Effect of feed grinding methods with and without expansion on prececal and total tract mineral digestibility as well as on interior and exterior egg quality in laying hens.

The grinding of cereals by various milling methods as well as thermal treatment of feed may influence mineral digestibility and egg quality. The present study investigated the effect of feed produced by disc mill (D) and wedge-shaped disc mill (WSD), as mash (M) or expandate (E) on apparent ileal absorption (AIA) and apparent total digestibility (ATD) of calcium, phosphorus, magnesium, zinc, manganese, copper and iron, as well as on egg quality in laying hens. A total of 192 hens (Lohmann Brown) aged 19 wk, were assigned using a randomized design with a 2 × 2 factorial arrangement. Four experimental diets were offered ad libitum. Eggs were analyzed for weight, shape index, area, shell weight per unit surface area, yolk color, air cell, blood spot, Haugh unit, albumen and yolk measures (index, weight, height, width and length), shell measures (surface area, stability, density, thickness and membrane weight), as well as percent contents of albumen, yolk, shell, and shell membrane. The ATD for phosphorus, manganese, and copper was higher in WSD compared with D treatment (P = 0.028, P = 0.028 and P = 0.016, respectively). The interaction between milling methods and thermal treatment influenced ATD of copper (P = 0.033), which was higher in WSD+M group (41.0 ± 20.2) compared with D+E group (-3.21 ± 25.1), whereas no differences were observed for D+M (1.90 ± 37.8) and WSD+E (8.02 ± 36.2) groups. Egg stability tended to be higher in E compared with M treatment (P = 0.055). Albumen weight, percentage albumen weight, and albumen: yolk were higher and percentage yolk weight was lower in D compared with WSD treatment (P = 0.043, P = 0.027, P = 0.024, and P = 0.041, respectively). Number of blood spots was higher in E than M treatment (P = 0.053). In conclusion, use of a wedge-shaped disc mill resulted in higher ATD for phosphorus, manganese, and copper than use of a disc mill; however, digestibility for majority of minerals as well as egg quality parameters was comparable. Therefore, feed produced by either disc mill or wedge-shaped disc mill as mash or expandate may be used for laying hens without negative effects on egg quality.

Effects of estradiol, progestogens, and of tibolone on breast proliferation and apoptosis.

AIM: To study the effects of estrogen therapy, alone or combined with progestogens, and of tibolone on the expression of proliferation and apoptosis markers in normal breast tissue. METHODS: Thirty 250-day-old Wistar rats were castrated and 3 weeks later received one of the following treatments by gavage for 5 weeks: (1) estradiol benzoate; (2) estradiol benzoate + medroxyprogesterone acetate; (3) estradiol benzoate + norethisterone acetate; (4) estradiol benzoate + dydrogesterone; (5) tibolone; (6) placebo. Following treatment, the expression of proliferating cell nuclear antigen (PCNA) and caspase-3 was analyzed by quantitative immunohistochemistry in the breast tissue, and proliferation and apoptosis were analyzed semiquantitatively by microscopic imaging. RESULTS: There was a statistically significant difference among the groups for PCNA, caspase-3 and the caspase-3 : PCNA ratio. Tibolone was associated with the lowest proliferative activity, followed by estradiol benzoate + dydrogesterone; however, estradiol benzoate + dydrogesterone showed the greatest rate of apoptosis. CONCLUSIONS: The various progestogens can have more or less proliferative and pro-apoptotic effects than estradiol alone. Among the treatment schemes analyzed, the estradiol + dydrogesterone combination resulted in a higher apoptosis rate in relation to the proliferation rate and tibolone was associated with the lowest proliferation.

Is expression of rat breast matrix components influenced by estrogen, progestins and tibolone?

