Excitable properties and voltage-sensitive ion conductances of horizontal cells isolated from catfish (Ictalurus punctatus) retina

External horizontal cells were enzymatically dissociated from intact catfish (Ictalurus punctatus) retina and pipetted onto a small chamber attached to the stage of an inverted phase-contrast microscope. Individual horizontal cells were recognized by their large size and restricted dendritic arborization. Low-resistance (3-12 M omega) patch-type electrodes were used to record intracellular potentials and to pass current across the cell membrane under either current or voltage-clamp conditions. The average resting potential of isolated horizontal cells was -67 V + 6.9 mV (mean +/- SD, n = 40). At the resting potential, the cell membrane appears to be mainly permeable to K. A depolarizing current step evoked an action potential in the cell. The maximum rate of rise of the action potential (dV/dt) in normal physiological solution was 6.5 +/- 1.8 V/s (means +/- SD, n = 24) and was reduced to 1.2 +/- 0.39 V/s (means +/- SD, n = 9) in 1-10 micron tetrodotoxin (TTX) and 3.2 +/- 1.4 V/s (means +/- SD, n = 6) in Ca-free solution. The maximum dV/dt was reduced in 10 mM extracellular K concentration [K]o to about half of that seen in standard saline, and values in 30 or 80 mM [K]o were similar to that measured in TTX. Following an action potential, the membrane potential reached a plateau potential of + 17.4 +/- 8.1 mV (means +/- SD, n = 17) and remained depolarized for variable periods of time lasting from less than a second to a few minutes. When the plateau potential was long lasting, the cell repolarized slowly and upon reaching zero rapidly repolarized to the original resting potential. The duration of the plateau potential decreased or was absent in saline containing one of the following calcium channel antagonists: La, Cd, Co, or Ni. The voltage-clamp technique was used to identify the membrane currents responsible for the membrane potential changes seen under current clamp. Experiments were carried out using either a single or two individual electrodes. Fast and steady-state inward currents were recorded from isolated horizontal cells in the voltage range between -20 and +20 mV. These currents were a result of increased membrane conductance to both Na and Ca ions. The Na channels are inactivated at depolarized potentials and are TTX sensitive. Ca channels are partially inactivated at depolarized potentials. The Ca conductance is decreased by Cd, Co, Ni, and La. Ba can substitute for Ca in the channel.(ABSTRACT TRUNCATED AT 400 WORDS)     

Determination of 99mTc-DTPA clearance by a single plasma sample method

To derive a formula for determination of 99mTc-DTPA clearance (Cl) from the radioactivity in a single plasma sample, the relation Cl = ECV/t was used, where ECV is the extracellular volume and t is the mean transit time of the tracer in the organism. By studying 99mTc-DTPA time activity curves of 45 consecutive patients we found that ECV could be estimated from the body surface area, and that t could be calculated from the radioactivity in a single plasma sample. Cl calculated by the single sample method was almost identical to Cl calculated by a standard multiple sample method (r = 0.987). It is concluded that the single sample method is accurate and that it may prove useful as a routine method provided that the method is not used in patients with 99mTc-DTPA clearance less than 30 ml min-1, since this investigation includes only patients with Cl greater than 30 ml min-1; the method is not used for children, since the test material did not contain any children; the method is only used for plasma samples drawn at 180 less than or equal to t less than or equal to 300 min.     

Pitfalls in the interpretation of leukocyte counts of newborn infants.

Some pitfalls in the interpretation of neonatal leukocyte counts are identified. The well-known variability in neonatal leukocyte counts was investigated by simultaneously sampling arterial, venous and capillary blood, and during periods of rest and mild and violent exercise. Venous blood leukocyte counts were 82% +/- 3.5 (mean +/- SE, P = less than .001) of counts in simultaneously drawn capillary blood from heel punctures; arterial blood counts were 77% +/- 5.3 (P less than .001) of capillary blood values. Following violent crying, capillary blood leukocyte counts increased to 146% +/- 6.1 (P less than .001) of baseline values, and a shift to the left occurred. Milder exercise induced an increase to 113% +/- 5.2 (P less than .05), without a leftward shift. Thus, counts from different vascular sources cannot be considered equivalent. Also, counts from vigorously crying babies may show leukocytosis and a leftward shift, and erroneously suggest bacterial infection. It is recommended that serial counts be obtained from a consistent vascular source in resting babies.     

DNA-aneuploidy in 46,XX hydatidiform moles

The DNA content of single villous cell nuclei in 14 normal pregnancies and 21 androgenetic moles with a 46,XX karyotype was measured by flow cytometry. Six molar specimens showed a low mean DNA content close to that of normal villi, whereas, 15 specimens were characterized by a high mean DNA content due to a large fraction of nuclei in the triploid to tetraploid region. The latter group presumably corresponds to the previously described "type II" moles in which the DNA pattern resembled the pattern seen in invasive moles and choriocarcinoma.     

The adaptation of the plaque reduction assay for measuring specific cytotoxic cells in limiting dilution cultures

The use of the original haemolytic plaque reduction technique to measure cytotoxic T lymphocytes (CTL) has been developed further as a rapid screening assay, particularly suitable for limiting dilution analyses. Using hybridoma cells as targets, the cytotoxicity has been measured by the loss of haemolytic plaque formation and by the reduction of the amount of haemolytic monoclonal antibody secreted from viable target cells into the assay supernatants. The assessment of large numbers of cytotoxic samples has been greatly facilitated by quantitating the amount of haemoglobin released in the assay with an automated microELISA multiscanner and by scoring visually using a modification of the spot test. Using these new techniques, relatively high frequency estimates of cytotoxic cell precursors in an allogeneic response (1 in 462 spleen cells) and an anti-fluorescein response (1 in 3970 spleen cells) were obtained.     

Taxonomy of 37-collar spined Echinostoma (Trematoda: Echinostomatidae) in studies on the population regulation in experimental rodent hosts

With reference to a recent taxonomic revision the species of the echinostomes used in key studies on the population regulation in infections with 37-collar-spined Echinostoma species in experimental rodent hosts were reconsidered. This was considered essential to prevent taxonomic problems blocking further fruitful progress within this field of experimental parasitology.