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Integrin-linked kinase (ILK), PINCH and Parvin proteins form the IPP-complex that has been established as a core component of the integrin-actin link. Our recent genetic studies on Drosophila parvin, reveal that loss of function mutant defects phenocopy those observed upon loss of ILK or PINCH in the muscle and the wing, strengthening the notion that these proteins function together in the organism. Our work identified that ILK is necessary and sufficient for parvin subcellular localization, corroborating previous data indicating a direct association between these two proteins. Further genetic epistasis analysis of the IPP-complex assembly at integrin adhesion sites reveals that depending on the cell context each component is required differently. At the muscle attachment sites of the embryo, ILK is placed upstream in the hierarchy of genetic interactions required for the IPP-complex assembly. By contrast, in the wing epithelium the three proteins are mutually interdependent. Finally, we uncovered a novel property for the CH1-domain of parvin: its recruitment at the integrin-containing junctions in an ILK-dependent manner. Apparently, this ability of the CH1-domain is controlled by the inter-CH linker region. Thus, an intramolecular interaction within parvin could serve as a putative regulatory mechanism controlling the ILK-Parvin interaction.

An experiment was conducted with 12 lactating dairy ewes and 12 goats with the objective to determine if there are differences between them in lipogenic genes [acetyl-CoA carboxylase (ACC), fatty acid synthetase (FAS), lipoprotein lipase (LPL), stearoyl-CoA desaturase (SCD) and leptin (LEP)] in their subcutaneous adipose tissue (SUBQ) under the same dietary treatments. The animals were fed with two diets (A, B) in different days of milk (DIM) due to the different milk yield, body weight, etc. in order to have the same food intake and to avoid dietary effects. Diet A with a forage/concentrate ( F / C ) ratio 53/47 on dry matter basis, was offered to each group of sheep and goats from DIM = 105 to DIM = 126, and from DIM = 63 to DIM = 84, respectively. Diet B with a F / C ratio 49/51 on dry matter basis, was offered to sheep and goats individually from DIM = 140 to DIM = 161 and from DIM = 98 to DIM = 119, respectively. The results showed that the mRNA of ACC was significantly lower in sheep SUBQ while the mRNA of LPL, SCD and LEP were higher compared with those of goats independently from the fed diet and the different DIM. In conclusion, there are significant species differences between sheep and goats in the lipogenic gene mRNA expression of SUBQ under the same dietary treatments.

In the laboratory under LD 12:12 h-conditions, mating behavior of D. oleae takes place during the last 3–4 h of the photophase and terminates abruptly with the onset of darkness. Mating behavior is independent of low light intensities, nor need it be graded. The acoustical components of sexual activity include ♂-courtship and strong wing-beat frequencies during copulatory attemps and pursuits in flight. Mating activity is a circadian event and in constant whitelight or redlight (600–700 nm) a free-running rhythm with τ < 24 h develops and persists for several cycles. However, the quantity of mating activity decreases rapidly. Zusammenfassung Uber den Begattungsrhythmus bei der Olivenfliege, Dacus oleae Gmel. Unter LD 12: 12 h-Bedingungen konzentriert sich das Begattungsverhalten von Dacus oleae auf die letzten 3–4 h der Lichtphase. Geringe oder graduell abnehmende Lichtintensitaten sind keine Voraussetzungen fur den Ablauf des Verhaltens. Akustische Verhaltensweisen im Rahmen der Sexualaktivitat umfassen die ♂-Balz, sowie verstarkte Flugelschlagfrequenzen wahrend der Kopulationsversuche und Verfolgungsfluge. Das Begattungsverhalten ist circadian gesteuert und hat unter konstanten Dauerlicht- und Rotlichtbedingungen eine Periode τ < 24 h. Der Rhythmus uberdauert mehrere Zyklen, aber die Anzahl der Begattungen nimmt rasch ab.

A number of studies have shown that the rumenic acid (RA = cis -9 trans -11 C 18:2 CLA) content of milk fat is usually higher in sheep than in goats, due partly to different dietary regimens. An experiment was conducted with 12 lactating dairy ewes and 12 goats with the objective to compare the two animal species (sheep/goats) fed diet with the same forage/concentrate ( F / C ) ratio, on their milk fatty acids (FA) profile with emphasis on RA and vaccenic acid (VA) production. The experiment was carried out in three consecutive phases, lasted 3 weeks each, immediately after weaning of lambs and kids. In phase I, the ewes and the goats were fed according to their maintenance and lactation requirements, with 14 kg alfalfa hay, 4 kg wheat straw and 12 kg concentrate the 12 ewes ( F / C ratio = 60/40), and with 14 kg alfalfa hay, 4 kg straw and 24 kg concentrate the 12 goats ( F / C ratio = 43/57). In phase II, 14 kg alfalfa hay, 4 kg straw and 14 kg concentrate were offered daily to each group of sheep and goats, with a F / C ratio = 56/44. In phase III, all ewes and goats were fed individually with 0.8 kg alfalfa hay, 0.2 kg wheat straw and 0.8 kg concentrate daily with a F / C ratio = 56/44. The results showed that the different F / C ratio between sheep and goats diets, in phase I, changed significantly the milk FA profile, with no significant difference in RA and VA milk fat content between sheep and goats in phase I. In phases II (group feeding) and III (individual feeding), where sheep and goats fed with the same amount of food of the same F / C ratio, the sheep milk fat had higher RA and VA content compared to goats. In conclusion, these findings support the hypothesis that there are species differences, as RA and VA production concerns, which needs further investigation.

An experiment was conducted with the objective to study the interaction between breed and diet on conjugated linoleic acid (CLA) and fatty acids profile (FA) content of milk fat of four pure of dairy sheep breeds (Awassi, Lacaune, Friesland and Chios). All sheep used in this study were kept in one flock under the same management. During the winter months (from October to March) all sheep were kept indoors and fed with alfalfa hay and concentrates without any grazing and from April onwards were grazing native pastures without any supplementary feed. A total of 160 individual milk samples were taken from the four breeds at two sampling times: in January (n = 80) and in May (n = 80), when the ewes were in average, on 62 and 185 days in milk. These milk samples were analyzed for FA profile. The results of this experiment showed that the sheep breed had no effect on milk FA profile or on atherogenicity index (AI). The only significant effect was on Δ−9 desaturase activity index expressed as C18:1/C18:0. On the contrary, the diet affected significantly the FA profile of all ewes milk fat, with pasture to cause lower proportions of saturated and higher proportions of unsaturated FA, compared with those of sheep kept and fed indoors. The interaction between breed and diet was significant for the FA C10:0, C14:0, C15:0, C16:0, C17:1, C18:1, C18:2n6c, C18:2n6t, C18:3n6 and CLA and for FA groups SCFA, MCFA, LCFA and MUFA. The S/U FA ratio and the Δ−9 desaturase indexes (C14:1/C14:0 and C18:1/C18:0) were also affected by the breed × diet interaction. In conclusion, pasture feeding when compared with supplementary diet, induce large variation in the milk FA profile. In addition pasture increases FA in milk fat which has potential interest for human health. Breed has small effect on milk FA profile, but it can be ignored as it is shown from the interaction breed × diet.

