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Centrifugal actions on amacrine and ganglion cells in the retina of the turtle.

1. An electrophysiological investigation of efferent synapses in the retina of the turtle was conducted by recording intracellularly from amacrine cells. These cells have been selected because in birds they have been shown to have direct anatomical connexions with centrifugal fibre terminals. 2. Amacrine cells could be easily distinguished from most other retinal cells, except ganglion cells, by their different photo-responses. Because both amacrine and ganglion cells may generate action potentials they were distinguished by their responses to optic nerve stimulation. 3. The response of ganglion cells to single shock stimulation of the optic nerve consists of an antidromic action potential followed by a late synaptic potential. 4. Cells which did not show antidromic responses but were electrically excitable, by passing direct current through the recording electrode, were considered to be amacrine cells. 5. Amacrine cells generate an e.p.s.p. in response to optic nerve stimulation. An analysis of the e.p.s.p. suggests that it may be due to a single afferent fibre terminating in the proximity of the cell soma. By analogy to the bird, it is concluded that the amacrine cells e.p.s.p.s result from the activation of centrifugal fibres.

Action Potentials↗

Influence of centrifugal pathways on forward masking of ventral cochlear nucleus neurons.

When responses to one part of a sequence of auditory signals reduce the responses to a subsequent portion of the signal, "forward masking" results. Although forward masking occurs in the auditory nerve, that observed in the ventral cochlear nucleus (VCN) more closely resembles psychophysical forward masking. In contrast to the auditory nerve in which the amount of forward masking is proportional to the amount of excitation produced by the masker, most VCN neurons show a poor correlation between forward masking and excitation produced by the masker, indicating a more complex interaction between responses to adjacent signals. This study tested the hypothesis that one component of forward masking is produced by inputs from centrifugal neural connections to the VCN. The centrifugal pathways were interrupted with knife-cut lesions medial to the CN. Responses of single units obtained 60 minutes after the lesions were compared to those obtained before the lesions. In primarylike, sustained chopper and on units the lesions resulted in a reduction in forward masking and enhanced recovery. In contrast, lesions resulted in increased masking in primarylike-notch and low-intensity chopper units. The relationship between masker-elicited excitation and forward masking became more monotonic for transient choppers and on units, approaching that observed for auditory nerve fibers. These effects are probably the result of removal of both inhibitory and excitatory inputs, ultimately reflecting a balance of excitation and inhibition to each neural population in the VCN.

Animals↗

Synaptic connections of the centrifugal fibers in the pigeon retina.

The centrifugal fibers in the pigeon retina end in the inner nuclear layer and form two kinds of terminals, convergent and divergent. In the inner nuclear layer the fibers synapse with amnacrine and displaced ganglion cells. Because of their great number and their even distribution these fibers appear to constitute a system for the localized centrifugal control of the retinal functions.

Animals↗

Viral contribution to dissolved DNA in the marine environment as determined by differential centrifugation and kingdom probing.

Dissolved or filterable (<0.2-(mu)m-pore-size filter) DNA is a ubiquitous component of the dissolved organic matter in the surface waters of this planet. In an effort to understand the composition and possible sources, we subjected dissolved DNA concentrated by vortex flow filtration from offshore and coastal environments to differential centrifugation and probing with 16S rRNA-targeted kingdom oligonucleotide probes. Initial studies with calf thymus soluble DNA and T2 phage particles indicated that high-speed ultracentrifugation (201,000 x g for 90 min), a method to separate viral particles from soluble DNA used by other investigators, resulted in pelleting of nearly all the DNA and virus particles. Lower-speed centrifugation (11,200 to 25,800 x g for 90 min) resulted in >99% of the virus particles being collected in the pellet and (equiv)65% of the calf thymus DNA remaining in the supernatant. Employing this approach, we estimate that approximately 50% of the filterable DNA from marine environments is truly soluble or free DNA and that the other half is composed of bound forms (viral particles and, potentially, colloids). Of the bound form, 17 to 30% could be accounted for by viral particles, by calculating the amount of viral DNA on the basis of viral abundance, leaving a portion of the bound form uncharacterized. Kingdom probing with universal, eubacterial, and eucaryotic probes indicated that dissolved DNA hybridized with all of these probes, while purified standard viral DNAs did not, or hybridized only slightly with the universal probe (tailed oligonucleotide only). Collectively, these data indicate that DNA in viral particles is a small component of the dissolved DNA, the majority being of eubacterial and eucaryotic origin.

