Control and eradication of animal diseases in New Zealand.

New Zealand is free from all the major epidemic (Office International des Epizooties List A) diseases of animals and other important diseases, such as rabies and the transmissible spongiform encephalopathies. The once endemic conditions of sheep scab (Psoroptes ovis), bovine brucellosis (Brucella abortus), hydatids (Echinococcus granulosus) and Aujeszky's disease have been eradicated. Anthrax (Bacillus anthracis) is no longer considered endemic and Pullorum disease (Salmonella Pullorum) has effectively been eradicated from commercial poultry flocks. There are current control programmes for bovine tuberculosis (Mycobacterium bovis), enzootic bovine leucosis in dairy cattle, infectious bursal disease, ovine epididymitis (Brucella ovis), and caprine arthritis encephalitis. Historically, incursions by three important non-endemic diseases, contagious bovine pleuropneumonia, classical swine fever and scrapie, have been successfully eliminated. Any new occurrence of a serious exotic disease would be dealt with swiftly using powerful legislative authorities available for the purpose.

Football protective gear and cervical spine imaging.

STUDY OBJECTIVE: We sought to assess the effect of football protective gear on the cervical spine radiographic evaluation of adult male subjects. METHODS: The study used a prospective, randomized, matched-pairs, observational design. Subjects served as their own control subjects, with cross-table lateral and open-mouth odontoid cervical spine radiographs. Radiographs were obtained with protective head and shoulder equipment (pads group) and without protective equipment (no pads group). Two emergency physicians and 2 neuroradiologists reviewed study radiographs. Physicians assessed radiographic views for adequate cervical spine visualization to the C7-T1 level and the odontoid and related structures. Comparison of radiographic readings for the pads and no pads groups used the McNemar exact test. A McNemar test of equality of paired proportions was used to estimate a population of 20 paired individuals to detect a significant outcome difference. RESULTS: Zero percent of the pads group's cross-table lateral structures were adequately visualized by all 4 reviewers (reviewer unanimity decision) compared with 25% of the no pads group's cross-table lateral films (between-group difference 25%; 95% confidence interval [CI] 6.0 to 44). When 3 of 4 reviewers noted adequate visualization (reviewer majority decision), 0% of the pads group's cross-table lateral structures were adequately visualized versus 40% of the no pads group's cross-table lateral radiographs (between-group difference 40%; 95% CI 19 to 62). With reviewer unanimity, 25% of the pads group's open-mouth odontoid structures were visualized versus 45% of the no pads group's open-mouth odontoid structures (between-group difference 20%; 95% CI -8.9 to 49). With reviewer majority analysis, 35% of the pads group's odontoid radiographs were adequately visualized versus 75% of the no pads group's open-mouth odontoid radiographs (between-group difference 40%; 95% CI 12 to 68). CONCLUSION: Football head and shoulder protective equipment appears to be an impediment to cervical spine radiographic visualization. Guidelines for players' cervical spine imaging should incorporate procedures for removal of equipment before initial radiographic evaluation.

Calcium channel current in rat dental pulp cells.

Voltage-gated Ca(2+) currents in early-passage rat dental pulp cells were studied using whole-cell patch-clamp techniques. With Ba(2+) as the charge carrier, two prominent inwardly-directed currents, I(f) and I(s), were identified in these cells that could be distinguished on the basis of both kinetics and pharmacology. I(f) was activated by membrane depolarizations more positive than -30 mV, and displayed fast inactivation kinetics, while I(s) was activated by steeper depolarizations and inactivated more slowly. At peak current, time constants of inactivation for I(f) and I(s) were approximately 17 vs. approximately 631 msec. Both I(f) and I(s) could be blocked by lanthanum. By contrast, only I(s) was sensitive to either Bay-K or nifedipine, a specific agonist and antagonist, respectively, of L-type Ca(2+) channels. I(s) was also blocked by the peptide omega-Conotoxin GVIA. Taken together, results suggested that I(f) was mediated by divalent cation flow through voltage-gated T-type Ca(2+) channels, whereas I(s) was mediated by L- and N-type Ca(2+) channels in the pulp cell membrane. The expression of these prominent, voltage-gated Ca(2+) channels in a presumptive mineral-inductive phenotype suggests a functional significance vis a vis differentiation of dental pulp cells for the expression and secretion of matrix proteins, and/or formation of reparative dentin itself.

