Control Points To Reduce Movement of Central Nervous System Tissue during Beef Slaughter.

Consumption of central nervous system tissue (CNST) from cattle with bovine spongiform encephalopathy (BSE) is thought to cause the human neurological disease, variant Creutzfeldt-Jacob disease. To identify points of cross-contamination of beef carcasses with CNST, 55 young beef cattle were slaughtered and processed through a federally inspected multispecies abattoir. The objectives of this study were to evaluate CNST spread following the placement of a plug in the penetration site of the skull after captive bolt stunning, to evaluate cross-contamination of carcasses before and after splitting, to compare the effects of hot water pasteurization (84°C for 10 s) versus cold water wash (10°C for 30 s) for reducing CNST on the carcass, and to examine other possible sources of cross-contamination in the abattoir. Results indicated that the use of a plastic plug reduced CNST contamination near the bolt penetration site. This study also confirmed that carcass splitting resulted in an increase in CNST contamination at various areas of the carcass. Hot water pasteurization appeared to be an effective means of removing CNST contamination from carcasses in most of the areas sampled.

Dietary vitamin E effects on the formation of heterocyclic amines in grilled lean beef.

The effects of tissue antioxidant levels on formation of heterocyclic amines (HAs) and their mutagenicity in grilled lean beef were studied. Meat from 54 feedlot steers fed different levels of vitamin E (340, 690, 1040 and 1740 IU/animal/day) for 120-days was used to provide beef with different levels of antioxidants (α-tocopherol). Prevalent HAs were then analyzed by HPLC using UV/Fluorescence detection. Five major HAs were found: 2-amino-3,8-dimethyl-imidazo(4,5-F)Quinoxaline (MeIQx), 2-amino-3,4,7,8-tetramethyl-imidazo(4,5-F)Quinoxaline (TriMeIQx), ß-Carboline-9H-Pyrido[3,4-b]indole (Norharmane), 1-Methyl-9H-pyrido[3,4-b]indole (Harmane) and 2-amino-1-methyl-6-phenylimidaza(4,5-B)pyridine (PhIP). Total content of HAs in grilled lean beef ranged from 9.57 ng/g to 11.59 ng/g. There was, however, a trend (P=0.097) found for reduced mutagenicity with increasing tissue levels of α-tocopherol. The increasing dietary vitamin E significantly increased the α-tocopherol level in lean beef (P<0.001), but it had no significant (P>0.05) inhibitory effects on the content of individual and total HAs.

Phylogeny and systematics of demospongiae in light of new small-subunit ribosomal DNA (18S) sequences.

The most diverse and species-rich class of the phylum Porifera is Demospongiae. In recent years, the systematics of this clade, which contains more than 7000 species, has developed rapidly in light of new studies combining molecular and morphological observations. We add more than 500 new, nearly complete 18S sequences (an increase of more than 200%) in an attempt to further enhance understanding of the phylogeny of Demospongiae. Our study specifically targets representation of type species and genera that have never been sampled for any molecular data in an effort to accelerate progress in classifying this diverse lineage. Our analyses recover four highly supported subclasses of Demospongiae: Keratosa, Myxospongiae, Haploscleromorpha, and Heteroscleromorpha. Within Keratosa, neither Dendroceratida, nor its two families, Darwinellidae and Dictyodendrillidae, are monophyletic and Dictyoceratida is divided into two lineages, one predominantly composed of Dysideidae and the second containing the remaining families (Irciniidae, Spongiidae, Thorectidae, and Verticillitidae). Within Myxospongiae, we find Chondrosida to be paraphyletic with respect to the Verongida. We amend the latter to include species of the genus Chondrosia and erect a new order Chondrillida to contain remaining taxa from Chondrosida, which we now discard. Even with increased taxon sampling of Haploscleromorpha, our analyses are consistent with previous studies; however, Haliclona species are interspersed in even more clades. Haploscleromorpha contains five highly supported clades, each more diverse than previously recognized, and current families are mostly polyphyletic. In addition, we reassign Janulum spinispiculum to Haploscleromorpha and resurrect Reniera filholi as Janulum filholi comb. nov. Within the large clade Heteroscleromorpha, we confirmed 12 recently identified clades based on alternative data, as well as a sister-group relationship between the freshwater Spongillida and the family Vetulinidae. We transfer Stylissa flabelliformis to the genus Scopalina within the family Scopalinidae, which is of uncertain position. Our analyses uncover a large, strongly supported clade containing all heteroscleromorphs other than Spongillida, Vetulinidae, and Scopalinidae. Within this clade, there is a major division separating Axinellidae, Biemnida, Tetractinellida, Bubaridae, Stelligeridae, Raspailiidae, and some species of Petromica, Topsentia, and Axinyssa from Agelasida, Polymastiidae, Placospongiidae, Clionaidae, Spirastrellidae, Tethyidae, Poecilosclerida, Halichondriidae, Suberitidae, and Trachycladus. Among numerous results: (1) Spirophorina and its family Tetillidae are paraphyletic with respect to a strongly supported Astrophorina within Tetractinellida; (2) Agelasida is the earliest diverging lineage within the second clade listed above; and (3) Merlia and Desmacella appear to be the earliest diverging lineages of Poecilosclerida.

