[Treatment of acetabular bone defects in revision hip arthroplasty using the Revisio-System]. / Defektadaptierte Versorgung azetabulärer Knochendefekte mit dem Revisio-System.

BACKGROUND: Many different systems for the management of primary and secondary acetabular defects are available, each with its inherent advantages and disadvantages. The Revisio-System is a press-fit oval mono-block implant that makes a defect-oriented reconstruction and restoration of the center of rotation possible. MATERIAL AND METHODS: In this study, we retrospectively reviewed the outcome of 92 consecutive patients treated with this oval press-fit cup due to periacetabular bone loss. The average follow-up was 58.2 months. Defects were classified according to D'Antonio. There were 39 type II, 38 Type III, and 15 type IV defects. After an average of 4.9 years, the implant survival rate was 94.6% with cup revision as the end point and 89.1% with revision for any reason as the end point. The Harris Hip Score increased from 41.1 preoperatively to 62.3 postoperatively. The mean level of pain measured with the Visual Analogue Scale (VSA) was reduced from 6.9 preoperatively to 3.8 postoperatively. RESULTS: The Revisio-System represents a promising toolbox for defect-orientated reconstruction of acetabular bone loss in revision hip arthroplasty. Our results demonstrate that the implantation of the Revisio-System can result in a good mid-term clinical outcome.

The adenovirus type 5 E1B-55K oncoprotein actively shuttles in virus-infected cells, whereas transport of E4orf6 is mediated by a CRM1-independent mechanism.

The E1B-55K and E4orf6 proteins of adenovirus type 5 are involved in viral mRNA export. Here we demonstrate that adenovirus infection does not inhibit the function of the E1B-55K nuclear export signal and that E1B-55K also shuttles in infected cells. Even during virus infection, E1B-55K was exported by the leptomycin B-sensitive CRM1 pathway, whereas E4orf6 transport appeared to be mediated by an alternative mechanism. Our results strengthen the potential role of E1B-55K as the "driving force" for adenoviral late mRNA export.

Investigation of nucleo-cytoplasmic transport using UV-guided microinjection.

Active nucleo-cytoplasmic transport is mediated by dynamic signal-mediated pathways. We investigated the effects of transcription inhibitors or fluorescent lectins on nuclear import mediated by nuclear localization signals (NLSs). Therefore, a novel experimental approach that allows the controlled sequential introduction of fluorescent substances into living cells was established. A microinjection system equipped with an UV-source enabled us to identify fluorescent-labeled cells for the subsequent introduction of additional fluorescent compounds, in order to study their interactions in vivo. Cells were initially labeled either by expression of autofluorescent proteins or by microinjection of fluorescent substances. Transcription inhibitors did not affect nuclear transport mediated by classical NLSs but inhibited import mediated by the M9-domain of hnRNPA1. Comparison of a mono- and bipartite NLS revealed that the bipartite signal was more active in import. Sequential injection of differentially labeled nuclear import and export substrates allowed monitoring of import and export simultaneously in the same living cell. The introduced experimental approach will also be useful to analyze a variety of biological processes in living mammalian cells.

Mature dendritic cells infected with herpes simplex virus type 1 exhibit inhibited T-cell stimulatory capacity.

Mature dendritic cells (DC) are the most potent antigen-presenting cells within the entire immune system. Interference with the function of these cells therefore constitutes a very powerful mechanism for viruses to escape immune responses. Several members of the Herpesviridae family have provided examples of such escape strategies, including interference with antigen presentation and production of homologous cytokines. In this study we investigated the infection of mature DC with herpes simplex virus type 1 (HSV-1) and the way in which infection alters the phenotype and function of mature DC. Interestingly, the T-cell-stimulatory capacity of these DC was strongly impaired. Furthermore, we demonstrated that HSV-1 leads to the specific degradation of CD83, a cell surface molecule which is specifically upregulated during DC maturation. These data indicate that HSV-1 has developed yet another novel mechanism to escape immune responses.

Inhibition of CD83 cell surface expression during dendritic cell maturation by interference with nuclear export of CD83 mRNA.

