Fisher's PLSD Research Articles

REGENERATIVE SIGNALS FOR INTESTINAL EPITHELIAL ORGANOID UNITS TRANSPLANTED ON SYNTHETIC BIODEGRADABLE POLYMER SCAFFOLDS FOR THE TISSUE ENGINEERING OF SMALL INTESTINE

101 Our laboratory is investigating the tissue-engineering of small intestine using intestinal epithelial organoid units seeded on biodegradable polymer tubes. Previous studies have demonstrated the formation of cysts with tissue morphology resembling small intestine. This study investigates methods of stimulation for optimizing neointestinal regeneration. Intestinal epithelial organoid units (4-6 × 104 units) harvested from neonatal Lewis rats were seeded onto synthetic biodegradable polymer tubes (n=34) created from nonwoven sheets of PGA fibers sprayed on the outer surface with 5% PLA. These organoid unit/polymer constructs were implanted into the omentum of adult male Lewis rats (175-250 grams) in the following conditions: 1). C: control, implantation alone (n=9); 2). SBR: after 75% small bowel resection (n=9); 3). PCS: after portacaval shunt (n=8); and 4). PH: after 75% partial hepatectomy (n=8). Ultrasonography was performed at 3, 7, and 10 weeks to document neointestinal cyst size and growth. The constructs were harvested at 10 weeks and examined by histology. Morphometric analysis of the neomucosa including villus number, villus height, mucosal surface area, crypt number, and crypt area was obtained using a video-microscope integrated computer program (NIH Image, version 1.59) by a blinded observer. Cyst development was noted in all animals. Ultrasonography demonstrated no significant differences in cyst size at 3 weeks. As shown in the table, at 7 and 10 weeks, cyst lengths and diameters were significantly greater in the SBR group compared to the other groups (*P<0.05, ANOVA, Fisher's PLSD). Histology revealed a neomucosa lining the lumen of the cyst with invaginations resembling crypt-villus structures. Morphometric analysis demonstrated a significantly greater villus number (18±2), villus height (133.9±22.6µm), and mucosal surface area (8533.3±977.6µm2) in the SBR group compared to the other groups (C: 11.4±3.6, 87.1±7.8µm, 5998.4±416.5 µm2; PCS: 12.3±1.8, 89.9±10.3µm, 6131.6±548.4 µm2; PH: 12±4, 97.3±9.6µm, 6000.1±777.1 µm2; P<0.01, ANOVA, Fisher's PLSD), and a significantly greater crypt number (23.3±2.2) and crypt area (68127.3±12431.5µm2) in the PCS group compared to control (14.6±5.7; 40150.8±16388.7µm2; P<0.01, ANOVA, Fisher's PLSD).TableIntestinal epithelial organoid units transplanted on biodegradable polymer tubes can successfully survive and regenerate into complex tissue resembling small intestine. Small bowel resection, and to a lesser extent, portacaval shunt, contribute significant regenerative stimuli for the morphogenesis and differentiation of tissue-engineered small intestine.

In vitro secondary caries inhibition around fluoride releasing materials

Objectives of the study were to compare the capacity of fluoride releasing materials to inhibit in vitro caries formation and to measure the width and height of the inhibition zones. Box-shaped cavities were prepared on bovine root dentine and restored with Fuji II, Fuji II LC, Vitremer, or Clearfil Liner Bond II system with a fluoride releasing composite. After immersion in a buffered demineralizing solution of 50 mmol/L acetic acid adjusted to pH 4.5 for 3 days, longitudinal sections were cut and imbibed in quinoline for analysis under a polarized light microscope (PLM). The contours of the lesions and inhibition zones were traced and their depth, width and height calculated. The data were analyzed by one-way ANOVA and Fisher's PLSD test at 95% level of confidence. The polarized light microscopy results showed a distinct inhibition zone adjacent to both conventional and resin-modified glass ionomer cements; however, it was not observed around the adhesive resin system. The width and height of this inhibition zone were significantly greater for Fuji II than for Fuji II LC and Vitremer. An inhibition zone was not observed adjacent to the adhesive resin system with fluoride releasing composite. The resin-modified glass ionomer cements produced less protection against in vitro secondary caries formation than the conventional glass ionomer cement. The recent bonding system that releases fluoride failed to produce an inhibition zone along the cavity wall adjacent to the restoration.

