculline-induced network oscillations many compounds alter oscillation frequencies or terminate activity in a substance-specific manner. Finally, the gp120 protein of the AIDS virus (at 1 microgram/ml) produces massive, unique paroxysmal discharges that may last as long as 2 min. These results indicate that cultured neuronal networks are practical systems that can be used for the detection and identification of a great variety of chemical substances. The concept of dynamic fingerprinting to identify specific compounds is discussed.     

The tryptase, mouse mast cell protease 7, exhibits anticoagulant activity in vivo and in vitro due to its ability to degrade fibrinogen in the presence of the diverse array of protease inhibitors in plasma

Mouse mast cell protease (mMCP) 7 is a tryptase of unknown function expressed by a subpopulation of mast cells that reside in numerous connective tissue sites. Because enzymatically active mMCP-7 is selectively released into the plasma of V3 mastocytosis mice undergoing passive systemic anaphylaxis, we used this in vivo model system to identify a physiologic substrate of the tryptase. Plasma samples taken from V3 mastocytosis mice that had been sensitized with immunoglobulin (Ig) E and challenged with antigen were found to contain substantial amounts of four 34-55-kDa peptides, all of which were derived from fibrinogen. To confirm the substrate specificity of mMCP-7, a pseudozymogen form of the recombinant tryptase was generated that could be activated after its purification. The resulting recombinant mMCP-7 exhibited potent anticoagulant activity in the presence of normal plasma and selectively cleaved the alpha-chain of fibrinogen to fragments of similar size as that seen in the plasma of the IgE/antigen-treated V3 mastocytosis mouse. Subsequent analysis of a tryptase-specific, phage display peptide library revealed that recombinant mMCP-7 preferentially cleaves an amino acid sequence that is nearly identical to that in the middle of the alpha-chain of rat fibrinogen. Because fibrinogen is a physiologic substrate of mMCP-7, this tryptase can regulate clot formation and fibrinogen/integrin-dependent cellular responses during mast cell-mediated inflammatory reactions.     

Systematic screening of the LDL-PLA2 gene for polymorphic variants and case-control analysis in schizophrenia

Systematic scans of the genome using microsatellite markers have identified chromosome 6p21.1 as a putative locus for schizophrenia in multiply affected families. There is also evidence from a series of studies for a role of abnormal phospholipid metabolism in schizophrenia. In light of these findings, and the role of platelet activating factor in neurotransmission and neurodevelopment, we have examined the LDL-PLA2 (plasma PAF acetylhydrolase, PAF-AH) gene, a serine dependent phospholipase that has been mapped by hybrid mapping to chromosome 6p21.1, as a positional candidate gene for schizophrenia. The gene was systematically screened using SSCP/HD analysis for polymorphisms associated with the disease. Four polymorphic variants were found within the gene and studied in a group of 200 schizophrenic patients and 100 controls. The variant in exon 7 (Iso195Thr) was found to be weakly associated with schizophrenia (p = 0.04) and the variant in exon 11 (Val379Ala) almost reached significance (p = 0.057). After correcting for multiple testing no significant associations were detected. Haplotype analysis combining pairs of polymorphisms also provided no evidence for association of this gene with schizophrenia in our sample of patients.     

Salmonella arizonae sepsis in a lynx

A 4.5-wk-old lynx (Felis lynx) was presented for necropsy with a history of poor growth, mild diarrhea, anemia, and lethargy. The liver was enlarged and had a 7 mm long fracture that resulted in severe intraabdominal hemorrhage and death. Microscopic lesions were indicative of severe ulcerative cystitis and septicemia. Pure cultures of Salmonella arizonae were isolated from the liver, kidney, and spleen. Based on differences in the chronicity of inflammation in the urinary bladder versus other organs, we speculate that chronic cystitis caused by S arizonae lead to septicemic infection.     