AIM: To study the effects of estrogen therapy, alone or combined with progestogens, and of tibolone on the expression of heparanase (HSPE), extracellular matrix metalloproteinases 2 and 9 (MMP-2 and MMP-9), perlecan and proliferating cell nuclear antigen (PCNA) in normal breast tissue. METHODS: Thirty 250-day-old Wistar rats were castrated and 3 weeks later received one of the following treatments by gavage for 5 weeks: (1) estradiol benzoate; (2) estradiol benzoate + medroxyprogesterone acetate; (3) estradiol benzoate + norethisterone acetate; (4) estradiol benzoate + dydrogesterone; (5) tibolone; (6) placebo. Following treatment, the expressions of mRNA for HSPE, MMP-2 and MMP-9 were analyzed by real-time PCR and the protein expressions of HSPE, MMP-2, MMP-9, perlecan and PCNA were quantified by immunohistochemistry. RESULTS: There was a statistically significant difference among the groups for the expression of HSPE mRNA due to high levels in the tibolone group. The groups differed in terms of PCNA, with lower levels found in the tibolone group followed by the estradiol benzoate + dydrogesterone group. A statistically significant positive correlation was observed for PCNA versus perlecan and MMP-9. CONCLUSIONS: There was no difference in the effects of combinations of estradiol and different progestogens on extracellular matrix components, and breast cell proliferation was associated with increases in perlecan and MMP-9.

Implication of milling methods, thermal treatment, and particle size of feed in layers on mineral digestibility and retention of minerals in egg contents.

Feed production with different milling methods, thermal treatment, and particle size may influence mineral digestibility and retention in eggs. The present study investigated the impact of roller (R) and hammer (H) mills, mash (M) and expandate (E) with fine (F) and coarse (C) particle sizes, on apparent ileal absorption (AIA) and apparent total digestibility (ATD) and retention of calcium, phosphorus, magnesium, zinc, manganese, copper and iron in yolk, albumen, and shell. A total of 384 hens (Lohmann Brown), 19 weeks old, were assigned using a randomized design with a 2×2×2 factorial arrangement. Eight experimental diets were offered ad libitum during the whole experimental period and one week before for diet adaption. The AIA of magnesium, zinc, copper, and iron was higher in treatment R in comparison with treatment H (P<0.01, P≤0.03, P<0.01 and P<0.01, respectively). The AIA of magnesium was higher in treatment M than treatment E (P<0.01). The AIA of magnesium was higher in treatment C in comparison with treatment F (P≤0.05) due to particle size. The ATD of copper and iron was higher in treatment R than treatment H (P<0.01 and P≤0.03, respectively). The ATD was higher for phosphorus and lower for iron in treatment F than treatment C (P≤0.05 and P≤0.02. respectively). The copper concentration in yolk and albumen was higher in treatment C than treatment F (P<0.01 and P≤0.03, respectively). Besides a few overall interactions, the AIA and ATD of copper and manganese were lower in H+M group than R+M group (P≤0.05). The ATD of iron was higher in the M+C group compared to the M+F group (P<0.01), whereas the albumen zinc concentration was higher in the E+C group than E+F group (P < 0.01). In conclusion, the feed produced by hammer mill had negative effects on AIA and ATD for trace elements in particular, but mineral concentrations in egg contents were mostly comparable for all treatments. Therefore, milling methods, thermal treatment, and particle sizes used in the present study can be used for layer feed formulation without negatively affecting egg quality.

Feed supplemented with organic acids does not affect starch digestibility, nor intestinal absorptive or secretory function in broiler chickens.

