Long-term results after semiconstrained distal radioulnar joint arthroplasty: A focus on complications.

Arthroplasty of the distal radioulnar joint (DRUJ) using a semiconstrained DRUJ implant yields good outcomes according to the literature. The aim of this study was to investigate the subjective, clinical and radiographic outcomes with a special focus on complications in nine patients with a mean follow-up of 6years and to compare them with our previously published 3-year follow-up results. No subjective or objective changes were seen between the 3-year and the 6-year follow-up. In the previous study, one implant loosening and two irritations of the superficial branch of the radial nerve occurred. We saw three complications that needed surgery in addition to the three complications already found 3years after surgery. One patient with a large ulna had loosening of the cemented ulnar stem and therefore the prosthesis was explanted. One patient had an allergic reaction to the metal alloy of the prosthesis, which also led to removal. One patient had an ulnar impaction syndrome caused by too-distal placement of the implant that needed revision. Prior studies reported low complication rates. In our study, six complications occurred in four out of nine patients, requiring reoperation including two revisions and two implant removals. A precise surgical technique is mandatory to avoid the otherwise frequent complications and potential implant failures. LEVEL OF EVIDENCE: IV.

Extending the limits of reconstructive microsurgery in elderly patients.

BACKGROUND: Population aging strongly affects the demographic development of industrialized countries. While microsurgical procedures were initially believed to be only feasible in patients of younger age because of the duration of the surgical procedure and the higher risk of vascular insufficiency due to age-related comorbidities, it has become evident that these procedures are beneficial even for patients at an advanced age. METHODS: We retrospectively investigated microsurgical procedures in a patient cohort (n = 25 with 27 free flaps) with a minimum age of 78 years with regard to patients' characteristics, flap survival, and postoperative surgical and medical complications. RESULTS: Median age was 81 years (IQR 6). Most defects were located in the head and neck region. The mean operation time was 384 min (standard deviation (SD) 131). Flap failure was observed in three cases (11%). The median length of hospital stay was 17 days (interquartile range (IQR) 8). The mean ASA score was 2.48. Patients' age and ASA group did not correlate. The mortality rate was 4%. Postoperative surgical complications were observed in 11 cases (41%), while 19 patients (70%) showed one or more medical complications. Higher ASA classes tended to show more postoperative complications. However, neither age nor operating time nor ASA status showed significant influence on the occurrence of postoperative medical or surgical complications. CONCLUSION: There is growing demand for structural and functional restoration using free tissue transfer in an aging population. If there are no alternative treatment options available promising similar structural and functional preservation, free tissue transfer is justifiably in very old patients despite a potentially increased flap failure. As such, free tissue transfer is used as a curative treatment concept aiming at a maximum of patients' independence and early ambulation. Occurrence of complications can be diminished by adequate patient selection and thorough perioperative care.

[Reduction of influenza-like infections by a phytoextract-vitamin-combination: a randomized, double-blind, placebo-controlled study]. / Reduktion grippaler Infekte durch eine Phytoextrakt-Vitamin-Kombination. Eine randomisierte, doppelblinde, placebokontrollierte Studie.