Dendritic cells (DCs), nature's adjuvant, must mature to sensitize T cells. However, although the maturation process is essential, it is not yet fully understood at the molecular level. In this study, we investigated the course of expression of the unique hypusine-containing protein eukaryotic initiation factor 5A (eIF-5A), which is part of a particular RNA nuclear export pathway, during in vitro generation of human DCs. We show that eIF-5A expression is significantly upregulated during DC maturation. Furthermore, an inhibitor of the hypusine modification, GC7 (N(1)-guanyl-1, 7-diaminoheptane), prevents CD83 surface expression by apparently interfering with nucleocytoplasmic translocation of the CD83 mRNA and, importantly, significantly inhibits DC-mediated T lymphocyte activation. The data presented suggest that CD83 mRNA is transported from the nucleus to the cytoplasm via a specific nuclear export pathway and that hypusine formation appears to be essential for the maturation of functional DCs. Therefore, pharmacological interference with hypusine formation may provide a new possibility to modulate DC function.

The adenovirus type 5 E1B-55K oncoprotein is a highly active shuttle protein and shuttling is independent of E4orf6, p53 and Mdm2.

The E1B-55K and E4orf6 oncoproteins of adenovirus type 5 are involved in the export of viral mRNAs. Previously, it was suggested that a complex composed of E1B-55K and E4orf6 serves as a nucleocytoplasmic transporter for viral mRNAs in which the E4orf6 protein directs both nuclear import and export. We now demonstrate that the E1B-55K protein itself shuttles efficiently in the absence of E4orf6. In addition, E1B-55K trafficking was independent of the defined shuttle proteins Mdm2 or p53, which interacts with E1B-55K. The identified N-terminal E1B-55K leucine-rich nuclear-export signal (NES) conferred rapid nuclear export even in a heterologous system in contrast to the postulated E4orf6NES. Interestingly, although shuttling was blocked by inhibitors of the CRM1 mediated export pathway, E1B-55K inhibited neither the activity nor the trafficking of the retroviral shuttle proteins HIV-1 Rev and HTLV-1 Rex. In contrast, Rev or Rex blocked the nuclear export of E1B-55K, most likely by competing for essential export factors. Our results provide new insights into the regulation of the adenovirus mRNA export system and the processes of adenovirus mediated transformation. Oncogene (2000) 19, 850 - 857.

Nuclear pore localization and nucleocytoplasmic transport of eIF-5A: evidence for direct interaction with the export receptor CRM1.

Eukaryotic initiation factor 5A (eIF-5A) is the only cellular protein known to contain the unusual amino acid hypusine. The exact in vivo function of eIF-5A, however, is to date unknown. The finding that eIF-5A is an essential cofactor of the human immunodeficiency virus type 1 (HIV-1) Rev RNA transport factor suggested that eIF-5A is part of a specific nuclear export pathway. In this study we used indirect immunofluorescence and immunogold electron microscopy to demonstrate that eIF-5A accumulates at nuclear pore-associated intranuclear filaments in mammalian cells and Xenopus oocytes. We are able to show that eIF-5A interacts with the general nuclear export receptor, CRM1. Furthermore, microinjection studies in somatic cells revealed that eIF-5A is transported from the nucleus to the cytoplasm, and that this nuclear export is blocked by leptomycin B. Our data demonstrate that eIF-5A is a nucleocytoplasmic shuttle protein.

Effect of halofuginone (Stenorol) on Chukar partridge (Alectoris chukar).