The current study aimed to determine the impact of acidified feed on apparent ileal starch digestibility, intestinal transport and barrier function and intestinal glucose transporter expression. The experiment included a control group and a treatment group with broilers fed a standard diet without or with 1.5% of a commercial organic acid product (64% formic acid, 25% propionic acid, 11% water). Broilers were fed with the experimental diets from hatching until days 32-35. Starch digestibility was determined using 0.2% titanium dioxide as ingestible marker. Gene expressions of the intestinal sodium glucose transporter 1 (SGLT-1) and glucose transporter 2 (GLUT-2) were analysed using qPCR analysis. Additionally, SGLT-1 function and chloride secretion were analysed in Ussing chamber experiments. Jejunal samples were sequentially exposed to 10 mm glucose, 100 µm phloridzin, 100 µm histamine and 100 µm carbachol. Apparent ileal starch digestibility (±SEM) of the control group (97.5 ± 0.35%) and the acid-treated group (97.0 ± 0.59%) did not differ (p = 0.674). The mean tissue conductance of intestinal samples obtained from the control group and the treatment group was similar [10.6 mS/cm(2) (±0.68) and 9.4 mS/cm(2) (±0.80) respectively (p = 0.147)]. The mean short-circuit currents (ΔIsc ) of the samples exposed to glucose, phloridzin, histamine and carbachol did not differ (p > 0.05). Additionally, no differences in the expression of SGLT-1 and GLUT-2 could be observed (p = 0.942, p = 0.413). Based on this study, the consumption of feed supplemented with organic acids was not associated with effects on ileal starch digestibility and functional traits of jejunal tissues, indicating that these additives have no major impact on the small intestinal function in broilers.

Carbohydrate coating reduces adhesion of biofilm-forming Bacillus subtilis to gold surfaces.

The growth of bacterial biofilms in pipes and food tanks causes severe problems in industry. Biofilms growing on medical implants or catheters are of great concern, as they can cause serious infections and decrease the functionality of the medical device. The prevention of bacterial adhesion--the first step in colonization and biofilm formation--is therefore very important. Current research comprises alterations in surface properties, the prevention of adhesin biosynthesis, inhibition with receptor analogs, or the development of anti-adhesive vaccines. We present a new approach that allows us to study bacterial adhesion with high sensitivity in real-time while testing several different surfaces in parallel. Using the cantilever-array technique we demonstrate that coating of gold surfaces with mono- or disaccharides results in a reduction of the bacterial adhesion of the biofilm-forming bacterium Bacillus subtilis NCIB 3610 to these gold surfaces. This reduction in bacterial adhesion is independent of the studied carbohydrate. Using several mutant strains, we investigate the underlying molecular interactions, and our results suggest that adhesion to gold surfaces is mediated by thiol groups present in proteins of the bacterial cell membrane or biofilm matrix proteins expressed at low levels by the wild-type strain. Furthermore, our data indicate that the adhesion of B. subtilis NCIB 3610 to carbohydrate-coated gold surfaces is facilitated by interactions between carbohydrates installed on the cantilever gold surface and an exopolysaccharide expressed by this strain. Understanding general and specific contributions of molecular interactions mediating bacterial adhesion will enable its prevention in the future.

The effects of different thermal treatments and organic acid levels on nutrient digestibility in broilers.

Poultry feed is a potential vector for pathogens. Heat processing and organic acid treatments may decontaminate feed and can affect bird performance as well as feed digestibility. The present study was performed to investigate the effect of different thermal treatments including pelleting (P), long-term conditioning at 85°C for 3 min (L), or expanding at 110°C (E110) and 130°C for 3 to 5 s (E130) without or with 0.75 and 1.5% organic acid supplementation (63.75% formic acid, 25.00% propionic acid, and 11.25% water) on performance, nutrient digestibility, and organ weights of broilers. In total, 960 one-day-old broiler chicks were randomly assigned to 8 replicates using a 3 × 4 factorial arrangement. Performance variables were determined, and the relative organ weights and ileal and total amino acid (AA) digestibilities were measured at d 35. The organic acid inclusion linearly improved feed efficiency in the first week (P ≤ 0.05). The acid inclusion levels and thermal treatments had no significant effect on the performance variables at later intervals of the growing period of the birds. The L group showed the lowest ileal AA and CP digestibility. The inclusion of organic acids had a quadratic effect on total and ileal digestibility of isoleucine (P ≤ 0.05), whereas it had no significant effect on the ileal digestibility of other AA and nutrients. The relative weights of the jejunum and small intestine were significantly higher in the E130 group compared with P and L (P ≤ 0.05). In conclusion, our study demonstrated that long-term heat conditioning can decrease ileal nutrient digestibility, whereas pelleting and expansion, independently of organic acid addition, seemed to have no negative impact on broiler performance and nutrient digestibilities. Moreover, adding a blend of organic acids to broiler diets had neither positive nor negative effects on nutrient digestibility and final broiler performance. This indicates the feasibility of short-term thermal treatment and acid supplementation for hygienization of broiler feed without negatively influencing performance.

