Brain morphology and immunohistochemical localization of the gonadotropin-releasing hormone in the bluefin tuna, Thunnus thynnus.

The present study was focused on the morphology of the diencephalic nuclei (likely involved in reproductive functions) as well as on the distribution of GnRH (gonadotropin-releasing hormone) in the rhinencephalon, telencephalon and the diencephalon of the brain of bluefin tuna (Thunnus thynnus) by means of immunohistochemistry. Bluefin tuna has an encephalization quotient (QE) similar to that of other large pelagic fish. Its brain exhibits well-developed optic tecta and corpus cerebelli. The diencephalic neuron cell bodies involved in reproductive functions are grouped in two main nuclei: the nucleus preopticus-periventricularis and the nucleus lateralis tuberis. The nucleus preopticus-periventricularis consists of the nucleus periventricularis and the nucleus preopticus consisting of a few sparse multipolar neurons in the rostral part and numerous cells closely packed and arranged in several layers in its aboral part. The nucleus lateralis tuberis is located in the ventral-lateral area of the diencephalon and is made up of a number of large multipolar neurones. Four different polyclonal primary antibodies against salmon (s)GnRH, chicken (c)GnRH-II (cGnRH-II 675, cGnRH-II 6) and sea bream (sb)GnRH were employed in the immunohistochemical experiments. No immunoreactive structures were found with anti sbGnRH serum. sGnRH and cGnRH-II antisera revealed immunoreactivity in the perikarya of the olfactory bulbs, preopticus-periventricular nucleus, oculomotor nucleus and midbrain tegmentum. The nucleus lateralis tuberis showed immunostaining only with anti-sGnRH serum. Nerve fibres immunoreactive to cGnRH and sGnRH sera were found in the olfactory bulbs, olfactory nerve and neurohypophysis. The significance of the distribution of the GnRH-immunoreactive neuronal structures is discussed.

Novel HPLC analysis of tocopherols, tocotrienols, and cholesterol in tissue.

Tocopherols and tocotrienols are being increasingly recognized to have an important role in the prevention of atherosclerosis. It has been reported that they protect low-density lipoprotein (LDL) and tissues from oxidative stress and that tocotrienols can reduce plasma cholesterol levels. Two isocratic high-performance liquid chromatography (HPLC) methods for simultaneous analysis of tocopherols, tocotrienols, and cholesterol in muscle tissue were developed. Method A involves basic saponification of the sample, but causes losses of the gamma- and delta-homologs of vitamin E. Method B does not involve saponification, thereby protecting the more sensitive homologs. Both permit rapid analysis of multiple samples and neither requires specialized equipment. These methods may provide techniques useful in simultaneous assessment of oxidative stress status (OSS) and cholesterol levels.

Capillary GC quantification of cholesterol oxidation products in plasma lipoproteins of fasted humans.

Cholesterol oxidation products have been hypothesized to be important factors in atherosclerosis, a process which can culminate in myocardial infarction. The relative importance of exogenous or in vivo sources of cholesterol oxidation products has not been determined. However, methodology used for cholesterol oxidation products analysis of foods is applicable to the determination of cholesterol oxidation products in human plasma lipoproteins. Such methodology, outlined in this report, permits numerous critical experiments to be conducted on the possible role of cholesterol oxidation products in coronary heart disease.

Atherogenic and anti-atherogenic factors in the human diet.

New atherosclerosis causative factors and preventive modalities have been identified. Atherogenic factors include lipid oxidation products, such as cholesterol oxidation products, malonaldehyde and other aldehydes; trans-fatty acids; some saturated fatty acids (lauric, myristic and possibly palmitic acids); and myristic acid plus cholesterol. Lipid oxidation products are well suited to induce arterial damage, based on their known cytotoxic effects; evidence also indicates the possibility of plaque promotion and stimulation of thrombogenesis. Anti-atherogenic factors include antioxidants, fish oils and other polyunsaturates (if protected from oxidation), fibre and trace minerals such as copper, manganese, selenium and zinc. Iron is unique, being considered as both a potential promoter of atherosclerosis (component of ferritin, conceivably inducing lipid oxidation) and a possible anti-atherogenic component (of antioxidant enzyme catalase). It is apparent that an entire new series of research challenges has been uncovered.

Cell-mediated biotransformation of S-nitrosoglutathione.