Epidemiological and experimental data support the hypothesis of ‘fetal programming’, which proposes that alterations in fetal nutrition and endocrine status lead to permanent adaptations in fetal homeostatic mechanisms, producing long-term changes in physiology and determine susceptibility to later disease. Altered hypothalamic–pituitary–adrenal (HPA) axis function has been proposed to play an important role in programming of disease risk. The aim of the present study was to examine the effects of maternal nutrient restriction imposed during different periods of gestation on the HPA axis function in sheep, at different ages postnatal. Pregnant ewes were fed a 50% nutrient-restricted diet from days 0–30 (group R1, n = 7), or from days 31–100 of gestation (group R2, n = 7) or a control 100% diet throughout pregnancy, (Control, n = 8). Blood samples were collected at 10-day intervals from day 40 of gestation to term. Lambs were born naturally and fed to appetite throughout the study period. At 2, 5.5, and 10 months of age lambs were given an i.v. injection of corticotrophin-releasing hormone (CRH) and blood samples were collected at −15, 0, 15, 30, 60, 120, and 180 min postinjection. Maternal cortisol levels were significantly higher (P < 0.05) in group R1 compared with the other two groups, whereas maternal insulin levels were lower (P < 0.05) in group R2 compared with control. Birth weight of lambs was not affected by the maternal nutritional manipulation. The area under the curve for ACTH and cortisol response to CRH challenge was greater (P < 0.05) in lambs of group R1 at two months of age, whereas no difference was detected at the ages of 5.5 and 10 months. However, significantly higher (P < 0.01) basal cortisol levels were observed in lambs of R1 group at 5.5 months of age. There was no interaction between treatment and sex for both pituitary and adrenal responses to the challenge. A significant sex effect was evident with females responding with higher ACTH and cortisol levels at the age of 5.5 months (P < 0.01, P < 0.001 respectively) and with higher cortisol levels (P < 0.01) at 10 months of age than males. It is concluded that the HPA axis is programmable by altered nutrition in utero. The sensitivity of the axis to exogenous stimulation is enhanced during early postnatal life and attenuated with age, suggesting a role for the postnatal influences in resetting of the HPA axis and emphasizing the importance of identifying the impact of maternal undernutrition at several time points after birth.

The cis-9, trans-11 C18:2 (CLA) content in milk fat of four pure sheep breeds (Awassi, Lacaune, Friesland and Chios) was examined. All sheep were kept indoors all year round under the same feeding practices, without any grazing at all. Sheep nutrition was based on alfalfa hay and concentrates. A total of 237 individual milk samples were collected at three sampling times (December, January and March) from sheep of different parity and different days in milk, for CLA determination. The results showed that: (a) there was a large variation in milk fat CLA content among individuals consuming the same diet, (b) the CLA content of milk fat was lower in Friesland breed, compared to the other three breeds but without any significant difference among the four sheep breeds, (c) the CLA content of milk fat was not affected by ewes parity or days in milk, (d) there was a negative but not significant correlation between milk fat and CLA concentration, (e) the CLA content of milk fat was not correlated with milk yield. In conclusion, no others factors, such as breed, parity, days in milk, etc., can affect CLA content in milk fat, which means that the dietary ones remain the sovereign factors explaining the highest proportion of CLA content variability in sheep milk fat.

The conjugated linoleic acid (CLA) content of grazing sheep and goat milk fat, throughout their lactation period, was examined. Six sheep and six goat representative farms were selected at random and milk samples were taken at monthly intervals for fatty acids profile determination. Sheep and goat nutrition was based on natural grazing and on supplementary feeding during the winter months. From April onwards, grazing native pastures was the only source of feed for sheep and goats. The University farm whose sheep are kept indoors all year round without any grazing, was also used as reference. Fifteen individual milk samples were also taken in April from a sheep and goat farm respectively, in order to see the variability of CLA inside the farm. The results showed that: a. the CLA content of grazing sheep and goat milk fat increased significantly in April–May (early growth stage of grass) and then declined while that of indoors kept sheep was more or less constant during the same period, b. the isomers cis-9, trans-11 and trans-10, cis-12 of CLA were found in grazing sheep milk fat, while in indoors kept sheep and goats' milk fat only the cis-9, trans-11 isomer was found, c. the CLA content of sheep milk fat was much higher than that of goats, d. a negative correlation between sheep milk fat and CLA content was found and e. there was considerable variation in milk fat CLA content between sheep and goat farms and inside the farms.

The mammalian focal adhesion kinase (FAK) family of non-receptor protein-tyrosine kinases has been implicated in controlling a multitude of cellular responses to the engagement of cell-surface integrins and G-protein-coupled receptors. The high level of sequence conservation between the mammalian proteins and the Drosophila homologue of FAK, Fak56,suggested that it would have similar functions. However, we show here that Drosophila Fak56 is not essential for integrin functions in adhesion,migration or signaling in vivo. Furthermore, animals lacking Fak56 are viable and fertile, demonstrating that Fak56 is not essential for other developmental or physiological functions. Despite this, overexpressed Fak56 is a potent inhibitor of integrins binding to the extracellular matrix, suggesting that Fak56 may play a subtle role in the negative regulation of integrin adhesion.

Τeτρακόσιοι νeοσσοί κρeοπαραγωγής, ηλικίας μιας ημέρας, χρησιμοποιήθηκαν για να μeλeτηθeί η eπίδραση του σeληνίου και της βιταμίνης E σe eπιλeγμένα σeληνιοένζυμα που σχeτίζονται μe τη λeιτουργία του θυροeιδούς αδένα. Υπήρχαν τέσσeρις διατροφικές eπeμβάσeις μe πέντe eπαναλήψeις η κάθe μία. Οι πeιραματικές ομάδeς πeριeλάμβαναν την ομάδα του μάρτυρα στην οποία χορηγήθηκe ένα βασικό σιτηρέσιο eπαρκές σe σeλήνιο και βιταμίνη E (0,3 και 80 mg/kg αντίστοιχα), την ομάδα Se+ στην οποία χορηγήθηκe eπιπρόσθeτα 1 mg/kg σeληνίου, την ομάδα E + στην οποία χορηγήθηκe eπιπρόσθeτα 350 mg/kg βιταμίνη E και τέλος την ομάδα Se+E+ στην οποία χορηγήθηκe συνδυασμός σeληνίου και βιταμίνης E σe συγκeντρώσeις 1 και 350 mg/kg αντίστοιχα. Τα σιτηρέσια ήταν ισοαζωτούχα και ισοeνeργeιακά. Για την eπιπλέον προσθήκη σeληνίου χρησιμοποιήθηκe σύμπλοκο της L-σeληνομeθeιονίνης μe ψeυδάργυρο, eνώ για την προσθήκη βιταμίνης η DL-α-τοκοφeρόλη. Το πeίραμα διήρκeσe 42 ημέρeς. Η συγκέντρωση σeληνίου στο πλάσμα αυξήθηκe στις eπeμβάσeις μe σeλήνιο, eνώ η eνeργότητα της υπeροξeιδάσης της γλουταθeιόνης στο αίμα αυξήθηκe σe όλeς τις eπeμβάσeις σe σχέση μe τον μάρτυρα. Η αποϊωδιάση της ιωδιοθυρονίνης (τύπου Ι) στο ήπαρ και οι συγκeντρώσeις των θυρeοeιδικών ορμονών στο αίμα δeν eπηρeάστηκαν από οποιαδήποτe eπέμβαση. Συμπeρασματικά, η συμπλήρωση μe σeλήνιο ή βιταμίνη E μeμονωμένα ή σe συνδυασμό πέραν των αναγκών του ζώου δeν eπηρέασαν τα eπίπeδα των θυρeοeιδικών ορμονών, ούτe την eνeργότητα της ηπατικής αποϊωδιάσης υπό τις συνθήκeς που eξeτάστηκαν.

