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At least 811 records · Page 45Linked to original sources
PubMed1991

Recurrent failure of in vitro fertilization: role of the hemizona assay in the sequential diagnosis of specific sperm-oocyte defects.

The results of predictive fertilization bioassays (hemizona assay, hamster ova-human sperm penetration assay), in vitro fertilization treatment, fertile donor cross-match tests with either sperm or oocytes, and oocyte micromanipulation for assisted fertilization were used to establish a pathophysiologic diagnosis in cases of recurrent failed fertilization in vitro. Disorders of sperm function manifested at the level of zona binding, zona penetration, oolemma fusion, and pronuclear decondensation as well as oocyte anomalies were considered to represent the specific gamete defects that led to abnormal sperm-oocyte interactions (i.e., failed fertilization). Our findings show that the sequential application of bioassays can elucidate specific sperm or oocyte defects that characterize functional abnormalities among sperm populations.

Acrosome↗
PubMed1985

Microchemical synthesis of the serotonin receptor ligand, 125I-LSD.

The synthesis and properties of 2-[125I]-lysergic acid diethylamide, the first 125I-labeled serotonin receptor ligand, are described. A novel microsynthesis apparatus was developed for this synthesis. The apparatus employs a micromanipulator and glass micro tools to handle microliter to nanoliter volumes on a microscope stage. This apparatus should be generally useful for the synthesis of radioligands and other compounds when limited amounts of material must be handled in small volumes.

Animals↗
PubMed1994

Biometric analysis of tooth migration after approximal contact removal in the rat.

Nineteen 8-week-old female Sprague-Dawley rats were used to observe macroscopically the direction of tooth movements following removal of approximal contacts. In 10 rats, under anaesthesia, approximal contact between the second and third right maxillary molars (M2-M3) was removed by grinding. These animals and a control group of nine rats were housed with normal diet and water ad libitum for 7 weeks. After killing, the skulls of all animals were removed, dried, and fixed in a standardized position on a Horsley-Clarke type stereotaxic frame. Using a micromanipulator graduated to 1/100 mm and mobile in three orthogonal directions, distances between the teeth (M1 or M3) and some bone structures that served as landmarks were measured. These measurements showed the direction of tooth movements in the experimental approximal space. After statistical analysis of the data, the results showed that the approximal space between M2 and M3 was closed by both a distal drift of M1-M2 and a mesial migration of M3. It was concluded that, in the rat, a mesial drift exists that can be induced by the loss of approximal contact. This movement is distinct from the distal physiological dental migration classically described in histological studies.

Animals↗
PubMed1980

Enzymatic hypermethylation of DNA in mouse-mouse somatic cell hybrids.

Somatic cell hybrids between mouse L fibroblasts (A9 cells) and Ehrlich ascites tumour cells were constructed by use of poly(ethyleneglycol). The hybrids were selected in hypoxanthine/aminopterin/thymidine medium and morphologically different syncaryons were isolated by a micromanipulation. They were simultaneously analysed for their tumourogeneity, a release of certain proteins into the culture medium, the activity of pyrimidine salvage pathways, and the extent of enzymatic DNA methylation. None of these hybrids gave a rise of tumours if transplanted into DBA/2 mice. Two of the hybrids released a protein immunologically cross-reacting with antibody against C-peptide of human proinsulin. Activities of pyrimidine salvage pathways as measured by incorporation of [14C]deoxycytidine into DNA cytosine and thymine, respectively, are different in the analyzed cells. Enzymatic DNA methylation in somatic cell hybrids was significantly elevated as compared to the parental cell lines.

Animals↗
PubMed1983

Catecholamine metabolism in the rat locus coeruleus as studied by in vivo differential pulse voltammetry. II. Pharmacological and behavioral study.

