Effects of air exposure, temperature and additives on fermentation characteristics, yeast count, aerobic stability and volatile organic compounds in corn silage.

Ensiling conditions strongly influence fermentation characteristics, yeast count, and aerobic stability. Numerous volatile organic compounds including esters are produced, which may negatively affect feed intake and animal performance and air quality. In addition to a farm survey, 3 laboratory experiments were carried out to study the effects of air (by delayed sealing or by air infiltration during anaerobic storage), temperature (20 and 35°C), and various types of additives [blends of either sodium benzoate and sodium propionate (SBSP) or of sodium benzoate and potassium sorbate (SBPS); buffered mixture of formic and propionic acids (FAPA); homofermentative inoculant (LAB)]. After additive treatment, chopped whole corn plants were packed into 1.5-L glass jars and stored for several months. For treatments with air infiltration, glass jars with holes in the lid and body were used. The farm survey in 2009 revealed large variation in lactate, acetate, ethanol, n-propanol, and 1,2-propanediol concentrations. Whereas ethyl esters were detected in all silages, the mean ethyl lactate concentrations were higher than those for ethyl acetate (474 vs. 38mg/kg of dry matter). In the ensiling experiments, few unequivocal effects of the tested factors on the analyzed parameters were observed due to many interactions. Delayed ensiling without additives decreased lactic acid production but, in one trial, increased acetic acid and had no effect on ethanol. The effect of delayed sealing on yeast counts and aerobic stability differed widely among experiments. Air infiltration during fermentation tested in one trial did not alter lactic acid production, but resulted in more acetic acid in delayed and more ethanol than in promptly sealed untreated silages. Greater ethanol production was associated with increased yeast numbers. Storage at high temperature resulted in lower lactic acid and n-propanol, and a trend toward reduced ethanol production was observed. The additive FAPA consistently caused increased ethanol and reduced n-propanol levels with no effect on yeast counts and aerobic stability. When the additives SBSP and SBPS decreased n-propanol and ethanol, reduced yeast counts were also found. Ethyl ester formation was strongly correlated with those of ethanol and to a lesser degree with those of the respective acid.

Molecular mechanism for feedback regulation of C4 biosynthesis in guinea pig peritoneal macrophage.

Previous reports have shown that regulation of local extrahepatic production of complement may not reflect the regulation of plasma concentrations of the corresponding proteins and, further, that alteration of the tissue microenvironment can affect local macrophage protein synthesis. This report describes the molecular basis for control of the biosynthesis and secretion of a class III major histocompatibility complex gene product, the fourth component of complement (C4), from guinea pig macrophages by extracellular native C4 protein. The effect is specific for C4 synthesis, since production of C2 and total secreted protein was unaffected by fluid phase C4. C4 synthesis by extracellular C4 is regulated at a pretranslational level, without an effect on posttranslational proteolytic cleavage, glycosylation, or secretion. Specific C4 and factor B cDNA probes were used to demonstrate, by dot hybridization and Northern blot analysis, a decrease in messenger RNA coding for C4 that paralleled the inhibition of C4 biosynthesis, while the amount of total RNA and mRNA specific for factor B remained constant. Inhibition of C4 biosynthesis and the disappearance of mRNA encoding C4 occurred between 4 and 6 h after exposure of the macrophages to biologically active or methylamine-inactivated C4 protein. These data demonstrate that regulation of C4 biosynthesis by guinea pig macrophages serves as a model for the study of the molecular mechanisms of macrophage activation as well as the control of production of a component of the inflammatory response.

Molecular basis of complement C3 deficiency in guinea pigs.

In experiments to ascertain the biochemical basis of a genetically determined deficiency of the third component of complement (C3) in guinea pigs, we found that C3-deficient liver and peritoneal macrophages contain C3 messenger RNA of normal size (approximately 5 kb) and amounts, that this mRNA programs synthesis of pro-C3 in oocytes primed with liver RNA and in primary macrophage cultures. In each instance, heterodimeric native C3 protein was secreted with normal kinetics but the C3 protein product of the deficient cells failed to undergo autolytic cleavage and was unusually susceptible to proteolysis. These data and a selective failure of C3 in plasma of deficient animals to incorporate [14C]methylamine suggested either a mutation in primary structure of the C3 protein or a selective defect in co- or postsynthetic processing affecting the thiolester bridge, a structure important for C3 function. A mutation in the primary structure of C3 was ruled out by comparison of direct sequence analysis of C3 cDNA generated from two C3 deficient and two C3 sufficient guinea pig liver libraries. Three base pair differences, none resulting in derived amino acid sequence differences were identified. Finally, restriction fragment length polymorphisms were identified in the C3 gene that are independent of the deficiency phenotype. This marker of the C3 gene permits testing of these hypotheses using molecular biological and classical genetic methods.

Cost-effectiveness analysis of telemedical devices for pre-clinical traffic accident emergency rescue in Germany.