Objectives The goal of this study was to investigate the dose-dependent effects of active cigarette smoking on endothelial nitric oxide (NO) and endothelin-1 (ET-1) biosynthesis. Background Limited studies have suggested that active cigarette smoking may be associated with a dose-dependent reduction of endothelium-dependent vasodilation (EDV). The underlying biochemical changes that cause this dose-specific effect, such as changes in the endothelial NO biosynthetic pathway and ET-1 production, have not been examined. Methods Flow- and nitroglycerin-mediated reactivity of the brachial artery were measured in eight nonsmokers, seven light smokers (≤1 pack/week) and eight heavy smokers (≥1 pack/day), and their sera were added to confluent (∼85%) monolayers of human umbilical endothelial cells (HUVECs) for 12 h. Basal and substance P-stimulated NO and basal ET-1 production were measured. The HUVECs used for measuring basal NO production were lysed, and both endothelial NO synthase (eNOS) protein expression and eNOS activity were determined. Results Serum cotinine level and pack-years of smoking were significantly lower in light smokers compared with heavy smokers (p Conclusions These results indicate light smoking may have similar detrimental effects on EDV and NO biosynthetic pathway as does heavy smoking. These data may have important implications concerning the amount of active cigarette exposure that imparts cardiovascular risk.

Developmental analysis of the tyrosine protein phosphorylation levels in larval integument and partial characterization of the endogenous protein tyrosine kinase activity (PTK) in Ceratitis capitata are described in this study. Larval integument contains high levels of PTK activity at the early stages of the third instar, which progressively declines to low levels in the white pupal stage. An integumental 90-kDa polypeptide was identified to have prominent endogenous PTK activity and follow a similar developmental pattern. The major integumental phosphotyrosine-containing polypeptides have apparent molecular weights of 30, 41, 44, 46, and 54 kDa, respectively. Polypeptides with molecular weights of 62 and 73 kDa were identified as Ser/Thr-containing phosphoproteins and were shown to exhibit high levels of phosphorylation at the middle stage of larval development. These differences are likely to be due to the higher activation state of the protein tyrosine kinase(s) at the early stages of larval development. Arch. Insect Biochem. Physiol. 50:9–20, 2002. © 2002 Wiley-Liss, Inc.

Για την eπιλογή της πλέον αποτeλeσματικής παγίδας και eλκυστικού μeταξύ ορισμένων από τους τύπους παγίδων και eλκυστικών ουσιών που χρησιμοποιούνται σήμeρα για την παρακολούθηση και καταπολέμηση της μύγας της Μeσογeίου, Ceratitis capitata, (Wiedemann) (Diptera: Tephritidae), συγκρίθηκαν σe πορτοκαλeώνeς eπτά τύποι παγίδων (δύο τύπου MePhail, Υαλοπλαστική, χάρτινο-δίπτυχο, χάρτινος φάκeλος-χαρτοπλαστική, διαφανής πλαστική φιάλη, παγίδα ξηρού τύπου), τέσσeρα τροφικά eλκυστικά ( Dacus bait 100, Entomela 12SL, όξινο ανθρακικό αμμώνιο και το μe κωδικό ZI), ένα eλκυστικό φύλου (Trimedlure) και ένας συνδυασμός τροφικού και eλκυστικού φύλου (Όξινο ανθρακικό αμμώνιο+Trimedlure). Τα αποτeλέσματα έδeιξαν ότι μeταξύ των παγίδων τύπου MePhail δeν υπάρχουν σημαντικές διαφορές. Η παγίδα τύπου πλαστικής φιάλης απeδeίχθη πολύ αποτeλeσματική όταν πληρούται μe το ZI (πρωτeϊνικό τροφικό eλκυστικό). Από τα δοκιμασθέντα eλκυστικά , τα πλέον αποτeλeσματικά ήταν το ZI και το Entomela. Ο συνδυασμός eλκυστικών τροφής και φύλου δeν έδeιξe σημαντική συνeργιστική δράση στην αποτeλeσματικότητα της παγίδας. Τα αποτeλέσματα eπιτρέπουν μια καλύτeρη eπιλογή μeταξύ των παραπάνω τύπων παγίδων και eλκυστικών, για παρακολούθηση και καταπολέμηση της μύγας της Μeσογeίου.

Background The mechanisms involved in the dysfunction of both endothelium-dependent vasodilatation (EDV) and NO biosynthesis related to smoking are unclear. In this study, EDV was assessed in healthy smokers and nonsmokers in vivo and, using serum from the same individuals, was related to the NO biosynthetic pathway in vitro. Methods and Results Flow-mediated EDV of the brachial artery was measured in 23 male patients (8 nonsmokers and 15 smokers). Serum was collected, added to confluent (≈85%) monolayers of human umbilical vein endothelial cells (HUVECs), and incubated for 12 hours. Basal and substance P–stimulated NO production was measured. The HUVECs used for measuring basal NO production were lysed, and both endothelial NO synthase (eNOS) protein expression and eNOS activity were determined. EDV was lower in smokers compared with nonsmokers ( P P P P P P r =0.54, P r =−0.60, P Conclusions This is the first study to combine an in vivo model with a near-physiological in vitro model to demonstrate an association between decreased NO production and reduced EDV. Cigarette smoking was associated with reduced EDV, NO generation, and eNOS activity in the presence of increased eNOS protein expression.

Integrin-linked kinase (ILK) was identified by its interaction with the cytoplasmic tail of human β1 integrin and previous data suggest that ILK is a component of diverse signaling pathways, including integrin, Wnt, and protein kinase B. Here we show that the absence of ILK function in Drosophila causes defects similar to loss of integrin adhesion, but not similar to loss of these signaling pathways. ILK mutations cause embryonic lethality and defects in muscle attachment, and clones of cells lacking ILK in the adult wing fail to adhere, forming wing blisters. Consistent with this, an ILK–green fluorescent protein fusion protein colocalizes with the position-specific integrins at sites of integrin function: muscle attachment sites and the basal junctions of the wing epithelium. Surprisingly, mutations in the kinase domain shown to inactivate the kinase activity of human ILK do not show any phenotype in Drosophila, suggesting a kinase-independent function for ILK. The muscle detachment in ILK mutants is associated with detachment of the actin filaments from the muscle ends, unlike integrin mutants, in which the primary defect is detachment of the plasma membrane from the extracellular matrix. Our data suggest that ILK is a component of the structure linking the cytoskeleton and the plasma membrane at sites of integrin-mediated adhesion.

Ten 4-month-old female sheep of the Karagouniko dairy breed were used to monitor the seasonal fluctuations of strongyle infections in sheep during the first year of grazing under the traditional production system in the region of Trikala, Greece, where control of nematodes is currently based exclusively on the frequent use of anthelmintics which might contribute to the appearance of anthelmintic resistance. The sheep grazed communal pasture plots and did not receive any anthelmintic treatment during the entire study period. Faecal egg counts (epg), plasma pepsinogen levels, blood values, and genera of parasitic strongyles recovered from coprocultures were recorded monthly. Mean epg for strongyle-type eggs were significantly higher during summer. Mean plasma pepsinogen levels were significantly higher during spring and summer. The genera of parasitic strongyles recovered from the faecal cultures were Haemonchus, Ostertagia, Trichostrongylus, Cooperia, Nematodirus, Oesophagostomum, and Chabertia. The percentages of larvae for Haemonchus, Ostertagia, Cooperia, and Oesophagostomum were significantly higher during winter. The percentages of larvae for Trichostrongylus were significantly higher during summer. During summer, levels of RBC, HGB, and HCT were depressed and levels of MCV, MCH, and MCHC were elevated. Mean epg for strongyle-type eggs had a significant inverse correlation with RBC, HGB, HCT, and positive correlation with MCH, MCHC, and percentages of eosinophils in differential leucocyte counts. The seasonal pattern of infection observed in the present study indicates that it is possible to decrease the number of treatments to one per year, thus, reducing the possibility for the appearance of anthelmintic resistance.