Differential pulse voltammetry used in combination with electrochemically treated carbon fiber electrodes allowed us to detect catechols in the locus coeruleus (LC) of conscious freely moving rats. A micromanipulator cemented on the rat skull was designed in order to implant carbon fiber electrodes without anesthesia. Voltammograms were recorded every 2 min for 5 h. After the in vivo experiments electrodes were tested in 3,4-dihydroxyphenylacetic acid (DOPAC), 3,4-dihydroxyphenylglycol (DOPEG) and noradrenaline (NA) solutions. The catechol peak recorded from LC was suppressed by pargyline treatment and slightly reappeared after inhibition of the NA reuptake by desipramine (DMI). This reappearing signal was attributed to NA and estimated at a concentration 50 nM NA. Various drug treatments (piperoxane, haloperidol, clonidine, DMI and reserpine) allowed us to further support the conclusion of part I of this study: the catechol peak recorded from LC is mainly due to DOPAC synthesized by LC noradrenergic neurons. This DOPAC signal corresponded to a DOPAC concentration which reached 23 microM when the whole active part of the electrode was implanted in the LC. In addition to this pharmacological study, data from stress experiments pointed out a striking parallel between the variations of the DOPAC signal and those of the activity of LC noradrenergic neurons as revealed by reported electrophysiological studies.

3,4-Dihydroxyphenylacetic Acid↗
PubMed1985

Receptive field size of hair follicle afferents in rats treated neonatally with capsaicin.

Receptive field parameters of G- and D-hair units were investigated in adult rats given capsaicin as neonates. An air jet stimulator mounted on a two-way micromanipulator was used for accurate mapping of receptive fields. Diameters of G- and D-hair unit receptive fields were similar in both control and treated groups averaging 2.14 and 2.44 mm respectively with a 1.5 X threshold stimulus. It is concluded that the reported expansion of rat spinal cord cutaneous receptive fields following capsaicin treatment does not have any significant peripheral component.

Animals↗
PubMed1988

The role of CD4 in antigen-independent activation of isolated single T lymphocytes.

The membrane molecule CD4 (L3T4) is thought to facilitate activation of Class II H-2-restricted T cells by binding to Ia determinants on antigen-presenting cells. Recent reports suggest that CD4 can also contribute to antigen-independent activation by anti-T cell receptor (TCR) antibodies. An assay which measures the secretion of two lymphokines, granulocyte-macrophage colony-stimulating factor and interleukin 3 (IL-3), by single T cells activated with an anti-TCR antibody, F23.1, was used to analyze the effects of anti-CD4 antibodies on antigen-independent T cell activation. Single cells of a CD4+F23.1+ clone were micromanipulated into wells to which F23.1 had been immobilized, and their lymphokine secretion was measured 24 hr later. The frequency of lymphokine-secreting cells was consistently reduced up to 10-fold in the presence of soluble anti-CD4 antibody (GK1.5) but only up to 2.5-fold by an antibody to the cell adhesion molecule, LFA-1. In both bulk and single-cell cultures, responses to suboptimal concentrations of F23.1 were more susceptible to inhibition by GK1.5 than responses to optimal F23.1. The failure of GK1.5 to inhibit IL-2-stimulated lymphokine synthesis in bulk cultures suggested that CD4 ligation did not deliver a negative signal to the clone. By contrast, when either anti-CD4 or anti-LFA-1 was immobilized on the same surface as F23.1, the frequency of lymphokine-secreting cells could be increased up to 10-fold. It is concluded that anti-CD4 antibodies can act directly on the responding T cell to affect TCR-dependent activation, in the absence of interaction with antigen-presenting cells or any other cell type.

Antibodies, Monoclonal↗
PubMed1990

Target-dependent induction of secretory capabilities in an identified motoneuron during synaptogenesis.

