Processed fava bean as a substitute for rapeseed meal with or without rumen-protected methionine supplement in grass silage-based dairy cow diets.

Fava bean offers a sustainable home-grown protein source for dairy cows, but fava bean protein is extensively degraded in the rumen and has low Met concentration. We studied the effects of protein supplementation and source on milk production, rumen fermentation, N use, and mammary AA utilization. The treatments were unsupplemented control diet, and isonitrogenously given rapeseed meal (RSM), processed (dehulled, flaked, and heated) fava bean without (TFB) or with rumen-protected (RP) Met (TFB+). All diets consisted of 50% grass silage and 50% cereal-based concentrate including studied protein supplement. The control diet had 15% of crude protein and protein-supplemented diets 18%. Rumen-protected Met in TFB+ corresponded to 15 g/d of Met absorbed in the small intestine. Experimental design was a replicated 4 × 4 Latin square with 3-wk periods. The experiment was conducted using 12 multiparous mid-lactation Nordic Red cows, of which 4 were rumen cannulated. Protein supplementation increased dry matter intake (DMI), and milk (31.9 vs. 30.7 kg/d) and milk component yields. Substituting RSM with TFB or TFB+ decreased DMI and AA intake but increased starch intake. There were no differences in milk yield or composition between RSM diet and TFB diets. Rumen-protected Met did not affect DMI, or milk or milk component yields but increased milk protein concentration in comparison to TFB. There were no differences in rumen fermentation except for increased ammonium-N concentration with the protein-supplemented diets. Nitrogen-use efficiency for milk production was lower for the supplemented diets versus control diet but tended to be greater for TFB and TFB+ versus RSM. Protein supplementation increased plasma essential AA concentration but there were no differences between TFB diets and RSM. Rumen-protected Met clearly increased plasma Met concentration (30.8 vs. 18.2 µmol/L) but did not affect other AA. Absence of differences between RSM and TFB in milk production together with limited effects of RP Met suggest that TFB is a potential alternative protein source for dairy cattle.

Effects of faba bean, blue lupin and rapeseed meal supplementation on nitrogen digestion and utilization of dairy cows fed grass silage-based diets.

There is increasing interest in using locally produced protein supplements in dairy cow feeding. The objective of this experiment was to compare rapeseed meal (RSM), faba beans (FBs) and blue lupin seeds (BL) at isonitrogenous amounts as supplements of grass silage and cereal based diets. A control diet (CON) without protein supplement was included in the experiment. Four lactating Nordic Red cows were used in a 4 × 4 Latin Square design with four 21 d periods. The milk production increased with protein supplementation but when expressed as energy corrected milk, the response disappeared due to substantially higher milk fat concentration with CON compared to protein supplemented diets. Milk protein output increased by 8.5, 4.4 and 2.7% when RSM, FB and BL were compared to CON. The main changes in rumen fermentation were the higher propionate and lower butyrate proportion of total rumen volatile fatty acids when the protein supplemented diets were compared to CON. Protein supplementation also clearly increased the ruminal ammonia N concentration. Protein supplementation improved diet organic matter and NDF digestibility but efficiency of microbial protein synthesis per kg organic matter truly digested was not affected. Flow of microbial N was greater when FB compared to BL was fed. All protein supplements decreased the efficiency of nitrogen use in milk production. The marginal efficiency (amount of additional feed protein captured in milk protein) was 0.110, 0.062 and 0.045 for RSM, FB and BL, respectively. The current study supports the evidence that RSM is a good protein supplement for dairy cows, and this effect was at least partly mediated by the lower rumen degradability of RSM protein compared to FB and BL. The relatively small production responses to protein supplementation with simultaneous decrease in nitrogen use efficiency in milk production suggest that economic and environmental consequences of protein feeding need to be carefully considered.

Effects of different barley and oat varieties on methane production, digestibility, and fermentation pattern in vitro.

