Ptychographic characterisation of polymer compound refractive lenses manufactured by additive technology.

The recent success in the development of high-precision printing techniques allows one to manufacture free-standing polymer structures of high quality. Two-photon polymerization lithography is a mask-less technique with down to 100 nm resolution that provides full geometric freedom. It has recently been applied to the nanofabrication of X-ray compound refractive lenses (CRLs). In this article we report on the characterization of two sets of CRLs of different design produced by two-photon polymerization-induced lithography.

Ptychographic analysis of the photorefractive effect in LiNbO3:Fe.

We present light induced refractive index changes in iron doped lithium niobate detected with a novel microscopy technique called ptychography. This method determines the change of the refractive index together with the intensity distribution of the writing beam from a single scan with a reconstructed spatial resolution of 3 µm and a sensitivity of the refractive index change of 10-5. We show that the light induced refractive index change is strongly connected to the intensity shape of the writing beam and that it shows the expected nonlocal behaviour. Applying the novel method to the investigation of the photorefractive effect offers an excellent opportunity to study this nonlocal response to the spatial distribution of the writing beam.

The effect of selenium supplementation before calving on early-lactation udder health in pastured dairy heifers.

Selenium (Se) deficiency has been associated with lowered resistance to mastitis in dairy cattle. However, little published data exists on the effect of Se supplementation before calving on udder health of pastured dairy heifers. Further, the relative efficacy of injectable barium selenate and oral organic Se for improving udder health in cows has not previously been tested. The objectives of this study were to determine the effects of precalving Se supplementation and type of supplementation on the blood activity of glutathione peroxidase and measures of udder health immediately after calving and during the first month of lactation in pastured dairy heifers. One hundred forty pregnant Chilean Holstein-Friesian heifers were fed a basal diet containing, on average, 0.15 mg of Se/kg of dry matter. One month before predicted calving, heifers were allocated to 1 of 3 groups. Group 1 (n=49) received no supplementary Se, group 2 (n=46) received a single subcutaneous injection of Se (1 mg/kg of live weight, as barium selenate), and group 3 (n=45) was fed Se yeast (3 mg/heifer/d until calving). Heifers supplemented with barium selenate had a higher glutathione peroxidase activity from 14 d in milk onwards. Selenium supplementation, irrespective of source, tended to reduce the prevalence of intramammary infection (IMI) and decrease the prevalence of quarters with high somatic cell count (SCC) at calving. Overall, Se supplementation did not result in a reduction of the incidence of new IMI or clinical mastitis or in decreased SCC during the balance of the first month of lactation. However, in pasture-based heifers injected with barium selenate before calving, and fed diets with 1.3 and 2.5 mg of Se/d precalving and during lactation, respectively, no cases of clinical mastitis were observed in the first month of lactation.

Barium selenate supplementation and its effect on intramammary infection in pasture-based dairy cows.

A significant proportion of cattle receive inadequate dietary Se because of its low content in soils and pastures of various regions of the world. Several economically important diseases in dairy cows, such as mastitis, have been associated with Se deficiency. The objective of this study was to evaluate the effect of a single injection of a long-acting form of Se at drying off on the risk and incidence rate of new intramammary infections and on milk somatic cell count in the subsequent lactation in pasture-based dairy cows. Forty-nine Chilean Holstein-Friesian cows were fed a diet containing <0.05 mg of Se/kg of ration dry matter. During the dry period, cows were allocated to 1 of 2 groups, a supplemented (n=24) group treated with a single subcutaneous injection of barium selenate 2 mo before calving and a control group (n=25) that remained unsupplemented. Duplicate foremilk samples were aseptically collected within 6 d after calving and every 2 wk until drying-off for bacteriological culture. Milk samples were also collected monthly for somatic cell count evaluation. Blood samples were collected before treatment and at 30, 90, 180, and 270 d after treatment for analysis of blood glutathione peroxidase (GPx) activity. The activity of glutathione peroxidase was higher in supplemented cows 30 d after the injection until the end of the study. The risk and incidence rate of new intramammary infections was not affected by supplementation. A progressive increase in somatic cell count was observed throughout lactation, but there was no effect of supplementation. In conclusion, a one-time injection of barium selenate 2 mo before calving in these pasture-based dairy cows did not affect udder health in the subsequent lactation, indicating that Se basal intake was adequate for preventing subclinical mastitis in pasture-based cows in southern Chile.

Somatic cell count in milk of selenium-supplemented dairy cows after an intramammary challenge with Staphylococcus aureus.

