Phosphofructokinase and Malate Dehydrogenase Participate in the In Vitro Maturation of Porcine Oocytes

Oocyte maturation depends on the metabolic activity of cumulus–oocyte complex (COC) that performs nutritive and regulatory functions during this process. In this work, the enzymes [phosphofructokinase (PFK) and malate dehydrogenase (MDH)] were tested to elucidate the metabolic profile of porcine COCs during the in vitro maturation (IVM). Enzymatic activity was expressed in U/COC and U/mg protein (specific activity) as mean ± SEM. In vitro maturation was performed with 2‐oxoglutarate (5, 10 and 20 mm ) or hydroxymalonate (30, 60 and 100 mm ) inhibitors of PFK and MDH, respectively. The PFK and MDH activities (U) remained constant during maturation. For PFK, the U were (2.48 ± 0.23) 10^?5 and (2.54 ± 0.32) 10^?5, and for MDH, the U were (4.72 ± 0.42) 10^?5 and (4.38 ± 0.25) 10^?5 for immature and in vitro matured COCs, respectively. The specific activities were significantly lower after IVM, for PFK (4.29 ± 0.48) 10^?3 and (0.94 ± 0.12) 10^?3, and for MDH (9.08 ± 0.93) 10^?3 and (1.89 ± 0.10) 10^?3 for immature and in vitro matured COCs, respectively. In vitro maturation percentages and enzymatic activity diminished with 20 mm 2‐oxoglutarate or 60 mm hydroxymalonate (p < 0.05). Viability was not affected by any concentration of the inhibitors evaluated. The U remained unchanged during IVM; however, the increase in the total protein content per COC provoked a decrease in the specific activity of both enzymes. Phosphofructokinase and MDH necessary for oocyte IVM would be already present in the immature oocyte. The presence of inhibitors of these enzymes impairs the meiotic maturation. Therefore, the participation of these enzymes in the energy metabolism of the porcine oocyte during IVM is confirmed in this study.     

Infectivity and virulence of Australian strains of Moraxella bovis for the murine and bovine eye in relation to pilus serogroup sub-unit size and degree of piliation

The degree of piliation of 29 haemolytic and 4 non-haemolytic Australian strains of Moraxella bovis representing 7 different pilus antigen groups was determined. The infectivity and virulence for the eye was measured in steroid-treated mice and in cattle. Non-piliated strains failed to infect the murine eye. Most moderately or heavily piliated strains reproducibly produced the highest infectivity and virulence scores in mice when compared with lightly or very lightly piliated strains (p less than 0.05). Non-haemolytic, piliated strains were infective and in one instance virulent for mice. Almost similar levels of infectivity and virulence were observed for 7 representative haemolytic strains tested in both cattle and mice. The relative molecular weight of pilin sub-units was compared using sodium dodecyl-sulphate polyacrylamide gel electrophoresis. Three classes of pili, alpha, beta and gamma of ascending sub-unit size were identified among the 7 pilus antigen serogroups. Pilin sub-unit size bore no relationship to the degree of piliation but most strains that were highly virulent in mice and cattle expressed alpha and gamma sub-units. Some strains appeared capable of switching from alpha to beta or form beta to gamma sub-unit production.     

Tenderness, sensory, and color attributes of two muscles from the M. quadriceps femoris when fabricated using a modified hot-boning technique

The M. quadriceps femoris from USDA Choice (n = 12) and USDA Select (n = 12) carcasses were fabricated traditionally (COLD) or innovatively (HOT), in which the seams it shares with the top round and bottom round were separated prerigor to evaluate positional and locational effects on Warner-Bratzler shear force (WBSF), sensory attributes, and objective color. At slaughter, paired USDA Choice and USDA Select carcasses were alternately assigned either the HOT or COLD treatment. At 48 h postslaughter, subprimals were removed, vacuum-packaged, and aged for an additional 5 d. After aging, the M. quadriceps femoris was cut into 2.54-cm-thick steaks and allowed to bloom 1 h. For the M. rectus femoris (REC) and M. vastus lateralis (VAL), L* values significantly (P < 0.050) decreased when moving from the proximal to distal position within the muscle. Similarly, a* and b* values decreased in the VAL when moving from the proximal to the distal aspect. After color measurement, steaks were vacuum-packaged and frozen (-26 degrees C) until shear and sensory data were collected. Significant position (proximal to distal) and location effects (cranial to caudal) were noted for both muscles. However, treatment did not affect WBSF of the VAL. Although intramuscular variation existed, WBSF and sensory panel tenderness ratings were acceptable for the REC. Although WBSF values were greater and tenderness ratings were less than the REC, the VAL were not extremely tough and therefore could be used in enhancement applications.     