BACKGROUND: Recurrent infections are signs of a weakened immune system, and can be traced to a lack of vitamins and minerals. The objective of this study was to investigate the effect of a nutriological combination of vitamins, trace elements and phytochemicals on the incidence and duration of influenza-like infections. METHODS: Healthy subjects at risk of increased infection exposure took part in a randomized, double-blind, placebo controlled study, each involving a supplementation period of 12 weeks. The occurrence and severity of influenza-like infections and resort to study medication was documented by diary entries. SF-36 questionnaires to measure the health-related quality of life were completed at the start and end of the study. At final examination compliance was examined with respect to returned study medication, diaries and a questionnaire administered to evaluate the effect, safety, and tolerability of the test substance. RESULTS: Of 100 randomized participants, 80 (38 test treatment and 42 placebo) completed the study according to the protocol. In the test treatment group there were 60 influenza-like infections (1.6 infections/12 weeks), significantly fewer (p = 0.0003) compared with the placebo group with 113 episodes (2.7 infections/12 weeks). Further, the average number of days that the infection lasted was shorter in the test treatment group at 10.0 +/- 6.7 days, which was significantly shorter (p = 0.0003) than the placebo group at 28.5 +/- 13.6 days. The influenza-like symptom score and the mean duration of sick leave did not differ statistically between groups. Participants in the test treatment group assessed the global effectiveness of the test substance significantly better (p < 0.0001) than the participants of the placebo group. At the beginning of the study there was no difference between groups with respect to SF-36 results. However, at the completion of the study 6 of the 8 scales of SF-36 in the test treatment group were significantly better compared with the placebo group. CONCLUSION: In summary, consumption of the complex nutriological test preparation was associated with a significant reduction in the incidence of influenza-like infections, as well as their duration, and it was associated with significantly improved vitality, social functioning, and psychological well-being.

Regulation of melatonin 1a receptor signaling and trafficking by asparagine-124.

Melatonin is a pineal hormone that regulates seasonal reproduction and has been used to treat circadian rhythm disorders. The melatonin 1a receptor is a seven- transmembrane domain receptor that signals predominately via pertussis toxin-sensitive G-proteins. Point mutations were created at residue N124 in cytoplasmic domain II of the receptor and the mutant receptors were expressed in a neurohormonal cell line. The acidic N124D- and E-substituted receptors had high-affinity (125)I-melatonin binding and a subcellular localization similar to the neutral N124N wild-type receptor. Melatonin efficacy for the inhibition of cAMP by N124D and E mutations was significantly decreased. N124D and E mutations strongly compromised melatonin efficacy and potency for inhibition of K(+)-induced intracellular Ca(++) fluxes and eliminated control of spontaneous calcium fluxes. However, these substitutions did not appear to affect activation of Kir3 potassium channels. The hydrophobic N124L and N124A or basic N124K mutations failed to bind (125)I-melatonin and appeared to aggregate or traffic improperly. N124A and N124K receptors were retained in the Golgi. Therefore, mutants at N124 separated into two sets: the first bound (125)I-melatonin with high affinity and trafficked normally, but with reduced inhibitory coupling to adenylyl cyclase and Ca(++) channels. The second set lacked melatonin binding and exhibited severe trafficking defects. In summary, asparagine-124 controls melatonin receptor function as evidenced by changes in melatonin binding, control of cAMP levels, and regulation of ion channel activity. Asparagine-124 also has a unique structural effect controlling receptor distribution within the cell.

Association between ancillary services and clinical and behavioral outcomes among HIV-infected women.

The purpose of this study was to evaluate the association between ancillary services, including case management, and clinical and behavioral outcomes for human immunodeficiency virus (HIV)-infected women. Data were obtained from databases systematically maintained by Family Advocacy, Care and Education Services (FACES) and the HIV Outpatient Program (HOP) in New Orleans. HIV-infected women receiving primary care from HOP and ancillary services from FACES between January 1, 1997 and December 31, 1998 were eligible. Data were analyzed using generalized estimating equations (GEE) with STATA software. The majority of women included in the study were African American (86.7%), infected heterosexually (78.8%), and had absolute CD4 counts greater than 200 (58.6%). After adjusting for age, time, entry time into HOP, pregnancy, CD4 count, substance abuse status, and social and clinical stressors, receipt of more than four combined case manager contacts or ancillary services per month was significantly associated with being prescribed a protease inhibitor, improved adherence and retention in primary care, and enrolling on a research protocol. Receiving more than one transportation service per month was significantly associated with improved adherence, improved retention, one or more emergency room visits per month, and one or more hospitalizations per month. Receiving more than one contact with case managers per month was associated with improved retention in primary care. Findings suggest that receipt of case management and ancillary services is associated with improvements in multiple outcomes for HIV-infected women. A client-centered approach to providing ancillary services appears to be effective in improving behavioral and utilization characteristics in this population of low-income, high-risk women.