Usefulness of left ventricular mass in predicting recovery of left ventricular systolic function in patients with symptomatic idiopathic dilated cardiomyopathy.

Prognosis of idiopathic dilated cardiomyopathy (IDC) is variable. We determined the prognostic value of left ventricular (LV) mass and systolic and diastolic function in patients with IDC of <12 months duration. Clinical and echocardiographic assessment was performed at baseline and at 8+/-6 months follow-up in 25 patients (47+/-13 years) with IDC and an LV ejection fraction (LVEF1) of <40% (22+/-7%). Based on a follow-up LVEF (LVEF2) of < or >40%, patients were divided into unimproved (n = 13, LVEF2 = 21+/-9%) and improved groups (n = 12, LVEF2 = 51+/-11%). There was no difference in the LVEF1 (22+/-8% vs. 22+/-6%), LV end-systolic (5.7+/-0.8 vs. 5.8+/-0.9 cm) or end-diastolic (6.5+/-0.6 vs. 6.6+/-0.9 cm) dimension, wall stress (102+/-26 vs 99+/-28 g/cm2), end-systolic (1.7+/-0.3 vs. 1.8+/-0.2) or end-diastolic (1.7+/-0.3 vs. 1.6+/-0.1) sphericity, dp/dt (582+/-163 vs. 678+/-222 mm Hg/s), or right ventricular fractional shortening (20+/-9% vs. 27+/-7%, p = 0.06) in unimproved and improved groups. LV mass was lower (1.00+/-0.21 vs. 1.38+/-0.27 g/ml, p = 0007) and mitral inflow E-wave deceleration time shorter (97+/-42 vs. 164+/-58 ms, p = 0007) in the unimproved versus the improved group. On Pearson correlation analysis, LV mass (r = 0.62, p = 0.001), deceleration time (r = 0.68, p = 0.0002), wall motion score index (r = -0.47, p = 02), and dp/dt (r = 0.52, p = 03) were the significant predictors of LVEF2. There was correlation between LV mass (grams per milliliter) and deceleration time (r = 0.61, p = 0.001). During follow-up, death occurred in 1, and readmission for worsening heart failure in 4 patients in the unimproved group versus no hospitalization in the improved group. Thus, in patients with recent onset IDC, LV mass and diastolic function determine late outcome.

A novel method to isolate odontoblasts from rat incisor.

Historically, odontoblasts have been isolated from rat incisor using a surgical curette to separate these cells from the dentin. Isolation of odontoblasts using this approach typically resulted in cells with membrane properties that made the application of patch-clamp electrophysiological techniques prohibitive. The studies here describe a new procedure for isolating mature odontoblasts from adult rat incisor to obtain enriched populations of intact, viable odontoblasts that can be readily studied using patch-clamp methodologies. Identification of isolated cells as odontoblasts was confirmed using in situ mRNA hybridization for expression of dentin sialoprotein, osteocalcin, bone sialoprotein, and type I collagen, and calcium flux was monitored in these cells by means of fura-2 microfluorometry. We suggest that either single odontoblasts or clusters of these cells isolated by this new method would be an ideal preparation for the study of odontoblast properties using electrophysiological techniques, in situ hybridization and/or microfluorometry.

Extracellular Ca2+ increases cytosolic free Ca2+ in freshly isolated rat odontoblasts.

Recent evidence suggests that extracellular Ca2+ may modulate cell function in mineralized tissue. To determine whether dentinogenic cells, in particular, are sensitive to extracellular Ca2+, fura-2 microfluorometry was used to monitor intracellular calcium levels in odontoblasts freshly isolated from rat incisor. In response to applications of 0.5-4.0 mM extracellular calcium (CaCl2), most odontoblasts (84%; 107/128) showed an increase in intracellular calcium. For the majority of these cells (70%; 75/107), the typical response was biphasic; there was an initial, transient increase in intracellular calcium which reached peak levels within 30-50 s and decayed rapidly, followed by a slower (> 300 s) recovery toward basal levels. In general, the response of these cells to calcium was repeatable and the mean calcium concentration for the half-maximal response was approximately 1.3 mM. This effect could be partially blocked by either 200 microM lanthanum, a nonspecific blocker of Ca2+ channels, or 20 microM dantrolene, a potent inhibitor of Ca2+ release from internal stores. Used in combination, lanthanum, and dantrolene nearly abolished the calcium response completely. In addition, this response was sensitive to the dihydropyridine-sensitive calcium channel blocking agent nicardipine (60 microM), indicating a role for voltage-gated calcium channels during these events. These results show that odontoblasts respond to external calcium through mechanisms involving both influx of external calcium as well as release of calcium from internal stores and suggest a role for extracellular calcium in regulating the function of these cells.