Microhardness of in vitro caries inhibition zone adjacent to conventional and resin-modified glass ionomer cements

Objective. This study was conducted to correlate Knoop and triangular hardness numbers by measuring the microhardness of in vitro caries-inhibited and -demineralized dentin adjacent to a conventional and two resin-modified glass ionomer cements. Methods. Box-shaped cavities were prepared on bovine root dentin and restored with either Fuji II, Fuji II LC, or Vitremer. The teeth were then decalcified in an acid buffered solution of 50 mmol l −1 acetic acid adjusted to pH=4.5 for 3 days. Knoop and triangular microhardness indentations were performed perpendicular to the surface and parallel to the cavity wall, in the demineralized lesion and inhibition zone. Calcium and phosphorous contents of the outer lesions and inhibition zones were compared using energy-dispersive X-ray spectrometry (EDS). The correlation between Knoop and triangular hardness was analyzed by correlation coefficient. The statistical significance of hardness data was analyzed by one-way ANOVA and Fisher's PLSD test ( p<0.05). Results. Triangular hardness (HT) correlated well with Knoop hardness number (KHN) ( r 2=0.81, p<0.05). The microhardness of the inhibition zone created by Fuji II was of 59.2±3.8 HT and was statistically significantly higher than the zone produced by Fuji II LC and Vitremer. Fuji II LC and Vitremer produced inhibition zones with similar microhardness [48.3±3.5 HT and 44.0±7.6 HT, respectively ( p>0.05)]. Calcium and Phosphorous were present in the inhibition zone, but did not exist in the demineralized lesion. Significance. Knoop and triangular hardness numbers correlated significantly ( p<0.05), and the latter seems to be a promising alternative method for measuring very narrow surfaces. Despite the fact that all glass ionomer materials used in this study were effective in producing an acid-resistant layer, microhardness and intensity of these layers were material dependent.

Effects of macrophage migration inhibitory factor and macrophage migration stimulatory factor on function and survival of foetal dopaminergic grafts in the 6-hydroxydopamine rat model of Parkinson's disease.

Activated microglia play an important role in the rejection of intracerebral grafts and the degeneration of axotomized neurones. We studied the effect of macrophage migration stimulatory factor (MSF) or macrophage migration inhibitory factor (MIF) on allogeneic foetal mesencephalic dopaminergic grafts transplanted into the striatum of 6-hydroxydopamine-lesioned rats. Rotation testing revealed a significant compensation of lesion-induced motor asymmetry 3 weeks post-grafting in animals treated with MIF and vehicle-treated controls compared with pre-graft values (Student's t-test, P < or = 0.005) and MSF-treated animals (ANOVA, post hoc Fisher PLSD test, P < or = 0.05). The MSF group showed no significant compensation. Graft recipients with MIF application (1452.06 +/- 164.32 tyrosine hydroxylase-positive ventral mesencephalic cells) and controls (1753.21 +/- 165.51 tyrosine hydroxylase-positive neurones) displayed good graft survival. Animals with MSF application showed a significant reduction of tyrosine hydroxylase-positive grafted cells (MSF 570.36 +/- 209.49 cells) and graft volumes compared with the MIF and the control group (ANOVA, post hoc Fisher PLSD test, P < or = 0.05). The proportional area of microglia was significantly reduced in MIF animals compared with control animals (ANOVA, post hoc Fisher PLSD test, P < or = 0.001). Activated microglia and macrophages were reduced by half in the MIF-treated group compared with MSF animals and controls. We conclude that intrastriatal injections of MSF result in impaired function and survival of allogeneic ventral mesencephalon (VM) grafts 3 weeks after transplantation. MIF can reduce the number of microglia and macrophages in allogeneic foetal VM grafts. A reduction of microglia via MIF application did not enhance graft function and survival.

