Short communication: Assessment of disbudding pain in dairy calves using nonlinear measures of heart rate variability.

The purpose of this study was to evaluate whether pain-related stress caused by disbudding could be detected using nonlinear measures of heart rate variability (HRV). Twenty-five female Holstein calves (4-7 wk of age) were randomly assigned to 1 of 3 treatments: (1) sham disbud (SHAM; n = 9), (2) disbud with lidocaine-meloxicam pain mitigation (MED; n = 8), or (3) disbud without pain mitigation (NoMED; n = 8). Heart rate variability (sample entropy, percentage determinism, percentage recurrence, or mean length of diagonal lines in a recurrence plot) was recorded on d -1, 0, 1, 3, and 5 relative to the experimental procedure, with disbudding taking place on d 0. The short-term detrended fluctuation analysis scaling exponent was greater in MED calves than in SHAM calves, indicating a greater stress response to the disbudding procedure regardless of pain mitigation. These results indicate that calves in the MED group may have experienced pain-related stress as a result of the disbudding procedure. The remaining nonlinear HRV measures did not differ by treatment. Future research on this topic should address additional potential confounding factors, such as the effect of pain-mitigating drugs on autonomic function or the influence of the autonomic nervous system on wound healing, that may prohibit HRV measurement as an indicator of disbudding pain severity.

Efficacy of Red Cell Salvage Systems in Open Acetabular Surgery.

Over the past 50 years, treatment of displaced acetabular fractures has moved away from conservative treatment with bedrest to operative intervention to achieve anatomic reduction, stable fixation, and allow early range of motion of the hip. However, operative fixation is not without complications. Internal fixation of traumatic acetabular fractures has been coupled with large volume of blood loss both at the time of injury and surgery. This often results in the need for allogenic blood products, which has been linked to increase morbidity (Vamvakas and Blajchman, 2009). In an attempt to avoid the risk associated with allogenic blood transfusion numerous techniques and methods have been devised. Red blood cell salvage (CS) is an intraoperative blood salvage tool where blood is harvested from the operative field. It is washed to remove the plasma, white blood cells, and platelets. The red cells are resuspended in a crystalloid solution. If the hematocrit of the resuspended red blood cells is sufficient, it is transfused to the patient intravenously. The benefits of CS in major spine surgery, bilateral knee replacement, and revision hip surgery are well established (Goulet et al. 1989, Gee et al. 2011, Canan et al. 2013). However, literature reviewing the use of cell saver in orthopedic trauma surgery, specifically acetabular surgery is limited. Our institute performed a retrospective review of 63 consecutive operative acetabular fractures at a level one trauma center. Our study revealed that patients with blood loss of less than 400 mL were 13 times less likely to receive autologous blood, and patients with hemoglobin less than 10.5 were 5 times less likely to receive autologous transfusion (p < 0.05). We also found that no patients with a hemoglobin level less than 10.5 and EBL less than 400 mL received autologous blood return. Autologous blood transfusion had no effect on volume or rate of allogenic blood transfusion. We believed that if a patient's preoperative hemoglobin is less than 10.5 or expected blood loss is less than 400 mL, then CS should have a very limited role, if any, in the preoperative blood conservation strategy. We found ASA greater than 2, BMI greater than 24 and associated fracture type to be a risk factor for high blood loss.

Nonlinear analysis of heart rate variability for evaluating the growing pig stress response to an acute heat episode.