Selenium-enriched sprouts. A raw material for fortified cereal-based diets.

The selenium supply in almost all European countries, including Austria and Germany, is below the recommended daily intake. In these countries, selenium fortification of foods and the use of selenium supplements are quite popular to compensate for low Se intake from diets. In general, wheat (Triticum aestivum) is known to be a good source for bioavailable selenium, and many studies have been performed to enrich selenium in wheat by selenium fertilization of the soil. In the present work, the process of sprouting was investigated as an alternative to enrich selenium in wheat. Sprouting was chosen because it additionally improves the nutritional value of seeds, for example, by a higher vitamin content, a better quality of protein, and some other parameters. Wheat, alfalfa (Medicago sativa), and sunflower (Helianthus annuus) seeds were germinated for 5 and 7 days in solutions containing selenate. The selenium sensitivity of the sprouts was tested by measuring visible germination levels and seedling development. Uptake rates were studied by determination of total selenium using inductively coupled plasma mass spectrometry (ICP-MS). Metabolism of the absorbed selenium was analyzed by determination of selenium species in extracts of the sprouts using anion exchange HPLC coupled to ICP-MS. It was shown that sunflower sprouts were the most resistant and had the highest uptake rates (up to 900 mg/kg), but almost 100% of the selenium was extracted with water and found to be nonmetabolized selenate. Wheat and alfalfa were less resistant and enriched selenium up to concentrations of 100 and 150 mg of Se/kg of dry mass, respectively. The metabolism of the selenate was inversely related to the total uptake rates. At low Se enrichment (approximately 1-2 mg of Se/kg), <20% of the total selenium content within the sprouts remained as inorganic selenium, indicating a high metabolism rate. With increasing uptake the amount of selenate increased to approximately 40-50%. However, with the method used it is possible to produce sprouts containing certain amounts of selenium, which might provide substantial proportions of bioavailable selenium. In combination with the generally high nutritional value of sprouts, they might serve for production of improved cereal-based diets.

Molecular characterization of a novel microneme antigen in Neospora caninum.

The apical complex of the parasites belonging to the phylum Sporozoa is believed to be critically involved in the events leading to host cell invasion. The characterization of the components of this subcellular structure is therefore an important step towards understanding how these parasites achieve host cell entry. Affinity-purification of an anti-Neospora caninum antiserum on a reactive protein band of approximately 40 kDa following Triton-X-114 extraction of parasite proteins, SDS-PAGE and Western blotting, yielded an immunoglobulin fraction which, by immunofluorescence, stained predominantly the apical portion of N. caninum tachyzoites. Following immunoscreening of a N. caninum tachyzoite lambdagt22 cDNA expression library, the respective full length cDNA sequence was determined. This sequence was found to encode a protein of 362 amino acids, with a calculated Mr of 38086. This protein is encoded by a single copy gene which produces a transcript of 2.4 kb. Sequence analysis showed that it contains a N-terminal putative signal peptide sequence and two potential membrane spanning regions. Four consecutive epidermal growth factor like domains were identified, as well a conserved sequence motif for binding of ATP/GTP (P-loop). The full length cDNA was expressed as a recombinant poly-histidine fusion protein in Escherichia coli, and antibodies affinity purified on this protein labelled exclusively a 38 kDa band on immunoblots of N. caninum extracts. In addition, specific labeling of a 45 kDa band in Toxoplasma gondii tachyzoite extracts was observed. By immunofluorescence, these antibodies stained predominantly the apical portion of both N. caninum and T. gondii tachyzoites, but the protein was absent from the parasite surface. Immunogold localization in LR-White embedded N. caninum tachyzoites demonstrated staining of predominantly the apically located micronemes, as well as of dense granules located at the posterior end of the tachyzoites. As evidenced by immunohistochemistry, this Neospora microneme antigen and its immunoreactive counterpart in Toxoplasma appeared to be expressed in both tachyzoite and bradyzoite stages.

Detection of surface-associated and intracellular glycoconjugates and glycoproteins in Neospora caninum tachyzoites.