The objective of this study was to evaluate the effect of selenium (Se) supplementation on milk somatic cell count (SCC) in dairy cows. Twelve multiparous Holstein-Friesian cows were fed a diet containing a suboptimal Se concentration (<0.05 ppm, dry basis) starting 2 months before calving. Supplemented cows (n=6) received a single s.c. injection of barium selenate (1 ml/50 kg BW) 45 days prior to calving, whereas control group was kept unsupplemented. Twenty weeks after calving, two mammary quarters (right side) of each cow were challenged with 205,000 cfu/ml of Staphylococcus aureus (strain Newbould 305). Blood was collected bi-weekly until day 150 of lactation for the analysis of blood glutathione peroxidase (GPx1; EC 1.11.1.9) activity. To re-isolate the challenging pathogen and to evaluate SCC, aseptic milk samples were collected daily starting on the day of challenge, and finishing 7 days after inoculation. Unsupplemented cows had a lower activity of GPx1 through the experiment (P<0.001). Natural log SCC (lnSCC) was higher in unsupplemented than Se-supplemented cows (P=0.04), showing evidence of significance after 5 days. Selenium supplementation of dairy cows fed a diet containing a suboptimal Se concentration, resulted in higher blood activity of GPx1, and lower mean lnSCC after an intramammary challenge with Staph. aureus.

Effect of grazing pasture with a low selenium content on the concentrations of triiodothyronine and thyroxine in serum, and GSH-Px activity in erythrocytes in cows in Chile.

AIM: To determine the effect of grazing pasture that had a low selenium (Se) concentration on serum concentrations of triiodothyronine (T3) and thyroxine (T4), and erythrocyte glutathione peroxidase (GSH-Px) activity in dairy cows. METHODS: Forty pregnant Friesian cows were grazed on pasture that contained 0.03-0.04 ppm Se on a dry matter (DM) basis. Two months before parturition, 20 cows were randomly selected and treated with 1 mg Se/kg bodyweight subcutaneously, as barium selenate (Group Se-S). The other group (Se-D) was not supplemented. Blood samples were taken before supplementation (-60 days) and 30, 60, 90, 180 and 270 days after parturition, for determination of concentrations of T3 and T4 in serum, and GSH-Px activity in erythrocytes. RESULTS: Erythrocyte GSH-Px activity in the Se-D group was <60 U/g haemoglobin (Hb) throughout the experiment. Supplementation increased (p<0.05) activities to >130 U/g Hb throughout lactation. Mean serum concentrations of T4 in Se-D and Se-S cows increased from 23.7 (SEM 0.7) and 23.4 (SEM 0.8) nmol/L, respectively, in the prepartum period to 69.6 (SEM 0.1) and 67.6 (SEM 0.2) nmol/L, respectively, at 180 days of lactation (p<0.01), and no effect of Se supplementation was evident. Serum concentrations of T3 in Se-D cows decreased (p<0.05) from 1.6 (SEM 0.1) nmol/L prepartum to 1.0 (SEM 0.2) nmol/L at the beginning of lactation, and remained lower (p<0.05) than those in the Se-S cows which did not decrease after calving and ranged from 1.9 (SEM 0.1) to 2.4 (SEM 0.2) nmol/L throughout lactation. CONCLUSIONS: Serum T3 concentrations decreased during early lactation in unsupplemented cows grazing pastures low in Se (0.03-0.04 ppm) and both serum T3 and erythrocyte GSHPx activities were consistently lower throughout lactation compared with Se-supplemented cows. Se supplementation had no effect on serum T4 concentrations.

Lilliputian: an AF4/FMR2-related protein that controls cell identity and cell growth.

Members of the AF4/FMR2 family of nuclear proteins are involved in human diseases such as acute lymphoblastic leukemia and mental retardation. Here we report the identification and characterization of the Drosophila lilliputian (lilli) gene, which encodes a nuclear protein related to mammalian AF4 and FMR2. Mutations in lilli suppress excessive neuronal differentiation in response to a constitutively active form of Raf in the eye. In the wild type, Lilli has a partially redundant function in the Ras/MAPK pathway in differentiation but it is essential for normal growth. Loss of Lilli function causes an autonomous reduction in cell size and partially suppresses the increased growth associated with loss of PTEN function. These results suggest that Lilli acts in parallel with the Ras/MAPK and the PI3K/PKB pathways in the control of cell identity and cellular growth.

Multicenter laboratory validation of susceptibility testing of Mycobacterium tuberculosis against classical second-line and newer antimicrobial drugs by using the radiometric BACTEC 460 technique and the proportion method with solid media.