Use of a resin-ionomer in the treatment of mechanical root perforation: report of a case.

A perforated root represents a difficult challenge to the clinician, and treatment of such defects often involves surgical and/or advanced restorative techniques. This typically requires a series of lengthy, often stressful appointments, may compromise esthetics, and invariably involves additional costs to the patient. This case report describes the nonsurgical treatment of separate mechanical perforations that resulted from the removal of a failed prefabricated post. The tooth was restored in a minimum number of visits through the use of a novel application of a resin-ionomer material originally designed for routine restorative procedures.

Potassium and chloride channels in freshly isolated rat odontoblasts.

It has been suggested that understanding the physiological properties of odontoblasts may be important in understanding the mechanisms underlying both metabolic and transductive processes in dental pulp. Because ion flux(es) may play a critical role in these events, it is of particular interest to understand ionic mechanisms in odontoblast cells. Thus, the aim of this study was to use patch-clamp recording techniques to examine the properties of resident ion channels in freshly dissociated odontoblasts. In recordings made in potassium-rich solutions, cells displayed at least three distinct channel amplitudes, with conductances of 130 +/- 18 pS, 52 +/- 4 pS, and 25 +/- 2 pS, respectively. Channel activity persisted in the presence of potassium salts of impermeant anions, and could be abolished by barium, a non-specific potassium channel blocker. In addition to the potassium conductances, we saw two separate anion channels in the odontoblast membrane. These channels were predominantly chloride-selective, weakly permeable to both acetate and aspartate, and had conductances of 391 +/- 64 pS and 24 +/- 3 pS. While questions remain regarding the functional role of these and other ion channels that presumably reside in the odontoblast membrane, our results demonstrate that it is possible to study ionic mechanisms of the odontoblast at the level of the single cell.

Membrane electric properties by combined patch clamp and fluorescence ratio imaging in single neurons.

An experimental method has been established to measure the electric properties of a cell membrane by combination of patch clamp and dual-wavelength ratio imaging of a fluorescent potentiometric dye, 1-(3-sulfonatopropyl)-4-[beta[2-(di-n-octylamino)-6-naphthyl]vinyl ]pyridinium betaine (di-8-ANEPPS). Pairs of fluorescence images from the dye-stained membrane of neuroblastoma N1E-115 cells excited at two wavelengths were initially obtained to calculate ratio images corresponding to the resting transmembrane potential. Subsequently, a whole-cell patch was established and the membrane potential clamped to levels varying from -100 to +60 mV; at each voltage, a pair of dual-wavelength images were acquired to develop a calibration of the fluorescence ratio. Using this method, the resting potentials could accurately be measured showing that the differentiated cells were 17 mV more polarized than undifferentiated cells. The combination of electrical and optical methods can also follow changes in other membrane electric properties, such as dipole potential, and thus permit a detailed analysis of the membrane electrical properties underlying the voltage regulation of ion channels.

Continuous mechanical loading alters properties of mechanosensitive channels in G292 osteoblastic cells.

G292 osteoblastic cells were cultured in dishes made with a flexible base of polytetrafluoroethylene (PTFE) and stretched ( approximately 1% strain level) continuously for 48 hours. Patch-clamp recording techniques were then used to monitor single channel currents of mechanosensitive ion channels in these cells. To stimulate mechanosensitive channels, we applied suction to the membrane, expressed as -cm Hg, directly through the patch pipette. GigaOhm seals were obtained on a total of 33 osteoblasts that contained a high-conductance ( approximately 180 pS) mechanosensitive channel, all in the cell attached configuration. Of these, 18 were obtained from cells that had been stretched for either 1 (n = 6), 24 (n = 4), or 48 (n = 8) hours, and 15 were obtained in control (nonstretched) cells at either 1 (n = 2), 24 (n = 5), or 48 (n = 8) hours. For unstrained cells, applied pressures ranging from -1 to -5 cm Hg increased the probability of channel opening (Popen) from 0.05 +/- 0. 01 (mean + SEM) to 0.12 +/- 0.07. By contrast, for the same values of applied pressure in stretched cells, Popen ranged from 0.06 +/- 0. 01 to 0.49 +/- 0.15. Our results suggest that intrinsic properties of mechanosensitive ion channels in the G292 osteoblastic cell may be modulated by continuous mechanical loading of the cell itself.