The surface-associated molecules of the invasive stages of apicomplexan parasites such as Neospora caninum and Toxoplasma gondii are most likely crucially involved in mediating the interaction between the parasite and its host cell. In N. caninum, several antigens have recently been identified which could participate in host cell adhesion and/or invasion. These are antigens which are either constitutively expressed on the outer plasma membrane, or antigens which are only transiently localised on the surface as they are expulsed from the secretory vesicles either prior, or after host cell invasion. Some of these proteins have been characterised at the molecular level, and it has been shown that they are, with respect to protein sequences, closely related to homologous counterparts in T. gondii. Nevertheless, there is only a low degree of cross-antigenicity between the two species. In microbial interactions it has been shown that carbohydrates could also play a crucial role in host cell recognition and immunological host parasite interactions. In this study we present data which strongly suggest that the surface of N. caninum tachyzoites is glycosylated. In SDS-PAGE, glycoproteins comigrated largely with glycosylphosphatidylinositol-anchored proteins which were identified using in vivo [3H]ethanolamine labelling followed by autoradiography. The lectin Con A reacted strongly with the surface of these parasites, binding of which is indicative for the presence of N-glycans. Additional surface binding was observed, although only in a subpopulation of all tachyzoites, for wheat germ agglutinin and Jacalin. Intracellular binding sites for Con A were mainly associated with the parasite dense granules. By lectin labelling of Western blots of N. caninum protein extracts, glycoproteins were identified which reacted specifically with the lectins Con A, wheat germ agglutinin, Jacalin and soy bean agglutinin.

Malignant fibrous histiocytoma of bone: a retrospective EMSOS study of 125 cases. European Musculo-Skeletal Oncology Society.

In an effort to learn more about malignant fibrous histiocytoma (MFH) of bone and its prognosis with different treatment approaches, the European Musculo-Skeletal Oncology Society (EMSOS) initiated a retrospective survey among its members. Data requested included patient and treatment variables and outcome. The information on all patients with histologically proven, primary, localized osseous extremity MFH was analyzed if surgical tumor removal was performed and disease status was documented for at least one follow-up date. 125 such patients were evaluable (74 male, 51 female; median age 34 years; tumor site femur 81, tibia 26, humerus 12, other 6). Local treatment was surgery only (110) or surgery plus radiotherapy (15). Chemotherapy was used in 97/125. On last follow-up, 85 patients remained in remission, 33 had developed metastases, 6 a local recurrence, and 1 a combined relapse. With a median follow-up of 3.9 years for patients at risk, actuarial 5-year disease-free survival was 59%. In univariate analyses, younger age and the use of chemotherapy were associated with a more favorable outcome, as was limb-salvage surgery. 23 of 66 tumors with information on response to preoperative chemotherapy responded well (> 90% necrosis). Among these 23, only one relapsed.

The antigenic composition of Neospora caninum.

Neospora caninum is an apicomplexan parasite which causes neosporosis, namely stillbirth and abortion in cattle, and neuromuscular disease in dogs. Although N. caninum is phylogenetically and biologically closely related to Toxoplasma gondii, it is antigenically clearly distinct. In analogy to T. gondii, three stages have been identified. These are: (i) asexually proliferating tachyzoites; (ii) tissue cysts harbouring slowly dividing bradyzoites; and (iii) oocysts containing sporozoites. The sexually produced stage of this parasite has only recently been identified, and has been shown to be shed with the faeces from dogs orally infected with N. caninum tissue cysts. Thus dogs are definitive hosts of N. caninum. Tachyzoites can be cultivated in vitro using similar techniques as previously described for T. gondii. Methods for generating tissue cysts containing N. caninum bradyzoites in mice, and purification of these cysts, have been developed. A number of studies have been undertaken to identify and characterise at the molecular level specific antigenic components of N. caninum in order to improve serological diagnosis and to enhance the current view on the many open questions concerning the cell biology of this parasite and its interactions with the host on the immunological and cellular level. The aim of this paper is to provide an overview on the approaches used for detection of antigens in N. caninum. The studies discussed here have had a great impact in the elucidation of the immunological and pathogenetic events during infection, as well as the development of potential new immunotherapeutic tools for future vaccination against N. caninum infection.