Journal Article↗

Isolation of Mycobacterium chelonei with the lysis-centrifugation blood culture technique.

Mycobacterium chelonei was isolated from a patient by the lysis-centrifugation and the conventional two-bottle blood culture methods. The lysis-centrifugation method was significantly more sensitive and rapid than the conventional method in detecting and isolating this organism; quantitations done by this method were useful for monitoring response to therapy.

Adult↗

Improvement of growth of Chlamydia pneumoniae on HEp-2 cells by pretreatment with polyethylene glycol in combination with additional centrifugation and extension of culture time.

The following adaptations led to improved growth of Chlamydia pneumoniae on HEp-2 cells compared to that by the standard method: monolayer preincubation with 7% polyethylene glycol (PEG), extension of incubation time to 7 days, and extension of incubation to 7 days in combination with centrifugation on days 3, 4, and 5. These adaptations resulted in approximate increases in numbers of inclusion-forming units (IFU) of 2-, 5-, and 69-fold, respectively. A combination of preincubation with PEG, prolonged incubation, and centrifugation on days 3, 4, and 5 increased the numbers of IFU >300-fold. This is therefore recommended as the optimal method for culturing C. pneumoniae.

Bacteriological Techniques↗

Isolation of Legionella pneumophila by centrifugation of shell vial cell cultures from multiple liver and lung abscesses.

A 7-year-old girl was admitted to the hospital with acute lymphoblastic leukemia and was treated with allogenic cord blood transplantation. At day 30 after graft, she developed a fever and multiple nodular lesions disseminated in the liver and lungs. All bacterial cultures attempted on liver and lung biopsy specimens and blood remained sterile on standard axenic media. However, inoculation of liver and lung biopsy specimens on eukaryotic cell monolayers by the centrifugation-shell vial technique (M. Marrero and D. Raoult, Am. J. Trop. Med. Hyg. 40:197-199, 1989) led to the recovery of a strain of Legionella pneumophila serogroup 1, identified by 16S rRNA gene amplification and sequencing and serotyping. Our findings demonstrate that the centrifugation-cell culture method, which has previously been useful for the isolation of other strictly or facultatively intracellular bacteria, can also serve as a method for the recovery of L. pneumophila from clinical material.

Biopsy↗

Role of centrifugal pathways to cochlear nucleus in determination of critical bandwidth.

Atropine sulfate was applied locally to the cochlear nucleus of three unanesthetized cats in order to block the cholinergic centrifugal innervation of the nucleus. The critical bandwidth was measured behaviorally from the masked thresholds of 1-kHz tone pips in different bandwidths of masking noise. Atropine increased the critical bandwidth markedly. This suggests that normally the centrifugal pathways influence the critical bandwidth. Saline solutions alone had no effect, and if the cannula was not accurately implanted over the cochlear nucleus, atropine had no effect either.

Acoustic Stimulation↗

Efferents and centrifugal afferents of the main and accessory olfactory bulbs in the snake Thamnophis sirtalis.

This study reinvestigates the efferent projections of the main and accessory olfactory bulbs and describes for the first time the centrifugal projections to the main and accessory olfactory bulbs in the snake Thamnophis sirtalis, using the intraaxonal transport of the anterograde and retrograde tracer biotinylated dextran amine and the retrograde tracer horseradish peroxidase. The olfactory projection consists of three tracts: the lateral olfactory tract, which projects bilaterally to the lateral cortex and the rostral amygdala, crossing the midline through the stria medullaris-habenular commissure system; the intermediate olfactory tract, which projects ipsilaterally to the olfactory tubercle and contributes to the contralateral projection; and the medial olfactory tract, which projects ipsilaterally to the dorsomedial retrobulbar formation. The vomeronasal projection is formed by a single tract, the accessory olfactory tract, that projects ipsilaterally to the nucleus of the accessory olfactory tract, the medial amygdala and the nucleus sphericus. The centrifugal projections to the main and accessory olfactory bulb are composed of two components: one that arises in areas that receive the olfactory or vomeronasal input (neurons in the olfactory tubercle, retrobulbar formation and lateral cortex project to the main olfactory bulb; and neurons in the nucleus of the accessory olfactory tract, the medial amygdala and the nucleus sphericus project to the accessory olfactory bulb), and another that arises in areas not directly implicated in processing the chemosensory information (the nucleus of the diagonal band of Broca and the dorsal cortex). These data allow the recognition of the general pattern of organization of the reptilian olfactory and vomeronasal systems.