Impact of thermal and organic acid treatment of feed on apparent ileal mineral absorption, tibial and liver mineral concentration, and tibia quality in broilers.

Minerals play an important role for growth and bone stability in broilers. Thermal treatment and inclusion of organic acids in feed may affect the mineral absorption and tibial quality in broilers. The study was conducted to investigate the effect of thermal processing of feed including pelleting (P), long-term conditioning at 85°C (L), and expanding at 130°C (E) without and with 1.5% of an acid mixture containing 64% formic and 25% propionic acid on the apparent ileal absorption (AIA) of calcium, phosphorus, magnesium, potassium, sodium, iron, copper, manganese, and zinc, their concentrations in liver and tibia, as well as various tibial quality parameters in broilers. In total, 480 one-day-old Cobb broiler chicks were assigned using a completely randomized design with a 3 × 2 factorial arrangement. The ileal digesta, liver, and tibia were collected at d 35. The AIA of calcium and sodium was improved in group E compared with L (P ≤ 0.02 and P ≤ 0.01). Group P and E showed higher AIA for potassium than L (P ≤ 0.01). Bone ash content was increased in group E compared with L (P ≤ 0.04). The BW to bone weight ratio was lower and tibial zinc content was higher in group P compared with E (P ≤ 0.05). Tibial iron content was higher in group L than E (P ≤ 0.03). Acid addition did not affect AIA, mineral content in tibia, or tibial quality parameters. Thermal and acid treatment did not affect mineral concentrations in the liver, except an inconsistent interaction effect for DM content and sodium (P ≤ 0.03 and P ≤ 0.04, respectively). In conclusion, long-term thermal treatment reduced AIA of some minerals compared with short-term thermal treatments, but had no impact on tibia composition. Acid inclusion had no effect on AIA of minerals and tibia quality. Thermal treatment and the use of organic acids can therefore be considered as safe with regard to their impact on bone development in broilers.

Effects of grinding method, particle size, and physical form of the diet on gastrointestinal morphology and jejunal glucose transport in laying hens.

Several studies illustrated that the structure of feed, i.e., the particle size, particle-size distribution, and the physical form of the diet, affects the avian gastrointestinal function and health leading to changes in productive performance. However, investigations concerning the effects of feeding differently processed diets on laying hens are limited and primarily concentrated on bird performance. The current study examines the effect of feed processing on the gastrointestinal morphology and on the jejunal glucose transport of laying hens. In 8 replicates, a total of 384 hens (Lohmann Brown) aged 20 wk were randomly allocated to 8 different groups and fed over a period of 21 d in a 3-factorial design. Diets differed in 1) grinding method, either hammer or roller mill; 2) physical form, either mash or expandate; and 3) particle size, either coarsely or finely ground. During the experimental trial, the laying performance of each feeding group was recorded daily and the feed intake and BW determined weekly. After slaughtering, the weights of the pancreas, proventriculus, gizzard, and small intestine were measured. Villus lengths and crypt depths of the duodenum, jejunum, and ileum were determined. The jejunal electrogenic glucose transport was studied in Ussing chambers. Hens that received mash instead of expandate had higher proventriculus (P = 0.011), gizzard (P < 0.001), and pancreas (P = 0.019) weights, whereas the feeding of coarsely instead of finely ground diets led to higher gizzard weights (P < 0.001). Mash-fed hens showed longer duodenal (P < 0.001) and shorter ileal villi (P = 0.047) and increased duodenal villus height-to-crypt depth ratios (P < 0.001) than those given the expandate. Mash-fed hens had higher glucose transport rates than expandate-fed hens (P < 0.001). In conclusion, the feeding of coarsely ground as well as mash diets had stimulating effects on the development of the gastrointestinal organs. Moreover, the feeding of mash influenced the intestinal microstructure of the epithelium that was accompanied by higher glucose transport capacities.