HYPOVOLEMIA CAUSES DIFFERENT EFFECTS ON HEART RATE AND BLOOD PRESSURE VARIABILITIES IN DOGS

S134 Spectral analysis of heart rate variability (HRV) and blood pressure variability (BPV) resulted in a characteristic power spectrum with two main regions: a low frequency peak (<0.15Hz) and a high frequency peak centered at the respiratory frequency. The shape of HRV and BPV power spectra may be markedly altered by hypovolemia, since hypovolemia modifies autonomic nervous control of the cardiovascular system. The aim of the present study was to evaluate the effects of hypovolemia on HRV and BPV simultaneously in dogs. METHODS: Seven dogs weighing 13-16kg were anesthetized with thiamylal(10mg/kg), maintained with alpha-chloralose(15mg/kg/hr) and vecuronium(0.1mg/kg/hr) and mechanically ventilated with room air at 14 breaths/min. Catheters were placed in the femoral artery for measurement of arterial blood pressure and in the left femoral artery for removing blood. Splenectomy was done. We induced hypovolemia by removing blood in graded stages (0%, 10%, 20%, 30%, 40% of the estimated blood volume; EBV). After blood pressure was stabilized in each stage, heart rate and mean blood pressure were continuously recorded for 10 min. The areas of the spectral peaks within each measurement were calculated as follows: LF (0.04-0.15 Hz) and HF (0.15-0.4 Hz). The data were analyzed using repeated measurement of ANOVA followed by Fisher PLSD test. P < 0.05 was considered statistically significant. RESULTS: HR did not change during hypovolemia while mean BP decreased significantly after 30% of EBV hemorrhage. LF of HRV significantly increased after 20% of EBV hemorrhage, but HF of HRV did not change. Both LF and HF of BPV increased significantly in proportion to cumulative withdrawal of the blood. DISCUSSION: LF of HRV and BPV during hypovolemia increased and HF of HRV did not changed, indicating the shift of the autonomic balance towards sympathetic dominant. An excellent quantitative correlation between the HF and LF of BPV and the degree of hypovolemia was shown during graded hemorrhage. Even small degree of hypovolemia was detected by changes in LFs of HRV and BPV and HF of BPV. HF of HRV was not useful to detect hypovolemia. Our study suggests that the spectral analysis of HRV and BPV during graded hemorrhage shows different characteristics in the quantitative evaluations of autonomic nervous system during hypovolemia. (Table 1)Table 1: Power spectrum of HRV and BPV during graded hemorrhage

Influence of weight and gonadal status on total and regional bone mineral content and on weight-bearing and non-weight-bearing bones, measured by dual-energy X-ray absosorptiometry

Objective: To evaluate the influence of weight on total body bone mineral content (BMC TB) and regional body bone mineral content (head, arms, trunk and legs). This was studied in accordance with gonadal status and the weight-bearing or non-weight-bearing status of each region. Methods: The study included 94 postmenopausal women (mean age 60.6 ± 10.5 years), 36 perimenopausal women (mean age 49.0 ± 2.3 years) and 60 premenopausal women (mean age 36.1 ± 6.9 years). Full-body bone densitometry (DXA), for measuring total body bone and regional bone mineral content, was carried out in all the women. Results: Among these groups, the influence of 1 kg of body weight on total and regional bone mineral content (percent) did not differ (paired test P ns). In the overall group of women, paired comparison showed differences between the head and other zones measured ( P=0.036 – 0.004). In the overall group of women, no differences were found in the percent influence of 1 kg body weight on bone mineral content in any study zone (by ANOVA, Fisher's PLSD post hoc test and the Kruskal-Wallis test). In the overall group of women, Fisher's r to z test revealed a non-significant relationship between weight and the bone mineral content of the head ( r = 0.49, P ns) but in every other region the relationship between weight and the bone mineral content was significant ( r = 0.36 – 0.54, P < 0.0001 in all). Conclusions: The effect of body weight on BMC TB and regional did not differ significantly with either gonadal status or weight-bearing (legs) and non-weight-bearing bones (arms).

