A Collection of cDNA Clones with Specific Expression Patterns in Mouse Brain

A total of 950 cDNA clones were randomly selected from mouse cerebellar cDNA libraries, and of these, about 130 clones were found to correspond to mRNAs which were expressed unequally between the cerebellum and other parts of mouse brain. Their distribution patterns in adult mouse brain were analysed by in situ hybridization, and eight clones were found restricted to specific regions of the brain, including four clones specific to cerebellar granule cells and one clone specific to Purkinje cells. Another 27 clones were preferentially expressed in a diverse, but distinctive subpopulation of brain cells. Among them seven clones were especially abundant in specific nuclei, and three in specific fibre bundles. These clones will be useful in defining new subpopulations of brain cells characterized by different gene expression.     

Distribution of Galanin mRNA Containing Cells and Galanin Receptor Binding Sites in Human and Rat Hypothalamus

The distribution of cells containing galanin mRNA and that of galanin receptor binding sites were investigated using in situ hybridization histochemistry and receptor autoradiography in male rat hypothalamus and in postmortem hypothalamic tissues from control human brains. Oligonucleotide probes labelled with 32P were used for hybridization experiments. The specificity of the hybridization signal was ascertained using several probes, competition assays and Northern blot analysis. High levels of hybridization were found in the paraventricular, supraoptic and arcuate nuclei of rat and human hypothalamus. Human intermediate nuclei and scattered cells of the posterior perifornical nucleus also contained galanin mRNA. Galanin mRNA was also found in the dorsomedial nucleus of the rat. The distribution of galanin receptor sites was investigated by receptor autoradiography using 125I-labelled porcine galanin. The specificity of the binding was assessed by competition with different neuropeptides. While galanin blocked the binding at nanomolar concentrations, the other neuropeptides examined were ineffective at 10-7 M concentrations. The highest densities of galanin binding sites were seen in the preoptic area, ventromedial and lateral nuclei, of rat and human hypothalamus. In contrast, very low densities of binding sites were observed in the paraventricular, supraoptic and arcuate nuclei. Our results show that the distribution of neurons expressing galanin is complementary to that of galanin receptors in the rat and human hypothalamus. This suggests that receptors for galanin are not located on the cell bodies of galaninergic neurons, but are probably presynaptic on or postsynaptic to the processes of these cells.     

The effect of flow and mass transport in thrombogenesis

The paper presents a mathematical analysis of the contributions of flow and mass transport to a single reactive event at a blood vessel wall. The intent is to prepare the ground for a comprehensive study of the intertwining of these contributions with the reaction network of the coagulation cascade. We show that in all vessels with local mural activity, or in “large” vessels (d>0.1 mm) with global reactivity, events at the tubular wall can be rigorously described by algebraic equations under steady conditions, or by ordinary differential forms (ODEs) during transient conditions. this opens up important ways for analyzing the combined roles of flow, transport, and coagulation reactions in thrombosis, a task hitherto considered to be completely intractable. We report extensively on the dependence of transport coefficient k_L and mural coagulant concentration C_w on flow, vessel geometry, and reaction kinetics. It is shown that for protein transport, k_L varies only weakly with shear rate in large vessels, and not at all in the smaller tubes (d<10⁻² mm). For a typical protein, k_L∼10⁻³ cm s⁻¹ within a factor of 3 in most geometries, irrespective of the mural reaction kinetics. Significant reductions in k_L (1/10–1/1,000) leading to high-coagulant accumulation are seen mainly in stagnant zones vicinal to abrupt expansions and in small elliptical tubules. This is in accord with known physical observations. More unexpected are the dramatic increases in accumulation which can come about through the intervention of an autocatalytic reaction step, with C_w rising sharply toward infinity as the ratio of reaction to transport coefficient approaches unity. Such self-catalyzed reactions have the ability to act as powerful amplifiers of an otherwise modest influence of flow and transport on coagulant concentration. The paper considers as well the effect on mass transport of transient conditions occasioned by coagulation initiation or pulsatile flow. During initiation, instantaneous flux varies with diffusivity and bulk concentration, favouring the early adsorption/consumption of proteins with the highest abundance and mobility. This is akin to the ‘Vroman effect’ seen in narrow, stagnant spaces. The effect of flow pulsatility on k_L has the potential, after prolonged cycling, of bringing about segregation or accumulation of proteins, with consequences for the coagulation process.     

Häufigkeit pathologischer Veränderungen des oberen Gastrointestinaltraktes bei Patienten vor Herzoperationen

Resümee Schwerwiegende gastroduodenale Erkrankungen sind bei fast der Hälfte aller Patienten, die sich einer Operation am offenen Herzen unterziehen müssen, auch bei Fehlen von Symptomen nachweisbar. Das erhebliche Überwiegen von Magenläsionen spricht dafür, daß die arteriosklerotisch bedingte Perfusionsminderung der Schleimhaut die entscheidende Ursache hierfür ist. Routinemäßige präoperative Ösophago-Gastro-Duodenoskopien können die nicht unerhebliche durch gastrointestinale Komplikationen bedingte postoperative Mortalität senken helfen.