Fabrication of plastic microfluid channels by imprinting methods

Microfluidic devices have been fabricated on poly(methyl methacrylate) substrates by two independent imprinting techniques. First-generation devices were fabricated using a small-diameter wire to create an impression in plastics softened by low-temperature heating. The resulting devices are limited to only simple linear channel designs but are readily produced at low cost. Second-generation devices with more complex microchannel arrangements were fabricated by imprinting the plastic substrates using an inverse three-dimensional image of the device micromachined on a silicon wafer. This micromachined template may be used repeatedly to generate devices reproducibly. Fluorescent analtyes were used to demonstrate reproducible electrophoretic injections. An immunoassay was also performed in an imprinted device as a demonstration of future applications.     

Intracrine and autocrine effects of basic fibroblast growth factor in vascular smooth muscle cells

In order to elucidate the effects of the different basic fibroblast growth factor (bFGF) isoforms on vascular smooth muscle, we examined aorta-derived vascular smooth muscle cells from transgenic mice expressing the human isoforms of bFGF. Four cell lines were examined from mice in which transgene expression was driven by the ubiquitous phosphoglycerate kinase promoter. Overexpression and cellular localization was confirmed by Western blot analysis in vascular smooth muscle cells from mice expressing: all four human bFGF isoforms (24, 22, 21, and 18 kDa); all three nuclear targeted isoforms (24, 22, and 21 kDa); only the 24 kDa isoform; and the only secreted/non-nuclear targeted isoform, 18 kDa. All lines showed approximate four-fold increases in bFGF expression, nuclear localization of all nuclear targeted bFGF isoforms, and cytosolic localization of only the 18 kDa bFGF. Measurement of [3H]thymidine incorporation into quiescent cells stimulated with increasing concentrations of serum, showed increased DNA synthesis in cell lines expressing any bFGF isoform when compared to non-transgenic control cells, and a further increase in DNA synthesis in cells expressing the nuclear targeted isoforms (24, 22, and 21 kDa) over the 18 kDa bFGF expressing cell line at any concentration of serum. All cells showed equal label incorporation when stimulated with 10 ng/ml of platelet-derived growth factor confirming an equal potential for DNA synthesis. Neutralizing the bFGF antibody markedly decreased serum-stimulated DNA synthesis, but only in the cell lines overexpressing the secreted/non-nuclear targeted 18 kDa isoform. These results suggest amplification of DNA synthesis through synergistic intracrine and autocrine effects of the nuclear targeted and non-nuclear targeted bFGF isoforms in vascular smooth muscle cells.     

Dosimetry of copper-64-labeled monoclonal antibody 1A3 as determined by PET imaging of the torso

We present biodistribution and dosimetry results for 64Cu-benzyl-TETA-MAb 1A3 from 15 human subjects injected with this tracer as determined by serial PET imaging of the torso. METHODS: PET imaging was used to quantify in vivo tracer biodistribution at two time points after injection. Absorbed dosimetry calculated using MIRD-11 and the updated MIRDOSE3 was compared with estimates obtained using rat biodistribution data. RESULTS: By measuring activity concentrations in the torso, and extrapolating for the whole body using standard organ and tissue volumes, we were able to account for 93% of the injected radiopharmaceutical over a range of imaging times from 0 to 36 hr postinjection. Based on PET imaging and the MIRD-11 schema, the liver and spleen are the critical organs with average absorbed doses of 0.12 and 0.10 mGy/MBq (0.44 and 0.39 rad/mCi). The revised MIRDOSE3 scheme yields similar values for these and other organs but also results in a dose of 0.14 mGy/MBq (0.53 rad/mCi) to the heart wall. In the rat, the large intestine is the critical organ at 0.14 mGy/MBq (0.52 rad/mCi), while liver and kidneys each receive 0.11 mGy/MBq (0.41 rad/mCi). Some disparities in absorbed doses determined by these methods are evident but are a result of dissimilar biodistributions in rats and humans. For most organs, rat extrapolated values are higher than the human measurements with PET. CONCLUSION: This study shows that torso PET imaging can quantitatively measure the whole-body biodistribution of a radiopharmaceutical as long as it has relatively slow pharmacokinetics.