Diet Knowledge and Actual Consumption of Food By Individuals Who Participated in Diet/Health Survey of CSFII-1989.

LEARNING OUTCOME:Will be able to compare the adequacy of diets of U. S. population as related to their diet knowledge level and age. The CSFII-1989 data on 1,505 individuals, who responded to the Diet/Health Knowledge Survey and had 2-day food intakes, were used. The mean age was 43.7 yrs (20–90 yrs); 51% were female, and 80% were Caucasians and 12.5% African-Americans. Weighted sample means were analyzed by ANOVA with Fisher's PLSD test. Ages were grouped into Young (20–24 yrs; n=313), Young-adult (25–50 yrs; n=598), Older-adult (51–64 yrs; n=272) and Elder (65–90 yrs; n=322). Mean intakes for all nutrients, except energy, Ca, Mg, Zn, and vit. B6 & E, exceeded the RDA. Correct response rates on the recommended daily servings for all 5 food groups were highest for Young and lowest for Elder. The correct response ranged 3–13% for grains, 38–70% for vegetables, 45–68% for meats, 60–87% for milk/cheese, and 78–98% for fruits. Young group had better knowledge on the recommended servings, but consumed only 35%, 56%, 65% and 81% of recommended fruits, vegetables, grains and meats, respectively, the lowest among all age groups; they exceeded only in intake of milk/cheese, at the level of 71%. Young-adult consumed most for meats and grains, 114% and 81% of recommendation; for vegetables, Older-adult consumed most at 92% of recommended amount. For each food group, those with a correct response consumed more servings (p<0.0001). Within each age group, servings consumed for milk/cheese, fruits and vegetables by those with correct answers were consistently greater than their counterparts with incorrect responses. In summary, the present study revealed that the US public was least informed on recommended daily servings of grains; Young group had most knowledge on recommended food servings but their compliance with the guidelines was least; and, within each age group, food servings consumed by those with correct responses were closer to the guidelines than for those with incorrect responses. This clearly demonstrates the necessity of nutrition information in improving diets by general public, and further points out that Young age group requires more focused dietary guidance.

Serotonin as a useful parameter for cold and warm ischemic injury in small bowel transplantation.

We investigated serotonin as a parameter of cold and warm ischemic injury prior to transplantation. Lewis rats were used as both donors and recipients, and the proximal 20 cm of jejunum served as the graft. The grafts were preserved in 4 degrees C lactated Ringer's solution for 0, 6, 12, 18, and 24 hr after harvest for cold ischemia (n=7/group). The superior mesenteric artery was clamped for 0, 15, 30, 60, and 120 min before harvest for warm ischemia (n=7/group). The serotonin concentration was measured in the luminal effluent and the preservation solution before transplantation, and total serotonin was calculated as the sum of these amounts. Finally, transplantation was performed heterotopically. Total serotonin increased significantly with both cold and warm ischemic time (P<0.01 by analysis of variance, Fisher's PLSD); however, between 18 hr and 24 hr of cold ischemic time only, there were no significant changes. Total serotonin levels correlated well with cold and warm ischemic time, as shown by linear regression analysis (cold ischemia: R2=80.2%, P<0.01; warm ischemia: R2=92.8%, P<0.01). We established the cutoff level of total serotonin to predict the graft survival at 2200 ng, and using this critical level, graft survival was predicted by total serotonin with a sensitivity of 71.4% and a specificity of 89.8%. Immunohistochemical staining with the serotonin antibody revealed that the number of serotonin-positive cells decreased with both cold and warm ischemic time. In conclusion, serotonin is a useful parameter of cold and warm ischemic injury before transplantation and can assist in predicting graft survival.