In a large multicenter study involving six major study sites in the United States, Canada, and Europe, the susceptibilities of 272 Mycobacterium tuberculosis strains to classical second-line antituberculosis (anti-TB) drugs (capreomycin, cycloserine, ethionamide, and kanamycin) and newer compounds (amikacin, clofazimine, ofloxacin, and rifabutin) were determined by the radiometric BACTEC 460 procedure and the conventional proportion method on Middlebrook 7H10 agar. Previously established critical concentrations for classical second-line anti-TB drugs were compared with several concentrations in liquid medium to establish equivalence. MICs of newer compounds determined in liquid medium were either the same or up to four times lower than those determined in agar medium. After establishing critical concentrations (breakpoints) in the extended testing of clinical isolates, we obtained an excellent overall correlation between the two systems, with no errors with amikacin, kanamycin, and ofloxacin and very few major or very major errors with the other drugs; however, for cycloserine, no breakpoint concentration could be recommended due to repeatedly inconsistent results by both methods. Based on these data we conclude that the BACTEC 460 procedure is a simple and rapid method requiring 4 to 8 days on average to generate accurate antimicrobial susceptibility testing (AST) results for eight anti-TB drugs other than those considered primary ones. These data not only fill a major gap of knowledge regarding the critical test concentrations of secondary anti-TB drugs but also provide a baseline for future evaluations of M. tuberculosis AST with the more recently developed, nonradiometric broth-based culture systems.

Use of metabolic profiles in dairy cattle in tropical and subtropical countries on smallholder dairy farms.

Metabolic profile testing has generally been used as part of a multidisciplinary approach for dairy herds in temperate climates. Our goal was to evaluate the effectiveness of the technique for identifying constraints on productivity in small herds in environments less favorable for milk production. Metabolites tested were chosen for stability in the sample after collection of blood, ease of analysis and practical knowledge of the meaning of the results. Blood levels of five different metabolites in low-producing dairy cows belonging to smallholders in tropical and subtropical environments were measured. The study involved 13 projects with 80 cows in each, carried out in six Latin American, six Asian, and one southern European countries. Data were also collected on feeding, body condition score (BCS) and weight change, parasitism, and reproduction. In Chile, Mexico, Paraguay, Philippines, Uruguay, and Venezuela, globulin levels were high in > 17% of cows sampled on each occasion. Globulin levels were also high in Turkey and Vietnam on one or more occasions. In Paraguay, 49% of cows had high globulin levels at two to three months after calving. These results suggest that inflammatory disease was present to a potentially important degree, although this was not always investigated and not always taken into account. In all countries except Mexico and Venezuela, high beta-hydroxybutyrate (BHB) levels before calving in many cows highlighted the presence of condition loss in late pregnancy, an important potential constraint on productivity and fertility. Fewer cows showed high BHB levels in lactation, whereas change in BCS and weight was more sensitive for measuring negative energy balance. Urea concentrations were low in only small numbers of cows suggesting that dietary protein shortages were not common. Albumin values were low mainly in cows where globulin values were high and, hence, did not generally provide additional information. The exception was in China where pregnant yaks over winter had high BHB and low albumin values, suggesting that they were seriously underfed. This observation stimulated a successful nutritional intervention in the following winter. Inorganic phosphate values were within the reference range in most countries a majority of the time suggesting, contrary to expectation, that this mineral was not commonly a constraint. The use of metabolic profile testing proved valuable in drawing attention to important potential constraints on productivity in dairy cows in tropical and subtropical environments and in confirming those which were not.

Bulk milk urea concentrations and their relationship with cow fertility in grazing dairy herds in southern Chile.

Milk urea determination is being used as a broad indicator of protein/energy imbalance in dairy herds. The main purpose of this study was to compare blood and bulk milk urea values in grazing herds, to evaluate their seasonal variation under South Chilean conditions, and to examine their potential relationships with herd fertility. The association between herd blood urea concentration (mean of seven lactating cows) and bulk milk urea concentration (tank containing milk from the previous 24 h) was determined in 21 diary herds. Reference values, seasonal and herd variance, and the frequency of herds with values outside a range of 2.5 to 7.3 mmol/l were determined in bulk milk samples obtained monthly for a period of one year from 82 suppliers at two creameries located in southern Chile. Finally, bulk milk urea was measured every two weeks in samples from 24 herds, and the first service conception rate (FSCR) from 2153 dairy cows was determined. Mean bulk urea concentration was highly correlated with mean herd blood urea concentration (r = 0.95; p < 0.01). Mean urea concentration in the bulk milk samples obtained during one year from 82 herds was 4.9 +/- 1.2 mmol/l, with a range of 1.5 to 11.6 mmol/l. The highest values were found during spring and the lowest values during the summer. There was a high seasonal variation (CV = 13-47%) and between-herd variation (CV = 20-31%). Out of a total of 984 samples, 5.4% had urea values > 7.3 mmol/l and 3.8% had values < 2.5 mmol/l. Of the 82 herds, 27% had values outside the reference interval (2.5-7.3 mmol/l) on two or more occasions. FSCR was lower in herds when the bulk milk urea was > 7.3 mmol/l (50.7%) than in cows, where the urea concentration was < 5.0 mmol/l (73.8%) at the time of insemination. The study concluded that bulk milk urea concentrations provided information similar to herd blood urea concentrations in local grazing dairy herds. There was a high frequency of herds with abnormal values, with large variations between herds and between seasons. Increased milk urea concentrations during spring were associated with lower conception rates.