Heart rate variability (HRV) is a proxy measure of autonomic function and can be used as an indicator of swine stress. While traditional linear measures are used to distinguish between stressed and unstressed treatments, inclusion of nonlinear HRV measures that evaluate data structure and organization shows promise for improving HRV interpretation. The objective of this study was to evaluate the inclusion of nonlinear HRV measures in response to an acute heat episode. Twenty 12- to 14-week-old growing pigs were individually housed for 7 days and acclimated to thermoneutral conditions (20.35°C ± 0.01°C; 67.6% ± 0.2% RH) before undergoing one of the two treatments: (1) thermoneutral control (TN; n = 10 pigs) or (2) acute heat stress (HS; n = 10 pigs; 32.6°C ± 0.1°C; 26.2% ± 0.1% RH). In Phase 1 of the experimental procedure (P1; 60 min), pigs underwent a baseline HRV measurement period in thermoneutral conditions before treatment [Phase 2; P2; 60 min once gastrointestinal temperature (Tg) reached 40.6°C], where HS pigs were exposed to heated conditions and TN pigs remained in thermoneutral conditions. After P2, all pigs were moved back to thermoneutral conditions (Phase 3; P3; 60 min). During each phase, Tg data were collected every 5 min and behavioural data were collected to evaluate the amount of time each pig spent in an active posture. Additionally, linear (time and frequency domain) and nonlinear [sample entropy (SampEn), de-trended fluctuation analysis, percentage recurrence, percentage determinism (%DET), mean diagonal line length in a recurrence plot] HRV measures were quantified. Heat stressed pigs exhibited greater Tg (P = 0.002) and spent less time in an active posture compared to TN pigs during P2 (P = 0.0003). Additionally, low frequency to high frequency ratio was greater in HS pigs during P3 compared to TN pigs (P = 0.02). SampEn was reduced in HS pigs during P2 (P = 0.01) and P3 (P = 0.03) compared to TN pigs. Heat stressed pigs exhibited greater %DET during P3 (P = 0.03) and tended to have greater %DET (P = 0.09) during P2 than TN pigs. No differences between treatments were detected for the remaining HRV measures. In conclusion, linear HRV measures were largely unchanged during P2. However, changes to SampEn and %DET suggest increased heat stress as a result of the acute heat episode. Future work should continue to evaluate the benefits of including nonlinear HRV measures in HRV analysis of swine heat stress.

Determining the effects of early gestation in utero heat stress on postnatal fasting heat production and circulating biomarkers associated with metabolism in growing pigs.

The study objective was to characterize effects of early gestation in utero heat stress (IUHS) on postnatal fasting heat production (FHP) and blood biomarkers associated with metabolism in growing pigs. Based on previous observation of increased postnatal core body temperature set point in IUHS pigs, we hypothesized that FHP would be altered during postnatal life because of IUHS. Pregnant first-parity gilts were exposed to thermoneutral (TN; = 4; 17.8 ± 0.1°C) or heat stress (HS; = 4; cyclical 28 to 38°C) conditions from d 30 to 60 of gestation. At weaning (21 d of age), 2 median-weight male pigs (1 barrow and 1 boar) were selected from each litter ( = 8 in utero TN [IUTN] and 8 IUHS pigs) and then housed in TN conditions based on age. Blood samples were collected at 8, 9, and 10 wk of age when pigs were in a fed state to analyze thyroxine (T4) and triiodothyronine (T3) concentrations. Pigs were trained to enter an indirect calorimeter from wk 8 through 10 of life and then acclimated over a 24-h period 1 wk prior to testing. At 12 wk of age, pigs were fasted for 24 h, and then indirect calorimetry was performed on individual pigs over a 23-h testing period to determine FHP and the respiratory quotient in 3 intervals (0900 to 1700 h, 1700 to 0000 h, and 0000 to 0800 h). Body weight was determined before and after testing and was similar for all pigs ( = 0.77; 37.0 ± 0.5 kg BW). Data were analyzed using PROC MIXED in SAS 9.4. No boar vs. barrow differences were observed with any analysis. Overall, FHP per kilogram BW was greater ( = 0.03; 12.1%) in IUHS pigs compared with IUTN pigs. Fasting heat production per kilogram BW was greater ( < 0.01; 19.8%) from 0900 to 1700 h compared with 1700 to 0000 h and 0000 to 0800 h and was greater (10.9%) from 1700 to 0000 h compared with 0000 to 0800 h. The RQ did not differ by in utero treatment ( = 0.51; 0.72 ± 0.01); however, the RQ was increased ( < 0.01; 13.0%) from 1700 to 0000 h compared with 0900 to 1700 h and 0000 to 0800 h. No other FHP and RQ differences were detected. Although no in utero treatment differences were observed for T4 ( = 0.11; 52.2 ± 6.2 ng/mL), T3 was greater overall ( = 0.04; 19.5%) in IUHS pigs than in IUTN pigs. In summary, FHP and circulating T3 were increased in IUHS pigs, and this may have implications for postnatal production efficiency in pigs gestated during hot summer months.

Evaluating Pekin duck walking ability using a treadmill performance test.