The effects of different thermal treatments and organic acid levels in feed on microbial composition and activity in gastrointestinal tract of broilers.

Thermal treatments of feed and organic acids are known to affect the gastrointestinal microbiota in chickens. The present study evaluated the effect of different thermal processes including pelleting (P), long-term conditioning at 85°C for 3 min (L), expanding at 110°C (E110), and 130°C for 3 to 5 s (E130) as well as organic acid (63.75% formic acid, 25.00% propionic acid, and 11.25% water) inclusion levels (0, 0.75, and 1.5%) on gastrointestinal microbiota in broilers. In total, 960 one-day-old chicks were randomly assigned to 8 replicates using a 3 × 4 factorial arrangement. At d 35, bacterial cell numbers in the crop, ileum, and cecum, and bacterial metabolites in the crop, gizzard, ileum, and cecum were determined. The inclusion of 1.5% organic acids increased cell numbers of all clostridial clusters in the crop. The organic acid supplementation increased the propionic acid concentration in the crop and gizzard and there was a decrease in lactic acid concentration. In the ileum, the 0% organic acid group had the highest numbers of Lactobacillus spp. and enterobacteria. Inclusion of 1.5% organic acids increased ileal acetate concentration. Increasing the feed processing temperature led to an increase of lactobacilli in the crop and ileum, whereas clostridia and enterobacteria seemed unaffected. Similarly, lactate concentrations increased in the ileum, but short-chain fatty acids remained identical. In the crop, an increase for acetate was found for the E130 group compared with all other thermal treatments. In conclusion, our study demonstrated that thermal treatments and organic acid supplementation to broiler diets more markedly influenced the bacterial status of the crop compared with the downstream segments and their effects decreased along the length of gastrointestinal tract. Whereas organic acids markedly modified bacterial composition and activity in the crop, expansion increased lactobacilli and lactate in the crop and ileum.

CSF neurofilament and N-acetylaspartate related brain changes in clinically isolated syndrome.

BACKGROUND: Axonal damage is considered a major cause of disability in multiple sclerosis (MS) and may start early in the disease. Specific biomarkers for this process are of great interest. OBJECTIVE: To study if cerebrospinal fluid (CSF) biomarkers for axonal damage reflect and predict disease progression already in the earliest stages of the disease, that is, in clinically isolated syndrome (CIS). METHODS: We assessed CSF levels of neurofilament heavy (NFH), neurofilament light (NFL) and N-acetylaspartate (NAA) in 67 patients with CIS and 18 controls with neuropsychiatric diseases of non-inflammatory aetiology (NC). Patients with CIS underwent baseline magnetic resonance imaging (MRI) at 3T, and a follow-up MRI after 1 year was obtained in 28 of them. RESULTS: Compared with NC, patients with CIS had higher NFH (p=0.05) and NFL (p<0.001) levels. No significant group differences were found for NAA. Patients' NFH levels correlated with physical disability (r=0.304, p<0.05) and with change in brain volume over 1 year of follow-up (r=-0.518, p<0.01) but not with change in T2 lesion load. CONCLUSION: Our results confirm increased neurofilament levels already in CIS being related to the level of physical disability. The association of NFH levels with brain volume but not lesion volume changes supports the association of these markers with axonal damage.

Interactions between risk assessors and risk managers during three major food incidents in Europe.