The objective of this in vitro study was to determine the effects of different barley and oat varieties on CH4 production, digestibility, and rumen fermentation patterns in dairy cows. Our hypothesis was that oat-based diets would decrease CH4 production compared with barley-based diets, and that CH4 production would differ between varieties within grain species. To evaluate this hypothesis, we conducted an in vitro experiment using a fully automated gas production technique, in which the total gas volume was automatically recorded by the system. The experiment consisted of triplicate 48-h incubations with 16 treatments, including 8 different varieties of each grain. The grain varieties were investigated as a mix with an early-cut grass silage (1:1 ratio of grain to silage on a dry matter basis) and mixed with buffered rumen fluid. We estimated predicted in vivo total gas production and CH4 production by applying a set of models to the gas production data obtained by the in vitro system. We also evaluated in vitro digestibility and fermentation characteristics. The variety of grain species did not affect total gas production, CH4 production, or fermentation patterns in vitro. However, in vitro-determined digestibility and pH were affected by variety of grain species. Grain species affected total gas and CH4 production: compared with barley-based diets, oat-based diets decreased total gas production and CH4 production by 8.2 and 8.9%, respectively, relative to dry matter intake. Grain species did not affect CH4 production relative to in vitro true dry matter digestibility. Oat-based diets decreased digestibility and total volatile fatty acid production, and maintained a higher pH at 48 h of incubation compared with barley-based diets. Grain species did not affect fermentation patterns, except for decreased molar proportions of valerate with oat-based diets. These results suggest that replacing barley with oats in dairy cow diets could decrease enteric CH4 production.

The effect of partial substitution of rapeseed meal and faba beans by Spirulina platensis microalgae on milk production, nitrogen utilization, and amino acid metabolism of lactating dairy cows.

Alternative protein sources such as microalgae and faba beans may have environmental benefits over rapeseed. We studied the effects of rapeseed meal (RSM) or faba beans (FB) as a sole protein feed or as protein feeds partially substituted with Spirulina platensis (spirulina) microalgae on milk production, N utilization, and AA metabolism of dairy cows. Eight multiparous Finnish Ayrshire cows (113 ± 36.3 d in milk; mean ± SD) were used in a balanced, replicated 4 × 4 Latin square with 2 × 2 factorial arrangement of treatments and 21-d periods. Four cows in one Latin square were rumen cannulated. Treatments were 2 isonitrogenously fed protein sources, RSM or rolled FB, or one of these sources with half of its crude protein substituted by spirulina (RSM-SPI and FB-SPI). Cows had ad libitum access to total mixed rations consisting of grass silage, barley, sugar beet pulp, minerals, and experimental protein feed. The substitution of RSM with FB did not affect dry matter intake (DMI) but decreased neutral detergent fiber intake and increased the digestibility of other nutrients. Spirulina in the diet decreased DMI and His intake. Spirulina had no effect on Met intake in cows on RSM diets but increased it in those on FB diets. Energy-corrected milk (ECM) and protein yields were decreased when RSM was substituted by FB. Milk and lactose yields were decreased in cows on the RSM-SPI diet compared with the RSM diet but increased in those on FB-SPI compared with FB. The opposite was true for milk fat and protein concentrations; thus, spirulina in the diet did not affect ECM. Feed conversion efficiency (ECM:DMI) increased in cows on FB diets with spirulina, whereas little effect was observed for those on RSM diets. The substitution of RSM by FB decreased arterial concentration of Met and essential AA. Spirulina in the diet increased milk urea N and ruminal NH4-N and decreased the efficiency of N utilization in cows on RSM diets, whereas those on FB diets showed opposite results. Met likely limited milk production in cows on the FB diet as evidenced by the decrease in arterial Met concentration and milk protein yield when RSM was substituted by FB. The results suggest the potential to improve milk production response to faba beans with supplementation of Met-rich feeds such as spirulina. This study also confirmed spirulina had poorer palatability than RSM and FB despite total mixed ration feeding and lower milk production when spirulina partially replaced RSM.

Effects of dry period energy intake on insulin resistance, metabolic adaptation, and production responses in transition dairy cows on grass silage-based diets.