Afferent Pathways↗

Correlation between macroscopic porosity location and liquid metal pressure in centrifugal casting technique.

Radiographic analysis of uniform cylindrical castings fabricated by the centrifugal casting technique has revealed that the macroscopic porosity is dependent on the location of the sprue attachment to the casting. This is attributed to the significant pressure gradient associated with the centrifugal casting technique. The pressure gradient results in different heat transfer rates at portions of the castings near and away from the free surface of the button. Consequently, the macroscopic porosity is invariably at portions of the casting close to the free surface of the button. In addition, some optimized sprue-reservoir combinations could be predicted and proved, based on this pressure gradient concept.

Chemical Phenomena↗

Experimental and computational studies of the relative flow field in a centrifugal blood pump.

The relative flow field within the impeller passage of a centrifugal blood pump had been examined using flow visualization technique and computational fluid dynamics. It was found that for a seven-blade radial impeller design, the required flow rate and static pressure rise across the pump could be achieved but the flow field within the blades was highly undesirable. Two vortices were observed near the suction side and these could lead to thrombus formation. Preliminary results presented in this article are part of our overall effort to minimize undesirable flow patterns such flow separation and high shear stress regions within the centrifugal blood pump. This will facilitate the future progress in developing a long-term clinically effective blood pump.

Computer Simulation↗

Application of a centrifugal shooting type polishing system to polish pure titanium.

Titanium is a difficult metal to grind, and thus mechanical polishing using conventional rotating tools takes a lot of time. The purpose of this study was to examine the polishing effect of a centrifugal shooting type polishing system on a pure titanium surface processed with a CAD/CAM system. The specimens' surfaces were polished for various times using this apparatus, then the center line average roughness (Ra) of each specimen was measured with surface roughness measuring apparatus. In addition, the surfaces after polishing were observed with a scanning electron microscope. After 50 sec of polishing, Ra decreased from 1.1 microns to less than 0.6 micron, and after 60 sec of polishing the cutting marks generated by the CAD/CAM system disappeared and a smooth surface was observed. Therefore, it appears that a centrifugal shooting type polishing system is effective at polishing pure titanium.

Computer-Aided Design↗

Leishmania donovani parasites in the nasal secretions, tonsillopharyngeal mucosa, and urine centrifugates of visceral leishmaniasis patients in Kenya.

In the early 1930s, investigators of visceral leishmaniasis stated that Leishman-Donovan bodies are found in body fluids of kala-azar patients, for example, in urine, feces, semen, and nasal and pharyngeal secretions. Based on this finding, we investigated the diagnostic potential of nasal secretions, tonsillopharyngeal mucosal swabs, and urine centrifugates inoculated into Schneider's Drosophila Medium (containing antibiotics and antifungal agents) as well as with Giemsa-stained smears. Consequently, 64 randomly selected patients with visceral leishmaniasis from Kenya (59 who were splenic culture or Giemsa stain positive and five who were culture negative but Giemsa stain positive) were tested by three noninvasive methods. These tests were all performed before the patients were treated with Pentostam. Cultures of nasal and tonsillopharyngeal swabs and urine centrifugates produced 28 positive samples representing 24 patients (37.5%). Moreover, a set of 25 Giemsa-stained slide smears made from the nasal and tonsillopharyngeal mucosa of 25 patients with visceral leishmaniasis who had not tested positive in cultures produced nine positives. Therefore, the overall total of patients who tested positive by all of the above methods was 33 or 51.6%. The cryopreserved Leishmania isolates were characterized by cellulose acetate electrophoresis using 20 enzyme systems. The isoenzyme profiles produced by the parasites were represented in five different L. donovani s.l. zymodemes. Representatives of these isolates were also characterized by DNA Southern blotting analysis, which corroborated the isoenzyme results.

Adolescent↗

An in vitro evaluation of an automatic clamp for use with centrifugal pumps.

The risk of air emboli is a concern for all perfusionists. A new clamping device for use with centrifugal pumps is designed to clamp both the arterial and venous lines at the first indication of air or retrograde flow, thereby allowing the perfusionist to evaluate the situation and correct the problem before entraining air into the arterial pump head. After evaluating this device in our lab, we conclude that this new safety device should be added to the heart lung machine by all perfusionists using centrifugal pumps.

Cardiopulmonary Bypass↗

Platelet function and hemolysis in centrifugal pumps: in vitro investigations.