Risk analysis consists of risk assessment (RA), risk management (RM), and risk communication (RC). In most countries, RA and RM of food safety are separated to achieve a high scientific integrity, and typically occur in sequential order. However, in case of a food safety incident, even though being separate processes, RA and RM are performed simultaneously due to great time pressure and expected high impacts. The aim of this study was to analyze and evaluate the observed interactions between RA and RM processes, during three major food incidents in Europe, and to provide suggestions for possible improvement. Based on the differences observed between the three cases, strengths and weaknesses of each system have been identified. The enterohemorrhagic Escherichia coli (EHEC) crisis in 2011 in Germany, the horsemeat scandal in 2013 in Ireland, and the fipronil incident in 2017 in the Netherlands were used as case studies. Timelines of these incidents and crisis management procedures in place in each of the three countries provided the basis for further analysis. First, results showed that details of the communication processes between RA and RM bodies were frequently lacking in crisis management protocols. Second, RA, RM, and RC processes differed for each incident, due to differences in estimated risk for public health, but also due to differences in the organization within a country. Based on our results, we recommend that crisis management protocols should contain a section on communication between RA, RM, and on communication between member states in the EU.

[U-HO1. A new cell line derived from a primary refractory classical Hodgkin lymphoma]. / U-HO1. Eine neue Zelllinie, abstammend von einem primär therapierefraktären klassischen Hodgkin-Lymphom.

The Hodgkin cell line U-HO1 was established from a malignant pleural effusion of a 23-yr-old male patient during the end stage of refractory nodular sclerosing classical Hodgkin lymphoma (cHL). Since its establishment in 2005, U-HO1 has maintained stable characteristics in vitro and has a doubling time of about 4 days under standard culture conditions. U-HO1 forms typical Reed/Sternberg cells in suspension, is EBV negative, lacks HLA-ABC- but expresses HLA-D- proteins/CD74 and surface exposes CD15 together with CD30 in the absence of CD19 and CD20. Karyotype analysis of U-HO1 revealed a hyperdiploid karyotype with multiple clonal aberrations. Most significant is an elongated chromosome 2, der(2)t(2;10)(q35;q16.1)add(2)(p13). CGH analysis revealed the following imbalances: ish cgh dim(1)(p13p31)(p12q21), enh(2)(p13p23), dim(4)(q31.3qter), enh(6)(q22q27), enh(12), enh(18),enh(20)(q13.1pter). FISH analysis showed about six-fold amplification of REL and BCL-11A, thus, U-HO1 is prototypical for cHL in every aspect tested so far. Compared to other HL cell lines, U-HO1 proved far less genetically aberrant suggesting that U-HO1's imbalances suffice to cause the full-blown phenotype of primary refractory cHL.

U-HO1, a new cell line derived from a primary refractory classical Hodgkin lymphoma.

The Hodgkin cell line U-HO1 was established from a malignant pleural effusion of a 23-year-old male patient during the end stage of refractory nodular sclerosing classical Hodgkin lymphoma (cHL). Since its establishment in 2005, U-HO1 has maintained stable characteristics in vitro and has a doubling time of about 4 days under standard culture conditions. U-HO1 forms typical Reed-Sternberg cells in suspension, is EBV negative, lacks HLA-A, -B, -C but expresses HLA-D proteins/CD74 and exposes CD15 together with CD30 in the absence of CD19 and CD20 on the cell surface. Karyotype analysis of U-HO1 revealed a hyperdiploid karyotype with multiple clonal aberrations. Most significant is an elongated chromosome 2, der(2)t(2;10)(q35; q16.1)add(2)(p13). CGH analysis revealed the following imbalances: ish cgh dim(1)(p13p31)(p12q21), enh(2)(p13p23), dim(4)(q31.3qter), enh(6)(q22q27), enh(12), enh(18), enh(20) (q13.1pter). FISH analysis showed about six-fold amplification of REL and BCL11A, thus, U-HO1 is prototypical for cHL in every aspect tested so far. As an outstanding feature compared to the existing HL cell lines, U-HO1 has high levels of microRNA transcripts of MIRN216 and MIRN217 located in the amplicon 2p16. Compared to other HL cell lines, U-HO1 proved far less genetically aberrant suggesting that U-HO1's imbalances suffice to cause the full-blown phenotype of primary refractory cHL.