Gait scoring is the most popular method for assessing the walking ability of poultry species. Although inexpensive and easy to implement, gait scoring systems are often criticized for being subjective. Using a treadmill performance test we assessed whether observable differences in Pekin duck walking ability identified using a gait scoring system translated to differences in walking performance. One hundred and eighty ducks were selected using a three-category gait scoring system (GS0 = smooth gait, n = 55; GS0.5 = labored walk without easily identifiable impediment, n = 56; GS1 = obvious impediment, n = 59) and the amount of time each duck was able to sustain walking on a treadmill at a speed of 0.31 m/s was evaluated. The walking test ended when each duck met one of three elimination criteria: (1) The duck walked for a maximum time of ten minutes, (2) the duck required support from the observer's hand for more than three seconds in order to continue walking on the treadmill, or (3) the duck sat down on the treadmill and made no attempt to stand despite receiving assistance from the observer. Data were analyzed in SAS 9.4 using PROC GLM. Tukey's multiple comparison test was used to compare differences in time spent walking between gait scores. Significant differences were found between all gait scores (P < 0.05). Behavioral correlates of walking performance were investigated. Video recorded during the treadmill test was analyzed for counts of sitting, standing, and leaning behaviors. Data were analyzed in SAS 9.4 using a negative binomial model for count data. No differences were found between gait scores for counts of sitting, standing, and leaning behaviors (P > 0.05). In conclusion, the amount of time spent walking on the treadmill corresponded to gait score and was an effective measurement for quantifying Pekin duck walking ability. The test could be a valuable tool for assessing the development of walking issues or the effectiveness of treatments aimed at promoting leg health.

Initial experience with larger laser sheaths for the removal of transvenous pacemaker and implantable defibrillator leads.

BACKGROUND: In a previous randomized trial, the 12F laser sheath removed pacing leads via the implant vein more successfully than traditional mechanical tools alone. Two larger sizes of laser sheath, the 14F and 16F models, were developed to extract defibrillator leads and large-diameter pacing leads implanted for the chronic. These devices use pulsed ultraviolet laser light to core though fibrotic tissue grown over the lead body to free the lead from the vasculature. A mandatory prospective registry studied the safety and effectiveness profiles of the larger laser sheaths vis-à-vis the 12F laser sheath. METHODS AND RESULTS: In this study, 863 patients underwent extraction of 1285 leads at 52 sites. Patients treated with the 14F device tended to have older leads than the 12F population; the 16F population, which comprised mostly defibrillator patients, were younger, had younger leads, and were more often male than the 12F population. Clinical success (extracting the entire lead or the lead body minus the distal electrode) was observed in 91% to 92% of cases for all device sizes. The overall complication rate was 3.6%, with 0.8% perioperative mortality. Incidence of complications was independent of laser sheath size. CONCLUSIONS: The 14F and 16F laser sheaths offer an extraction option for larger long-term transvenous pacemaker and defibrillator leads that is as safe and effective as the 12F laser sheath.

Pacemaker lead extraction with the laser sheath: results of the pacing lead extraction with the excimer sheath (PLEXES) trial.

OBJECTIVES: The purpose of this study was to evaluate the safety and effectiveness of pacemaker lead extraction with the excimer sheath in comparison to nonlaser lead extraction. BACKGROUND: Fibrotic attachments that develop between chronically implanted pacemaker leads and to the venous, valvular and cardiac structures are the major obstacles to safe and consistent lead extraction. Locking stylets and telescoping sheaths produce a technically demanding but effective technique of mechanically disrupting the fibrosis. However, ultraviolet excimer laser light dissolves instead of tearing the tissue attachments. METHODS: A randomized trial of lead extraction was conducted in 301 patients with 465 chronically implanted pacemaker leads. The laser group patients had the leads removed with identical tools as the nonlaser group with the exception that the inner telescoping sheath was replaced with the 12-F excimer laser sheath. Success for both groups was defined as complete lead removal with the randomized therapy without complications. RESULTS: Complete lead removal rate was 94% in the laser group and 64% in the nonlaser group (p = 0.001). Failed nonlaser extraction was completed with the laser tools 88% of the time. The mean time to achieve a successful lead extraction was significantly reduced for patients randomized to the laser tools, 10.1 +/- 11.5 min compared with 12.9 +/- 19.2 min for patients randomized to nonlaser techniques (p < 0.04). Potentially life-threatening complications occurred in none of the nonlaser and three of the laser patients, including one death (p = NS). CONCLUSIONS: Laser-assisted pacemaker lead extraction has significant clinical advantages over extraction without laser tools and is associated with significant risks.

