Identification of bacteria in total mixed ration silage produced with and without crop silage as an ingredient.

As a forage source for total mixed ration (TMR) silage production, locally produced crop silage is now used in addition to imported hay. This type of TMR ensiling is regarded as a two-step fermentation process; hence, a survey was carried out to determine whether the bacteria in crop silage affect the subsequent TMR ensiling. Fermentation product contents and bacterial community were determined for TMR silage and its ingredient silages collected in August, October and November. August product contained corn, sorghum and Italian ryegrass silages, October product had wheat silage exclusively and November product did not include any crop silages. Acetic acid, lactic acid, 2,3-butanediol and ethanol were predominant fermentation products in corn, sorghum, Italian ryegrass and wheat silages, respectively. Robust lactic acid fermentation was seen in TMR silage, even if acetate-type and alcohol-type silages were mixed as ingredients. The finding that bacterial community of the TMR silage appeared unrelated to those of ingredient silage supported this. Silages of various fermentation types can therefore be formulated without interfering with lactate-type fermentation in TMR silage.

Identification of bovine hibernation-specific protein complex and evidence of its regulation in fasting and aging.

Hibernation-specific protein (HP) is a plasma protein that regulates hibernation in chipmunks. The HP complex (HP20c) consists of three homologous proteins, HP20, HP25 and HP27, all produced by liver and belonging to the C1q family. To date, HP20c has not been identified in any mammalian species except chipmunk and ground squirrel hibernators. Here, we report a bovine HP20 gene isolated from liver tissue and aortic endothelial cells. Total homology between bovine and chipmunk variants was 63% at the amino acid level. Gene expression was highest in the liver. Western blot revealed HP20 protein in foetal, newborn, calf and adult serum, with highest concentrations in the adult. Similar proteins were detected in sera of other ruminants but not in humans, bears, mice or rats. Bovine HP20 protein was found mainly in ovaries, stomach, heart, kidneys, lungs, testes and prostate, but not in the skeletal muscle. Native HP20 was purified from bovine adult serum as a complex containing 25 and 27 kDa proteins. Mass spectrometry revealed that these proteins are orthologues of chipmunk HP25 and HP27, respectively. Interestingly, bovine HP20 was highly expressed in cattle serum after fasting. Native bovine HP20c may be a useful tool for investigating HP function.

Isolation of bovine milk-derived microvesicles carrying mRNAs and microRNAs.

By a series of centrifugation and ultracentrifugation, we could isolate microvesicles with approximately 100 nm in diameter from bovine milk. We also found that approximately 1700 and 1000 ng of total RNA, in which small RNAs were major components, was contained inside the microvesicles isolated from 6 ml of colostrum and mature milk, respectively, despite high RNase activity in the milk. Polyadenylated gene transcripts for major milk proteins and translation elongation factor-1alpha (EF-1alpha) were present in the microvesicles, and integrity of some transcripts was confirmed by real-time PCR targeting 5'- and 3'-ends of mRNA and by in vitro translation analysis. Moreover, a considerable amount of mammary gland and immune-related microRNAs were present in the milk-derived microvesicles. Acidification of milk to mimic gastrointestinal tract did not mostly affected RNA yield and quality. The milk related gene transcripts were detected in cultured cells when incubated with milk-derived microvesicles, suggesting cellular uptake of the microvesicle contents including RNA. Our findings suggest that bovine breast milk contains RNAs capable for being transferred to living cells and involved in the development of calf's gastrointestinal and immune systems.

Tc-99m MAA SPECT in pectus excavatum: assessment of perfusion volume changes after correction by the Nuss procedure.

PURPOSE: The Nuss procedure is a minimally invasive technique for the repair of pectus excavatum. This investigation attempted to assess the impact of this corrective protocol on pulmonary functional volume through analysis of distribution volume data derived from Tc-99m MAA SPECT. MATERIAL AND METHODS: SPECT was performed in 3 patients before and 6 months after completion of the Nuss procedure. Before the clinical application of SPECT, phantom experiments were conducted to establish the threshold level based on the volume calculation program. Haller's index, a quantitative index of chest-wall deformity, was also obtained by x-ray CT. RESULTS: A threshold level of 30% was the most accurate determinant of pulmonary volume. Preoperative and postoperative Tc-99m MAA distribution volumes (MAA volume in mL) were 2812 and 3107 (an increase of 10%), 2212 and 2410 (9%), and 2341 and 2872 (23%), respectively. In all patients, MAA volume increased after corrective intervention. Changes were most striking in the left lung, which is affected to a greater extent by dislocation of the heart. Morphologic improvement in the lungs was also demonstrated by Haller's CT index in all patients, which decreased from 4.35, 3.52, and 10.67, preoperatively, to 3.45, 2.75, and 4.25, respectively, postoperatively. CONCLUSION: MAA volume assessment is an easy, suitable approach for detection of favorable changes produced by the Nuss procedure in pectus excavatum.