[Neoadjuvant therapy for localized osteosarcoma of extremities. Results from the Cooperative osteosarcoma study group COSS of 925 patients]. / Neoadjuvante Therapie des lokalisierten Osteosarkoms der Extremitäten. Erfahrungen der Cooperativen Osteosarkomstudiengruppe COSS an 925 Patienten.

BACKGROUND: Owing to twenty years of multicentric interdisciplinary cooperation, the COSS group has been able to collect data on a large group of osteosarcoma patients treated by neoadjuvant therapy. This paper reviews results achieved in patients with localized extremity tumors. INCLUSION CRITERIA: Registration into a completed neoadjuvant COSS-Study. Histologically confirmed, primary, localized, high-grade, central osteosarcoma of an extremity; age < 40 years; no pretreatment; interval diagnosis to chemotherapy < or = 3 weeks; no severe comorbidity. Chemotherapy: HD-methotrexate +/- doxorubicin +/- cisplatin +/- ifosfamide +/- BCD. Scheduled local therapy: Surgery. RESULTS: 925 evaluable patients from 101 institutions. Median age 15 years, m:f 1.4:1. Primary site: femur 510, tibia 251, humerus 100, fibula 51, other 13. Tumor-size < 1/3 of the involved bone 616, > or = 1/3 304. Definitive surgery in 903/925 cases, 443 limb salvage procedures. Good response (> 90% necrosis) in 469/806 (58.2%) evaluated tumors. Median follow-up for surviving patients: 5.42 years. Actuarial survival after 5 and 10 years: 72.5% (95%-CI 69.3-75.7) and 66.3% (62.5-70.0), relapse-free 62.1% (58.7-65.4) and 59.4% (55.8-63.0). 683/925 alive (601 first remission), 242 deceased (212 tumor progression, 30 other causes). 66.2% (97.3%) of all relapses within 2 (5) years. Prognosis correlates with tumor-size (< vs. > or = 1/3: 69.9% vs. 58.3% at 10 years) and -site (tibia: 74.2%, humerus: 54.5%) and -response (good vs. poor: 78.2% vs. 52.5%) (all p < 0.01). Actuarial 10-year survival by response grading I-VI according to Salzer-Kuntschik 80.9%, 82.8%, 71.1%, 60.7%, 47.7%, 27.3%. COSS-studies with preoperative 4-drug therapy more efficacious than less aggressive protocols. No impact of doxorubicin scheduling (sequential: rapid vs. 48 h-continuous infusion) or cisplatin scheduling (randomized: 5 h vs. 72 h-infusion) on prognosis detected. CONCLUSIONS: Intensive multiagent chemotherapy and delayed surgery for localized extremity osteosarcoma led to excellent oncologic results in the COSS-studies. Tumor-size, -site, and -response as well as the intensity of upfront chemotherapy correlated with outcome. Giving doxorubicin and cisplatin by continuous infusions did not result in discernible prognostic disadvantages.

Early infections in patients undergoing bone marrow or blood stem cell transplantation--a 7 year single centre investigation of 409 cases.