OBJECTIVES: The purpose of this study is to assess the cost-effectiveness (net costs per life year gained) of telemedical devices for pre-clinical traffic accident emergency rescue in Germany. METHODS: Two equipment versions of a telemedical device are compared from a societal perspective with the baseline in Germany, i.e. the non-application of telemedicine in emergency rescues. The analysis is based on retrospective statistical data covering a period of 10 years with discounted costs not adjusted for inflation. Due to the uncertainty of data, certain assumptions and estimates were necessary. The outcome is measured in terms of "life years gained" by reducing therapy-free intervals and improvements in first-aid provided by laypersons. RESULTS: The introduction of the basic equipment version, "Automatic Accident Alert", is associated with net costs per life year gained of euro 247,977 (at baseline assumptions). The full equipment version of the telemedical device would lead to estimated net costs of euro 239,524 per life year gained. Multi-way sensitivity-analysis with best and worst case scenarios suggests that decreasing system costs would disproportionately reduce total costs, and that rapid market penetration would largely increase the system's benefit, while simultaneously reducing costs. CONCLUSION: The net costs per life year gained in the application of the two versions of the telemedical device for pre-clinical emergency rescue of traffic accidents are estimated as quite high. However, the implementation of the device as part of a larger European co-ordinated initiative is more realistic.

Secondary Ph-positive CML with a minority monosomy 7 clone.

We report a case of Ph-positive chronic myelogenous leukemia (CML) secondary to previous treatment for a lymphoma. At the time of original diagnosis of lymphoblastic lymphoma, chromosome studies of blood and bone marrow were normal. Following therapy and a clinical remission complicated by CNS relapse, the patient presented 16 months after treatment was discontinued with a WBC of 110,000 mm-3, consistent with CML. Blood and marrow cytogenetic studies at this time showed a Ph chromosome, t(9;22)(q34;q11) translocation, without other karyotypic alteration. A separate small clone with the karyotype 45,XY, -7 was found in the blood. His disease followed an aggressive course and he died 3 months later. The autopsy findings indicated CML in blast crisis. Molecular studies performed on cells replacing a lymph node revealed a rearrangement of the breakpoint cluster region (bcr) of chromosome #22 and of the immunoglobulin heavy chain locus. Taken together, it seems most likely that the patient's CML developed as a second neoplasm following successful elimination of his lymphoblastic lymphoma by therapy.

Adenocarcinoid tumor of the ampulla of Vater.

We report a rare case of adenocarcinoid tumor of the ampulla of Vater. The tumor contained an intermixture of adenocarcinoma and carcinoid tumor and was removed successfully by pancreaticoduodenectomy. The characteristics of these rare tumors are reviewed.

Extramedullary hematopoiesis during therapy with granulocyte colony-stimulating factor.

Granulocyte colony-stimulating factor is a glycoprotein that promotes the proliferation and differentiation of neutrophils. It also results in an increase in circulating hematopoietic progenitor cells. We describe two cases of extramedullary hematopoiesis in patients receiving granulocyte colony-stimulating factor with chemotherapy for metastatic breast cancer.

In vitro metabolism of fumonisin B1 by ruminal microflora.

Fumonisin B1 (FB1) is a mycotoxin produced by Fusarium moniliforme and F. proliferatum. Little is known of its metabolic fate after oral ingestion in ruminants, but these animals are reported to be tolerant towards FB1. The metabolism of this mycotoxin was evaluated following incubation (1 microg/ml) in ruminal fluid for up to 72 h, in the presence or absence of alfalfa as a substrate for microbial growth, using a model rumen (sealed flask, anaerobic conditions, exclusion of light, gentle agitation, 39 degrees C). The decrease in FB1 concentration and the production of short-chain fatty acids were determined. FB1 had no effect on SCFA production. After 72 h incubation, FB1 depletion was 12% and 18% in samples with and without alfalfa, respectively. No hydrolysed metabolites (aminopolyols or aminopentol) were detected. These results indicate that FB1 is poorly metabolized in the rumen and suggest that such metabolism is not the cause of the tolerance to this toxin displayed by ruminants.

Alveolar soft part sarcoma. A clinicopathologic and immunohistochemical study.