Hindwings of insects as concept generator for hingeless foldable shading systems.

Hingeless shading systems inspired by nature are increasingly the focus of architectural research. In contrast to traditional systems, these compliant mechanisms can reduce the amount of maintenance-intensive parts and can easily be adapted to irregular, doubly curved, facade geometries. Previous mechanisms rely merely on the reversible material deformation of composite structures with almost homogeneous material properties. This leads to large actuation forces and an inherent conflict between the requirements of movement and the capacity to carry external loads. To enhance the performance of such systems, current research is directed at natural mechanisms with concentrated compliance and distinct hinge zones with high load-bearing capacity. Here, we provide insights into our biological findings and the development of a deployable structure inspired by the Flexagon model of hindwings of insects in general and the hierarchical structure of the wing cuticle of the shield bug (Graphosoma lineatum). By using technical fibre-reinforced plastics in combination with an elastomer foil, natural principles have been partially transferred into a multi-layered structure with locally adapted stiffness. Initial small prototypes have been produced in a vacuum-assisted hot press and sustain this functionality. Initial theoretical studies on test surfaces outline the advantages of these bio-inspired structures as deployable external shading systems for doubly curved facades.

Nuclear membrane fusion in electrofused mammalian cells.

Fusion of nuclei was studied in electrofused cells using staining procedures and DNA flow cytometry. Homogeneous and heterogeneous electrofusion of Ehrlich ascites tumor cells. Muntjac cells and V79-S181 cells were performed in balanced-salt solutions at low temperature. Incubation of the cells subjected to electrofusion in fusion media for about 2 h was required to complete cell fusion and, in particular, nuclear membrane fusion. Under optimum electrofusion conditions it was found that fusion of nuclei is a very frequent event. Half of the fused cells (about 30 to 50% of the field-exposed cells) underwent nuclear membrane fusion. It is shown that the high frequency of nuclear membrane fusion in electrofused, unsynchronised cells resulted from intracellular dielectrophoresis occurring during cell alignment. In accordance with theory, maximum nuclear membrane fusion was observed using alignment fields of between 1 and 4 MHz (depending on the cell species), that is above the frequencies at which the plasmalemma capacity no longer shielded the cell interior from participation in the conduction process. In this frequency range a potential difference can be built up across the nuclear membrane leading to repositioning of the nuclei into the contact zone of the plasmalemmas of two attached cells. This intracellular dielectrophoresis apparently facilitated fusion of nuclei once intermingling of the plasma membranes had occurred. It was further demonstrated that exponentially growing cells showed higher cell fusion rates than cells taken from the unfed plateau phase. One, but not the only reason, might be the higher ATP content of exponentially growing cells compared to cells of the plateau phase. Addition of external ATP to plateau phase cells during electrofusion resulted, in accordance with this assumption, in an increase of fusion frequency, whereas ATP had apparently no effect on the fusion yield of exponentially growing cells. G1 cells obtained by mitotic selection after nocodazole-induced blockage in metaphase also showed higher cellular and nuclear membrane fusion yields than exponentially growing cells. Most importantly, it could be demonstrated both experimentally and theoretically that electrofusion of cells in a dielectrophoretically aligned chain is controlled by a simple law of probability resulting predominantly in fusion of two cells independent of the number of cells in the chain. The likelihood of fusion of various numbers of cells in a chain is given by the appropriate power of the probability of two-cell fusion.(ABSTRACT TRUNCATED AT 400 WORDS)

Okadaic acid stimulates ouabain-sensitive 86Rb(+)-uptake and phosphorylation of the Na+/K(+)-ATPase alpha-subunit in rat hepatocytes.