This study was conducted to assess the effect of the coccidiostat halofuginone (Stenorol) on growth, feed consumption, and survival of Chukar partridge. Halofuginone was fed to three replicates (14 chicks per replicate) of chukar chicks from 2 to 7 d of age at levels of 0, 1.5, 3.0, 6.0 and 12 ppm. Mortality from 2 to 7 d was 0, 0, 0, 11, and 21 birds, respectively, by treatment. Seven-day body weight showed a significant linear decrease with increasing halofuginone level (P < 0.01). On the 7th d, replicates receiving 6.0 and 12.0 ppm halofuginone were transferred to unmedicated feed for the remainder of the test due to excessive mortality. The other groups were continued until 6 wk of age. At 6 wk, chicks fed 6 or 12 ppm halofuginone from 2 to 7 d and then unmedicated feed did not differ in body weight from those fed the unmedicated control diet. A significant difference in mortality was not observed among the other three treatment groups to 6 wk of age. A linear depression in 3-, 4-, 5-, and 6-wk body weight with increasing halofuginone level was observed within the first three treatment levels (P < 0.05). It was concluded that 1.5 ppm halofuginone depressed growth of young chukars and that 6 ppm resulted in increased mortality.

Effect of diet on growth and plasma ascorbic acid in chicks.

Six experiments were conducted to study the effect of diet on growth and plasma ascorbic acid in chickens. D-Glucuronolactone failed to improve growth with either a crude yeast-fish meal diet or a purified diet based on casein and gelatin. With the purified diet, D-glucuronic acid and L-gulonolactone also failed to improve growth and did not influence plasma ascorbic acid levels. Dietary ascorbic acid improved growth of chicks with a purified diet in most cases, but not with a corn-soybean diet. Meat meal and fish meal caused slight increases in plasma ascorbic acid, whereas soybean meal, safflower meal, and cottonseed meal caused greater increases when used in a purified diet. Gulonolactone oxidase activity in the kidney was not different between chicks fed the purified or the corn-soybean diets, but was reduced by 0.1% dietary ascorbic acid. The mechanism for the increase in plasma ascorbic acid with the addition of soybean meal and other plant protein sources to the diet is not known.

Availability to chicks of biotin from dried egg products.

Two feeding experiments were conducted with duplicate groups of five chicks each to study the availability of biotin in spray-dried egg products. In the first experiment chicks that were fed diets containing 43% dried whole egg (DWE) grew poorly and developed perosis and dermatitis. The signs were prevented and growth improved progressively with supplementation of 0.5 and 1.0 mg biotin/kg diet. In the second experiment dried egg yolk (DEY) and dried egg white (DEW) were compared with DWE at equivalent levels of egg components. Signs of biotin deficiency and reduced growth were slightly more severe with DEW than with DWE, although liver biotin content was slightly lower at 0 and 0.5 mg biotin/kg with DWE than with DEW. Growth with DEY and no added biotin was not different from that with DEY and 500 or 1000 mg biotin/kg diet, although liver biotin was lower than when supplemental biotin was added. Liver fat was approximately five times greater in the groups receiving DWE and DEY than in the groups receiving DEW. The results show that the biotin contained in egg yolk is inadequate to counteract the deficiency of biotin caused by the avidin in egg white so that unheated dried whole egg is deficient in this vitamin.

Effects of dietary biotin and linoleate on polyunsaturated fatty acids in tissue phospholipids.

Two experiments with factorial designs were conducted to study the interaction between dietary biotin and linoleate in male broiler chicks. Chicks were fed a purified basal diet containing varying levels of d-biotin (0, 200, or 400 micrograms/kg diet) and linoleate. In Experiment 1, chicks were fed the basal diet containing three levels of added linoleate (.5, 3.1, or 4.1% of diet) for each biotin level and in Experiment 2, four levels of linoleate were fed (.27, .98, 2.00, or 2.20% of diet). The average body weights of chicks fed biotin at 200 micrograms/kg of diet and .27 or .5% added linoleate were not different (P greater than .05) from those of chicks fed higher levels of biotin or linoleate supplements to the basal diets. Linoleate deficiency resulted in elevated omega-9 fatty acids (18:1 omega 9 and 20:3 omega 9) in liver and heart phospholipids. For liver phospholipids, linoleate deficiency led to reduced 18:2 omega 6, 20:3 omega 6, and 20:4 omega 6 but for heart phospholipids only 20:4 omega 6 was lowered. Biotin deficiency resulted in elevated 18:3 omega 6 in liver and heart lipids and decreased 20:3 omega 6 in liver phospholipids. Liver microsomes from biotin-deficient chicks contained increased 18:2 omega 6 and reduced 20:3 omega 6 compared with those of biotin-adequate chicks.