Information concerning worm control practices of sheep and goat farmers in the region of Trikala (central Greece) was collected through a questionnaire survey by visiting farms and interviewing farmers. Questionnaires from 57 farmers residing in 23 rural communities were collected. Anthelmintics were used by 89% of the farmers. On average, lambs, kids and goats were treated once annually, while sheep were treated either once or twice annually. Only 2% of farmers reported treatment of animals with anthelmintics when moving to new pastures. The most common broad-spectrum anthelmintics used were those belonging to the benzimidazoles and probenzimidazoles. Fifty nine percent of the farmers used the same anthelmintic group for 3 or more years and 34% used two or more anthelmintic groups in the same year. Almost all farmers reported estimating live weights for calculating anthelmintic doses through visual perception on the basis of an average weight (96%). Tablets and boluses were the most preferred anthelmintic formulation used by 96% of farmers. The selection of an anthelmintic was based for 58% of farmers on recommendation by a veterinarian and for 39% of farmers on the cost of the drug. The most common occasions for deworming the animals were at turn out (86%) and after parturition (31%). Only 6% of farmers reported deworming new animals before introducing them onto the farm. Farmers preferred to seek information about the use of anthelmintics and worm control strategies from veterinarians (63%) and other farmers (37%).

In order to examine the effects of diets, with two hay to concentrate ratios and two forms of concentrates on the rumen fermentation of dry sheep and milk production of lactating ewes, three trials were conducted.In the first trial, four wether sheep, each fitted with a rumen cannula, were used in a 4 × 4 Latin-square design, by allocating them to one of four diets with different hay to concentrate ratio and concentrate type. The diets consisted of 0·5 kg hay and 0·75 kg either starchy (HRSC) or fibrous (HRFC) concentrates, or 0·25 kg hay and 1·0 kg either starchy (LRSC) or fibrous (LRFC) concentrates. The diets were designed to provide similar intakes of energy and crude protein. Volatile fatty acids (VFA) and ammonia concentrations, pH values and number of protozoa were determined in wether rumen fluid.Total VFA concentration was not significantly affected by diet but acetate and n-butyrate proportions were higher (P < 0·002) on the higher forage diets and propionate higher (P < 0·002) on the lower forage diets. pH values and number of protozoa in sheep rumen fluid were unaffected by treatment, while ammonia concentration was lower (P < 0·05) on the HRFC diet.In the second trial, 60 lactating dairy ewes were allocated to one of four diets for weeks 11 to 15 of lactation (period 1). Ewes were given food daily: 0·87 kg hay and 1·3 kg either HRFC or HRSC concentrates, or 0·4 kg hay and 1·6 kg either LRFC or LRSC concentrates. During weeks 16 to 20 of lactation (period 2), the ewes were given the same amount and type of concentrates but hay was supplied ad libitum and this period formed the third trial.Milk yield and milk composition were affected significantly by hay to concentrate ratio with the lower hay to concentrate ratio resulting in a significant increase in milk yield, milk protein efficiency, protein content and protein and lactose yields. Milk fat content and yield were reduced. Hay intake was proportionately greater (P < 0·002) by 0·15 to 0·20 with starchy than with fibrous concentrate diets in period 2. Problems such as acidosis or ‘off-food’ periods in ewes given the low hay to concentrate ratio diets were not observed, irrespective of concentrate type.

It is well known that activated prophenoloxidase (proPO) plays an important role in cuticular melanization and sclerotization. In addition, studies dealing with immune response of insects suggest that phenoloxidase (PO) is also critical in the defense reactions of insects against invaders. proPO is activated by elicitors derived from microbial cell wall components such as peptidoglycan, beta-1,3-glucan, and lipopolysaccharide (LPS). According to our recent studies we proposed a model clarifying the role of PO in both cellular and humoral immune responses. LPS triggers Ceratitis capitata hemocytes via induced protein tyrosine phosphorylation to release biologically active molecules, including p47 and proPO-activators. Furthermore, hemocytes in response to LPS facilitate clearance of LPS from the hemocoel of medfly. The effector molecules involved in the LPS clearance are hemocyte surface-associated p47 (mp47), soluble p47 (sp47), activated proPO, and tyrosine. A similar LPS clearance system in the integument of medfly in vitro was also demonstrated. According to our data, the proposed mechanism for LPS clearance from hemocoel and from integument is the crosslinking of LPS to p47 or certain integumental proteins via the intermediacy of reactive tyrosine derivatives generated by PO activity, as is the case for cuticular protein-chitin crosslinks during sclerotization. We also demonstrated that metabolites of the eumelanin biosynthesis and not melanin itself or N-acetyldopamine (NADA), the key precursor of sclerotizing agent, were necessary for the immune responses by hemocytes and integument.

The purpose of this study was to investigate the preservation time of fresh olive tree leaves (OTL) after treatment with ammonia (AM-OTL) solution, the improvement of its nutritive value and the effect on milk yield and milk composition of dairy ewes. The crude protein (CP) content of OTL treated with a 25% aqueous ammonia solution (5 1 per 100 kg fresh OTL) increased from 96 to 180 g kg−1 dry matter (DM) just after the bags were opened, and 164 and 155 g kg−1 DM 2 and 12 months after treatment, respectively. The non-protein nitrogen of AM-OTL was 312 g kg−1 CP compared with 100 g kg−1 CP of fresh untreated OTL (F-OTL). Results of a digestibility experiment showed that the apparent digestibility (g kg−1) of AM-OTL increased (P < 0.05) for CP, crude fibre (CF), neutral detergent fibre and acid detergent fibre by 47.4%, 19.4%, 13.3% and 10.5% and decreased for ether extractives (EE) by 10.7% in relation to F-OTL. The nutritive value was estimated to be 4.60 and 4.81 MJ of net energy of lactation (NEL) kg−1 DM for F-OTL and AM-OTL, respectively. In the second experiment 24 dairy lactating ewes of the Karagouniko breed (55 kg of average BW) were used and divided into two equal groups, immediately after weaning of their lambs. Using a cross-over design, ewes in Group 1 were fed a diet (A) with 1.2 kg AM-OTL + 0.25 kg wheat straw+ 0.88 kg concentrates. Ewes in Group 2 were fed a diet (B) with 0.8 kg alfalfa hay +1 kg concentrates. Diet (A) caused an increase (P < 0.05) in soluble CP and a reduction (P < 0.05) in casein content of milk. Fatty acid composition of milk fat was affected by AM-OTL with a reduction (P < 0.001) of short chain (C6 to C16:l) and an increase of long chain (C18, C18:1) fatty acids. The results showed that ammonia treatment preserves OTL, improves their nutritive value and allows their inclusion in ruminant diets by replacing other expensive roughages.

Integrin-linked kinase (ILK), PINCH and Parvin constitute the tripartite IPP-complex that maintains the integrin-actin link at embryonic muscle attachment sites (MASs) in Drosophila. Here we showed that parvin null mutations in Drosophila exhibit defects in muscle adhesion, similar to ILK and PINCH mutants. Furthermore, the identical muscle phenotype of the triple mutant, which for the first time in any organism removed the entire IPP-complex function, genetically demonstrated that parvin, ILK and PINCH function synergistically. This is consistent with the tight localization of the tripartite complex at sites of integrin adhesion, namely MASs in the developing embryo and focal contact-like structures in the wing epithelium. Parvin contains tandem unconventional Calponin-Homology (CH) domains separated by a linker sequence, and a less well conserved N-terminal region. In vivo structure-function analysis revealed that all the domains are essential for parvin function, whereas recruitment at integrin adhesion sites is mediated by two localization signals: one located within the CH2-domain as previously reported, and a second novel signal within the CH1 domain. Interestingly, this site is masked by the linker region between the two CH-domains, suggesting a regulatory mechanism to control parvin localization. Finally, whereas in muscles only ILK controls the stability and localization of both PINCH and parvin, in the wing epithelium the three proteins mutually depend on each other. Thus molecular differences exist in the assembly properties of IPP-complex in specific tissues during development, where differential modulation of the integrin connection to cytoskeleton is required.