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Pathological changes in the upper gastrointestinal tract in patients awaiting open heart surgery

Summary While waiting for open heart surgery, in 153 patients (104 male, 49 female, 22–76 years of age) without gastrointestinal symptoms and/or history esophago-gastro-duodenoscopy was performed. 124 patients suffered from coronary heart disease, 29 from valvular defect, aneurysm of the sinus of Valsalva or tumor of the heart.In 47.1% endoscopy revealed serious abnormal findings: in 16.3% gastric ulcer, in 20.9% erosive gastritis, duodenal ulcer and erosive duodenitis in 5.2%, respectively, 1 case of gastric carcinoma, 2 of large polyps and 3 of reflux esophagitis of higher degree (totally 3.9%).In patients with coronary artery disease, the relation of erosive and ulcerous gastric lesions as compared with those of duodenal origin was 41, in patients with other cardiac diseases it was 21, respectively (p<0,001).Compared with a normal population, the incidence of pathological gastric findings was 54-fold higher in our patients, and 1.7-fold concerning duodenal lesions, respectively (p<0.001).51 patients on acethylsalicylic acid (160 mg/ die) showed pathologic findings in 41.2%, and 96 patients without ulcer-inducing therapy in 51%. Thus, low-dose Aspirin does not seem to have serious gastric side effects.The results of the study stress the necessity of routinely performed endoscopy of the upper gastrointestinal tract in patients awaiting open heart surgery. This will lead to a lower incidence of serious gastrointestinal complications postoperatively, which are known to have a high mortality.

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Abkürzungen ASS Azetylsalizylsäure - J. Jahre - GI-Trakt Gastrointestinaltrakt     

Enteroenterale Invagination des Dünndarms beim Erwachsenen

Zusammenfassung Es wird über eine 43jährige Patientin berichtet, die mehrere Wochen lang über inkonstant auftretende, krampfartige Mittelbauchschmerzen klagte. Die Diagnose einer ileoilealen Invagination konnte bei der Patientin erst beim dritten stationären Aufenthalt gestellt werden. Folgende Fehldiagnosen waren während des 4wöchigen Verlaufs bei der Patientin gestellt worden: Nierenbeckenentzundung, acute Appendizitis, chronische Appendizitis und zuletzt psychosomatische Bauchschmerzen. Die Patientin wurde zunächst antibiotisch, zuletzt mit Psychopharmaka medikamentös behandelt. Schließlich wurde die Diagnose einer Invagination des Dünndarms mittels Sonographie vermutet und in der konventionellen fortlaufenden Magen-Darm-Passage röntgenologisch gesichert. Bei der Patientin wurde daraufhin eine Dünndarmsegmentresektion durchgeführt. Die enteroenterale Invagination ist im Erwachsenenalter ein sehr seltenes Ereignis, bei der meistens eine Ursache (oft Tumore) als Auslöser der Invagination ermittelt werden kann. Durch Peristaltik und Ingesta wird der nach aboral bewegte Tumor zum Motor der Invagination. Prädilektionsorte sind die Übergänge eines beweglichen zu einem retroperitoneal fixierten Darmabschnitts (z. B. Ileozökalregion). Die Sonographie des Abdomens ist die Methode der ersten Wahl bei der Diagnose einer enteroenteralen Invagination. Beim Erwachsenen ist die operative Beseitigung der Invagination und deren Ursache (meist Tumore) angezeigt.

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Entero-enteric invagination of the small intestine in adultsA rare cause of abdominal distress

The present paper reports on a 43-year-old female patient who complained over a number of weeks of paroxysms of crampy pain in the mesogastrium. The diagnosis of ileoileal invagination was only made after she had been admitted to hospital for the third time. The following false diagnoses had been made during the 4-week course of the condition: pyelonephritis, acute appendicitis, chronic appendicitis and, most recently psychosomatic abdominal distress. The patient was initially treated with antibiotics and finally with psychotropic drugs. Eventually ultrasound suggested the diagnosis of invagination of the small intestine, which was then verified by conventional barium follow-through radiography. The patient subsequently underwent resection of a segment of the small intestine. Entero-enteric invagination is a very rare event in adults, in which a single (often malignant) cause is identified as triggering the invagination. Peristalsis and ingested food push the tumor distad, thus acting as a motor for invagination. The preferred localizations are the junctions between freely moving segments to retroperitoneally fixed segments (e.g., ileocecal region). Ultrasound of the abdomen is the examination of choice for diagnosis of enteroenteric invagination. Surgical resection of the invagination and its cause (generally tumors) is indicated in adults.