Spontaneous release of nitric oxide (NO) from S-nitrosothiols cannot explain their bioactivity, suggesting a role for cellular metabolism or receptors. Using immortalised cells and human platelets, we have identified a cell-mediated mechanism for the biotransformation of the physiological S-nitrosothiol compound S-nitrosoglutathione (GSNO) into nitrite. We suggest the name "GSNO lyase" for this activity. GSNO lyase activity varied between cell types, being highest in a fibroblast cell line and lowest in platelets. In NRK 49F fibroblasts, GSNO lyase mediated a saturable, GSNO concentration-dependent accumulation of nitrite in conditioned medium, which was inhibited both by transition metal chelators, and by subjecting cells to oxidative stress using a combination of the thiol oxidant diamide and Zn2+, a glutathione reductase inhibitor. Activity was resistant, however, to both acivicin, an inhibitor of gamma-glutamyl transpeptidase (EC 2.3.2.2), and to ethacrynic acid, an inhibitor of Pi class glutathione-S-transferases (EC 2.5.1.18), thus neither of these enzymes could account for NO release. Although GSNO lyase does not explain the platelet-selective pharmacological properties of GSNO, cellular biotransformation suggests therapeutic avenues for targeted delivery of NO to other tissues.

Analyses of nuclear ldhA gene and mtDNA control region sequences of Atlantic northern bluefin tuna populations.

There has been considerable debate about whether the Atlantic northern bluefin tuna exist as a single panmictic unit. We have addressed this issue by examining both mitochondrial DNA control region nucleotide sequences and nuclear gene ldhA allele frequencies in replicate size or year class samples of northern bluefin tuna from the Mediterranean Sea and the northwestern Atlantic Ocean. Pairwise comparisons of multiple year class samples from the 2 regions provided no evidence for population subdivision. Similarly, analyses of molecular variance of both mitochondrial and ldhA data revealed no significant differences among or between samples from the 2 regions. These results demonstrate the importance of analyzing multiple year classes and large sample sizes to obtain accurate estimates when using allele frequencies to characterize a population. It is important to note that the absence of genetic evidence for population substructure does not unilaterally constitute evidence of a single panmictic population, as genetic differentiation can be prevented by large population sizes and by migration.

Occurrence of lipid oxidation products in foods.

Lipid oxidation products are ubiquitous in foods, although much variation exists in the levels present. Although these levels are generally low, the problem of lipid oxidation severely compromises the quality of some foods and limits the shelf-life of others. Lipid oxidation represents a key barrier in the development of new food products and processes, especially convenience items and processes required to manufacture them. Deleterious changes in foods caused by lipid oxidation include loss of flavour, development of off-flavours, loss of colour, nutrient value and functionally, and the accumulation of compounds which may be detrimental to the health of consumers. All foods that contain lipids are susceptible to oxidation but especially affected are foods which are dehydrated, subjected to high temperatures or cooked and subsequently stored, e.g. dehydrated eggs, cheeses and meats, foods fried in frying oils, and cooked (uncured) meats. Specific examples of compounds which are of health concern include lipid peroxides and the free radicals involved in their formation and propagation, malonaldehyde, and several cholesterol oxidation products. Coronary artery disease (CAD) may be in part caused by the consumption of lipid oxidation products.

Biological effects of oxysterols: current status.

A review of relevant literature on biological activities of oxysterols (OS) and cholesterol is presented. The data clearly demonstrate manifold biological activities, often detrimental, for OS compared with little or no such activity of a deleterious nature for cholesterol itself. Cholesterol is perhaps the single most important compound in animal tissue and, as such, it is difficult to imagine it as a toxin or hazard. In contrast, OS exhibit cytotoxicity to a wide variety of cells leading to angiotoxic and atherogenic effects; alter vascular permeability to albumin; alter prostaglandin synthesis and stimulate platelet aggregation, an important process facilitating atherosclerosis and thrombosis; alter the functionality of low density lipoprotein (LDL) receptors, possibly stimulating hypercholesterolaemia; modify cholesteryl ester accumulation in various cells, inducing foam cell formation; and enrich the LDL particle in cholesteryl esters, possibly increasing its atherogenicity. Furthermore, OS are mutagenic and carcinogenic, although some have been studied as antitumour agents based on their cytotoxic properties. Moreover, numerous studies have implicated OS in membrane and enzyme alterations that are interrelated with many of the foregoing effects. The authors find that OS deserve much more attention than cholesterol itself in terms of research activity but that unfortunately the reverse is true with regard to funding.

HPLC method for quantitation of cholesterol and four of its major oxidation products in muscle and liver tissues.