The effects of centrifugal pumps on blood components other than erythrocytes, namely platelets and their interaction with the coagulation system, are not very well known. In a comparative study with three centrifugal pumps (BioMedicus BP-80, St. Jude Isoflow, and Sarns Delphin) and the Stockert roller pump hemolysis, platelet counts, thromboplastin and partial thromboplastin times, as well as resonance thrombography (RTG) parameters for the assessment of platelet and coagulation function were evaluated in vitro. Normalized indices of hemolysis (NIH) with ACD anticoagulation after 360 minutes were 0.008+/-0.004 (Isoflow), 0.018+/-0.017 (BP-80), 0.085+/-0.051 (Delphin), and 0.049+/-0.010 g/1001 (roller pump). Plasmatic coagulation was activated in all circuits. Platelet function was severely inhibited by the BP-80, indicated by increase in RTG platelet time to 358%+/-150% of initial values compared to 42%+/-29% (Isoflow), 40%+/-20% (Delphin), and 12%+/-10% (roller pump). Fibrin polymerization was affected similarly. The large surface area of the BP-80 leads to an extensive activation of platelets and plasminogen.

Blood Coagulation↗

[Comparative study of two methods for salvaging red blood cells: centrifugation techniques (cell-saver) versus filtration through a 100,000-dalton filter].

OBJECTIVES: Some loss of blood occurs during blood salvage. We hypothesized that plasmapheresis filtering would damage blood much less than centrifugation techniques do, thereby allowing more red blood cells to be transfused. MATERIAL AND METHODS: Laboratory study in which 16 units of whole donor blood were distributed randomly in two groups and processed either by a conventional "cell-saver" method or by hemofiltration using recirculation through a 100,000 dalton filter. We analysed hemoglobin, hematocrit, free hemoglobin, extracellular potassium, platelets, leukocytes, protein and albumin in whole blood before and after processing, and in the waste bag in each group. RESULTS: The recovery of hemoglobin and red blood cell volume was about 80% with both methods. More free plasma hemoglobin was found in the waste bag with the filtration technique. Hemolysis in processed blood was low, less than 0.1% in both groups. Platelet recovery with conventional centrifugation and filtration was 11 and 49%, respectively. Albumin, total protein and extra-cellular potassium were recovered at a rate of about 20% with the filtration technique, whereas recovery of these elements was minimal with the cell saver method. CONCLUSIONS: Both methods of autotransfusion caused moderate loss of red blood cells and low plasma levels of free hemoglobin in processed blood. Recovery of platelets, albumin, total protein and potassium was better with filtration than with the "cell-saver" method.

Blood Donors↗

Disruption of the olfactoretinal centrifugal pathway may relate to the visual system defect in night blindness b mutant zebrafish.

We describe here a dominant mutation, night blindness b (nbb), which causes an age-related visual system defect in zebrafish. At 4-5 months of age, dark-adapted nbb(+/-) mutants show abnormal visual threshold fluctuations when measured behaviorally. Light sensitizes the animals; thus early dark adaptation of nbb(+/-) fish is normal. After 2 hr of dark adaptation, however, visual thresholds of nbb(+/-) mutants are raised on average 2-3 log units, and rod system function is not detectable. Electroretinograms recorded from nbb(+/-) mutants are normal, but ganglion cell thresholds are raised in prolonged darkness, suggesting an inner retinal defect. The visual defect of nbb(+/-) mutants may be likely caused by an abnormal olfactoretinal centrifugal innervation; in nbb(+/-) mutants, the olfactoretinal centrifugal projection to the retina is disrupted, and the number of retinal dopaminergic interplexiform cells is reduced. A similar visual defect as shown by nbb(+/-) mutants is observed in zebrafish in which the olfactory epithelium and olfactory bulb have been excised. Homozygous nbb fish display an early onset neural degeneration throughout the CNS and die by 7-8 d of age.

Age Factors↗

Evaluation of a centrifuge with rapid turnaround time for the preparation of plasma samples for measurement of common STAT markers on the ACS: 180 system.

Reported is the evaluation of a new centrifugation method, Statspin, that addresses both time and sample separation integrity. The method can successfully separate the plasma fraction from the cellular material in 2 minutes as compared to 20 minutes for the conventional centrifuge method. The Statspin, combined with the ACS:180 system, can generate test results in less than 30 minutes, exclusive of transport to the laboratory. This study demonstrated that the combined technologies offer timing-saving improvements for clinical laboratories offering STAT immunoassays for cardiac markers, endocrine molecules, and therapeutic drugs.

Biomarkers↗