Infections are a major cause of morbidity and mortality in patients undergoing high-dose therapy and subsequent autologous or allogeneic haemopoietic stem cell transplantation, despite the change from topical to systemic anti-infection prophylaxis and the introduction of growth factors and new antimicrobial drugs. We report our single centre experience with data from 409 patients treated at our unit from its opening in 1990 until May 1997. Three hundred and seventy-eight patients were transplanted for the first time, 12 patients were retransplanted or boosted and 19 patients were readmitted for miscellaneous reasons. 245 patients were allografted and 157 autografted. Antimicrobial prophylaxis was mainly quinolones, fluconazole plus amphotericin-B orally, aciclovir, and TMP/SMX or pentamidine. Three hundred and nineteen (78%) developed fever of significantly longer duration in the allogeneic setting with anti-CMV seropositivity. The most frequent infection was fever of unknown origin (50.6%), followed by septicaemia (12.5%) and pneumonia (11.0%). Pathogens isolated in 24.6% of the infections were mostly gram-positive bacteria (57.9%), followed by non-fermenting rods (11.2%), Aspergillus spp. and Candida spp. (10.3%, each). Cumulative response rate to antimicrobial therapy was 66.9%. Infections were responsible for 62.5% (25/40) of deaths after transplantation. Predominant pathogens were Aspergillus spp. (11), Candida spp. (four), and Pseudomonas spp. (three). None of the patients died from gram-positive bacterial infection. The risk of dying from infection was 11.2% after allografting and 0.8% after autotransplantation. Infections remain a major risk for early death after allogeneic transplantation of haemopoietic stem cells. Infection with gram-negative bacteria can be prevented by quinolone prophylaxis. Predominant pathogens are Aspergillus spp. Candida spp. and nonfermenting rods. Systemic infection with these pathogens is associated with a poor prognosis. Antimycotic prophylaxis and the therapy must be improved.

Long-term results of the co-operative German-Austrian-Swiss osteosarcoma study group's protocol COSS-86 of intensive multidrug chemotherapy and surgery for osteosarcoma of the limbs.

BACKGROUND: In an effort to intensify osteosarcoma therapy, systemic ifosfamide was added pre- and postoperatively to an already aggressive three-drug regimen. In a subgroup of patients, loco-regional treatment intensification was attempted by using the intraarterial route to give cisplatin. PATIENTS AND METHODS: Patients < or = 40 years at diagnosis of a localised, de novo high-grade central extremity osteosarcoma were eligible for inclusion into study COSS-86 if registered within three weeks from biopsy. Doxorubicin, high-dose methotrexate, and cisplatin were given to all patients. Patients who fulfilled one or more of three defined high-risk criteria received early systemic treatment intensification by adding ifosfamide as the fourth agent. Preoperatively, these high-risk patients received cisplatin either intraarterially or intravenously. RESULTS: 171 eligible patients were entered, of which 128 were stratified into the high-risk group. When all 171 were analysed by intention-to-treat, actuarial overall and event-free survival rates at ten years were 72% and 66%, respectively. No benefit of intraarterial cisplatin application was detected. Cumulative treatment toxicity was considerable. CONCLUSIONS: In a multicenter setting, intensive treatment of osteosarcoma according to protocol COSS-86 led to long-term disease-free survival for two thirds of patients. We saw no benefit of using the intraarterial route to administer cisplatin.

Differential expression of cell surface- and dense granule-associated Neospora caninum proteins in tachyzoites and bradyzoites.

Morphologically, the tachyzoites and the tissue cysts of Neospora caninum are difficult to distinguish from those of other cyst-forming apicomplexan parasites such as Toxoplasma gondii. Several stage-specific antigens have been identified in T. gondii tachyzoites and bradyzoites, and respective antibodies are useful tools for discriminating between the 2 stages during tachyzoite-bradyzoite interconversion in T. gondii infections. Whereas several cell surface- and dense granule-associated proteins have been identified and characterized in N. caninum tachyzoites, not much is known about antigenic components expressed in N. caninum bradyzoites. In this study, the differential expression of the 2 N. caninum surface proteins Nc-p43 and Nc-p36 and the dense granule protein Nc-p33 (NCDG1) within tachyzoites and bradyzoites of N. caninum has been investigated.

Identification and characterisation of a dense granule-associated protein in Neospora caninum tachyzoites.