Dietary biotin effects on polyunsaturated fatty acids in chick tissue lipids and prostaglandin E2 levels in freeze-clamped hearts.

Chicks were fed a purified diet with 0, 100, 200, 300, 400, or 500 micrograms/kg diet of added biotin to determine the effects of biotin deficiency on polyunsaturated fatty acids in tissue lipids. Body weight was reduced by biotin deficiency and liver and heart biotin levels varied with the biotin in the diet. Fatty acids in liver and lung from biotin-deficient chicks at 15 days contained elevated (P less than .03) 18:3 omega 3 and 18:2 omega 6 but prostaglandin precursors 20:3 omega 6 and 20:4 omega 6 were reduced (P less than .03) in liver lipids. Heart tissues from 15-day-old chicks fed the biotin-deficient diet were low (P less than .03) in 20:3 omega 6. Feeding acetylsalicylic acid in diets containing added biotin (0, 100, 400, and 500 micrograms/kg) did not significantly alter fatty acid levels in chick tissue lipids but significantly reduced plasma prostaglandin E2 (PGE2). Biotin deficiency reduced heart PGE2 levels in 22-day-old chicks. An 8-h fast reduced (P less than .04) 20:4 omega 6 in chick heart total fatty acids.

Nutritional qualities of stabilized and raw rice bran for chicks.

Rice bran either raw or processed in an extrusion cooker at 130 C was fed to meat strain chickens for 25 days after hatch. Either full fat or hexane-extracted rice bran was placed in the diet at the equivalent of 60% full fat bran. Raw full fat bran for one diet was stored at -23 C until fed, whereas rice bran for all other diets was stored at 32 C. Four experiments were conducted at 6-week intervals. Free fatty acid (FFA) content in oil from raw rice bran stored at the elevated temperature reached 81% by the start of the final experiment whereas FFA in stabilized bran oil remained at about 3%. Chickens fed stabilized rice bran made significantly greater gains than chickens fed raw bran diets. Feed efficiency was superior for chickens fed either full fat or extracted stabilized bran compared with full fat bran stored at either 32 or -23 C. Feed conversion for extracted raw bran was intermediate between stabilized bran and full fat raw bran. Raw bran stored at 32 C (with elevated FFA content) tended to produce lower gains than the frozen raw bran. Analysis of the combined data from all four trials indicated that raw bran held at 32 C produced the lowest gains among all of the diets.

Tissue lipid fatty acid composition of biotin-adequate and biotin-deficient chicks.

Male broiler chicks were fed a purified ingredient diet containing various levels of added biotin to study dietary biotin effects on polyunsaturated fatty acids in selected tissues. Chicks were fed a basal diet containing adequate or deficient levels of added biotin and an additional group was fed a natural ingredient practical diet as a control. No significant differences in average body weights were found in chicks at 21 days fed 200 micrograms or more of biotin or the stock mash diet. Liver and heart biotin levels were lower in chicks fed less than 200 micrograms of biotin (P less than .01). At 21 days of age, the liver fatty acids of biotin-deficient chicks contained greatly (P less than .05) elevated c16:l, 18:ln9, 18:2n6, 18:3n6, and 18:3n3, whereas 20:3n6 and 20:4n6 were significantly lowered (P less than .05). Heart fatty acids c16:l and 18:2n6 of 21-day-old biotin-deficient chicks were elevated (P less than .05) and 18:3n3, 20:3n6, and 20:5n3 were lowered (P less than .05). It appears that the impaired conversion of 18:2n6 to 20:4n6 in liver tissue from biotin-deficient chicks at 3 weeks can result in lowered prostaglandin precursors not only in the liver but in the heart and muscle lipids as well. Changes in the brain and lung lipids are less marked.

Use of plasma and egg yolk biotin of white Leghorn hens to assess biotin availability from feedstuffs.