Larval Ceratitis capitata phenoloxidases (POs) from hemocytes, serum, integument, and fat body were analyzed. Two types of PO were recorded: the tyrosinase type found in hemocytes, serum, integument, and fat body and the laccase type found in integument. Tyrosinase from all larval tissues and integumental laccase as well, showed similarity in molecular weight (93 KDa), activation by Escherichia coli at 5 mM Ca2+, and reactivity to antibodies raised against serum tyrosinase. However, the enzymes differed with respect to their glycosylation and adhesiveness. The serum and integumental enzyme forms contain concanavalin A reacting material, whereas hemocyte and integumental tyrosinase(s) are adhesive. These differences in enzyme forms, although not influencing their substrate specificity, seem to give advantages to performing their function, i.e., the adhesive enzyme form facilitates the adherence to E. coli cell wall and hemocyte surface (unpublished data) while the glycosylated form facilitated the secretion into serum. © 1994 Wiley-Liss, Inc.

Expression of a 54 kDa tyrosyl phosphorylated protein in epidermal cells during the third instar larval stage was followed. It was demonstrated that the 54 kDa protein moiety and its phosphorylated counterpart follow the same developmental profile. The system seems to be regulated only at the onset of the second moult, by an initial signal which regulates both the synthesis and phosphorylation of a 54 kDa protein. The continuous presence this protein in epidermal cells during the third instar stage, as well as during apolysis and histolysis, suggests that it might participate in cell activities taking place during this developmental period. However, the 54 kDa protein could no be involved in specific epidermal cell activities such as histolysis, melanization and sclerotization, since these activities occur only at specific times during the third instar stage.

Integrin-linked kinase (ILK) is an essential component of a multiprotein complex that links actin to the plasma membrane. Here, we have used a genetic approach to examine the molecular interactions that are essential for the assembly of this ILK-containing complex at Drosophila muscle attachment sites (MASs). We show that, downstream of integrins, talin plays a decisive role in the recruitment of three proteins: ILK, PINCH and paxillin. The accumulation of ILK at MASs appears to follow an amplification mechanism, suggesting that numerous binding sites are generated by minimal levels of the upstream integrin and talin effectors. This property suggests that ILK functions as an essential hub in the assembly of its partner proteins at sites of integrin adhesion. We found that PINCH stability, and its subcellular localization at MASs, depends upon ILK function, but that ILK stability and localization is not dependent upon PINCH. An in vivo structure–function analysis of ILK demonstrated that each ILK domain has sufficient information for its independent recruitment at embryonic MASs, whereas at later developmental stages only the kinase domain was effectively recruited. Our data strengthen the view that the ILK complex is assembled sequentially at sites of integrin adhesion by employing multiple molecular interactions, which collectively stabilize the integrin–actin link.

Presentation : This final round table has been organized to debate whether traditional knowledge in animal production is an obstacle or a source of innovation. I suggest you starting this discussion by a presentation of what "traditional knowledge" means. Then, the several chairmen would mobilize the mains results of their sessions to feed the exchanges: Pr. Zervas is questioned about the relationship between tradition and innovation in terms of technical efficiency of production systems and activities, Dr. Brunori on the management of knowledge and innovation processing, then Dr. Faye on the relationships between social demand and animal production sectors. To finish, Dr. Boutonnet will give us his vision on how to structure between innovation and tradition and will conclude this round table

During the late larval stage, the integument of Ceratitis capitata , contains an alkaline phosphatase activity based on hydrolysis of p -nitrophenyl phosphate. In the present work we identified that the integumental activity also dephosphorylates a 54 kDa tyrosyl-phosphorylated protein, isolated from the integument of the same stage of development. The isolation of this protein was performed by affinity chromatography using antiphosphotyrosine antibodies, raised against the synthetic hapten p -azobenzyl phosphonate which specifically crossreacts with phosphotyrosine. The enzyme activity is greater at alkaline region for pNP-P and at neutral region for phosphotyrosyl protein. This shift towards physiological pH indicates possible physiological significance. The chelator EDTA (2 mM) which is a potent inhibitor of enzyme activity when pNP-P was used as the substrate (80%) was slightly inhibitory when free phosphotyrosine or 54 kDa phosphoprotein was used as a substrate. In addition, the divalent cation Mg 2+ has stimulatory action on the enzyme activity with all the substrates tested. Finally, 2 mM Zn 2+ and 2 mM NaF exerted a sloghtly inhibitory action on the phosphatase activity for all the substrates used. These results suggest the existence of an alkaline phosphatase activity in the integument of C. capitata , based on hydrolysis of pNP-P. This enzyme behaved as phosphotyrosyl protein phosphatase at neutral pH, dephosphorylating the physiologically existed 54 kDa tyrosyl-phosphorylated protein. Furthermore, the coexistence of phosphorylating-dephosphorylating activities as well as the 54 kDa tyrosyl phosphorylated protein in the integument, of C. capitata calls for a physiological regulatory mechanism in the integument during the transition from larvae to pupae.

The objective of this work was to determine the apparent bioavailability of magnesium from four different types of calcined magnesites of Greek origin. These four types (A, B, C, D) were different in particle size fractions and production process. The solubility of the above four magnesium samples and their fractions were determined: (a) in the laboratory using ammonium nitrate solution after 24-h shaking and (b) in vivo using a nylon bag technique after 24-h incubation.Solubility was correlated with apparent bioavailability as determined in a balance experiment with five wether sheep in a 5×5 Latin-square design.Sheep were kept in metabolism cages and were given daily a basal diet (500 g hay + 700 g concentrate) plus 2 g magnesium supplements from types A to D. The magnesium bioavailability of the basal diet was used as a base for the determination of the bioavailability of magnesium in A, B, C and D supplements.Magnesium apparent bioavailability of the basal diet was found to be 0·203 and, for the supplements A, B, C and D, 0·381, 0·339, 0·315 and 0·292 respectively. The solubility of supplements A, B, C and D in ammonium nitrate solution and in vivo (using the nylon-bag technique) was found to be 0·812, 0·805, 0·784 and 0·773 or 0·294, 0·152, 0·102 and 0·200 respectively.The solubilities of the magnesium supplements in ammonium nitrate solution and in vivo both reflected dietary bioavailability differences up to a point, but neither was well enough correlated with the apparent bioavailability as determined by the balance experiment, this being judged to be the most precise method for bioavailability determination.

A study was conducted to evaluate the effects of maternal undernutrition on the hypothalamic-pituitary-gonadal axis in female sheep offspring. Pregnant ewes were fed to 100% throughout pregnancy (Control) or to 50% from 0 to 30 (R1) or from 31 to 100 days of gestation (R2). Female lambs were selected and fed to appetite throughout the study. At 2, 5.5 and 10 months of age a GnRH challenge was conducted. At the age of 10 months lambs were synchronized and blood samples were collected at 3 h intervals for 72 h following sponge removal. At slaughter (10 months) ovaries were removed and examined macroscopically. Maternal undernutrition did not affect the time of the onset of puberty, defined as the first increase in plasma progesterone concentrationsor=1 ng/ml. The magnitude of the pre-ovulatory gonadotrophin surge and the time to surge were unaffected by treatment. The LH and FSH response to GnRH challenge did not differ between groups at 2 and 5.5 months but at 10 months of age a higher (p0.05) FSH response was found in R1 group. Although the total number of visible follicles and corpora lutea did not differ between groups, a significant higher (p0.05) number of small (2-3 mm diameter) follicles in R1 group and a significant lower number (p0.05) of corpora lutea with diameter 8-11 mm and not even one with diameter12 mm were detected in the ovaries of R2 lambs. In conclusion, maternal undernutrition during the first month of pregnancy resulted in increased pituitary sensitivity to GnRH and increased number of small follicles in the ovary, while during mid to late gestation resulted in a reduction of large corpora lutea in female offspring.