Effect of abx, bx and pbx mutations on expression of homeotic genes in Drosophila larvae.

We have examined the patterns of expression of the homeotic gene Ubx in imaginal discs of Drosophila larvae carrying mutations in the abx, bx and pbx regulatory domains. In haltere discs, all five bx insertion mutations examined led to a general reduction in Ubx expression in the anterior compartment; for a given allele, the strength of the adult cuticle phenotype correlated with the degree of Ubx reduction. Deletions mapping near or overlapping the sites of bx insertions, including three abx alleles and the bx34e-prv(bx-prv) allele, showed greatly reduced Ubx expression in parts of the anterior compartment of the haltere disc; however, anterior patches of strong Ubx expression often remained, in highly variable patterns. As expected, the pbx1 mutation led to reduced Ubx expression in the posterior compartment of the haltere disc; surprisingly, pbx1 also led to altered expression of the en protein near the compartment border in the central region of the disc. In the metathoracic leg, all the bx alleles caused extreme reduction in Ubx expression in the anterior regions, with no allele-specific differences. In contrast, abx and bx-prv alleles resulted in patchy anterior reductions in third leg discs. In the larval central nervous system, abx but not bx alleles affected Ubx expression; the bx-prv deletion gave a wild-type phenotype, but it could not fully complement abx mutations. In the posterior wing disc, the bx-prv allele, and to a much lesser extent the bx34e chromosome from which it arose, led to ectopic expression of Ubx. Unlike other grain-of-function mutations in the BX-C, this phenotype appeared to be partially recessive to wild type. Finally, we asked whether the ppx transformation, which results from early lack of Ubx+ function in the mesothorax and is seen in abx animals, is due to ectopic Scr expression. Some mesothoracic leg and wing discs from abx2 larvae displayed ectopic expression of Scr, which was variable in extent but always confined to the posterior compartment.

Development of the olfactory bulb in the clawed frog, Xenopus laevis: a morphological and quantitative analysis.

The relationship between olfactory axons and the cells of the olfactory bulb during normal development was analyzed to determine whether olfactory afferent axons could play a role in the induction of olfactory bulb formation. The morphology of the olfactory bulb in Xenopus larvae from stages 26 to 58 and in adult frogs was analyzed with light and electron microscopy. Axons were first observed beneath the basal lamina of the neural tube at stages 30 and 32; at stage 32, neurons in this area of the neural tube began to differentiate. Synapses of olfactory axons onto young neuronal processes were observed as early as stages 36 and 38. By stage 44, all of the layers of the olfactory bulb were present. The basic structure of the mature form of the olfactory bulb was apparent as early as stage 48/49 and remained constant throughout late larval life and even into adulthood, with only the size increasing. To determine the numerical relationship between olfactory axons from both main and vomeronasal epithelia and mitral/tufted cells in the main and accessory olfactory bulbs, a quantitative study was also performed in which the number of olfactory axons and the number of mitral/tufted cells were estimated for larval stages (stages 50 to 58) and adults. The number of axons increased with stage, with a 16-fold increase between stage 58 and adulthood. The number of mitral/tufted cells increased with stage, with only a 2.3-fold increase between stage 58 and adults. There is a correlation between axon number and mitral/tufted cell number during larval development that is consistent with the hypothesis that olfactory axons influence olfactory bulb development. The convergence ratio of olfactory axons onto mitral/tufted cells was 5:1 in larvae and increased to 34:1 in adults; this increase probably results in increased olfactory sensitivity in adult frogs.

Morphological and quantitative evaluation of olfactory bulb development in Xenopus after olfactory placode transplantation.