ONTOGENY OF THE RESPONSE TO GROWTH HORMONE † 374

Growth hormone (GH) is essential for postnatal growth and regulation of fuel homeostasis. Its role in fetal and neonatal growth is less certain. The time at which GH begins to regulate perinatal growth in developing mammals is not established. Some models of GH deficiency show very early effects on growth. We have shown that the GH-responsive mRNAs, Spi 2.1 and 2.3, increase around birth in rat pups while GHR mRNA is stable at ≈10% of adult levels(A J Physiol 264:E973). To test the hypothesis that GH is active in perinatal life in promoting hepatic GH-responsive gene expression, we assessed GH action by measurement of hepatic Spi 2.1/2.3 gene expression in a model of neonatal GH deficiency. To determine if GH-responsive gene expression in the perinatal period is diminished by induction of GH deficiency and restored by GH administration, maternal/neonatal hypothyroidism (and consequent GH deficiency) was induced by administration of methimazole to timed pregnant female rats beginning at d 14 of gestation. Pups at days 2 and 7 of age were treated with GH 1.5 ug/g sq (GH-deficient pups injected with an equal vol of saline.) GH response was evaluated by measurement of hepatic Spi 2.1/2.3 mRNA, as expression of these genes is significantly increased by GH administration in adult animals. RNA levels were measured using Northern blots hybridized simultaneously to Spi 2.1/2.3 and GAPDH cDNAs. Intensity of hybridization was quantitated by phosphorimaging. Spi 2.1/2.3 values were normalized to GAPDH values. There was no significant difference between GH-deficient (def) and GH-treated (Rx) pups at d 2 (mean d 2 GH-def 0.40 +/- 0.01 (SEM) OD units, mean d 2 GH-Rx 0.44 +/- 0.04 OD units) but at d 7 GH administration resulted in significant (95% by Fisher PLSD) Spi 2.1/2.3 induction (mean d 7 GH-def 0.21 +/- 0.04 (SEM) OD units, mean d 7 GH-Rx 0.82 +/- 0.12 OD units). We conclude that the response to GH administration changes between days 2 and 7 of neonatal rat life, with relative insensitivity to GH administration at d 2 but clear response to GH by d 7. We speculate that this may be due to changes in post receptor responses to GH.

Hormonal changes related to eating behavior in oligomenorrheic women.

The aim of this study was to determine those hormonal alterations in the gonadotropin-ovarian axis that are related to eating behavior in oligomenorrheic patients. We studied 74 oligomenorrheic women aged 26.2 +/- 0.8 years, divided into group IA (N = 13) with eating disorders, group IB (N = 61) without eating disorders and 18 normally cycling controls aged 29.2 +/- 1.6 years (group II). No subject had ovarian failure, pituitary disease, thyroid dysfunction or was taking any drug. Blood samples were taken on days 3-6 after the last menses. Luteinizing hormone (LH) was measured in two plasma pools, each made up of three samples taken at 30-min intervals, starting at 15.00 h (LH-15h) and 09.00 h (LH-9h), which allowed the mean LH (mLH) and variability in LH (V-LH: percentage increase from the lower to the higher of the two LH values) to be calculated. Follicle-stimulating hormone (FSH), sex steroids, and gonadotropin-releasing hormone-stimulated LH (sLH) and -FSH (sFSH) were also evaluated. Eating behavior was evaluated with the EAT questionnaire; the EAT 26 score, the dieting score (DS) and bulimia score (BS) were calculated. Dietary intake was evaluated in 35 group IB patients based on food diaries analyzed with the REGAL program, to evaluate daily calorie intake (Cal) and calories provided by carbohydrates (Carb), lipids (Lip) and proteins (Prot). Comparisons between groups were done by analysis of variance (followed by the Fisher PLSD test) and the Kruskal-Wallis test. Groups IA, IB and II did not differ regarding age, body mass index, LH-9h, LH-15h, mLH, FSH, sLH, sFSH, estradiol or dehydroepiandrosterone sulfate; group IA had a higher V-LH than group II (p < 0.02) and a higher testosterone level than groups IB and II (p < 0.05). Positive correlations were found between V-LH and DS (p < 0.01) and BS (p < 0.05), and between testosterone and BS (p < 0.02) and DS (p < 0.05). The V-LH was negatively correlated with Cal and Carb, and testosterone was positively correlated with Cal and Lip. In patients referred for oligomenorrhea, it is concluded that testosterone levels and variability of LH levels are related to eating behavior.