A fast, sensitive, high performance liquid chromatographic method was developed for the quantitation of cholesterol and four of its major oxidation products: 3 beta-hydroxycholest-5-en-7-one (7-ketocholesterol), cholest-5-ene-3 beta, 7 alpha-diol (7 alpha-hydroxycholesterol), cholest-5-ene-3 beta, 7 beta-diol (7 beta-hydroxycholesterol), and cholest-5-ene-3 beta,25-diol (25-hydroxycholesterol). In this procedure 2:1 chloroform:methanol (v/v) extracts of tissue homogenate were combined, dried over anhydrous Na2SO4, filtered, evaporated to dryness under N2 and dissolved with a mobile phase of either 97:3 or 93:7 hexane:isopropanol (v/v). After membrane filtration and without further purification, aliquots were directly injected onto a 10-microns pore size, 30 X 0.39 cm mu-Porasil normal phase column. The separation of cholesterol and its oxidation products was monitored by a UV detector at 206 and 233 nm. This method was successfully applied to pork muscle as well as mouse liver tissues and was able to detect cholesterol oxidation products (COP) in the ppm range. The identity of the COP was confirmed by mass spectroscopy.

Porcine stress syndrome and postmortem muscle characteristics of two purebreds and three specific terminal crosses.

The incidence of Porcine Stress Syndrome (PSS), determined by halothane screening, and parameters of muscle quality and carcass leanness were studied in 108 pigs from a larger population of 658 pigs of Pietrain (P), Minnesota No. 1 (M), Hampshire (H) x (P x M), Yorkshire (Y) X (P X M) and P X (P X M) breeding. The larger population was also surveyed by the halothane screening procedure for incidence of PSS, and growth rate was measured. At 6 to 14 weeks of age, pigs were classified as PSS if they exhibited muscle rigidity within 5 min after the commencement of anesthetization with 3% halothane in oxygen. The incidences noted for the larger population were: H x (P x M) and Minnesota No. 1, 0%; Pietrain, 88%; Y x (P x M), 3%, and P x (P x M), 17%. Results demonstrated that the H x (P x M) group displayed excellent carcass meatiness combined with acceptable meat quality and freedom from PSS.

Effects of postexsanguination vascular infusion of cattle with a solution of saccharides, sodium chloride, phosphates, and vitamins C, E, or C+E on meat display-color stability.

Grain-finished, high-percentage Charolais steers (n = 36) were selected for uniformity. Immediately after jugular vein exsanguination, 27 steers were infused at 10% of live weight via the carotid artery with a solution developed by MPSC, Inc. (St. Paul, MN) consisting of 98.52% water, 0.97% saccharides, 0.23% sodium chloride, and 0.28% phosphate blend plus either 500 ppm vitamin C (MPSC+C; n = 9), 500 ppm vitamin E (MPSC+E; n = 9), or 500 ppm vitamin C + 500 ppm vitamin E (MPSC+C+E; n = 9). Uninfused controls (CON) were exsanguinated conventionally. Carcasses were fabricated at 48 h postmortem. Longissimus thoracis (LT), psoas major (PM), and semimembranosus (SM) muscles were removed, vacuum-packaged, and stored at 2 degrees C until 14 d postmortem. Then, steaks 2.54 cm thick were sliced from the three muscles, placed on foam trays, and overwrapped with polyvinyl chloride film. Ground beef (GB) was formulated from the quadriceps femoris to contain 20% fat, mounded into 0.45-kg portions, placed on styrofoam trays, and wrapped with polyvinyl chloride film. Steaks were visually evaluated for uniformity and initial color on display d 0. Instrumental color measurements of L*, a*, b* and trained sensory panel color evaluations were obtained daily for 4 d (PM and GB) or 5 d (LT and SM) of display. No display time x treatment interaction existed for L*, a*, or b* values. The LT from CON cattle had more uniform color (P < 0.05) and was more cherry red than that from all infused cattle on d 0. Visual scores indicated that GB from MPSC+E cattle was more red (P < 0.05) than that from MPSC+C infused cattle throughout display, and GB from MPSC+E cattle was more red (P < 0.05) than that from CON cattle for the last 3 d of display. The vascular infusion solutions generally did not improve color or display-color stability of steaks, but the infusion solution with vitamin E did improve display-color stability of GB.

Further evidence on the inheritance of halothane reaction in pigs.