Cholinergic neurons isolated from the buccal ganglia of Helisoma were plated into cell culture with a variety of defined target cells to study the specificity of synaptogenesis. Motoneuron B19 selectively formed chemical connections with single dissociated muscle fibers derived from its appropriate target, the supralateral radular tensor (SLT) muscle. B19 did not form such connections with novel neuronal targets. In contrast to neuron B19, cholinergic neuron B5 nonselectively formed chemical connections with novel muscle and neuronal targets. Target cells were micromanipulated into contact with presynaptic neurons to examine the latent period until the onset of functional synaptic transmission. Neuron B5 formed chemical connections within the first minutes of contact with ACh-sensitive neurons and muscle while B19 required sustained periods of muscle-specific contact to induce the acquisition of a functional excitation-secretion coupling mechanism. These different latent periods from the onset of target contact suggest that neuron B5 acquires presynaptic secretory function before target contact, while B19 must receive a specific signal(s) from its appropriate target to induce the transformation of its terminal into a secretory state.

Action Potentials↗
PubMed1990

Analysis of the hotfoot (ho) locus by creation of an insertional mutation in a transgenic mouse.

Hotfoot (ho) mutation is a recessive trait in mice, characterized by motor disorder and male sterility, that maps to chromosome 6. We have identified a transgenic mouse pedigree with a similar trait. Using genetic and molecular approaches, we have demonstrated that the foreign DNA element is located in or near the ho locus. This new allele, designated hoJwg and presumably created by insertional mutagenesis, should make it possible to clone the ho gene. Male infertility in hoJwg male homozygotes was determined to be due to inability of sperm to penetrate the zona pellucida. This was demonstrated by rescuing mutant males by a new technique of gamete micromanipulation, zona pellucida drilling. These findings show that zona drilling is useful both for analysis and preservation of animals with reduced male fertility.

Animals↗
PubMed1983

Microdissection of the mouse egg.

A rapid and efficient method for microdissection of the mouse egg is described. The dissection is carried out in hanging drops of medium surrounded by heavy liquid paraffin oil at room temperature. Eggs are first deformed into a cylindrical shape and then dissected at a predetermined site with a glass needle on a Leitz micromanipulator. The survival rate of the dissected fragments is 75-90% and between 20 and 30 eggs can be dissected in an hour. Development of the dissected eggs is at least as good as that described after other types of manipulation. Cytoplasts and karyoplasts of various sizes can be prepared, as well as gynogenetic and androgenetic eggs with different amounts of cytoplasm. This procedure may help to examine nuclear-cytoplasmic interactions in eggs reconstituted from a variety of fragments.

Animals↗
PubMed1984

Microinjection in the chick blastoderm. An improved method to study the extracellular matrix in the living organism.

A microinjection technique for the chick blastoderm is described. With a micropipette attached to a de Fonbrune micromanipulator, 25-45 nl of a reagent was injected into the entophyllic crescent of a chick blastoderm explanted in vitro according to New [7]. This procedure offers the advantage of eliminating the concentration variability which was observed after subblastodisc injection, and in contrast to the in ovo techniques, it allows one to stage the blastoderms properly. To check its applicability, testicular hyaluronidase was injected. On the basis of morphological and histochemical observations we ascertained that the experimental procedure itself did not interfere with the results. This method may provide a reliable experimental procedure with which to study the interactions between several macromolecules and the tissues during morphogenesis.

Animals↗
PubMed1991

Spectrometric measurements in the anterior eye vasculature of the albino rabbit--a study with the EMPHO I.