Effect of lipophilic vehicles on in vivo skin penetration of methyl nicotinate in different races

Studies on racial differences in percutaneous penetration have mainly focused on measurements of either the pharmacodynamic response of a drug or skin reactivity to irritants. In order to clarify the contribution of the barrier stratum corneum to the overall drug penetration rate in different races, an in vivo permeability study was conducted with methyl nicotinate as the model drug. Four races (Caucasians, Hispanics, Blacks, Asians) were investigated. The drug was dissolved in various lipophilic vehicles (dimethicone 100, light mineral oil, isopropyl myristate, medium chain triglycerides) at concentrations that provide equal drug escaping tendencies. Drug solutions were applied to the upper arms of 48 subjects (four races with 12 subjects each) with a glass chamber system. To avoid drug depletion, drug disappearance rates were measured under steady-state conditions by the so-called difference method. Drug flux among the four races increases in the following order: Blacks < Asians < Caucasians < Hispanics. The effect of race almost reaches significance level ( P = 0.067). Fisher's PLSD post-hoc procedure indicated a significant difference between drug penetration in Hispanics and Blacks ( P = 0.009) suggesting structural or functional differences in the stratum corneum. Enhancement factors were calculated from the steady-state flux values, (i.e. drug disappearance rates per area unit) for each race. Significant enhancing effects ( P = 0.0001) were observed with the vehicles isopropyl myristate and mineral oil with all races.

Cytotoxic effects of kainate ligands on HEK cell lines expressing recombinant kainate receptors

Exposure of neurons either for prolonged periods of time or to high concentrations of excitatory amino acids (EAA), such as glutamate, results in neuronal death. Kainate also causes cell toxicity through the glutamate receptors. However, it is unclear whether the kainate receptor itself mediates any of the toxic responses. In the present study, HEK cells expressing the GluR6 ± KA2 receptor subunit(s) were studied for their susceptibility to toxicity through the kainate receptor by kainate ligands. The natural ligand, glutamate, did not result in toxicity to the recombinant cell lines over that observed with the untransfected HEK cells, whereas kainate produced a 2–3-fold increase in LDH in both the HEK/GluR6 (ANOVA, P = 0.0001) and HEK/GluR6 + KA2 (ANOVA, P = 0.0002) cell lines following treatment with various dosages, but did not affect the HEK cells. Similar 2–3-fold increases in LDH activity were detected in both recombinant cell lines following treatment with 100 nM of SYM2081 ((2S,4R)-4-methylglutamic acid), a dose at which agonistic activity is elicited. The rank order potencies for eliciting toxicity are consistent with the previously reported EC 50 values (SYM2081 > kainate ⋙ glutamate). Surprisingly, the kainate antagonist, NBQX, was the most toxic of the compounds tested although it had an affinity for the kainate receptor similar to glutamate. Treatment with as little as 10 nM elicited a dramatic increase in toxicity (6–10-fold) in the recombinant cell lines. At 1 μM, NBQX was significantly more toxic (Fisher PLSD, P < 0.05) than any of the other compounds tested. Thus, it appears that cell toxicity can be mediated via kainate receptor through two independent mechanisms: activation and blockage of the kainate receptor.