High energy intake in the dry period has reportedly had adverse effects on mobilization of body reserves, dry matter intake, and productivity of dairy cows. We investigated whether grass silage (GS) fed ad libitum (high energy intake, HEI; 141% of daily metabolizable energy requirements) in an 8-wk dry period affects metabolic adaptation-specifically, peripheral insulin resistance-compared with a total mixed ration consisting of GS, wheat straw, and rapeseed meal (55/40/5%; controlled energy intake, CEI; 108% of metabolizable energy/d) fed ad libitum. Multiparous Ayrshire dairy cows (n = 16) were used in a randomized complete block design until 8 wk after parturition. Commercial concentrates were fed 1 and 2 kg/d during the last 10 to 6 and 5 to 0 d before the expected calving date, respectively. Postpartum, a similar lactation diet with ad libitum access to GS and increasing concentrate allowance (maximum of 16 kg/d) was offered to all. The HEI group gained more body weight and had higher plasma insulin, glucose, and ß-hydroxybutyrate concentrations than the CEI group prepartum. Postpartal plasma glucose tended to be higher and milk yield was greater from wk 5 onward for HEI compared with CEI cows. An intravenous glucose tolerance test (IVGTT) was performed at -13 ± 5 d and 9 ± 1 d relative to calving. The HEI cows had greater insulin response to glucose load and smaller area under the response curve for glucose than CEI cows in prepartal IVGTT. Thus, compensatory insulin secretion adapted to changes in insulin sensitivity of the peripheral tissues, preserving glucose tolerance of HEI cows. Higher insulin levels were needed in HEI cows than in CEI cows to elicit a similar decrement of nonesterified fatty acid concentration in prepartal IVGTT, suggesting reduced inhibition of lipolysis by insulin in HEI cows before parturition. In conclusion, high energy intake of moderately digestible GS with low concentrate feeding in the close-up dry period did not have adverse effects on metabolic adaptation, insulin sensitivity, and body mobilization after parturition. Instead, this feeding regimen was more beneficial to early-lactation performance than GS-based total mixed ration diluted with wheat straw.

Effect of incremental amounts of camelina oil on milk fatty acid composition in lactating cows fed diets based on a mixture of grass and red clover silage and concentrates containing camelina expeller.

Camelina is an ancient oilseed crop that produces an oil rich in cis-9,cis-12 18:2 (linoleic acid, LA) and cis-9,cis-12,cis-15 18:3 (α-linolenic acid, ALA); however, reports on the use of camelina oil (CO) for ruminants are limited. The present study investigated the effects of incremental CO supplementation on animal performance, milk fatty acid (FA) composition, and milk sensory quality. Eight Finnish Ayrshire cows (91d in milk) were used in replicated 4×4 Latin squares with 21-d periods. Treatments comprised 4 concentrates (12kg/d on an air-dry basis) based on cereals and camelina expeller containing 0 (control), 2, 4, or 6% CO on an air-dry basis. Cows were offered a mixture of grass and red clover silage (RCS; 1:1 on a dry matter basis) ad libitum. Incremental CO supplementation linearly decreased silage and total dry matter intake, and linearly increased LA, ALA, and total FA intake. Treatments had no effect on whole-tract apparent organic matter or fiber digestibility and did not have a major influence on rumen fermentation. Supplements of CO quadratically decreased daily milk and lactose yields and linearly decreased milk protein yield and milk taste panel score from 4.2 to 3.6 [on a scale of 1 (poor) to 5 (excellent)], without altering milk fat yield. Inclusion of CO linearly decreased the proportions of saturated FA synthesized de novo (4:0 to 16:0), without altering milk fat 18:0, cis-9 18:1, LA, and ALA concentrations. Milk fat 18:0 was low (<5g/100g of FA) across all treatments. Increases in CO linearly decreased the proportions of total saturates from 58 to 45g/100g of FA and linearly enriched trans-11 18:1, cis-9,trans-11 18:2, and trans-11,cis-15 18:2 from 5.2, 2.6, and 1.7 to 11, 4.3, and 5.8g/100g of FA, respectively. Furthermore, CO quadratically decreased milk fat trans-10 18:1 and linearly decreased trans-10,cis-12 18:2 concentration. Overall, milk FA composition on all treatments suggested that one or more components in camelina seeds may inhibit the complete reduction of 18-carbon unsaturates in the rumen. In conclusion, CO decreased the secretion of saturated FA in milk and increased those of the trans-11 biohydrogenation pathway or their desaturation products. Despite increasing the intake of 18-carbon unsaturated FA, CO had no effect on the secretions of 18:0, cis-9 18:1, LA, or ALA in milk. Concentrates containing camelina expeller and 2% CO could be used for the commercial production of low-saturated milk from grass- and RCS-based diets without major adverse effects on animal performance.

Effects of replacing rapeseed meal with fava bean at 2 concentrate crude protein levels on feed intake, nutrient digestion, and milk production in cows fed grass silage-based diets.