Composition and stability of bacterial communities associated with granular activated carbon and anthracite filters in a pilot scale municipal drinking water treatment facility.

Granular activated carbon (GAC) is an alternative filter substrate for municipal water treatment as it provides a high surface area suitable for microbial colonization. The resulting microbial growth promotes biodegradation of organic materials and other contaminants from influent waters. Here, the community structure of the bacteria associated with three GAC and two anthracite filters was examined over 12 months to monitor changes in community composition. Nearly complete 16S rRNA genes were polymerase chain reaction amplified for terminal restriction fragment length polymorphism (T-RFLP) analyses. The identity of commonly occurring peaks was determined through the construction of five representative 16S rRNA clone libraries. Based on sequence analysis, the bacterial communities associated with both anthracite and GAC filters appear to be composed of environmentally derived bacteria, with no known human pathogens. Analysis of similarity tests revealed that significant differences in bacterial community structure occurred over time, with filter substrate playing an important role in determining community composition. GAC filters exhibited the greatest degree of bacterial community variability over the sampling period, while anthracite filters showed a lower degree of variability and less change in community composition. Thus, GAC may be a suitable biologically active filter substrate for the treatment of municipal drinking water.

Potential of solid phase extraction disks to aid determination of dislodgeable foliar residues of chlorpyriphos, malathion, diazinon, and acephate.

The utility of solid phase extraction (SPE) for concentrating four organophosphate insecticides from solutions of water and sodium dioctyl sulfosuccinate, a surfactant, was evaluated. Reverse phase (C18, octadecyl bonded silica) sorbent in the form of a disk was the SPE medium evaluated. Chlorpyriphos, malathion, and diazinon, but not acephate, were retained on and eluted from the SPE disks. For pesticides that were retained on SPE disks, recoveries from the disks were equal to or higher than recoveries achieved by solvent partitioning. Dislodgeable foliar residues of acephate were successfully concentrated for analysis by lyophilization of water-surfactant solutions. Recoveries of pesticides from SPE disks stored at -15 degrees C for one week were equal to or higher than those of pesticides stored in water-surfactant for one week at -15 degrees C. Malathion- and diazinon-fortified samples in watersurfactant and on SPE disks were prepared in one state and shipped for analysis in another state. Pesticides in the water-surfactant samples were concentrated by solvent partitioning and were underestimated by 41% (diazinon) and 16% (malathion). Conversely, diazinon samples on the SPE disks were on average underestimated by 3% and malathion was overestimated by an average of 55%. The overestimation of malathion was attributed to a matrix effect during analysis associated with the presence of surfactant, which was retained on and subsequently eluted from the SPE disks. The retention of surfactant by the SPE disks and its subsequent elution may considerably limit their usefulness in determination of dislodgeable foliar residues.

Analysis of 4-hydroxyproline using 4-chloro-7-nitrobenzo-2-oxa-1,3-diazol derivatization and micellar electrokinetic chromatography combined with laser-induced fluorescence detection.

Micellar electrokinetic chromatography (MEKC) with laser-induced fluorescence (LIF) detection was used for internal standard (pyrrolidinol) based quantification of 4-hydroxyproline (Hyp) in muscle hydrolysates. Hydrolysates were first derivatized with o-phthaldialdehyde to reduce primary amine interference and then 4-chloro-7-nitrobenzo-2-oxa-1,3-diazol (NBD) to enable selective detection of secondary amines. This method allows for rapid and sensitive detection of hydroxyproline in dilute hydrolysates using commercially available capillary electrophoresis equipment.

Mitochondrial phylogeny of extant Hawaiian tree snails (Achatinellinae).