     

Evidence for feeding elicited through antihistaminergic effects of tricyclic antidepressants in the rat hypothalamus

We studied central mechanisms of antidepressants that affect feeding behavior in rats. The tricyclic compounds amitriptyline, doxepin and imipramine significantly induced feeding after their infusion into the third cerebral ventricle in the light phase, but the tricyclic, desipramine, and the dicyclic zimelidine, did not. Drinking was not affected by any compound tested. The relative order of potency in eliciting feeding was: amitriptyline and doxepin > imipramine > desipramine and zimelidine. To clarify the involvement of neuronal histamine in antidepressant-induced feeding, alpha-fluoromethylhistidine (FMH), a suicide inhibitor of histidine decarboxylase, was intraperitoneally administered before infusion of amitriptyline. FMH attenuated the amitriptyline's effect. Bilateral microinfusion of amitriptyline into the ventromedial hypothalamus or the paraventricular nucleus verified that these are loci for the modulation of feeding by amitriptyline. In the lateral hypothalamus, amitriptyline was less effective. These findings indicate that tricyclic antidepressants directly facilitate feeding, which is, at least in part, mediated by histamine in the hypothalamus.     

Molecular genetic studies of early breast cancer evolution

Summary In the past few years there has been an explosion in the number of patients diagnosed with hyperplastic breast disease andin situ breast cancer. Based on epidemiological data, these morphologically defined lesions may be categorized as those with little malignant potential (e.g. typical hyperplasia or proliferative disease without atypia [PDWA]), those with significant malignant potential which may already be initiated (e.g. atypical ductal hyperplasia [ADH]), and early transformed lesions which are malignant but not yet invasive (e.g. ductal carcinomain situ [DCIS]). They may represent sequential evolutionary stages in the ontogeny of invasive breast cancer, with each morphologically defined stage resulting from accumulating genetic changes culminating in a transformed clonal lineage capable of invasion and metastasis. Using loss-of-heterozygosity (LOH) analysis, we are studying the genetic changes associated with these lesions in archival tissue samples. 50% (6/12) of the proliferative lesions (PDWA and ADH) and 80% of the DCIS shared their LOH patterns with more advanced lesions from the same breast, strongly supporting a precursor/product relationship between these lesions and the cancers they accompany.     

Potential use of embryo coculture with human in vitro fertilization procedures

Purpose This review was designed to outline potential uses of an embryo co-culture system in human assisted reproduction programs to improve embryo quality and pregnancy rates.Results The various cell types used in embryo co-culture were reviewed in addition to the use of co-culture for both animal and human embryos. Co-culture provides a method to enhance embryo development in an inadequate in vitro environment without compromising embryo quality. Human IVF laboratories have used various types of helper cells to improve rate of development, reduce cell fragmentation rate and in some instances increase pregnancy and implantation rates.Conclusion In conjuction with several assisted reproduction procedures such as IVF, microsurgical fertilization, cryopreservation and genetic evaluation, co-culture may increase the number of viable embryos for replacement and improve pregnancy rates.Presented at the First Asian Symposium on Micromanipulation and Co-culture, April 1, 1993, Singapore.     

Effect of platelet-activating factor on secretion of mucous glycoprotein from chinchilla middle ear epithelial cells in vitro

Platelet-activating factor (PAF) is a naturally occurring phospholipid that acts as a pleiotropic mediator and mediates cell-cell reactions under physiological and pathological conditions. Recently, it has been shown that PAF is a strong secretagogue of mucous glycoprotein in the airways, suggesting its role in mucous glycoprotein secretion and the pathogenesis of otitis media with effusion. In the current study, we examined the effect of PAF on mucous glycoprotein secretion in cultured chinchilla middle ear epithelial cells. PAF at 1 M significantly stimulated mucous glycoprotein secretion from cultured chinchilla middle ear epithelial cells. This action was concentration-dependent, with secretions reaching near maximum when the cells were incubated with PAF at 100 M. In a time-dependent study, PAF demonstrated an initial rapid stimulation of mucous glycoprotein secretion, followed by a gradual increase thereafter. A six-fold increase was seen in the first 2 h compared with controls. Cycloheximide, a protein synthesis inhibitor, demonstrated an inhibitory effect on PAF-stimulated mucous glycoprotein secretion in this study. These findings suggest that PAF plays an important role in the pathogenesis of otitis media with effusion by stimulating mucous glycoprotein secretion in vitro.Supported by NIH grant P0I-D000133 from the National Institute on Deafness and Other Communication Disorders.     

Fertilization and pregnancies following intracytoplasmic injection of testicular spermatozoa

Purpose Intracytoplasmic injection (ICSI) with testicular sperm was performed in 16 couples. All men had ductal obstruction and failed previous attempts of epididymal sperm microaspiration.Methods Testis tissue was obtained by excisional biopsies and incubated in HEPES buffered EBSS medium over 24 h at 37C. Motile sperm (Grade 1 to 2) were recovered in 13 patients and fertilized a total of 62 oozytes. Four pregnancies were achieved.Results One healthy boy and two girls (twin pregnancy) were born.Conclusions The ongoing pregnancies revealed no fetal abnormalities on ultrasound scanning.Presented at the IXth World Congress on In Vitro Fertilization and Alternate Assisted Reproduction, April 3–7, 1995, Vienna, Austria.