One thousand six hundred forty-one Pietrain, 163 Minnesota No. 1 and 158 Pietrain X Minnesota No. 1 crosses and their reciprocals were tested for porcine stress syndrome susceptibility using halothane gas between September 1975 and July 1981. The frequency of reactors in the Pietrain breed was 93.9%. Matings of reactor males to reactor females within the Pietrain breed resulted in 632 reactor and 14 nonreactor offspring. A boar, judged to be nonpenetrant on the basis of the halothane reaction of his parents and littermates, was shown by progeny test to be homozygous for the halothane allele. These data indicate that halothane sensitivity is due to a single autosomal recessive gene with a penetrance of about 98% and a frequency of .98 in this Pietrain herd. No halothane reactors were found in the 163 Minnesota No. 1 pigs tested. Only one Pietrain X Minnesota No. 1 gilt reacted positively to halothane and later, the same gilt produced both positive and negative offspring, indicating that she was most likely heterozygous. Blood group typing of 107 crossbred pigs provided insufficient information to predict accurately the halothane reaction, although some associations were observed between the A and H loci and halothane sensitivity. The time taken by Pietrain pigs to react to halothane was measured and recorded. Analysis of these data showed that progeny of some sires had significantly faster reaction times than others and that reaction time had decreased over the years. These results as well as other data presented here indicate the existence of certain modifier genes that influence halothane reaction.

Effects of postexsanguination vascular infusion of carcasses with calcium chloride or a solution of saccharides, sodium chloride, and phosphates on beef display-color stability.

Hereford x Angus crossbred steers (n = 36) were stunned, exsanguinated, and infused via the carotid artery either with an aqueous solution containing 98.52% water, 0.97% saccharides, 0.23% sodium chloride, and 0.28% phosphates (MPSC; n = 12) or with 0.3 M CaCl2 (n = 12). The remaining 12 steers served as noninfused controls. At 48 h postmortem, the quadriceps muscles and subcutaneous fat were removed from the carcasses, frozen, and later made into ground beef (18 to 20% fat). The longissimus lumborum (LL), semimembranosus, and psoas major (PM) also were removed, vacuum packaged, aged until 14 d postmortem, and then one steak was sliced from each muscle for visual and instrumental color evaluations. The inside (ISM) and outside (OSM) portions of the SM were evaluated separately. The LL and OSM steaks from MPSC-infused carcasses had a lighter red (P < 0.05) initial appearance than steaks from the other treatments. The LL steaks from noninfused carcasses had the most (P < 0.05) uniform color; the MPSC treatment was intermediate, and the CaCl2 treatment was the most two-toned. Steaks from both infusion treatments had higher (P < 0.05) L* values for the LL, ISM, and OSM muscles compared with noninfused carcasses. In general, the LL from CaCl2-infused carcasses had lower (P < 0.05) a* values, saturation indices, and 630 nm to 580 nm reflectance values, and had larger (P < 0.05) hue angles. Infusion with MPSC increased (P < 0.05) hue angles in the LL and OSM. Display color stability was lowest (P < 0.05) for LL steaks from CaCl2-infused carcasses, whereas steaks from MPSC-infused carcasses were lighter red in initial color, but otherwise had display color stability similar to those from noninfused carcasses. No differences (P > 0.05) due to infusion were found for any color traits for the PM muscle and ground beef. Carotid artery vascular infusion of carcasses with CaCl2 resulted in undesirable meat colors, whereas the MPSC solution lightened loin and inside round color in a desirable way, but the color stability was slightly less compared to muscle from noninfused carcasses. Infusion effects were not consistent among muscles, and further research will be needed to determine what caused these differences.

Effects of vascular infusion with a solution of saccharides, sodium chloride, and phosphates with or without vitamin C on carcass traits, Warner-Bratzler shear force, flavor-profile, and descriptive-attribute characteristics of steaks and ground beef from Charolais cattle.

Two groups of 18 grain-finished steers were utilized. Nine from one group were infused via the carotid artery immediately after jugular vein exsanguination with an aqueous solution containing saccharides, NaCl, and phosphates (MPSC; MPSC, Inc., Eden Prairie, MN, USA). Nine steers served as non-infused controls (CON). An additional 18 steers were infused with either MPSC (n=9) or MPSC plus 1000 ppm vitamin C (MPSC+C, n=9) solutions. Steers infused with MPSC had higher dressing percentages and organ weights than CON steers. Vascular infusion with MPSC had no effects on USDA yield or quality grade traits, descriptive-attribute sensory panel evaluations, or Warner-Bratzler shear force of longissimus lumborum and semitendinosus muscles. Vascular infusion with MPSC resulted in some significant, but inconsistent effects on flavor-profile characteristics of cooked beef. The addition of vitamin C to the MPSC solution did not provide any benefit.