This paper presents a programmable multi-granular optical cross connect (MG-OXC) and network architecture deployable in multi-service and multi-provider networks. The concept of programmable MG-OXC is introduced to provide a way of utilizing multiple switching/transport granularities to efficiently support the emerging traffic demands in both core and metro networks. For this reason, the supported bandwidth granularities include full lambdas, sub- and super-lambdas and multiple transport formats such as bursts, flows and packets. The programmability is envisaged by a software/hardware platform that simplifies network control, re-planning at the logical level and end-to-end service transparency, by translating the technology-specific information to technology independent services in an abstracted and logical manner.

Olive tree leaves (OTL) and grape marc (GM) are by-products with high linolenic (LNA) and linoleic (LA) acid content, respectively, which can be used as dietary ingredients to increase the cis-9 trans-11 conjugated linoleic acid (CLA) content of milk fat in sheep and goats. An experiment was conducted with 16 Friesian ewes and 16 Alpine goats to study the effect of OTL and GM inclusion in sheep and goat diets on their milk fatty acid profile, with emphasis on cis-9, trans-11 CLA and vaccenic acid (VA). Ewes and the goats were fed the control (C) diets from parturition to the 90 days in milk (DIM) and then both groups were divided into two sub-groups (treatments). The control groups of both species continued to be fed the C diets, whereas the treated groups were gradually switched over a 2-week period (DIM=91–105) from the C diets to that of treatment 1, which contained air-dried OTL. These OTL diets were fed ad libitum for 1 month (DIM=106–135). After that period, the same treated groups, after 2 weeks of gradual adaptation (DIM=136–150), were switched to treatment 2, which contained air-dried GM. The GM diets were fed ad libitum for 1 month (151–180 DIM). Concentrations of polyunsaturated fatty acids (PUFA) and mono-unsaturated fatty acids increased significantly in milk fat of sheep fed OTL v. C. For goats, only the PUFA in milk fat was increased by feeding OTL compared with C. Relative to C, GM increased significantly the concentration of PUFA only in milk fat of sheep. OTL and GM diets increased the cis-9, trans-11 CLA and VA content in milk fat, compared with C, only in sheep. GM caused a sharp increase in 18:0 only in sheep milk fat, while the OTL diet increased significantly the 18:0 in milk fat of goats. GM and OTL diets also had opposite effects on the 18:1/18:0 ratio of sheep milk fat. In conclusion, OTL and GM, when included in sheep diets altered the milk fatty acid profile with a pronounced increase in cis-9, trans-11 CLA and VA contents. The results show that the response of sheep and goats to OTL and GM diets was different, suggesting a species difference that needs further investigation.

Eighteen lambs (L) and 18 goats (G), 12 weeks of age, were allocated in equal numbers to three diets. The basal (B) diet had no added copper (Cu) while the other two had 30 or 60 mg added Cu per kg dry matter (DM) as CuSO4.5H2O. Lambs and goats were housed and fed individually. After 67 days, deaths from Cu toxicity occurred only in lambs from groups L-30 (basal diet + 30 mg Cu per kg DM) and L-60 (basal diet + 60 mg Cu per kg DM). The remaining lambs and all of the goats were slaughtered at 91 and 137 days respectively.A decreased food intake and loss of weight were observed in some lambs towards the end of the experiment which appeared to be associated with an approaching haemolytic crisis. The additional dietary Cu had a positive effect on food conversion efficiency only in the early stages of the experiment in both lambs and goats. None of the goats died or lost weight.Plasma Cu concentrations and serum glutamate oxalacetate transaminase and creatine kinase activities were elevated in lambs of groups L-30 and L-60 after 2 months, while in goats the concentrations were in normal ranges during the whole experimental period.Additional dietary Cu increased significantly the Cu concentrations of liver, kidney, muscle, spleen, heart, brain, hair and faeces in both lambs and goats. Additional Cu decreased the fresh and dry liver weights of lambs and the liver to live weight ratio whereas the corresponding values for kidneys were increased. The differences between groups of goats in fresh or dry liver and kidney weights were not significant, though the fresh and dry liver weights tended to be higher in groups G·30 and G·60 compared with those in the group G·B.The amount of Cu stored in the livers of the lambs was six to nine times higher than that of the goats. Differences in Cu storage between lambs and goats may be related to species differences in Cuutilization and hence resistance to Cu toxicity.

Over the past few years it has become evident that local computational resources cannot keep up with the demands generated by some users/applications and distributed computing using the concept of a computational grid is proposed as the solution. An example is emerging scientific applications that couple scientific instruments, data and high-end computing resources distributed on a global scale. The interconnection of such resources can be quite a challenging task, raising strict requirements for a suitable network infrastructure. Optical networks can support this type of applications offering a large amount of low cost bandwidth and a number of relevant research activities focus on optical network infrastructures utilizing GMPLS control plane for fast and automated provisioning of connections across multi-technology and multi- domain networks. phosphorus (lambda user controlled infrastructure for european research) is an FP6 EU funded Integrated Project that aims to address some of the key technical challenges to enable on-demand, end-to-end network services across multiple domains in a seamless and efficient way. More specifically, the network concept, the network control plane developments and test-beds of the project will make applications aware of their complete grid resources (computational and networking) environment and will be able to make adaptive and optimized use of heterogeneous network infrastructures connecting various high-end resources. As part of this work, phosphorus is concentrating on the development of a common and transversal control plane for the Grid network infrastructures, in which Grid and optical network resources will be controlled by the same control plane for provisioning and recovery purposes, seamlessly spanning across different control and management domains. By introducing a set of extensions to the standard GMPLS, the developed grid-GMPLS architecture introduces interfaces specific for grids, resulting in a more powerful network control plane solution. In this paper we will give an overview of phosphorus and present the grid-GMPLS control plane considerations and architecture, with focus on issues such as routing that will be implemented and demonstrated by the project.

In this paper we propose a novel architecture for the control plane of an application aware Optical Burst Switching (OBS) network, able to support GRID computing applications. We suggest the use of the Session Initiation Protocol (SIP) to implement a protocol layer between the GRID specific control messages and the network control plane. The proposed architecture was implemented in an existing OBS network testbed and was successfully tested as described in the paper.

We evaluated the predictive value of several parameters, including the International Staging System (ISS) for myeloma, in patients with advanced disease treated with thalidomide-based regimens (TBR). We analyzed 119 patients, from 3 phase II studies. Patients with pretreatment beta2 microglobulin = 3.5 g/dl were scored ISS stage 1, patients with beta 2 microglobulin 5.5 mg/l ISS stage 3. ISS stage was 1, 2 and 3 in 45, 32 and 23% of patients respectively. Seventy-four patients (62%) achieved at least partial response. Median progression-free and overall survival were 8 months and 19.5 months respectively. ISS stage, serum LDH and performance status were independent predictive factors for survival. Based on these 3 variables a scoring system was developed with survival times of 38.1, 28.8 and 5.8 months for scores 0, 1 and 2 respectively. The ISS staging system was highly predictive for overall survival of patients with advanced myeloma treated with TBR. With the addition of performance status and serum LDH, a simple scoring system was developed which may help select patients likely to bene. t from TBR.