The Effects of Music Therapy Intervention on Agitation Behaviors of Alzheimer's Disease Patients

The purpose of this study was to examine the effects of live music therapy on agitation behavior of Alzheimer's Disease (AD) patients during and after music therapy intervention. A second purpose was to determine if there was a difference in the effect of music therapy between those patients who had a musical background and those who did not. A third purpose was to examine if there was a difference between music therapists' and caregivers' post-music therapy agitation scores. Agitation behavior in this study was defined as overt behavior that indicates restlessness, hyperactivity, or subjective distress. The dependent measures included: (a) scores on the Agitation Behavior Scale, and (b) number of dosages of PRN medication. Subjects for this study were 20 residents, 17 females and 3 males, from four different facilities in the Northwest. Their age range was 70 to 96(M = 82, SD = 6.57). The criteria for selection were: (a) A primary diagnosis of dementia with strong medical and behavioral indications that a post mortem examination would indicate the patients' presenting problems were the result of Alzheimer's disease, (b) the presence of agitation, (c) sufficient verbal ability to answer simple social and activity questions, and (c) the written consent of the patient's guardian or representative. Subjects, grouped by three or four, participated twice a week for 30 minutes for a total of five music therapy sessions. Caregivers having close contact with patients rated subjects' agitation behavior based on their observation of the subjects' behavior during the morning just prior to the beginning of music therapy (baseline). Music therapy sessions were videotaped for post-hoc agitation behavior analysis during music therapy, and for reliability purposes. Once music therapy sessions were over, subjects' behavior was observed for the next 20 minutes by a music therapist and a caregiver. In addition, subjects' charts were reviewed weekly to note the number of dosages of PRN medication administered each day of the week. The results of a two-factor analysis of variance with repeated measures show no effects with respect to music background (F1 = 1.79, p = .20), but identify significant main effects of agitation behavior (F3 = 16.33, p = .0001). Subsequent Fisher PLSD tests indicate that subjects appeared significantly more agitated before music therapy (M = 11.46) than during either of the two music therapy observations (M = 7.68 & M = 7.52, respectively) and after music therapy sessions (M = 8.37). No two-way interaction between music background and agitation behavior (F3 = 1.34, p = .28) was noted. Results of an independent t test between music therapists and caregivers post-music therapy agitation scores indicate no significant differences between scores [t(78) = .67, p = .50]. Implications for music therapy practice are discussed.

Safety of bronchoalveolar lavage and bronchial biopsies in patients with asthma of variable severity.

The safety of fiberoptic bronchoscopy, bronchoalveolar lavage (BAL), and bronchial biopsies has been questioned in asthma, and current recommendations indicate that bronchoscopies should only be performed in mild to moderate asthma. Moreover, in most studies patients receive premedication with nebulized bronchodilators that may enhance the safety of the procedures. The purpose of this study was to determine (1) whether the overall safety of fiberoptic bronchoscopy, BAL, and bronchial biopsies in mild to moderate asthma could be extended to patients with more severe asthma and (2) whether these procedures are safe without premedication with nebulized bronchodilators. A group of 50 patients with asthma of variable severity (FEV1 ranging from 37 to 107% of predicted values) and 25 healthy volunteers were studied. Bronchoscopy, BAL (250 ml), and four bronchial biopsies were performed in a standardized manner, without premedication with a nebulized bronchodilator, by the same investigator. Safety was assessed by clinical follow-up, continuous recording of arterial oxygen saturation during the procedure with a digital oximeter, and measuring FEV1, FEF25-75, and FVC just before and 5 min after bronchoscopy. Arterial oxygen saturation decreased in asthmatic patients from 97% (range 91 to 99%) (T1) to 92% (range 79 to 98%) (T8) (ANOVA, Fisher's PLSD) and in control subjects from 97% (range 94 to 99%) (T1) to 93% (range 88 to 98%) (T8) (ANOVA, Fisher's PLSD). The fall in arterial oxygen saturation was not significantly different between asthmatic and normal subjects, and there was no correlation between arterial oxygen desaturation and the severity of asthma.(ABSTRACT TRUNCATED AT 250 WORDS)

TCDD causes stimulation of c-ras expression in the hepatic plasma membranes in vivo and in vitro.