A linear relationship (r greater than .98) was found between the level of dietary biotin and plasma and egg-yolk biotin in White Leghorn hens after feeding the experimental diets for 2 weeks. This shows high reliability of these parameters as indicators of biotin availability. When plasma and egg yolk biotin were used to assess biotin availability in feedstuffs, they indicated low values for wheat (0%) and sorghum (10 to 20%) and high values for corn (75 to 100%), soybean meal (100%), and meat and bone meal (85%). When these parameters were used to assess commercial poultry diets, they indicated values for a "layer" diet of 75 to 100% and for a "broiler" diet of 81 to 92%. Pelleting gave an increase in biotin availability of 10% over the mash diet. This method for assessing biotin availability promises to be simple, economic, rapid, and reliable.

Drinking water treatment with a commercial preparation of a concentrated Lactobacillus culture for broiler chickens.

Three hundred male broiler chicks were used to determine the effects of supplying Lactobacillus in the drinking water. The product used, Biomax 40TM, was a fresh-frozen, pure culture of Lactobacillus containing 40 X 10(9) cfu/ml. Water treatments consisted of continuous lactobacilli dosing (CD) and skip-a-day lactobacilli dosing (SAD). A control group (C) received no lactobacilli. Each treatment and control contained two pens of 50 chicks each and were fed for a duration of 7 weeks. At the termination of the experiment, weights of the wet viscera, and wet and dry small intestines of the broilers revealed no significant differences between treatments and control. Surface pH readings taken from crop and duodenum showed that there was no significant difference (P greater than .05). Microbiology performed on duodenal contents revealed higher numbers of lactobacilli for CD and SAD broilers than for C broilers. The liver biotin contents of the lactobacilli-treated broilers were not significantly different from those of the controls. Body weights were not changed by either treatment, although they were greater for the CD broilers than for the SAD broilers. No difference was observed in feed conversion (feed/gain).

Furazolidone residues in chicken and swine tissues after feeding trials.

Broiler chickens and swine fed furazolidone in their diet were sacrificed, and samples of liver, kidney, skin/fat and muscle were harvested and analyzed for furazolidone residue. Chickens fed 200 g of furazolidone/ton of feed were withdrawn from treatment 21, 14, 7, 5, 3, or 0 days before slaughter. Birds withdrawn from medication more than 5 days prior to slaughter had no residues in any of the tissues sampled. One of the 12 birds in each of the 5 day and 3 day withdrawal groups had detectable residues in the skin/fat. Seven of the 12 birds in the 0 day withdrawal group had residues of less than 2 ppb in skin/fat samples. Chickens fed 400 g furazolidone/ton of feed were withdrawn from treatment 0 days before slaughter. Residues of 0.7 to 3.5 ppb were found in the skin of these birds; residues were not found in other tissues. Swine were fed 300 g furazolidone/ton of feed for 2 weeks or 150 g/ton for 5 weeks. They were withdrawn from treatment 10, 7, 5, 3, or 0 days before slaughter. Tissue samples taken from these swine did not contain detectable furazolidone residues.

Failure of dietary leucine to influence the tryptophan-niacin pathway in the chicken.

Three experiments were conducted with young chicks to determine whether dietary leucine affects the conversion of tryptophan to niacin. Either tryptophan or niacin improved growth and reduced perosis when chicks were fed a purified diet marginal in tryptophan and deficient in niacin. Addition of 4.8% L-leucine to the diet did not alter the growth and perosis prevention response obtained with tryptophan. Liver weight was slightly increased by the addition of 5.4% L-leucine to the diet. Plasma insulin was slightly reduced by leucine and by isoleucine and valine. Picolinic carboxylase in the kidney was reduced in chicks fed 0.2% tryptophan with no niacin and was also reduced when isoleucine and valine were added to the diets. Liver picolinic carboxylase activity was not influenced by diet. Plasma isoleucine and valine were reduced by the addition of leucine to the diet and were increased again when isoleucine and valine were added to the diet. Plasma leucine was increased by the addition of leucine but was not altered by valine and isoleucine. Plasma tryptophan was not influenced by dietary supplements of leucine or isoleucine and valine. The results show that in the chick there is no evidence for an effect of leucine on the tryptophan to niacin pathway.