Localized upregulation of a new expansin gene predicts the site of leaf formation in the tomato meristem.

Expansins are extracellular proteins that increase plant cell wall extensibility in vitro and are thought to be involved in cell expansion. We showed in a previous study that administration of an exogenous expansin protein can trigger the initiation of leaflike structures on the shoot apical meristem of tomato. Here, we studied the expression patterns of two tomato expansin genes, LeExp2 and LeExp18. LeExp2 is preferentially expressed in expanding tissues, whereas LeExp18 is expressed preferentially in tissues with meristematic activity. In situ hybridization experiments showed that LeExp18 expression is elevated in a group of cells, called I1, which is the site of incipient leaf primordium initiation. Thus, LeExp18 expression is a molecular marker for leaf initiation, predicting the site of primordium formation at a time before histological changes can be detected. We propose a model for the regulation of phyllotaxis that postulates a crucial role for expansin in leaf primordium initiation.

Evolution of fluoride concentrations in cattle and grass following a volcanic eruption.

This study determined fluoride concentrations of forage, water and bovine serum 2-y after the cessation of a volcanic eruption. Three farms within the polluted area were selected, and water and green grass were collected from April 1990 to February 1992. Samples of hay were collected twice from each farm. Five cows exposed during the 13-mo eruption and 6 young bulls not exposed during the volcanic eruption had blood collected monthly or bimonthly. Serum fluoride concentrations and alkaline phosphatase activities were determined and dental conditions observed in all animals. Grass fluoride remained low except during spring of the second year; high fluoride was in the hay. Serum fluoride of the cows remained near acceptable reference values, although some cows had higher values; serum fluoride of the bulls markedly increased during the spring of the second year. Severe incisor attrition occurred in the cows, while the young bulls had less severely eroded teeth.

The effect of 1 alpha-hydroxycholecalciferol on the concentrations of calcium, magnesium and inorganic phosphate in the plasma of parturient cows.

135 Cows which were expected to produce a second or subsequent calf within 4--11 received an intramuscular injection of 500 micrograms 1 alpha-hydroxycholecalciferol (1 alpha-OHD3). In those animals in which the interval between injection and calving was at least 7 d there was, at 7 d after injection of 1 alpha-OHD3, a significant increase (P less than 0.001) in the blood plasma concentrations of Ca and Pi and a significant reduction (P less than 0.05) in that of Mg, compared with those in control animals. Within 24 h of calving there was a significant increase (P less than 0.001) in the concentration of Pi but no change in Ca or Mg compared with pre-injection levels, whilst the control animals showed a significant fall (P less than 0.001) in the concentration of Ca and a rise in those of Pi (P less than 0.001) and Mg (P less than 0.05). There was a significant reduction (P less than 0.001) in the number of animals with hypocalcaemia (defined as a plasma Ca concentration less than 1.9 mmol/l) in all animals injected with 1 alpha-OHD3; this reduction was particularly marked when the interval between injection and calving was between 1 and 8 d. The previously reported reduction in the incidence of clinical milk fever is considered to be a consequence of the effect of 1 alpha-OHD3 on the concentration of Ca in plasma.

1alpha hydroxycholecalciferol in the prevention of milk fever.

During the 12 months commencing July 1976 the animals in a herd of 400 Friesian cows which were carrying their second or later calf were injected intramuscularly with either microgram of 1alpha-hydroxycholecalciferol in 5 ml aqueous carrier or with 5 ml of carrier. The injections were randomised so that 135 of the 269 injected cows received the drug and 134 received the carrier solution. Ten of the experimental group (7.4 per cent) and 22 of the control group (16.4 per cent) had to be treated for milk fever, (P less than 0.08). The protective effect was greatest, (P less than 0.001), when the cows calved between one and eight days after injection of the drug.