Neospora caninum is an apicomplexan parasite which is morphologically and ultrastructurally very similar to Toxoplasma gondii. In order to identify molecules involved in host cell entry and subsequent modification of the parasitophorous vacuole, a polyclonal antiserum directed against N. caninum tachyzoites was raised in a rabbit. Subcellular fractionation of tachyzoites was performed using the non-ionic detergent Triton-X-114. Membrane fractions were analysed by immunoblotting using the polyclonal antiserum. One of the immunoreactive protein bands had a mol. wt of 33,000 and was subsequently named Nc-p33. Affinity-purified anti-Nc-p33 antibodies were used to characterise this polypeptide using SDS-PAGE, isoelectric focusing, Western blot analysis and immuno-EM. Nc-p33 was found in two isolates of N. caninum (NC-1 and Liverpool), but could not be detected in T. gondii tachyzoites. Immunogold EM revealed that Nc-p33 constituted a dense granule-associated protein, and Western blotting demonstrated that Nc-p33 was most likely identical to the recently described antigen NCDG1. Shortly after invasion, this dense granule protein was targeted to the parasitophorous vacuole membrane, and, at later timepoints after infection, was also found on the parasitophorous vacuolar network. This suggested that Nc-p33 could play a functional role in the modification of the parasitophorous vacuole and its membrane.

Tacrolimus impairs wound healing: a possible role of decreased nitric oxide synthesis.

BACKGROUND: The effect of the immunosuppressant tacrolimus on wound healing is not known. Tacrolimus has been shown to decrease nitric oxide synthesis. The systemic inhibition of wound nitric oxide synthesis leads to impaired healing. METHODS: We studied the effect of systemic tacrolimus treatment on wound-breaking strength and collagen deposition 10 days after wounding in rats and to correlate the outcome of healing with wound nitric oxide synthesis. Beginning at the day of wounding, rats were treated once daily by intraperitoneal injections with 0.5, 1.0, or 2.0 mg tacrolimus/kg body weight. Nitrite and nitrate were measured in wound fluid as an index of wound nitric oxide synthesis. Expression of inducible nitric oxide synthase in the wound was investigated by immunohistochemistry. Splenic lymphocytes were tested for proliferative activity. Tacrolimus levels in blood and wound fluid were measured by enzyme-linked immunosorbent assay. RESULTS: Systemic tacrolimus treatment was well tolerated by all rats. Tacrolimus accumulated in wound fluid. Tacrolimus levels in wound fluid were found to be approximately 10-fold higher than blood levels (P < 0.001). Tacrolimus (2.0 mg/kg/day) reduced wound-breaking strength (P < 0.01) and collagen deposition (P < 0.05). This was paralleled by decreased wound nitrite + nitrate levels (P < 0.001) and wound-inducible nitric oxide synthase expression. Splenic lymphocyte proliferative activity was significantly decreased by 1.0 and 2.0 mg tacrolimus/kg body weight/day (P < 0.05), indicating that the tacrolimus doses used were immunosuppressive. CONCLUSION: Our data show for the first time that tacrolimus impairs wound healing, and this is reflected by diminished wound nitric oxide synthesis.

The major 36 kDa Neospora caninum tachyzoite surface protein is closely related to the major Toxoplasma gondii surface antigen.

The tachyzoites and the tissue cysts containing bradyzoites of Neospora caninum and Toxoplasma gondii, respectively, are difficult to distinguish morphologically. Specific antigens have been identified in T. gondii tachyzoites and bradyzoites, some of which are stage-specifically expressed, and different functions have been attributed to some of them. A tachyzoite stage-specifically expressed surface protein is the major surface antigen 1 (SAG1) which has been shown to be involved in host cell attachment and invasion. Previously we have identified a cell surface-associated glycoprotein (p36) in N. caninum tachyzoites. The full length coding sequence of the cDNA coding for p36 was determined, and analysis of the deduced amino acid sequence demonstrated that p36 is closely related to SAG1. p36 is encoded by a single copy gene which produces a transcript of 1.4 kb. Immunogold labeling of resin-embedded parasites using polyclonal antibodies affinity-purified on a recombinant p36 fusion protein expressed in Escherichia coli showed that this protein is located exclusively on the tachyzoite cell surface. As SAG1 in T. gondii, p36 is expressed in the tachyzoite stage, but is absent from bradyzoites. p36 is recognized by antibodies present in sera of cows experimentally infected with N. caninum tachyzoites.