Intracapillary haemoglobin oxygenation (oxygen saturation) and haemoglobin concentration (corresponding with blood volume) were measured in different regions of the albino rabbit anterior eye segment using the Erlangen micro-lightguide spectrophotometer (EMPHO I). The tip of the microlightguide (active diameter 210 microns, cover diameter 0.5 mm) was placed in the vitreous by a goniometric mounting and controlled by a micromanipulator. After stepwise elevation of intraocular pressure (IOP) the haemoglobin oxygenation decreased slightly in the iris at an IOP-level of 60 mmHg, whereas in the three regions of the ciliary processes and in the peripheral choroid haemoglobin oxygenation did not decrease until values of 80 mmHg were applied. In contrast, haemoglobin concentration decreases when the IOP increases in all regions except in the pars plana where the haemoglobin concentration increased at 40 and 60 mmHg. The most pronounced reactive hyperaemia was found in the major ciliary processes. Quite passive changes of haemoglobin concentration were seen in the peripheral choroid. Effective regulatory mechanisms must exist which result in IOP-independent (20-60 mmHg) haemoglobin oxygenation, whereas pronounced changes in intracapillary haemoglobin concentration were observed. The decay of both parameters at IOP 80 mmHg is indicative of an exhaustion of their regulatory capacity. Epinephrine topically applied onto the conjunctival sac or injected into the common carotid artery led to a short-term decrease of haemoglobin concentration and, later, a hyperaemic response. A short-term haemoglobin deoxygenation was found especially in the major ciliary processes.

Animals↗
PubMed1988

Plasmodium falciparum: induction of resistance to mefloquine in cloned strains by continuous drug exposure in vitro.

A genetically homogeneous population of Plasmodium falciparum prepared by a single erythrocyte micromanipulation technique was used to produce lines of P. falciparum resistant to mefloquine hydrochloride in vitro. Parasites were maintained in a culture medium containing gradually increased concentrations of mefloquine hydrochloride (CMP-mef) starting with 2 ng/ml. One of the mefloquine-resistant culture lines (W2-mef) was obtained after 96 weeks of continuous culture in CMP-mef, the last 4 weeks in medium containing 40 ng/ml of mefloquine hydrochloride. The W2-mef was four to six times more resistant to mefloquine than was the parent clone W2. Means of multiple determinations of 50% inhibitory concentrations (IC-50) of mefloquine hydrochloride against W2-mef and clone W2 were 20.39 +/- 5.08 ng/ml and 4.50 +/- 1.94 ng/ml, respectively.

Animals↗
PubMed1992

Stimulation of acid secretion increases the gastric gland luminal pressure in the rat.

The gastric mucosal gland luminal pressure was measured in vivo with a pressure-sensitive microelectrode technique (servo-null) in anesthetized rats. A microelectrode was inserted into a gland lumen by means of a micromanipulator at an angle of 30 degrees to the mucosal surface. Acid secretion was estimated by measuring the pH in the solution covering the mucosa. During control conditions, when the mucosa was secreting acid spontaneously, gland luminal pressure was 12.3 +/- 1.2 mm Hg. At about 9 minutes after starting pentagastrin administration, the luminal pressure stabilized at 17.2 +/- 1.7 mm Hg. In the rats given impromidine (500 micrograms.kg-1.h-1) luminal pressure gradually increased (during 9-10 minutes) from a control level of 9.0 +/- 1.9 to 17.3 +/- 2.6 mm Hg. During the majority of experiments, the luminal pressure oscillated at 3-7 cycles per minute. The results show that intraluminal pressure increases during stimulated acid secretion, indicating that a resistance to the volume secretion exists in the upper part of the gastric crypts. This hydrostatic pressure may well be the driving force for creating channels for acid and pepsin to cross the mucus layer covering the mucosal surface.

Animals↗
PubMed1994

Effectiveness of infertility treatments: choice and comparative analysis.