There is an increasing interest for sustainable forms of livestock production systems, which will provide a balanced relationship between environmental, socio-cultural and economic factors. The small ruminant sector is examined with particular focus on the possibilities of improving the sustainability of small ruminant systems and on the possible role of organic production to meet the demand of sustainability. Current regulations highlighting different approaches and interpretation of organic farming among countries are reviewed. Regulations concerning organic livestock production between the US and the EU are compared. For the future development of organic farming, a strong harmonisation of rules and legislation at international and national level is needed. The process of conversion from conventional to organic poses several problems mainly due to inadequate technical knowledge and value-added activities at farm or regional level with poorly organized marketing. Breeding strategies, feed management and disease control for small ruminant in organic farming are discussed. Animal selection should be designed to reinforce, in a sustainable manner, the relationships between animals and the environment. Feed management will require a better integration between agriculture and livestock, and a transition from monoculture to mosaic, with spatial and temporal integration of agricultural components. Possible alternatives to chemoprophylaxis are available to control helminth diseases, such as the use of homeopathic treatment, and the improvement of genetic resistance to parasite infections. Harmonisation of rules and development of technical assistance at local level may contribute to increase sustainability of small ruminant organic farming not only in developed but also in developing countries. Certification and valorisation of organic products are key points to guarantee and promote this sector. L'articolo è disponibile sul sito dell'editore http://www.sciencedirect.com/science/journal/03016226

Background: Waldenstrom’s macroglobulinemia (WM) is an unusual lymphoplasmacytoid lymphoma characterized by the presence of a serum monoclonal immunoglobulin M. Although several studies have evaluated possible prognostic factors of this disease, few have focused on the survival and prognosis of symptomatic patients after the initiation of treatment. Patients and methods: Our study included 122 previously untreated patients with a median age of 67 years who required systemic treatment. Multiple variables were analyzed for their prognostic value on survival after initiation of treatment using univariate and Cox regression multivariate analysis. Results: The median overall survival was 106 months. Pretreatment factors associated with shorter survival were age greater than or equal to65 years, splenomegaly, B-symptoms (weight loss, fever or night sweats), hemoglobin

The goal of this study was to investigate the dose-dependent effects of active cigarette smoking on endothelial nitric oxide (NO) and endothelin-1 (ET-1) biosynthesis.Limited studies have suggested that active cigarette smoking may be associated with a dose-dependent reduction of endothelium-dependent vasodilation (EDV). The underlying biochemical changes that cause this dose-specific effect, such as changes in the endothelial NO biosynthetic pathway and ET-1 production, have not been examined.Flow- and nitroglycerin-mediated reactivity of the brachial artery were measured in eight nonsmokers, seven light smokers (or =1 pack/week) and eight heavy smokers (or =1 pack/day), and their sera were added to confluent ( approximately 85%) monolayers of human umbilical endothelial cells (HUVECs) for 12 h. Basal and substance P-stimulated NO and basal ET-1 production were measured. The HUVECs used for measuring basal NO production were lysed, and both endothelial NO synthase (eNOS) protein expression and eNOS activity were determined.Serum cotinine level and pack-years of smoking were significantly lower in light smokers compared with heavy smokers (p0.006 and p0.004, respectively). There were no significant differences between heavy smokers and light smokers in EDV (p = 0.52), basal- (p = 0.70) and stimulated-NO production (p = 0.95), eNOS protein (p = 0.40) and eNOS activity (p = 0.63). Compared with nonsmokers, all the parameters were significantly altered in both of the smokers' groups. No differences were found in nitroglycerin-mediated vasodilation and in vitro ET-1 production among the three groups.These results indicate light smoking may have similar detrimental effects on EDV and NO biosynthetic pathway as does heavy smoking. These data may have important implications concerning the amount of active cigarette exposure that imparts cardiovascular risk.

Information concerning worm control practices of sheep and goat farmers in the region of Trikala (central Greece) was collected through a questionnaire survey by visiting farms and interviewing farmers. Questionnaires from 57 farmers residing in 23 rural communities were collected. Anthelmintics were used by 89% of the farmers. On average, lambs, kids and goats were treated once annually, while sheep were treated either once or twice annually. Only 2% of farmers reported treatment of animals with anthelmintics when moving to new pastures. The most common broad-spectrum anthelmintics used were those belonging to the benzimidazoles and probenzimidazoles. Fifty nine percent of the farmers used the same anthelmintic group for 3 or more years and 34% used two or more anthelmintic groups in the same year. Almost all farmers reported estimating live weights for calculating anthelmintic doses through visual perception on the basis of an average weight (96%). Tablets and boluses were the most preferred anthelmintic formulation used by 96% of farmers. The selection of an anthelmintic was based for 58% of farmers on recommendation by a veterinarian and for 39% of farmers on the cost of the drug. The most common occasions for deworming the animals were at turn out (86%) and after parturition (31%). Only 6% of farmers reported deworming new animals before introducing them onto the farm. Farmers preferred to seek information about the use of anthelmintics and worm control strategies from veterinarians (63%) and other farmers (37%).

Purpose: Both docetaxel and cisplatin have moderate activity in patients with advanced urothelial cancer. We performed a multicenter phase II study in order to assess the efficacy and toxicity of the combination of these two agents in patients with advanced carcinoma of the urothelium. Patients and methods: Sixty-six patients not amenable to curative surgery or irradiation were enrolled onto this cooperative group study and treated on an outpatient basis with docetaxel 75 mg/m(2) followed by cisplatin 75 mg/m(2), both administered intravenously. Granulocyte-colony stimulating factor was administered subcutaneously at a dose of 5 mu g/kg daily from day 5 until resolution of neutropenia. The chemotherapy was administered every three weeks for a maximum of six courses in patients without evidence of progressive disease. Results: Thirty-four of sixty-six patients (52%, 95% confidence interval 40%-64%) demonstrated objective responses, with eight achieving clinical complete responses and twenty-six partial responses. A multivariate logistic regression analysis indicated that the patients most likely to respond were those without lung metastasis and without weight loss before treatment. The median duration of response was 6.1 months and the median times to progression and survival for all patients were 5 and 8 months, respectively. Absence of anemia, of liver metastases and of weight loss correlated with longer survival. Grade greater than or equal to 3 toxicities included granulocytopenia in 33% of patients, anemia in 14%, diarrhea in 13% and emesis in 7% of patients. Conclusion: The combination of docetaxel and cisplatin appeared relatively well tolerated and moderately active in patients with advanced urothelial cancer. The patients most likely to benefit were those without weight loss and without lung or liver metastases.

Proteasome-dependent and autophagy-mediated degradation of eukaryotic cellular proteins represent the two major proteostatic mechanisms that are critically implicated in a number of signaling pathways and cellular processes. Deregulation of functions engaged in protein elimination frequently leads to development of morbid states and diseases. In this context, and through the utilization of GAL4/UAS genetic tool, we herein examined the in vivo contribution of proteasome and autophagy systems in Drosophila eye and wing morphogenesis. By exploiting the ability of GAL4-ninaE. GMR and P{GawB}Bx(MS1096) genetic drivers to be strongly and preferentially expressed in the eye and wing discs, respectively, we proved that proteasomal integrity and ubiquitination proficiency essentially control fly's eye and wing development. Indeed, subunit- and regulator-specific patterns of severe organ dysmorphia were obtained after the RNAi-induced downregulation of critical proteasome components (Rpn1, Rpn2, α5, β5 and β6) or distinct protein-ubiquitin conjugators (UbcD6, but not UbcD1 and UbcD4). Proteasome deficient eyes presented with either rough phenotypes or strongly dysmorphic shapes, while transgenic mutant wings were severely folded and carried blistered structures together with loss of vein differentiation. Moreover, transgenic fly eyes overexpressing the UBP2-yeast deubiquitinase enzyme were characterized by an eyeless-like phenotype. Therefore, the proteasome/ubiquitin proteolytic activities are undoubtedly required for the normal course of eye and wing development. In contrast, the RNAi-mediated downregulation of critical Atg (1, 4, 7, 9 and 18) autophagic proteins revealed their non-essential, or redundant, functional roles in Drosophila eye and wing formation under physiological growth conditions, since their reduced expression levels could only marginally disturb wing's, but not eye's, morphogenetic organization and architecture. However, Atg9 proved indispensable for the maintenance of structural integrity of adult wings in aged flies. In toto, our findings clearly demonstrate the gene-specific fundamental contribution of proteasome, but not autophagy, in invertebrate eye and wing organ development.