We found previously that the number of olfactory axons is correlated with the number of mitral/tufted cells (output neurons of the olfactory bulb) during normal larval development. To examine the significance of this quantitative relationship, we evaluated the effects of transplanting an extra olfactory placode on the development of the larval olfactory bulb. We found that the transplanted tissue retained the normal, pseudostratified, columnar appearance and had the same cell types as normal olfactory epithelium, and the olfactory bulbs had the same laminar organization as control bulbs. With gross examination of the olfactory bulb, the side innervated by the transplant appeared slightly larger than the contralateral side in animals analyzed at a young larval stage (stage 50) and in 2 of the 9 animals examined at late larval stages (57/58). Tissue sections and area measurements, however, revealed that the volume of the olfactory bulbs in animals with a transplant was not significantly different from control values. Our quantitative analysis also showed that in stage-50 animals with a transplant, the total number of olfactory axons (in nerves from the transplanted and host olfactory organs) appeared to be greater than in control animals, but not to a statistically significant level. The number of mitral/tufted cells was not different from controls. In animals examined at stage 57/58, there was no difference from controls in either the total number of olfactory axons, total number of mitral/tufted cells, or convergence ratio of olfactory axons onto mitral/tufted cells. Thus, in the late-stage larvae, the quantitative relationship between olfactory axons and mitral/tufted cells was not altered by the experimental manipulation. These results suggest that the olfactory bulb can regulate the number of afferent fibers.

Organization of the olfactory system in the adult zebrafish: histological, immunohistochemical, and quantitative analysis.

The zebrafish, Danio rerio, is becoming an important model system for developmental studies. We have used a variety of histological techniques to characterize the adult structure of the olfactory system in this teleost to form a base for future developmental work. The olfactory epithelium in this fish contains ciliated and microvillar sensory neurons, microvillar supporting cells, secretory goblet cells, and basal cells, and the adjacent nonsensory epithelium contains ciliated supporting cells. The olfactory bulb is a diffusely organized structure with four laminae: olfactory nerve, glomerular, mixed mitral cell/plexiform, and granule cell layers. These structures and the synapses observed in the olfactory bulb are typical of what is found in other vertebrates. We also examined the distribution of several neurotransmitter markers (tyrosine hydroxylase, neuropeptide Y, dopamine-beta-hydroxylase, and serotonin) in the olfactory bulb. Antibodies to neuropeptide Y, dopamine-beta-hydroxylase, and serotonin labeled fibers in the olfactory bulb and cell bodies in caudal regions of the brain in distributions comparable to other species. Tyrosine hydroxylase immunoreactivity was observed in a set of intrinsic bulb neurons with extensive processes in the glomerular layer. In addition, the structural proteins glial fibrillary acidic protein and vimentin have distributions similar to those in the olfactory bulbs of other animals. Thus, the adult olfactory structures are analogous to the structures in other vertebrate animals in morphology and chemical neuroanatomy. This similarity, along with its numerous advantages for developmental studies, makes the zebrafish a good model for studies of olfaction and forebrain maturation.

Extra-adrenal myelolipomas compared with extramedullary hematopoietic tumors: a case of presacral myelolipoma.

A large presacral myelolipoma associated with increasing constipation is reported in a 70-year-old obese diabetic female. The resected tumor was encapsulated and dark red to yellow. Light and electron microscopy revealed mature and immature hematopoietic tissue, lymphoid aggregates and mature adipocytes. Myelolipomas arising in extra-adrenal sites are rare. Considered in the differential diagnosis are reactive extramedullary hematopoietic tumors that usually occur in patients with severe chronic anemias. Patients with extramedullary hematopoietic tumors are characterized by anemia, frequent hepatosplenomegaly and abnormal peripheral blood smears, and may be any age. The tumors are usually multiple and usually located in the mediastinum or epidural space. In contrast, patients with extra-adrenal myelolipomas are usually older than 40 years, have normal blood studies, absent hepatosplenomegaly, and usually have chronic debilitating diseases or endocrinopathies. Extra-adrenal myelolipomas are single and usually occur within the abdomen. Extramedullary hematopoietic tumors have a predominance of the hematopoietic component, with erythroid hyperplasia of that component. Extra-adrenal myelolipomas may have a predominance of either the hematopoietic or fatty component, chiefly the latter, and generally have a more conspicuous lymphocyte population. Although the components of these tumors are the same, we consider them separable clinically, pathogenetically and, in many cases, pathologically.

Neurogenesis in the olfactory bulb of adult zebrafish.