Choice of infertility treatments usually depends on a balance of the chances of conceiving with or without treatment, and with more or less complex and costly treatments, and on other factors such as duration of infertility and the woman's age. Pregnancies often occur independent of treatment and prospective controlled trials are needed. Comparability between reported results requires pregnancy and birth rates to be calculated in a time-specific or cycle-specific way. Cumulative rates are preferable to account for the usual tendency of fecundity to fall progressively. This review focuses on such published data in order to assess the relative effectiveness of treatments, both conventional and assisted conception methods. The main conclusions are: (1) The only treatments that can achieve a normal chance of pregnancy are the ovulation induction methods in cases of oligomenorrhea/amenorrhea, and the assisted conception methods for other female causes and unexplained infertility. (2) Tubal/pelvic infective damage and endometriosis require new severity classifications which are sensitive to functional potential before and after surgery, and in vitro fertilization (IVF) would often be indicated as the primary choice. (3) Duration of unexplained infertility determines the need and therefore benefit of any of the treatments used, of which the assisted conception methods are by far the most effective. (4) Interpretation of reported results of treatments for 'male factor' infertility is critically affected by the diagnostic accuracy of defining sperm dysfunction. (5) In cases of well-defined sperm dysfunction there is little or no therapeutic benefit to the chance of natural conception, nor by intrauterine insemination; there is moderate success by IVF, but no proven benefit over standard IVF by any micromanipulative method except probably intracytoplasmic sperm injection.

Female↗
PubMed1978

Classical and alloimmune anaphylactic degranulation of isolated single mast cells.

Viable mast cells, directly isolated by micromanipulation from a mouse peritoneal cell suspension, were deposited on the bottom of microtiter-plate wells and submitted to histamine release. Conventional antigen-induced anaphylactic degranulation as well as direct allogeneic anaphylactic degranulation were strongly inhibited when these mast cells were settled on normal tissue culture plastic surfaces. Nevertheless, normal degranulation could be recovered by pretreatment of the experimental surface with a multipositive charged molecule (poly-L-lysine). Under these conditions, we demonstrate that the degranulation of one isolated mast cell is possible and consequently, as regards the direct allogeneic anaphylactic degranulation, confirm the "self-triggering mechanism" in which the recognition of histocompatibility antigens on the membrane of the mast cell itself is the trigger to the secretory response. The technique of monocellular degranulation described in this paper provides a new tool which leads us to think that the problem of detection of anaphylactic antibody-secreting cells can be solved.

Anaphylaxis↗
PubMed1986

Human T lymphocyte clones with killer or natural killer activity.

We describe a reliable method for obtaining a significantly higher frequency of human cloned T lymphocytes with killer and/or NK-like activity. Human peripheral blood mononuclear cells were treated with recombinant interferon-gamma (rIFN-gamma) and recombinant interleukin-2 (rIL-2) in a culture medium containing autologous serum and were then cloned by single cell micromanipulation. The cloned T lymphocyte populations were tested simultaneously for their ability to proliferate in response to exogenous IL-2, to exhibit lectin-dependent cytolysis and to kill the tumor cell line K562. Results indicate that the cloning technique allowed each isolated T lymphocyte to undergo cell expansion. Furthermore when T cells were pretreated with rIFN-gamma and rIL-2, 88% of the T cell clones were capable of mediating cytotoxicity in the presence of PHA. Moreover one third of the clones which exhibited lectin-dependent lysis were able to kill K562 target cells.

Clone Cells↗
PubMed1986

A study of the effects of buspirone, BMY 13805, and 1-PP on dopaminergic metabolism in the nucleus accumbens using in vivo voltammetry in freely moving rats.

The effects of buspirone, a buspirone analogue (BMY 13805) and a buspirone metabolite (1-PP) on dopaminergic metabolism in the nucleus accumbens were investigated using in vivo voltammetry. Differential pulse voltammetry coupled with electrochemically pretreated carbon fiber electrodes was used to provide a continuous and selective measure of the 3,4-dihydroxyphenylacetic acid (DOPAC). An implanted micromanipulator enabled the use of freely moving animals. Buspirone injections induced a marked and rapid increase of the DOPAC peak in the nucleus accumbens. Buspirone was 10 times more potent when injected subcutaneously than intraperitoneally. BMY 13805 and 1-PP were without effect on dopaminergic metabolism in the nucleus accumbens. In conclusion, the anxiolytic properties of these drugs and their effects on dopaminergic metabolism do not appear related.

3,4-Dihydroxyphenylacetic Acid↗