Effects of aging on beef chuck and loin muscles enhanced with ammonium hydroxide and salt

The objective of this study was to determine whether aging would alter the beneficial effects (tenderness, juiciness, and flavor) of enhancing beef chuck and round steaks with a 20% solution of water, ammonium hydroxide, salt, and carbon monoxide. A randomized, complete block design was used, with 2 treatments [0% (control) and 20% (pump)], 3 aging periods (1, 2, and 3 wk), 3 muscles (M. triceps brachii from the clod heart, M. biceps femoris from the sirloin cap, and M. rectus femoris from the knuckle), and 3 replications. There were a total of 12 subprimals per treatment per aging period (n = 72 each). Individual steaks were cut to a thickness of 2.54 cm and packaged in a high-oxygen modified-atmosphere package (80% oxygen, 20% carbon dioxide). At the end of the aging period, steaks were vacuum packaged and frozen. Steaks were used to determine Warner-Bratzler shear force (WBS) and consumer sensory ratings. For all muscles, WBS values were lower for pump steaks than control at every aging period (P < 0.050). In addition, as aging period increased, WBS values for all steaks increased. Consumer taste panels revealed more desirability for tenderness, juiciness, flavor, and overall acceptability for pump steaks than control steaks. In addition, steaks aged for 1 d were more desirable than steaks aged for 7 and 14 d (P < 0.050). These data indicate that aging does not decrease the benefits (tenderness, juiciness, and flavor) of enhancement.     

Heat Shock Protein 70 and Sex Steroid Receptors in the Follicular Structures of Induced Ovarian Cysts

The purpose of this study was to estimate the expression and relative amounts of estrogen (ER) and progesterone receptors (PR) and their isoforms as well as heat shock protein 70 (HSP70) in ovaries of rats with induced cystic ovarian disease (COD). Primary, secondary, tertiary, atretic and cystic follicles were evaluated by immunohistochemistry and total ovarian proteins were analyzed by Western blot. In the granulosa layer, growing and cystic follicles in the treated group have a higher expression of ERα than growing follicles of control individuals. In the theca interna layer, tertiary follicles presented a significantly higher expression of ERα in the treated group. An increase in total ERα protein was detected in the treated group. Granulosa cells of all growing, atretic and cystic follicles show a lower expression of ERβ in animals with COD, and the total protein expression of ERβ was lower in this group. The expression of PR was lower in the granulosa cell layer of tertiary and cystic follicles in treated animals, and theca interna layer had less intense immunostaining in this group. Although there were no differences in the expression of PR-B by Western blotting, the expression of PR-A was higher and the expression of PR-C was smaller in the treated group. An intense HSP70 immunostaining was observed in the cells of cystic follicles. By Western blotting, higher protein expression of HSP70 was detected in the ovarian samples of the control group than those of the treated ones. Ovaries of animals with COD exhibited an altered steroid receptor expression and subtype balance as compared with control animals, and an increase in HSP70 immunoexpression.     

Testing Usability of Butylated Hydroxytoluene in Conservation of Goat Semen

The objective of this study was to investigate whether butylated hydroxytoluene (BHT) could be used as a suitable supporter or alternative of egg yolk during preservation of goat spermatozoa. Three in vitro experiments and a fertility test were conducted to evaluate the effect of BHT on viability of chilled‐stored semen as well as motility and kidding rate of frozen‐thawed spermatozoa. In the first two experiments, ejaculates (n = 30/experiment) were collected from 10 bucks, split, diluted with egg yolk‐based and egg yolk‐free extenders supplemented with or without 0.3, 0.6, 2, 5 and 8 mm BHT and stored at 5°C for 168 h. In the third experiment, 30 ejaculates were collected from the above‐mentioned bucks, split and diluted with egg yolk‐free extenders supplemented with or without 0.3, 0.6 and 0.9 mm BHT and egg yolk‐based extenders supplemented with or without 5 mm BHT. Diluted semen was cooled to 5°C over a period of 4 h, frozen and thawed in the form of 0.3‐ml pellets. In the fertility test, 75 ejaculates were collected from two proven fertile bucks, split, diluted with egg yolk‐free extenders containing 0.6 mm BHT and egg yolk‐based extenders supplemented with or without 5 mm BHT, frozen and thawed as described above. An insemination volume of 0.6 ml containing 120–140 × 10⁶ progressively motile spermatozoa was used for a single cervical insemination of cloprostenol‐synchronized does (n = 230). The results showed that addition of 5 mm BHT to egg yolk‐deficient (2.5%) extenders significantly improved viability of chilled‐stored semen together with motility (48.5%) and fertility (62.5%) of frozen‐thawed spermatozoa. Replacement of egg yolk in semen extenders by 0.6 mm BHT could sustain not only viability of chilled‐stored semen but also post‐thaw motility (47.5%) and fertility (53.75%) of frozen‐thawed spermatozoa. In conclusion, supplementation of semen diluents with BHT can ameliorate preservability of goat sperm.     