Cell genesis in the adult brain of zebrafish, with specific reference to the olfactory bulbs, was examined using bromodeoxyuridine immunocytochemistry. Mature fish were exposed to a 1% solution of the thymidine analog 5-bromo-2'-deoxyuridine for 1 h and then killed after short (4-h) or long (3-4-week) survival periods. A monoclonal antibody to bromodeoxyuridine allowed visualization of cells that incorporated the drug during the S phase of mitosis. Four hours after administration of the drug, antibody-labeled cells were found almost exclusively in the proliferative zones around the ventricles and in the cerebellum. Very few labeled nuclei were seen in other locations in the brain, indicating that cell genesis occurs in discrete regions in adults. The few labeled profiles in the olfactory bulbs were located in the olfactory nerve layer; these profiles had the morphology of glial nuclei and did not stain with a neuronal marker, the Hu antibody. After longer survival times, labeled cells were present throughout the layers of the olfactory bulb, and many of the immunoreactive profiles in the internal cell layer were also labeled with the Hu antibody, indicating that they are likely adult-formed interneurons. Thus, neurogenesis continues in the olfactory bulb of adult zebrafish. Understanding the process of the generation of new neurons in the brain of adult animals can lead to important insights into neural regeneration and adult plasticity.

Factors responsible for immunizations referrals to health departments in North Carolina.

BACKGROUND: Despite the existence of Medicaid and other programs designed to eliminate cost as a barrier to immunization in physicians' offices, referrals to local health departments for immunizations are common. Many children leave their physicians' offices without receiving needed immunizations. PURPOSE: To determine: 1) the frequency and determinants of immunization referrals to health departments in North Carolina, and 2) the factors associated with private physicians' decisions to immunize Medicaid children in their offices and participate in the state-funded vaccine replacement program. METHODS: The 2537 pediatricians and family physicians licensed in North Carolina were surveyed by mail using a 23-item, self-administered questionnaire. RESULTS: Seventy-two percent of physicians responded; 93% referred at least some children to local health departments for immunizations. Concern regarding parents' ability to pay for immunizations was the most important reason for referral for 93% of respondents. Forty percent referred all or some of their Medicaid patients; excessive paperwork, inadequate reimbursement, and parental preferences were the most common reasons. Only 33% of physicians had participated in the state's vaccine replacement program. Family physicians, and physicians in solo or two-physician practices in rural counties, and in practices caring for a small number of children on Medicaid were most likely to refer children covered by Medicaid and not participate in the state's existing vaccine replacement program. CONCLUSIONS: Medicaid and North Carolina's vaccine replacement program are not preventing large numbers of immunization referrals to health departments. Future programs designed to increase the proportion of children immunized in physicians' offices will not succeed if more effective incentives for physician participation are not developed.

Intravascular techniques for extraction of permanent pacemaker leads.

Intravascular techniques were used to extract 226 leads from 124 patients. Indications for lead extraction were life-threatening septicemia (30%), complications of free-floating leads (2%), abandonment of pockets (40%), and replacement of malfunctioning leads (28%). Extraction tools included flexible, telescoping sheaths advanced over the lead to dilate scar tissue and apply countertraction, deflection catheters, and wire basket snares. Countertraction is defined as the direct force of traction on the lead countered by the circumference of an extraction sheath. One hundred sixty-four leads were extracted through the superior vena cava by advancing the sheaths over the lead to the myocardial wall. Most of these leads passed through the subclavian vein. An approach through the inferior vena cava was used for the remaining 62 leads. The countertraction sheaths were passed from the femoral vein into the right atrium. A maneuvering catheter and an extracting snare were placed inside the sheaths. The lead was positioned by the maneuvering catheter, entangled in the extracting snare, and the sheaths advanced over the snare and lead to the myocardium. An atriotomy by means of a limited surgical approach was required to free one lead. One infected lead broke 5 cm from the electrode and was removed through a median sternotomy, ventriculotomy, and retrograde extraction. All patients had unremarkable recoveries. Intravascular countertraction techniques proved to be a viable alternative, minimizing the risks and morbidity of lead removal.

Technique for the surgical extraction of permanent pacing leads and electrodes.

When a pacing lead becomes infected, the most expedient and successful therapy is its removal. A new surgical extraction technique, combining the use of fluoroscopy and a limited surgical dissection of the right atrium, is presented for the removal of a lead entrapped in the heart or venous system.

Addition of new cells to the olfactory bulb of adult zebrafish.