The objective of this study was to evaluate the production and physiological responses of dairy cows to the substitution of fava bean for rapeseed meal at 2 protein supplementation levels in grass silage-based diets. We used 6 primiparous and 6 multiparous Finnish Ayrshire cows in a cyclic changeover trial with a 2×3 factorial arrangement of treatments. The experimental diets consisted of formic acid-treated timothy-meadow fescue silage and 3 isonitrogenous concentrates containing either rapeseed meal, fava bean, or a 1:1 mixture of rapeseed meal and fava bean at low and high inclusion rates, resulting in concentrate crude protein (CP) levels of 15.4 and 19.0% in dry matter. Silage dry matter intake decreased linearly when rapeseed meal was replaced with fava bean, the negative effect being more distinct at the high CP level than the low (-2.3 vs. -0.9kg/d, respectively). Similarly, milk and milk protein yields decreased linearly with fava bean, the change tending to be greater at the high CP level than the low. Yield of milk fat was lower for fava bean compared with rapeseed meal, the difference showing no interaction with CP level. Especially at the high CP level, milk urea concentration was higher with fava bean compared with rapeseed meal indicating better utilization of protein from the rapeseed meal. The apparent total-tract organic matter digestibility did not differ between treatments at the low CP level, but digestibility was higher for fava bean than for rapeseed meal at the high CP level. Plasma concentrations of essential amino acids, including methionine and lysine, were lower for fava bean than for rapeseed meal. Compared with rapeseed meal, the use of fava bean in dairy cow diets as the sole protein supplement decreased silage intake and milk production in highly digestible formic acid-treated grass silage-based diets.

Effects of plant species, stage of maturity, and level of formic acid addition on lipolysis, lipid content, and fatty acid composition during ensiling.

Forage type and management influences the nutritional quality and fatty acid composition of ruminant milk. Replacing grass silage with red clover (RC; L.) silage increases milk fat 18:3-3 concentration. Red clover has a higher polyphenol oxidase (PPO) activity compared with grasses, which has been suggested to decrease lipolysis and . The present study characterized the abundance and fatty acid composition of esterified lipid and NEFA before and after ensiling of grass and RC to investigate the influence of forage species, growth stage, and extent of fermentation on lipolysis. A randomized block design with a 2 × 3 × 4 factorial arrangement of treatments was used. Treatments comprised RC or a mixture of timothy ( L.) and meadow fescue ( Huds.) harvested at 3 growth stages and treated with 4 levels of formic acid (0, 2, 4, and 6 L/t). Lipid in silages treated with 0 or 6 L/t formic acid were extracted and separated into 4 fractions by TLC. Total PPO activity in fresh herbage and the content of soluble bound phenols in all silages were determined. Concentrations of 18:3-3 and total fatty acids (TFA) were higher ( < 0.001) for RC than for grass. For both forage species, 18:3-3 and TFA content decreased linearly ( < 0.001) with advancing growth stage, with the highest abundance at the vegetative stage. Most of lipid in fresh RC and grass herbage (97%) was esterified, whereas NEFA accounted for 71% of TFA in both silages. Ensiling resulted in marginal increases in TFA content and the amounts of individual fatty acids compared with fresh herbages. Herbage total PPO activity was higher ( < 0.001) for RC than grass (11 vs. 0.11 µkatal/g leaf fresh weight). Net lipolysis during ensiling was extensive for both forage species (660 to 759 g/kg fatty acid for grass and 563 to 737 g/kg fatty acid for RC). Formic acid application (0 vs. 6 L/t) resulted in a marked decrease ( = 0.026) in net lipolysis during the ensiling of RC, whereas the opposite was true ( = 0.026) for grass. In conclusion, results suggest that formic acid addition during the ensilage of RC decreases lipolysis . For both plant species, total PPO activity was not associated with the extent of lipolysis . However, bound phenols formed via PPO activity appear to have a role in protecting lipid and protein against degradation in grass and lowering proteolysis of RC during ensiling.

Sense of coherence and health behaviour in adolescence.

AIM: To explore associations of Sense of coherence (SOC) with health behaviour and social competence among 15-year-old adolescents. METHODS: Study population is a prospective cohort of a randomized cluster sample of families and their first-born children from south-western Finland in 1986-1987. In this study, cross-sectional data of the 15-year-olds were used. The present data were based on mailed, pretested questionnaires. The outcome variable, SOC, was based on the 13-item scale of Antonovsky's Orientation to Life Questionnaire (OLQ). The principal explanatory variables were health behaviour, including experienced oral health, and social competence. The statistical analysis was performed using linear regression modelling. RESULTS: Strong SOC of adolescents associated significantly with lighter use of alcohol, being a non-smoker, better care of oral health and better social competence compared with the others. CONCLUSION: Sense of coherence is a useful tool for identifying adolescents in need of extra support and motivation for their health behaviour.