Chemical and sensory properties of beef of known source and finished on wet distillers grains diets containing varying types and levels of roughage

Beef knuckles (n = 160) were obtained from source-verified cattle finished on 30% wet distillers grains plus solubles enriched with varying levels of alfalfa hay (4 or 8%), corn silage (6 or 12%), or corn stalks (3 or 6%) based on NDF. Proximate analysis, pH, oxidation-reduction potential, fatty acid composition, and sensory analysis were conducted on the rectus femoris muscle to determine if roughage inclusion, in conjunction with wet distillers grains plus solubles and cattle source, affects beef flavor with particular interest in liver-like off-flavor. Proximate analysis, fat content, and oxidation-reduction potential were unaffected (P ge;0.129) by diet or source. For s.c. adipose tissue, cattle from Nebraska (NE) had greater amounts of MUFA (P = 0.048) and unsaturated fatty acids (P = 0.068) but less SFA (P = 0.065) when compared with cattle from South Dakota. Diet affected s.c. adipose tissue levels of 15:0, 17:0, and n-3 fatty acids in which cattle from NE finished on the low corn stalk diet had (P < or =0.050) lower levels. Cattle from NE had (P < or = 0.049) greater i.m. adipose proportions of 13:0 and CLA. Dietary effects (P < or = 0.050) were observed for i.m. adipose tissue proportions of 16:0, 18:1(n-9), 18:2(n-6), 20:4(n-6), 22:5(n-3), MUFA, PUFA, and n-6 fatty acids. Sensory analysis revealed that cattle from NE were (P < or = 0.023) less juicy and had less bloody notes when compared with cattle from South Dakota. Cattle finished on the low alfalfa diet were (P < or = 0.014) more tender and juicy but had more bloody notes. No (P ge; 0.670) dietary or source effects were noted for liver-like off-flavor. Subcutaneous amounts of 18:2(n-6 trans) (r = -0.17) were inversely related to the incidence of liver-like off-flavor, whereas 20:1(n-9) (r = 0.21), CLA cis-9, trans-11 (r = 0.16) were directly related. Data from this study indicate that type and level of roughage inclusion and cattle source have minimal effects on fatty acid profiles and sensory properties of the musculus rectus femoris. However, individual fatty acids of s.c. and i.m. adipose tissue were significantly correlated with liver-like off-flavor.     

A "Schrodinger Cat" Superposition State of an Atom

A "Schrodinger cat"-like state of matter was generated at the single atom level. A trapped 9Be+ ion was laser-cooled to the zero-point energy and then prepared in a superposition of spatially separated coherent harmonic oscillator states. This state was created by application of a sequence of laser pulses, which entangles internal (electronic) and external (motional) states of the ion. The Schrodinger cat superposition was verified by detection of the quantum mechanical interference between the localized wave packets. This mesoscopic system may provide insight into the fuzzy boundary between the classical and quantum worlds by allowing controlled studies of quantum measurement and quantum decoherence.     

基于3D-CDF（2,2）和3D SPIHT的序列图像压缩编码研究

研究了3D DWT和3D SPIHT算法,用CDF（2,2）双正交小波为帧内小波变换的小波基,考虑到边界延拓效应,时间维小波变换也选用CDF（2,2）双正交小波为时间维小波变换的小波基,实验结果表明算法对于视频序列图像压缩是非常有效的,其压缩效果明显优于基于3D-DCT的压缩编码算法.