We have been examining patterns of cell proliferation in the brain of adult zebrafish. Understanding this process in fish may lead to important insights due to the tremendous regenerative capabilities of these animals. Fish were exposed to a 1% solution of the thymidine analog 5-bromo-2'-deoxyuridine (BrdU) for 1 hr before being returned to small aquaria that received frequent water changes. Animals were overanesthetized and perfused with Bouin's fixative solution after two survival periods (4 hr or 3-4 weeks). Paraffin immunohistochemistry using a monoclonal antibody against BrdU was used to visualize the newly generated cells. Quantitative analyses were performed on serial, 10-micron sections from 4 animals for each survival group. Statistical determinations were based on the nonparametric Mann-Whitney U test. The average number of BrdU-labeled nuclear profiles in the bulbs, from analysis of every 5th section, was significantly different between the two groups (22.0 +/- 6.8 [SEM] for the 4-hr group and 136.7 +/- 11.3 for the 4-wk group; p < 0.05). The volumes of the bulbs, however, were not different between the two groups (p > 0.5). These data indicate that either cells divided repeatedly during the longer survival period or cells migrated into the bulb from other brain regions. To examine this phenomenon further, the location of the new cells was analyzed in three mid-bulb sections (20 microns apart) from each animal. Both the area and number of labeled nuclei in each lamina were measured to obtain an average profile density. Comparison of the 4-hr and the 4-wk groups showed that density was significantly greater in all bulb layers in the long survival group (p < 0.05 for all). In the 4-hr survival group, cells were found mainly in the olfactory nerve layer. When examined after 4 wk, proportionately more labeled cells were found in the internal cell layer. This addition of new cells could be a result of neurogenesis, gliogenesis, and/or angiogenesis. We are currently performing double-labeling experiments to determine the types of cells that are added to the adult bulb. In addition, our future plans include investigating the origin of these cells and the signals that direct their formation.

Long-term results of "simple" thrombectomy for thrombosed Björk-Shiley aortic valve prostheses.

During the past two years 8 patients were seen with thrombosis of their Bjork-Shiley aortic valves. Six patients were from our series, an incidence of 4% and 2 patients had their original valve implanted at another institution. All patients had substantial problems with anticoagulation therapy. Three died prior to operation. Early detection and emergency surgical intervention is mandatory for survival. At operation debridement of all thrombotic material from the valve provided satisfactory immediate hemodynamic improvements as well as freedom from complications for up to two years. Therapeutic levels of anticoagulation with warfarin are the only apparent protection from thrombosis of the Bjork-Shiley aortic valve.

Noninvasive cardiac evaluation (NICE) of intracardiac shunts using inhaled radioactive carbon dioxide.

A totally new, noninvasive technique is presented for the detection and analysis of left-to-right cardiac shunts. Five millicuries of oxygen 15-labeled carbon dioxide is administered by a single-breath inhalation technique. The resultant lung washout curve accurately reflects the presence or absence of a shunt. The quantitative studies are promising, but the method must be compared with standard techniques in a statistically significant series. This work is in progress.

Deafferentation-induced changes in the olfactory bulb of adult zebrafish.

The influence of the olfactory organ on maintenance of olfactory bulb structure was examined in zebrafish, using peripheral deafferentation. This fish provides a model in which the olfactory organ is easily accessible for removal, the animals easily survive the surgery, and the olfactory bulbs are small enough to allow rigorous analysis of the resulting effects. Unilateral olfactory organ ablations were performed on anesthetized adult zebrafish using a small-vessel cautery iron. Fish were allowed to survive for 1, 3, or 6 weeks following the procedure. Analysis of deafferented animals revealed that most, if not all, of the olfactory organ was missing on the ablated side, and the structure did not regenerate. The morphology of the olfactory bulb was affected notably by the removal of its primary afferent innervation. The olfactory nerve layer was diminished at 1 week and absent by 3 weeks post-deafferentation. At all of the survival times the deafferented bulb appeared significantly smaller at the gross level, and there was a statistically significant effect on bulb size and cell number after 6 weeks. Tyrosine hydroxylase expression, as revealed by immunohistochemistry, was decreased noticeably on the ablated side. In conclusion, the olfactory organ is important in the preservation of normal olfactory bulb anatomy and neurochemistry in adult zebrafish. Thus, the influence of the periphery does not end with the formation of the mature olfactory bulb.