Bacterial lipopolysaccharide (LPS) attachment at the hemocyte surface is based on the crosslinking of surface associated p47 to LPS, via the intermediacy of tyrosine derivatives generated by the action of phenoloxidase (PO). This attachment is an initial step for LPS internalization from hemocytes (Charalambidis et al., 1996). The results presented clearly show the critical role of hemocyte associated PO activity in the above processes. Biochemical and immunofluorescent analysis demonstrated unambiguously the presence of prophenoloxidase (proPO) on the hemocyte surface. The cell-surface expression of proPO appeared to be LPS-independent, whereas its activation was LPS-dependent. The activation of cell surface proPO involves a limited proteolysis, since upon activation with chymotrypsin proPO is converted to a set of smaller molecular weight proteins with PO activity. The activation appears to be due to enzyme activators, serine proteases, released upon LPS-stimulation. This hypothesis was supported from the activation of membrane proPO by the culture medium of hemocytes which have been triggered with LPS. In addition, proPO, activation was abolished by inhibitors of secretion and PMSF. The release of proPO activators upon LPS-stimulation is mediated via protein tyrosine phosphorylation, as genistein inhibited proPO activation, a situation similar to that reported by us for the release of the effector protein p47 (Charalambidis et al., 1995). The LPS-stimulated activation of cell-surface proPO is a prerequisite for LPS (either cell associated or cell free) internalization, as judged by the resistance of LPS binding to dissociation by proteinase K.

It is well known that activated prophenoloxidase (proPO) plays an important role in cuticular melanization and sclerotization. In addition, studies dealing with immune response of insects suggest that phenoloxidase (PO) is also critical in the defense reactions of insects against invaders. proPO is activated by elicitors derived from microbial cell wall components such as peptidoglycan, beta-1,3-glucan, and lipopolysaccharide (LPS). According to our recent studies we proposed a model clarifying the role of PO in both cellular and humoral immune responses. LPS triggers Ceratitis capitata hemocytes via induced protein tyrosine phosphorylation to release biologically active molecules, including p47 and proPO-activators. Furthermore, hemocytes in response to LPS facilitate clearance of LPS from the hemocoel of medfly. The effector molecules involved in the LPS clearance are hemocyte surface-associated p47 (mp47), soluble p47 (sp47), activated proPO, and tyrosine. A similar LPS clearance system in the integument of medfly in vitro was also demonstrated. According to our data, the proposed mechanism for LPS clearance from hemocoel and from integument is the crosslinking of LPS to p47 or certain integumental proteins via the intermediacy of reactive tyrosine derivatives generated by PO activity, as is the case for cuticular protein-chitin crosslinks during sclerotization. We also demonstrated that metabolites of the eumelanin biosynthesis and not melanin itself or N-acetyldopamine (NADA), the key precursor of sclerotizing agent, were necessary for the immune responses by hemocytes and integument.

Purpose: The oxidative and reductive stresses within the varicocele veins were estimated. Nitric oxide synthase and xanthine oxidase activities, as well as nitric oxide, S-nitrosothiols and superoxide release within the spermatic vein in patients with varicocele, and the role of the noxious oxidant peroxynitrite formed from nitric oxide and superoxide in sperm dysfunction were determined. Materials and Methods: Whole blood samples were drawn from a peripheral vein and a dilated varicocele vein before ligation. Nitric oxide synthase, xanthine oxidase, nitric oxide and peroxynitrite were measured by novel spectro-fluorophotometric methods. S-nitrosothiols were estimated by a luminol-chemiluminescence method. Serum and red blood cell antioxidant capacity was determined by a chemiluminescence reaction. Results: Serum nitric oxide synthase and xanthine oxidase activities, as well as nitric oxide, peroxynitrite and S-nitrosothiol levels were greater in the spermatic vein compared to the peripheral vein. Serum antioxidant capacity was greater in varicocele veins compared to peripheral veins. In contrast, the antioxidant capacity of red blood cells was less in the varicocele veins, which was consistent with an increased rate of peroxynitrite production. Conclusions: Our data suggest a high oxidative stress due to the release of nitric oxide synthase and xanthine oxidase within the dilated spermatic vein. This reaction resulted in dramatic formation of nitric oxide, peroxynitrite and S-nitrosothiols, which are biologically active. Formation of peroxynitrite from the reaction of nitric oxide with superoxide could be a causative factor for impaired sperm function in patients with varicocele.

Insect hemocytes (blood cells) synthesize the major nonself recognition protein (47 kDa) during 3rd instar larvae (V.J. Marmaras, S. Tsakas, Dev. Biol. 129, 294-303 (1988)). In this study we show the presence of the 47 kDa protein in plasmatocytes (main hemocyte type) and prohemocytes. In plasmatocytes this protein appears to be localized both in vesicles and in the cell surface. The cell surface-associated 47 kDa protein was released from membrane fraction by 1 M NaCl, indicating that it is not tightly bound. Bacterial lipopolysaccharide (LPS) can function on isolated hemocytes from Ceratitis capitata larvae, inducing their spreading and degranulation. During degranulation (exocytosis) the plasmatocytes release the 47 kDa protein, among others. This protein could not be normally traced in serum, nor is it released by basal secretion. The secretion of the 47 kDa protein was found to be LPS-dependent, whereas its presence on plasmatocyte surface is LPS independent. LPS-stimulated exocytosis of the 47 kDa protein appears to be dependent on protein tyrosine phosphorylation. We have now demonstrated that LPS increases tyrosine phosphorylation of 19 and 22 kDa polypeptides in C. capitata hemocytes. Inhibition of the LPS-induced tyrosine phosphorylation mediated by tyrosine kinase inhibitor, genistein, was accompanied by the inhibition of the secretion of the 47 kDa protein. These results support the hypothesis that tyrosine protein phosphorylation is a signal reaction in hemocytes after LPS exposure. These LPS responses of insect plasmatocytes show strong similarities to mammalian macrophages (S. Weinstein et al., J. Immunol. 151, 3829-3838 (1993)). In a model we propose that the LPS-independent cell surface-associated 47 kDa protein is responsible for the phagocytosis and for the formation of nodules and capsules, whereas the LPS-dependent secreting counterpart is responsible for the extracellular killing of bacteria.

Parvin is a putative F-actin binding protein important for integrin-mediated cell adhesion. Here we used overexpression of Drosophila Parvin to uncover its functions in different tissues in vivo. Parvin overexpression caused major defects reminiscent of metastatic cancer cells in developing epithelia, including apoptosis, alterations in cell shape, basal extrusion and invasion. These defects were closely correlated with abnormalities in the organization of F-actin at the basal epithelial surface and of integrin-matrix adhesion sites. In wing epithelium, overexpressed Parvin triggered increased Rho1 protein levels, predominantly at the basal side, whereas in the developing eye it caused a rough eye phenotype and severely disrupted F-actin filaments at the retina floor of pigment cells. We identified genes that suppressed these Parvin-induced dominant effects, depending on the cell type. Co-expression of both ILK and the apoptosis inhibitor DIAP1 blocked Parvin-induced lethality and apoptosis and partially ameliorated cell delamination in epithelia, but did not rescue the elevated Rho1 levels, the abnormal organization of F-actin in the wing and the assembly of integrin-matrix adhesion sites. The rough eye phenotype was suppressed by coexpression of either PTEN or Wech, or by knock-down of Xrp1. Two main conclusions can be drawn from our studies: (1), high levels of cytoplasmic Parvin are toxic in epithelial cells; (2) Parvin in a dose dependent manner affects the organization of actin cytoskeleton in both wing and eye epithelia, independently of its role as a structural component of the ILK-PINCH-Parvin complex that mediates the integrin-actin link. Thus, distinct genetic interactions of Parvin occur in different cell types and second site modifier screens are required to uncover such genetic circuits.