Hawaiian tree snails in the endemic subfamily Achatinellinae display a staggering variety of shell colors and banding patterns. Despite numerous attempts to classify this morphological variation, a conclusive phylogeny has not been proposed. To improve conservation efforts, we sought to better understand the species identities and phylogenetic relationships among the extant species of Achatinella and Partulina using partial mitochondrial 16S ribosomal DNA sequences. The reconstructed phylogeny showed a high degree of support for more recent branches, but gave little support to deeper nodes. The most confident branches challenge previous systematic arrangements of these snails, grouping species that previously had been placed into different subgenera. High levels of sequence divergence within some species may reflect the long-term isolation of subpopulations. Rapid rates of sequence divergence may have saturated base substitutions and contributed to the lack of resolution of higher-order relationships. We did not find support for the monophyly of the Achatinella species, nor thus for a single colonization of Oahu from Maui Nui.

Emotional and psychological consequences of sexual harassment: a descriptive study.

The emotional and psychological consequences of sexual harassment were investigated. On the basis of previous empirical evidence concerning the correlates of sexual harassment, the role of the working relationship between harasser and target, type of harassment, gender composition of the work group, duration of the harassment, and gender were examined in relation to two psychological states: feelings about work and emotional/physical condition.

Volatile shell-investigation cues of land hermit crabs: Effect of shell fit, detection of cues from other hermit crab species, and cue isolation.

Land hermit crab responses to volatile shell-investigation cues from land hermit crabs and from marine hermit crabs are analogous to the responses of marine hermit crabs to shell-investigation cues from marine crabs and from snails. Land hermit crabs attracted to shell cues are in worse-fitting shells and are more likely to investigate conspecifics' shells than are crabs attracted to feeding cues. Moving land hermit crabs from worse shells to better shells decreases the number of crabs investigating shells, while moving crabs from better shells to worse shells increases the number of crabs investigating shells. Gravid females have better-fitting shells than nongravid females or males. Crabs from two different populations in Panama have different shell fits and show different levels of responses to shell-investigation cues. Land hermit crabs respond to volatile shell-investigation cues from both land and marine hermit crabs, but marine hermit crabs do not respond to cues from land hermit crabs. A cue detection system for volatile cues most likely evolved in land hermit crabs during their transition from a marine to a terrestrial existence. Thus, the cues found in land hermit crabs and marine hermit crabs may be chemically similar. Volatile compounds collected from hermit crabs onto Tenax columns can be eluted with ethanol and act as shell-investigation cues in field assays.

2,3-Dihydrofarnesoic acid, a unique terpene from trichomes ofLycopersicon hirsutum, repels spider mites.

Lycopersicon hirsutum, a wild relative of the tomato, is highly resistant to arthropod herbivores. Both botanic forms ofL. hirsutum, L. hirsutum f.glabratum (C.H. Mull.) andL. hirsutum f.typicum (Humb. & Bonpl.), are resistant to two-spotted spider mites,Tetranychus urticae Koch. However, leaves and trichome secretions from f.typicum repel mites more so than those from f.glabratum. We have previously demonstrated that trichome secretions from LA 1363 and LA 1927, accessions of f.typicum, repelled mites. In this paper we report the identification of the primary component of trichome secretions responsible for repellency. Leaflet washes having compositions similar to trichome secretions were collected and separated into neutral and acid fractions; repellency was mainly associated with the acid fraction, which, when applied to nonrepellent leaflets of f.glabratum, rendered them repellent. Separation of leaflet washes by HPLC allowed purification and subsequent identification by gas chromatography-mass spectrometry and nuclear magnetic resonance of 2,3-dihydrofamesoic acid (3,7,11-trimethyl-6, 10-dodecadienoic acid) as the primary chemical component responsible for repellency. Application of this acid to leaflets ofL. esculentum rendered them repellent. Other volatile compounds present in minor amounts in the acid fractions were farnesoic acid and 16∶0, 16∶3, 18∶0, 18∶2, and 18∶3 fatty acids. This is the first report of the natural occurrence of 2,3-dihydrofarnesoic acid.

Affirmative action after the Supreme Court's 1988-1989 term: what employers need to know.

Three recent Supreme Court decisions have aroused fears about the future of a key weapon against employment discrimination: the affirmative action plan. The author reviews these cases and pronounces affirmative action alive and well--but warns that thoughtful planning is required to ward off challenges by nonminority groups claiming adverse impact.

Effect of ascorbic acid and vitamin E on biochemical changes associated with vitamin E deficiency in rats.