Dental fear: one single clinical question for measurement.

A new dental fear measurement instrument, the Short Dental Fear Question (SDFQ), was developed and tested for clinical practice purposes. The correlations of the SDFQ with the Dental Anxiety Scale (DAS) and the Dental Fear Survey (DFS) were tested in 15-16-year-old adolescents. The Spearman correlations (rs) between the dental fear measurement instruments were: SDFQ - DFS: r(s) = 0.79, n = 26; DFS - DAS: r(s) = 0.72, n = 26; SDFQ- DAS: r(s) = 0.69, n = 27. DAS and DFS mean scores were clearly higher in the SDFQ fear group than SDFQ in the relaxed group. The SDFQ is a short and compact instrument which might be convenient for the measurement of dental fear in clinical practice.

Screening and selection of lactic acid bacteria strains suitable for ensiling grass.

AIM: Lactic acid bacteria (LAB) strains shown to have broad-spectrum antimicrobial activity were screened for potential as grass silage inoculants. The strains capable of rapidly lowering the pH of the grass matrix and with low proteolytic activity were assessed in laboratory-scale silos in a grass matrix containing natural microbial flora. METHODS AND RESULTS: Screening of nine candidate strains was performed first in a grass extract medium. The four most promising strains were selected on the basis of growth rate in the medium, capacity to reduce pH and ability to limit the formation of ammonia-N. The efficiency of the selected strains was further assessed in a laboratory-scale ensiling experiment. Untreated (no additive) and formic acid served as controls. All tested inoculants improved silage quality compared with untreated. With one exception (Pediococcus parvulus E315) the fermentation losses in the inoculated silages were even lower than in the acid-treated control silage. Pure lactic acid fermentation was obtained in the timothy-meadow fescue silage with all inoculants. The results obtained in the ensiling experiments were consistent with those of the screening procedure, which appeared to predict correctly the potential of LAB as silage inoculants. The strains with a low ammonia production rate in the grass extract medium behaved similarly in the silage. Especially in this respect the strain Lactobacillus plantarum E76 was superior to the other candidates. CONCLUSIONS: The screening method using grass extract proved to be useful in strain selection. SIGNIFICANCE AND IMPACT OF THE STUDY: The rapid screening method developed for the LAB strains provides a useful tool for more systematic product development of commercial inoculant preparations. Time consuming and laborious ensiling experiments can be limited only to the most promising strains.

Effects of replacing grass silage with barley silage in dairy cow diets.

This study examined the effects of gradually replacing grass silage with whole-crop barley silage on feed intake, ruminal and total tract digestibility, and milk yield in lactating dairy cows. Four dairy cows in early lactation, equipped with rumen cannulas, were fed 4 diets over four 21-d periods. The diets consisted of 4 forage mixtures of grass silage and whole-crop barley silage supplemented with 8.9 kg/d of concentrates [dry matter (DM) basis]. The proportion of barley silage in the forage was adjusted to 0, 0.20, 0.40, and 0.60 kg/ kg of DM. Ruminal nutrient metabolism was measured on the basis of digesta flow entering the omasal canal. Ammonia concentrations and volatile fatty acid profiles were determined in the rumen fluid. Ruminal digestion and passage kinetics were assessed by the rumen evacuation technique. Replacement of grass silage with barley silage had no effect on DM, digestible organic matter, or neutral detergent fiber (NDF) intake, but starch intake increased, whereas nitrogen and digestible NDF (dNDF) intake decreased. Increases in the proportion of barley silage linearly decreased milk yield, and the molar proportion of acetate in the rumen, and increased that of propionate, butyrate, and valerate. Decreases in milk yield due to inclusion of barley silage were attributed to decreases in diet digestibility and nutrient supply to the animal. Barley silage linearly decreased organic matter digestibility in the total tract and NDF and dNDF digestibility in the rumen and the total tract, and decreased nonammonia N flow entering the omasal canal. No significant differences between diets were noted in the digestion rate of dNDF or passage rate of indigestible NDF from the rumen. Decreases in organic matter and NDF digestibility were attributed to the higher indigestible NDF concentration of barley silage compared with that of grass silage and to the smaller pool size of dNDF in the rumen.