A series of in vivo and in vitro experiments were conducted to determine the effects of 2, 3, 7, 8-tetrachlorodibenzo-p-dioxin (TCDD) administered on the expression of c-ras. Differences in c-ras expression between control and TCDD treated groups were determined by immunoassay of p21ras protein, or indirectly measured by the specific binding of 3H-GTP to hepatic plasma membrane preparations. Intraperitoneal injection of sublethal doses of TCDD significantly elevated (P less than 0.05, Student t test) levels of hepatic p21ras protein in Sprague-Dawley rats and TCDD sensitive C57BL/6J mice. Such an increase occurred at an early stage of poisoning in the C57BL/6J mice. The earliest increase was detectable 6 hr after dosing, and the difference became statistically significant by 12 and 24 hr after dosing. In contrast, TCDD tolerant DBA/2J mice had only a marginal increase in hepatic p21ras protein which did not become statistically significant even at 24 hr host-dosing. TCDD evoked increases in hepatic p21ras protein of C57BL/6J mice were accompanied by the increase in the specific binding of GTP to hepatic plasma membranes. Column chromatography of solubilized rat hepatic membrane proteins on sephadex G-50 showed TCDD administration increased levels of a 3H-GTP binding protein with MW of approximately 21 Kd. 3H-GTP binding in total hepatic membranes was also elevated (P less than 0.05, Fisher PLSD multiple comparison test) 6 hr and 24 hr after dosing of C57BL/6J mice, but as expected the effect of TCDD was not as conspicuous as that found in the plasma membrane. TCDD treatment increased levels of a 21 Kd protein found in the in vitro translation products of RNA purified from guinea pig liver. This protein was identified as a c-ras protein based upon its ability to bind GTP, precipitation by a polyclonal antibody against the rasHa and Ki proteins and subsequent SDS-PAGE which showed a single protein band of approximately 21 Kd.

Subfornical organ lesions in rats abolish hyperdipsic effects of isoproterenol and serotonin

Isoproterenol (300 μg/ml/kg) and serotonin (2 mg/ml/kg) given SC to rats ( n = 27) caused significant drinking (Fisher PLSD, Scheffe F test, Dunnett t) in the 1 and 2 hours after injection. Such drinking was completely prevented in rats later shown to have complete lesions of their subfornical organs ( n = 7). In contrast a response not significantly different from the prelesion response was found in rats later given cortical lesions ( n = 11) or other lesions which did not damage the subfornical organ ( n = 7). We conclude that drinking evoked by SC injection of serotonin and isoproterenol is brought about by peripheral production of angiotensin II. Blood borne angiotensin II in turn stimulates neurons in subfornical organ which initiate the neural organization of a drinking response.

Nicotinamide enhances skin flap survival

The effects of nicotinamide in an abdominal island pedicle skin flap were examined. A 7 x 7 cm island pedicle skin flap ligating the left inferior neurovascular pedicle was created on 50 male Sprague Dawley rats (250-275 grams) that were divided into five groups. Animals received either 0.6 cc of saline or doses of nicotinamide for 16 days (14 days preoperatively and 2 days postoperatively): 25 mg b.i.d., 50 mg b.i.d., 100 mg b.i.d. or 200 mg b.i.d. Forty-eight hours postoperatively each animal received 25 mg of Fluorescein via the tail vein. The area of necrosis was visualized and quantified and is presented as % survival. A one factor Fisher PLSD test was performed with a statistical significance of p less than 0.05 with the results as follows: saline 58.8%, 25 mg 68.6%, 50 mg 82%, 100 mg 80.8%, and 200 mg 86%. From this data it would appear that the angiogenic factor nicotinamide may increase random flap survival.