Faster voltage-dependent activation of Na+ channels in growth cones versus somata of neuroblastoma N1E-115 cells.

Kinetics of voltage-gated ionic channels fundamentally reflect the response of the channels to local electric fields. In this report cell-attached patch-clamp studies reveal that the voltage-dependent activation rate of sodium channels residing in the growth cone membrane differs from that of soma sodium channels in differentiating N1E-115 neuroblastoma cells. Because other electrophysiological properties of these channels do not differ, this finding may be a reflection of the difference in intramembrane electric field in these two regions of the cell. This represents a new mechanism for channels to attain a range of activities both within and between cells.

NaF potentiates a K(+)-selective ion channel in G292 osteoblastic cells.

Clinical studies have established that NaF increases mineral content in bone, although the cellular mechanisms underlying its osteoinductive effects remain unclear. Because metabolic effects of fluoride have been linked to ion flux and alterations in membrane potential, we used patch-clamp recording techniques to examine the electrophysiological response of osteoblastic cells to NaF. In these experiments, we show that NaF increased the amplitude and P(open) of a 73 pS potassium-selective ion channel. The effect of NaF depended on extracellular Ca2+ and could be blocked by a combination of calcium-channel blocking agents, suggesting that potentiation of channel activity was dependent on external calcium. Because all patches were in the cell-attached configuration, the effect of NaF was presumable indirect. Although the underlying cellular mechanisms remain unclear, our findings suggest that activity of calcium and/or potassium-selective channels via second messenger cascades may mediate many of the early events involved in the response of bone cells to inorganic fluoride.

Description of an in vitro platelet function analyzer--PFA-100.

A new in vitro system for the detection of platelet dysfunction, PFA-100, has been developed. It provides a quantitative measure of platelet function in anticoagulated whole blood. The system comprises a microprocessor-controlled instrument and a disposable test cartridge containing a biologically active membrane. The instrument aspirates a blood sample under constant vacuum from the sample reservoir through a capillary and a microscopic aperture cut into the membrane. The membrane is coated with collagen and epinephrine or adenosine 5'-diphosphate. The presence of these biochemical stimuli, and the high shear rates generated under the standardized flow conditions, result in platelet attachment, activation, and aggregation, slowly building a stable platelet plug at the aperture. The time required to obtain full occlusion of the aperture is reported as the "closure time." We have found that impairment of von Willebrand factor, or inhibition of platelet receptors glycoprotein Ib or IIb/IIIa with monoclonal antibodies or peptides, resulted in abnormal closure times. An antifibrinogen antibody, in contrast, failed to show any effect. The test appears to be sensitive to platelet adherence and aggregation abnormalities. The PFA-100 system has potential applications in routine evaluation of platelet function in the clinical setting because of its accuracy, ease of operation, and rapid turnaround of results.

Neural form of voltage-dependent sodium current in human cultured dental pulp cells.

Intradental, i.e. pulpal, cells may play an important part in sensory transduction in teeth, although the cellular mechanisms and the identity of the specific cell types involved are still unclear. Because the majority of cells in dental pulp are derived from neural crest, it seemed likely that these might have the membrane properties of other neural-derived cells found in the peripheral or central nervous system. The patch-clamp recording technique was used to show that cells in explant cultures from human dental pulp contain a voltage-gated, tetrodotoxin-sensitive inward current. Mean activation potential of the current was -42 +/- 2.5 mV and the voltage at half-inactivation was -79.4 +/- 5.3 mV, suggesting a neural-like sodium conductance. In addition, these cells were immunoreactive to glial acidic fibrillary protein, growth-associated protein (GAP-43), and vimentin, further suggesting that dental pulp contains a population of cells with membrane properties similar to neuronal satellite cells. These cells may contribute, either directly or indirectly, to somatosensation in teeth.

Amyloid beta-peptide (A beta P) potentiates a nimodipine-sensitive L-type barium conductance in N1E-115 neuroblastoma cells.