Ca(2+)-mobilizing and cAMP-dependent hormones rapidly increase sodium, potassium-dependent adenosine triphosphatase (Na+/K(+)-ATPase)-mediated transport in rat hepatocytes. To explore the possible role of protein phosphatases in these responses we used a protein phosphatase inhibitor, okadaic acid. Okadaic acid stimulation of ouabain-sensitive 86Rb(+)-uptake was maximal between two and three minutes and displayed an EC50 of 41 +/- 1 nM. Inhibition of Na+/H+ exchange with an amiloride analog abolished the response to insulin, but had no effect on okadaic acid-mediated stimulation of Na+/K(+)-ATPase transport. In hepatocytes metabolically-radiolabeled with 32Pi, okadaic acid stimulated the incorporation of radioactivity into several 95 kDa peptides, one of which reacted with anti-LEAVE peptide antisera, that recognizes Na+/K(+)-ATPase alpha-subunits. In other experiments Na+/K(+)-ATPase was immunoprecipitated from detergent-solubilized membrane fractions of metabolically-radiolabeled cells with an antisera to purified rat kidney Na+/K(+)-ATPase. A 95 kDa phosphoprotein was immunoprecipitated using anti-Na+/K(+)-ATPase antisera, but not by preimmune serum. Okadaic acid stimulated incorporation of radioactivity into this band by 220 +/- 28%. These findings provide support for the hypothesis that rapid stimulation of hepatic Na+/K(+)-ATPase by hormones may be related to protein kinase/phosphatase-mediated changes in the phosphorylation state of the Na+/K(+)-ATPase alpha-subunit.

Plasma ammonia response to sprint swimming.

BACKGROUND: To study the plasma ammonia response after sprint crawl swimming. METHODS: Nine sprinters (S) and ten non-sprinters (NS) completed a 15-, a 25- and a 50-m crawl at maximal intensity with a 10-min and a 15-min resting period in-between. Capillary blood samples were collected before and at regular intervals after each effort for plasma ammonia determination. RESULTS: Ammonia kinetics differed among distances, but not between groups, with peak values (observed 2-8 min postexercise) being higher after 50 m as compared to shorter distances. Significant differences between S and NS were found in peak ammonia after 50 m (124.5 +/- 58.2 vs 98.7 +/- 6.3 mumoL-1) and in the change of ammonia relative to swim time (delta NH3/delta t) after 25 m (2.66 +/- 1.87 vs 1.49 +/- 0.84 mumol L-1 s-1) and 50 m (1.87 +/- 1.33 vs 1.01 +/- 0.49 mumol L-1 s-1). delta NH3/delta t was highest after 15 m (3.33 +/- 2.53 in S, 3.92 +/- 1.67 mumol L-1 s-1 in NS). CONCLUSIONS: These differences in the plasma ammonia response to sprint swimming according to duration and athlete seem to be connected to distinctions in muscle fiber profile and energy providing processes.

[Mortality in the Swiss and foreign immigrants in the Canton of Ticino]. / La mortalità negli svizzeri e negli stranieri immigrati nel Cantone Ticino.

The Canton Ticino is one of the Swiss cantons with the highest number of foreigners (mainly Italians): roughly 25% of the entire canton's population. The aim of this work, which is part of a wider study on the immigration problem in Switzerland, is to evaluate whether the immigrant population in the Canton Ticino presents differences in death rates, both in general and according to cause of death, with respect to the Swiss population. Crude and standardized death rates were calculated for autochthonals, Italians and other foreigners, using data on those deceased in the Canton during 1991 and 1994. Higher death-rates (both sexes and all age groups, except 0-19 years) were calculated for almost all causes of death, of the Swiss group with respect to Italians and other foreigners. The low death-rates found in the immigrants may be due, at least partially, to the "healthy emigrant" effect. This effect seems to be stronger than the effect of the often negative factors to which immigrants are exposed in the host country.

Draft Genome Sequence of the Biofilm-Producing Bacillus subtilis Strain B-1, Isolated from an Oil Field.

We report here the draft genome sequence of the Bacillus subtilis strain B-1, a strain known to form biofilms. The biofilm matrix mainly consists of the biopolymer γ-polyglutamate (γ-PGA). The sequence of the genome of this strain allows the study of specific genes involved in biofilm formation.