Weanling male Sprague Dawley rats were fed a vitamin E and C-free basal diet with or without supplementation of 100 IU vitamin E per kg diet. After 20 weeks, the vitamin E-deficient rats were divided into four groups, six in each group, and received supplemental ascorbic acid and/or vitamin E by tube feeding daily for 7 days: Group I, 30 mg ascorbic acid/100 g body wt.; Group II, 0.03 mg RRR-alpha-tocopheryl acetate/100 g body wt.; Group III, 30 mg ascorbic acid and 0.03 mg RRR-alpha-tocopheryl acetate/100 g body wt.; and Group IV, placebo. The six control rats (Group V) received placebo. The rats were sacrificed, blood and liver samples were collected for biochemical determinations. Vitamin E deficiency significantly increased erythrocyte (RBC) spontaneous hemolysis, liver thiobarbituric acid (TBA) value, activities of glutamateoxaloacetate transaminase (GOT), pyruvate kinase (PK), and creatine phosphokinase (CPK) in plasma, and significantly lowered plasma vitamin E levels and glutathione peroxidase (GPX) activities. Tube-feeding ascorbic acid for 7 days produced partial reversal effect on liver TBA values, activities of plasma PK, GOT, CPK, and plasma vitamin E levels but not on RBC hemolysis and plasma GPX activity. Tube feeding both ascorbic acid and vitamin E showed similar partial reversal effect as feeding vitamin E alone on all the parameters stated above. The results suggest that ascorbic acid may spare the metabolism of vitamin E and partially reverse the changes in some of the biochemical parameters characteristic of vitamin E deficiency.

Vitamin C partially reversed some biochemical changes produced by vitamin E deficiency.

Feeding a basal diet free of vitamins E and C to weanling male rats for 8 months resulted in biochemical changes characteristic of vitamin E deficiency. These included increased liver thiobarbituric acid values; decreased blood GSH levels, plasma vitamin E levels, and glutathione peroxidase activities; and increased activities of plasma pyruvate kinase, glutamic-oxaloacetic transaminase, creatine kinase, lactic dehydrogenase, and malic dehydrogenase. Tube-feeding vitamin C for 21 days resulted in partial reversal effects on the above parameters except activities of glutathione peroxidase, lactic dehydrogenase, and malic dehydrogenase. The results suggest that vitamin C may spare in part the metabolism of vitamin E through its antioxidant property.

Lower levels of vitamin C and carotenes in plasma of cigarette smokers.

Plasma levels of vitamins A, C, and E, selenium, and carotenes were determined in 125 male cigarette smokers and 125 age- and race-matched nonsmokers. The smokers had a mean daily consumption of 30.6 cigarettes and a cumulative consumption of 22.8 pack years. Plasma levels of vitamin C and total carotenes were significantly (p less than 0.05) lower in smokers than those of nonsmokers, while levels of vitamin A, selenium, and vitamin E were not significantly different between these two groups. Similar results were found when only those subjects not taking any form of dietary supplements were included for analysis. Except for negative correlation between vitamin A and pack-year, no significant correlates were observed between plasma levels of these micronutrients and indices of smoking status or cigarette consumption in smokers. These data suggest that chronic cigarette smoking is associated with depressed levels of plasma vitamin C and carotenes; however, the relationship between smoking and these plasma micronutrients is still unclear.

Effects of dietary vitamin E and high level of ascorbic acid on iron distribution in rat tissues.

The effects of dietary vitamin E and high-level supplementation of ascorbic acid on iron distribution in rat tissues were studied. Weanling male Sprague-Dawley rats, fed ad libitum a vitamin E and ascorbic acid free basal diet, were divided into four groups. They were supplemented with 0 or 45 IU/kg diet of vitamin E, and O or 0.2% ascorbic acid in a 2 X 2 complete factorial design. After 12 weeks, rats were killed; blood, liver, spleen, heart and skeletal muscle were collected for analysis. Vitamin E deficiency resulted in significantly decreased plasma iron levels and total iron binding capacity. The total iron and nonheme iron contents of the liver and spleen were significantly higher in the vitamin E-deficient groups compared with control groups. Vitamin E or ascorbic acid supplementation had no effect on iron content of the heart. Non-heme iron levels on per gram tissue were highest in the skeletal muscle of the group to which no vitamin E or ascorbic acid were supplemented. It appears that vitamin E and ascorbic acid interactively affect the iron distribution in rat tissues.

Dietary vitamin E and pulmonary biochemical responses of rats to cigarette smoking.

