Intermuscular variation in tenderness: association with the ubiquitous and muscle-specific calpains

The biochemistry of intermuscular variation in tenderness is not fully understood. To investigate the role of the calpains in this process we performed two experiments using bovine and ovine species. In the bovine experiment, two distinct muscles, longissimus thoracis et lumborum (LT) and psoas major (PM), were used. In the ovine experiment, four muscles, LT, PM, semimembranosus (SM), and semitendinosus (ST), were used. Muscles were sampled at death for the determination of the steady-state mRNA level of calpains and calpastatin and the activities of calpain 1, 2, and calpastatin. Muscles were also sampled to determine the temporal changes in pH, tenderness, and the activity of the ubiquitous calpain system during postmortem aging. The results of the relative rate of tenderization in both species was found to be related to muscle type; LT had the highest value in both species. Within species, the mRNA steady-state levels of calpain 1 and calpastatin were similar in various bovine and ovine muscles. Bovine calpain 2 mRNA level was significantly lower in the LT than in the PM. Ovine calpain 2 mRNA level was lower, but not significantly different, in the LT compared to the other muscles. The mRNA level of bovine calpain 3 was significantly higher in the LT muscle than in the PM. In the ovine, the mRNA level of calpain 3 was highest in the LT, followed by SM, PM, and ST. Results on the activity of the ubiquitous calpain system in various muscles at death were dependent on muscle type and species. Temporal changes in the activity of calpains and calpastatin during the first 24 h of postmortem aging were similar in the muscles studied: calpain 1 and calpastatin declined significantly and calpain 2 remained relatively unchanged. The temporal changes in muscle pH in both experiments indicated that the extent and rate of pH decline during aging was related to muscle type. Correlation analysis between the relative rate of tenderization and mRNA expression of calpains revealed a strong relationship with calpain 3 in both species.     

Production of Lambs After the Transfer of Fresh and Cryopreserved in vitro Produced Embryos from Prepubertal Lamb Oocytes and Unsorted and Sex-sorted Frozen-thawed Spermatozoa

Cumulus-oocyte complexes from hormone-stimulated 3-4-week-old (n=43) and 6-7-week-old (n=12) prepubertal lambs were matured in vitro and incubated with unsorted, or X- or Y-spermatozoa separated with a high-speed cell sorter (SX MoFlo)frozen-thawed. Presumptive zygotes were then cultured to the blastocyst stage, and transferred to recipients fresh or after cryopreservation (frozen). Oocyte cleavage was higher (p <0.05) with unsorted (515/926, 55.6%) than X- or Y-spermatozoa (261/672, 38.8% and 229/651, 35.2%, respectively) and blastocyst formation (% zygotes) by Day 9 of in vitro culture was lower (p <0.05) for X- (102/261, 39.1%) than unsorted spermatozoa (249/515, 48.3%), but did not differ between Y-spermatozoa (103/229, 45.0%) and unsorted spermatozoa, or between X- and Y-spermatozoa (p >0.05). For fresh embryos, survival to term was 50.0% (3/6) for unsorted, 0.0% (0/6) for X- and 16.7% (1/6) for Y-spermatozoa-derived embryos (p >0.05), and for frozen embryos was 4.0% (2/50) for unsorted, 9.1% (2/22) for X- and 2.9% (1/34) Y-spermatozoa-derived embryos (p >0.05). Of the two lambs born from X-spermatozoa-derived embryos, one was female (50%), and from the two Y-spermatozoa-derived lambs, both were male (100%), demonstrating that lambs can be produced after the transfer of fresh and cryopreserved IVP embryos derived from prepubertal lamb oocytes and frozen-thawed sex-sorted sperm.     

Direct observations of structural phase transitions in planar crystallized ion plasmas

Laser-cooled 9Be+ ions confined in two-dimensionally extended lattice planes were directly observed, and the images were used to characterize the structural phases of the ions. Five different stable crystalline phases were observed, and the energetically favored structure could be sensitively tuned by changing the areal density of the confined ions. The experimental results are in good agreement with theoretical predictions for the planar (infinite in two dimensions) one-component plasma. Qualitatively similar structural phase transitions occur, or are predicted to occur, in other experimentally realizable planar systems.     

Improving the health of the global poor

We analyzed the technical basis for a major global program to reduce disease among the poor. Effective interventions exist against the few diseases which most account for excess mortality among the poor. Achieving high coverage of effective interventions requires a well-functioning health system, as well as overcoming a set of financial and nonfinancial constraints. The annual incremental cost would be between $40 billion and $52 billion by 2015 in 83 low-income and sub-Saharan African countries. Such a program is feasible and would avoid millions of child, maternal, and adult deaths annually in poor countries.     

Localization of Transforming Growth Factor Beta3 in Squamous Metaplasia of the Porcine Oviduct: a Case Report

Squamous metaplasia of the oviduct epithelium is a rare disorder of reproductive organs. We noted squamous metaplasia of the oviduct epithelium in a sow routinely slaughtered at day 2 of the oestrous cycle. Expression of transforming growth factor beta3 (TGF beta3) in the metaplastic epithelia was evaluated by immunohistochemistry, because TGF beta3 appears to play a key role as regulator of a variety of tissue remodelling events. Our results show that TGF beta3 immunostaining was specifically localized to foci of squamous metaplasia of the epithelial linings. Non‐metaplastic epithelial cells of the oviduct were not immunostained with anti‐TGF beta3 antibody. At the subcellular level, TGF beta3‐labelled cells occasionally showed signs of apoptotic cell death. It is concluded that signals produced by TGF beta3 in metaplastic lesions of the oviduct are potentially involved in pathophysiological processes.     

The effect of organophosphate insecticides on adrenal corticosterone formation

In vitro and in vivo experiments with Sprague-Dawley rats showed that the three organophosphate insecticides tested (see below) depressed endogenous corticosterone synthesis and blocked corticosteroidogenesis in response to ACTH and cAMP stimulation of a suspension of adrenal cells. Pregnenolone stimulation of adrenal cells was not inhibited at the insecticide concentrations which blocked the ACTH and cAMP stimulation of corticosteroidogenesis. It was concluded that the insecticides act beyond the site of action of ACTH and at or beyond the level of cAMP metabolism and prior to the metabolism of pregnenolone.Insecticides tested were: dichlorvos (O,O-dimethyl-O,2,2-dichlorovinyl phosphate), Dursban (phosphorothioic acid: O,O-diethyl O-3,5,6-trichloro-2-pyridyl ester), and Diazinon (phosphorothioic acid: O,O-diethyl O-(2-isopropyl-6-methyl-4-pyrimidinyl ester).