The neurodegenerative pathology observed in Alzheimer's Disease (AD) has been partially attributed to the neurotoxic effects of the amyloid beta-peptide (A beta P), although the mechanisms underlying this neurotoxicity are unknown. Since A beta P is capable of forming cation channels in lipid bilayers, it is possible that the neurotoxic effects on neurons may be mediated by a cation flux. We have used patch-clamp recording techniques to study the effects of A beta P on cation currents in differentiated mouse N1E-115 neuroblastoma cells. In whole-cell recordings, incubation of cells with A beta P for 24 h significantly increased the median peak inward current from -201.8 pA to -352.0 pA, and shifted the voltage at peak current (Vpeak) and that of current activation (Vact) towards more positive potentials. For untreated cells, median Vpeak was 1.7 mV and Vact was -28.9 mV, vs. 10.5 mV and -24.7 mV in A beta P-treated cells. Incubation with the reverse sequence A beta P(40-1) or A beta P(25-35) did not produce significant changes in the amplitude or kinetic behavior of the inward current. At the single channel level, A beta P added to the pipette increased the open probability of cation-conducting ion channels. As determined by cell viability counts, both A beta P(1-40) and the A beta P(25-35) fragment had neurotoxic effects; within 24 h, addition of A beta P reduced the number of viable cells by more than 50%. It is suggested that the neurotoxic effects of A beta P may be mediated by its ability to form cation channels de novo and/or alter the activity of cation channels already present in the cell membrane.

Temporal changes in myocardial uptake on serial bone scans in two cases of presumed senile cardiac amyloidosis.

Two unusual examples of cardiac uptake in elderly patients who had received serial bone scans, one with a "waxing" and the other with a "waning" pattern of myocardial tracer uptake, are presented. An apparent temporal discordance between echocardiographic and scintigraphic findings is noted. Neither of the two patients had clinical, electrocardiographic, or other laboratory evidence of old or new myocardial infarction. Possible causes for the temporal changes are described. Presumptive diagnoses of senile cardiac amyloidosis were made based on clinical findings, noninvasive imaging, and comparison with a historic cohort at the Veterans Administration Medical Center in Houston.

Potassium currents in cells derived from human dental pulp.

Patch-clamp recording methods were used to monitor ion currents in tissue-cultured cells derived from human dental pulp. Recordings were made in excised, outside-in or whole-cell patches. In single-channel experiments, the majority of patches contained a high-conductance (approx. 140-180 pS) K(+)-selective ion channel. The probability of the channel being in an open state was dependent on membrane potential, internal calcium and negative pressure applied to the cell membrane. Whole-cell recordings were consistent with these findings; in response to step-wise depolarizations of the cell membrane, most displayed a family of outwardly rectifying, barium-sensitive currents. In addition, a number of patches contained a second class of potassium channel of intermediate (approx. 85-100 pS) conductance, which was largely voltage insensitive and independent of calcium concentration. These results suggest that pulp cells contain a high-conductance potassium channel which probably underlies the outwardly rectifying current found at the whole-cell level. Further, the existence of mechanosensitive channels in these cells raises the possibility that the response to mechanical perturbation of dental pulp may be mediated, in part, by direct effects on odontogenic cells.

Membrane stretch activates a high-conductance K+ channel in G292 osteoblastic-like cells.

A high-conductance K(+)-selective ion channel was studied in excised membrane patches from human G292 osteoblast-like osteosarcoma cells. Channel conductance averaged approximately 170 pS in symmetric solutions of 153 mM KCl, and approximately 135 pS when the pipette was filled with standard saline (150 mM NaCl). The probability of the channel being in an open state (Popen) increased with membrane potential, internal calcium, and applied negative pressure. At pCa7, channel activity was observed at membrane potentials greater than approximately 60 mV, while at pCa3, channel activity was seen at approximately 10 mV. Likewise, in the absence of applied pressure, channel openings were rare (Popen = 0.02), whereas with -3 cm Hg applied pressure, Popen increased to approximately 0.40. In each case, i.e., voltage, calcium concentration, and pressure, the increase in Popen resulted from a decrease in the duration of long-closed (interburst) intervals and an increase in the duration of long-open (burst) intervals. Whole-cell responses were consistent with these findings. Hypotonic shock produced an increase in the amplitude and conductance of the outward macroscopic current and a decrease in its rise time, and both single-channel and whole-cell currents were blocked by barium. It is suggested that the voltage-gated, calcium dependent maxi-K+ channel in G292 osteoblastic cells is sensitive to membrane stretch and may be directly involved in osmoregulation of these cells. Further, stretch sensitivity of the maxi-K+ channel in osteotrophic cells may represent an adaptation to stresses associated with mechanical loading of mineralized tissues.