Electron-spin-resonance instability in two-dimensional atomic hydrogen gas.

We study by electron-spin-resonance spin-polarized atomic hydrogen adsorbed on the surface of superfluid helium at temperatures T(S) from 50 to 110 mK. The average dipolar field in this 2D system shifts the electron-spin-resonance peak of the adsorbed atoms relative to that of bulk atoms. The shift is directly proportional to surface density. The role of longitudinal magnetization relaxation is played by particle exchange between the 2D and the 3D phases, which diminishes exponentially with decreasing T(S). Therefore at T(S) less, similar 80 mK an excitation field of 0.1 mG disturbs the equilibrium surface density and leads to a magnetization instability observed as sawtooth shaped resonance lines.

Digestive processes of dairy cows fed silages harvested at four stages of grass maturity.

The objective of this experiment was to quantify ruminal digestive processes that could help to identify factors limiting DMI when silages differing in grass maturity were fed to dairy cows. Four silages were harvested at 1-wk intervals from a primary growth of a timothy-meadow fescue sward, resulting in feeds with digestible OM content in DM (D-value) of 739, 730, 707, and 639 g/kg in the order of succeeding harvest date. Four ruminally cannulated dairy cows were given ad libitum access to these silages supplemented with 7 kg concentrate per day in a 4 x 4 Latin square design. Rumen function was clearly affected by decreasing digestibility of silage fed. Passage rate of digestible NDF (DNDF) and indigestible NDF (INDF) increased, but it could not prevent the accumulation of DM, NDF, DNDF, and INDF into the rumen when silages of progressing grass maturity were fed. The greatest proportional increases in rumen pool were found in INDF and in medium particles (separated by wet sieving and measuring 315 to 2,500 microm). The passage of medium INDF particles decreased (P < 0.01) linearly (from 0.0365/h to 0.0281/h) with increasing maturity of grass ensiled, and it was slower than passage of small (80 to 315 microm) particles (on average 0.0524/h). Particle size reduction of large INDF particles to medium INDF particles was slower (P < 0.001) in the early cut silages (0.0216/h to 0.0484/h) but reduction of medium INDF particles to small INDF particles was faster (P < 0.001) in early cut silages (0.0436 to 0.0305). Passage of medium size particles and(or) rate of medium particle breakdown to small particles were potential intake-constraining properties of low digestibility forages, whereas large particle reduction to medium particles seemed not to be limiting. The increased feed intake of the early-cut silages was accompanied by decreased rumen fill, suggesting that rumen fill was not at least solely responsible for feed intake control.

Adsorption and two-body recombination of atomic hydrogen on 3He-4He mixture films.

We present the first systematic measurement of the binding energy E(a) of hydrogen atoms to the surface of saturated 3He- 4He mixture films at temperatures 70-400 mK. E(a) is found to decrease almost linearly from 1.14(1) K down to 0.39(1) K, when the population of the ground surface state of 3He grows from zero to 6x10(14) cm(-2), yielding the value 1.2(1)x10(-15) K cm(2) for the mean-field parameter of H- 3He interaction in 2D. Measuring by ESR the rate constants K(aa) and K(ab) for second-order recombination of hydrogen atoms in hyperfine states a and b, we find the ratio K(ab)/K(aa) to be independent of the 3He content and to grow with temperature.

Detection of prostatic cells in peripheral blood: correlation with serum concentrations of prostate-specific antigen.

We have studied the expression of prostate-specific antigen (PSA) mRNA by reverse transcriptase-polymerase chain reaction in peripheral blood of 25 patients with cancer of the prostate (CAP), four with benign prostatic hyperplasia (BPH), two with renal stones, three with other types of cancer, and six healthy male and three female controls. Expression of mRNA specific for a certain tissue in peripheral blood is thought to indicate the presence of circulating cancer cells and metastatic spread of a tumor originating from this tissue. We detected PSA mRNA in 9 of 18 CAP patients with metastatic disease but in none of 7 patients without metastases. Negative results in patients with metastatic disease were associated with successful endocrine therapy and low concentrations of serum PSA, and the correlation between serum concentrations of PSA and the presence of PSA mRNA in peripheral blood was statistically significant. PSA mRNA was not found in patients with BPH, other types of cancer, or in healthy controls. Thus the occurrence of PSA mRNA in peripheral blood is associated with metastatic CAP.