Malignant hyperthermia-triggering liability of selected inhalant anesthetics in swine.

Twenty pigs, known to be malignant hyperthermia (MH)-susceptible by previous MH-positive [MH(+)] reactions to halothane testing, were given 5 inhalant anesthetics. The number of MH(+) responses evoked by the different anesthetics varied. In response to halothane, isoflurane, and enflurane, MH(+) reactions were exhibited by 12, 11, and 7 pigs, respectively. The number of MH(+) reactions to nitrous oxide (0) and methoxyflurane (1) was significantly (P = 0.02) less. In the present study, all 4 halogenated-hydrocarbon volatile liquid inhalant anesthetics were capable of evoking a MH(+) reaction in susceptible pigs.

Porcine malignant hyperthermia susceptibility: erythrocytic osmotic fragility.

Erythrocyte osmotic fragility was determined in 27 Pietrain swine which were susceptible to malignant hyperthermia (MH), 29 Yorkshire swine which were resistant to MH (controls), and 50 crossbred swine (Pietrain x Yorkshire), half of which were MH susceptible. Halothane challenge tests and blood creatine kinase activity were used as criteria for determining MH susceptibility. Mean values for osmotic fragility of erythrocytes in concentrations of NaCl between 60 and 120 mM were significantly different for the 3 groups (P less than 0.001). Hemolysis (50%) of erythrocytes occurred at NaCl concentrations of 90 mM for Pietrains, 85 mM for crossbreds, and 78 mM for controls. Increased fragility values occurred in 96% of the Pietrains, 3% of the controls, and 42% of crossbred swine that were halothane test-positive, and 58% of halothane test-negative crossbreds (P less than 0.05). The mean time of onset of signs of MH in response to halothane challenge testing was twice as long in the crossbreds as in Pietrains (P less than 0.01). Reticulocyte counts were moderately high in blood samples from both the Pietrains (P less than 0.001) and the crossbreds (P less than 0.05). Of the swine which were tested for erythrocyte selenium-dependent glutathione peroxidase activity, values were within acceptable laboratory limits in 18 of 20 Pietrains, 14 of 14 halothane test-negative crossbreds, and 8 of 8 halothane test-positive crossbreds. In 2 of 20 Pietrains, a 35% deficiency of this enzyme was found. Heinz bodies were not detected in erythrocytes examined from 21 Pietrains, 20 crossbred swine (8 halothane test positives), and 12 controls.(ABSTRACT TRUNCATED AT 250 WORDS)

The role of tendon stiffness in development of equine locomotion with age.

REASONS FOR PERFORMING STUDY: The flexor tendons support the metacarpophalangeal (MCP) and distal interphalangeal (DIP) joints during stance phase and since tendon stiffness and strain changes with age, it is likely that kinematics are also age-dependent. HYPOTHESIS: Maximum MCP and DIP angles decrease in the young horse, plateau in the mature horse and increase towards senescence. METHODS: The distal limbs of 57 walking horses age 3-212 months were filmed and digitised with an automated tracking system. Maximum MCP and DIP angles during stance phase were used to calculate strain in the superficial and deep digital flexor tendons. Horses were divided into 3 age groups; young (3-35 months), mature (36-99 months) and older horses (100-212 months). Pearson's correlation coefficients were calculated to determine the relationship between age and kinematics. RESULTS: Tendon strain decreased in young horses, stayed constant in mature horses and increased in older horses. Joint angles showed significant negative correlation in young horses, with coefficients of -0.88 (MCP) and -0.81 (DIP). In mature horses, correlations were not significant (P = 0.2 for MCP; P = 0.5 for DIP). In older horses, angles showed significant positive correlation, with coefficients of 0.62 (MCP) and 0.48 (DIP). CONCLUSIONS: Joint angles decreased in the young horse as tendon stiffness increases, remained constant in the mature horse where tendon stiffness is constant and increased in older horses as tendons weakens and stiffness decreases. Strain patterns were similar to those found in vitro. POTENTIAL RELEVANCE: Changing tendon stiffness appeared to influence the development and degeneration of gait. This has implications for studying musculoskeletal development, especially for identification of normal and pathological development.