The effect of dietary vitamin E on cellular susceptibility to cigarette smoking was studied in rats. Young male rats maintained on a basal vitamin E-deficient diet with or without 100 ppm vitamin E supplementation for 4 or 5 weeks were exposed to either sham or cigarette smoke for up to 7 days. Higher animal mortality rate was observed in the animals fed the vitamin E-deficient diet than in the supplemented group when they were subjected to acute levels of cigarette smoking. Relative to the respective sham groups, a greater alteration of biochemical parameters, such as reduced glutathione (GSH) and related enzymes, was found in the lungs of smoked rats fed the deficient diet than in the supplemented group. Animal lungs exhibited a greater biochemical response to whole smoke than the gaseous phase of smoke. The results suggest that the nutritional status of vitamin E may influence the cellular susceptibility of rats to cigarette smoking.

Effect of post preparation on the apical seal.

Apical leakage was analyzed quantitatively in extracted teeth with varied levels of remaining gutta-percha. The effect of the method of gutta-percha removal on the apical seal was also evaluated. Ninety teeth were prepared so that 30 teeth had 3 mm of gutta-percha, 30 teeth had 5 mm, and 30 teeth had 7 mm. An additional 75 teeth were divided into three groups of 25 each. A different method of gutta-percha removal was used for each group: hot instrument, mechanical rotary instrument, and chemical solvent. An electrochemical method was used to analyze apical microleakage. Measurements of leakage of each sample were obtained at 24-hour intervals for 30 days. The data suggest that the mechanical method is the most desirable for gutta-percha removal in post preparation. As the level of gutta-percha increased to 7 mm the degree of leakage decreased. At least 5 mm of gutta-percha is necessary for an adequate apical seal.

Plasma creatine kinase isozyme electrophoretic pattern of long-term vitamin E-deficient rats.

Weanling male Sprague Dawley rats were fed a vitamin E-deficient basal diet with or without supplementation of 100 IU/kg diet of vitamin E for 12 months. The isozyme pattern of creatine kinase (CPK) in plasma was determined electrophoretically. The CPK activity of the plasma of control rats averaged 43.2 nmoles/min/ml and was predominantly CPK3 isozyme, while that of the plasma of vitamin E-deficient rats averaged 100.8 nmoles/min/ml and consisted of CPK2 and CPK3 isozymes. The results suggest that the increased CPK activity in the plasma of vitamin E-deficient rats may originate from heart and skeletal muscle degeneration.

The prevalence of auscultatory crackles in subjects without lung disease.

Crackles heard during auscultation of the lungs are generally considered an abnormal physical finding. This study was done to see if crackles could be induced to occur in the lungs of normal subjects. We studied 52 nonsmoking normal subjects by listening at the right and left posterior bases and over the lower right anterior chest during inspiration from functional residual capacity (FRC) and then during inspiration from residual volume (RV). Crackles were recorded on magnetic tape for subsequent time-expanded waveform analysis. No crackles were heard at any of the three sites during inhalation from FRC. Of 52 subjects, 26 had crackles over the anterior site when inhaling from RV. These crackles were often profuse, loud, and easily recorded. Waveform analysis revealed these crackles in normal subjects to be similar to those occurring in patients with interstitial lung diseases. We conclude that crackles, heard over the anterior chest during inspiration from low lung volumes, are not necessarily adventitious sounds.

Dietary selenium and levels of L-ascorbic acid in the plasma, livers, and lungs of polychlorinated biphenyls-treated rats.

Administration of polychlorinated biphenyls (PCB) (500 mg Aroclor, 1254/kg body weight) intraperitoneally significantly increased the levels of L-ascorbic acid in the plasma and livers, but not in the lungs of one-month-old male rats maintained on a basal low selenium diet with or without 2.0 ppm selenium (as sodium selenite) supplementation for 19 weeks prior to PCB treatment. The levels of L-ascorbic acid were not significantly altered by dietary selenium. In another experiment, 40 days or 15-month-old male rats were fed the same basal selenium diet with or without 1.0 ppm selenium for 1 month prior to PCB treatment. The plasma levels of ascorbic acid were found to increase significantly by PCB treatment, but not by the status of dietary selenium or by animal age. As expected, the activity of selenium dependent glutathione peroxidase was markedly decreased in the plasma, livers, and lungs of rats fed the low selenium diet. However, the enzyme activity was not significantly altered by PCB treatment in plasma, livers, and lungs of animals in both dietary groups.