Influence of grass maturity and diet on ruminal dry matter and neutral detergent fibre digestion kinetics.

The effects of, and interactions between, the diet and maturity of grass were studied in cattle in a 6 x 6 Latin square experiment with a 2 x 3 factorial arrangement of treatments. Barn-dried grass or direct-cut silage from the same sward were fed together with 250 (L), 500 (M) or 750 (H) g concentrate dry matter (DM) kg-1 total DM. Six timothy-meadow fescue grasses, cut at 7 d intervals, were incubated in nylon bags in the rumen for 6, 12, 24, 48, 72 and 96 h, and the kinetics parameters for DM and neutral detergent fibre (NDF) digestion were estimated. Ruminal NDF digestibility was calculated using a value of 0.02 for the rate of passage. With increasing maturity of grass, crude protein content and in vitro digestibility decreased with associated increases in the contents of cell wall constituents. Both the rate and extent of DM and NDF digestion decreased with maturity. The changes were curvilinear with an increasing depression in the extent of digestion and a decreasing depression in the rate of digestion with maturity. Ruminal NDF digestibility averaged over the diets decreased from 61.2 to 39.7% with the maturity. DM and NDF disappearance and calculated NDF digestibility decreased as the proportion of concentrate in the diet increased. This adverse effect of concentrate increased more rapidly as the proportion of concentrate increased from M to H and rumen pH dropped from 6.2 to 6.0 than as the proportion of concentrate increased from L to M with a consequent reduction in rumen pH from 6.4 to 6.2. The minimum rumen pH explained more of the variation in NDF digestibility than mean pH, duration or summation (pH x time) of the pH depression below 6.2, 6.0 or 5.8. Predicted NDF digestibility was higher in cattle given the dried grass diets than in those given the silage diets. The adverse effect of the concentrate level on NDF digestibility increased with the maturity of the grass. The decrease was 0.54, 0.59, 0.72% units per one day delay in cutting for low, medium and high levels of concentrate, respectively.

Hyperglycemic glucose concentrations up-regulate the expression of type VI collagen in vitro. Relevance to alterations of peripheral nerves in diabetes mellitus.

Electron microscopy of peripheral nerves obtained from two diabetic patients revealed large deposits of microfibrils and the presence of Luse bodies in the vicinity of perineurial cells. Microfibrils were found to accumulate also in the sciatic nerves of diabetic BB rats; these microfibrillar deposits were shown to contain type VI collagen by immunoelectron microscopy. Connective tissue cells cultured from rat sciatic nerves were exposed to high glucose concentrations. High glucose concentrations up-regulated the mRNA steady-state levels of alpha 1(VI), alpha 2(VI), and alpha 3(VI) chains of type VI collagen and caused accumulation of type VI collagen-containing fibrils in the cultures. Immunostaining and in situ hybridizations demonstrated that perineurial cells, Schwann cells, and fibroblasts expressed type VI collagen at the mRNA and protein levels. The results suggest that the turnover and supramolecular assembly of type VI collagen are perturbed in diabetic nerves and that glucose per se increases the expression of type VI collagen in vitro.

Amplification of fgfr4 gene in human breast and gynecological cancers.

We have investigated gene amplification of fibroblast growth factor receptor-4 (FGFR4) gene in 30 primary breast tumor samples and 15 gynecological tumor samples. Ten percent of the breast tumors showed 2- to 4-fold amplification. Amplification was found more frequently in estrogen- and progesterone-receptor-positive tumors and in tumors with high lymph-node involvement. Breast tumor samples were also analyzed for the amplification of fgfr3 and erbB2 genes and the chromosome 11q13 located genes hst1/int2/bcl1/sea. erbB2 gene was amplified 2- to 13-fold in 13% of the cases, but no amplification of int2/hst1/bcl1/sea amplicon was found. Gynecological tumors were also analyzed for the amplification of fgfr4 and fgfr3 genes and for int2 and hst1 oncogenes. Eleven of the 15 gynecological tumors were ovarian neoplasms including 2 benign tumors; the remainder comprised 1 ovarian metastasis of breast cancer; 1 endometrial cancer; 1 uterine leiomyosarcoma and 1 carcinosarcoma of the fallopian tube. In gynecological tumors, fgfr4 gene was found to be amplified in 2 ovarian tumors. Amplification of hst1 was found in 1 benign ovarian tumor. Thus, the fgfr4 gene may be involved in breast and ovarian tumorigenesis.