The histogenesis of alveolar soft part sarcoma (ASPS) has been investigated since its description. Twenty ASPS cases were analyzed for immunohistochemical content, with emphasis directed toward the paraganglial, Schwann cell, and muscle theories of histogenesis. In addition, the cases were examined for possible prognostic clinical features. The clinical characteristics of the patients were similar to those reported previously concerning average age (23 years); male:female ratio (1:1); and predominant primary site (lower extremity, nine cases). Despite a local recurrence rate of 20% and a metastatic rate of 68% (including four at presentation), the natural history was often indolent and relapse commonly occurred very late. The average follow-up period was 10.1 years. While the overall 5-year survival was 67%, only seven of 18 patients were alive without disease at last follow-up (1.7-32 years), and one patient died of tumor after a 28-year disease-free interval. Neither tumor size nor site appeared to affect prognosis. The tumors were analyzed immunohistochemically for neurofilament, S-100 protein, met-enkephalin, leu-enkephalin, acetylcholinesterase, alpha 1-antichymotrypsin, Factor VIII-related antigen, serotonin, lysozyme, neuron-specific enolase, myoglobin, cytokeratins, desmin, and vimentin. Except for weak vimentin immunoreactivity, no other antigenic expression was detected despite multiple repeated experiments with several antibodies. S-100 protein which is present in virtually all granular cell tumors was absent in the cases of ASPS. The lack of detectable expression of neurofilament, met-enkephalin and leu-enkephalin, and neuron-specific enolase is interpreted as evidence against the paraganglial theory of histogenesis. Similarly, the repeated absence of the muscle proteins, desmin and myoglobin, in contrast to a previous report, is interpreted as evidence against a myogenic origin.

Prevention ofPenicillium roqueforti - associated aerobic deterioration of maize silage by various additives.

Application of benzoic acid (0.2%) is a technological measure to prevent aerobic deterioration of maize silages caused by yeasts and moulds, particularly byP. roqueforti. Furthermore, benzoic acid inhibits undesired butyric acid fermentations.

C1s-induced vascular permeability in C2-deficient guinea pigs.

Normal guinea pigs that have been intradermally injected with C1s exhibit increased vascular permeability at the injection site. Guinea pigs that are genetically deficient in complement component C2 do not exhibit increased vascular permeability when given a similar injection. The C2-deficient guinea pigs respond normally to injections of bradykinin and kallikrein, suggesting that these animals can respond to kinins and have a normal kininogen pathway. When the C2-deficient guinea pigs are given guinea pig C2 before C1s injection, increased vascular permeability is observed. These results demonstrate a definite requirement for complement component C2 in the generation of C1s-induced vascular permeability.

Malignant mixed Müllerian tumors of the uterus. An immunohistochemical study.

Malignant mixed müllerian tumors (MMMT) of the uterus have been subdivided into two types: those with heterologous sarcomatous elements (e.g., rhabdomyosarcoma, and chondrosarcoma) and those with only homologous elements (e.g., stromal sarcoma). The distinction, which may have prognostic significance, was based on the identification by light microscopy of cells that exhibited definite cross-striations, cartilage, or osteoid production. We studied 32 cases of uterine MMMT to assess the value of immunohistochemical markers in delineating the sarcomatous and epithelial components. Of 32 cases, 11 showed heterologous sarcoma (6, rhabdomyosarcoma, and 5, chondrosarcoma), and the remaining 21 were homologous MMMT. Six antigens--desmin, myoglobin, S-100, alpha 1-antichymotrypsin (ACT), epithelial membrane antigen (EMA), and monoclonal cytokeratin (AE1 and AE3), (to test for possible myogenic, chondroid, fibrohistiocytic, and carcinomatous differentiation)--were analyzed by the avidin-biotin-peroxidase method. EMA was found in neoplastic cells in all cases; 31 of 32 cases showed keratin. Desmin reactivity was detected in 14 of 32 cases, whereas myoglobin was present in 10 of 32. Three cases exhibited S-100 positivity (two in areas of chondrosarcoma, and one in some stromal sarcoma cells). Twenty-two cases (69%) exhibited ACT reactivity. Several cases displayed a malignant fibrous histiocytoma pattern. These demonstrated ACT positivity in both the neoplastic spindle and giant cells. We conclude that immunohistochemical staining for the above mentioned antigens is a useful diagnostic aid in delineating the sarcomatous and carcinomatous elements in MMMT.

Prospective controlled study of home and hospital therapy of cystic fibrosis pulmonary disease.

A controlled prospective study was undertaken to compare the efficacy and benefits of home and hospital treatment for patients with exacerbations of pulmonary disease caused by cystic fibrosis. A total of 41 home and 41 hospital treatments were analyzed. Home and hospital patients were matched according to sex, age, pulmonary function tests, and arterial blood gas values. Both home and hospital treatments resulted in statistically significant improvement in pulmonary function. A comparison of these values did not show any statistically significant difference between groups at admission or discharge. Furthermore, the mean number of treatment days for both groups, individually determined by the primary physician, was equivalent (home 17.7 +/- 1.1 days, hospital 18.1 +/- 4.1). The mean charge for a home treatment was approximately $10,000, and for a hospital treatment $18,000. Sixty-five percent of home care patients and 68% of hospital patients required retreatment for pulmonary exacerbations within the study period; the interval between pulmonary exacerbations for the two groups was not significantly different. In addition, 85% of patients receiving treatment at home were able to maintain at least some of their school or work activities. These data indicate that home therapy for cystic fibrosis patients with pulmonary exacerbations is less costly and is as effective as in-hospital therapy.

Sarcoidosis in an adult with cystic fibrosis.

Sarcoidosis in an adult patient with cystic fibrosis lung disease was diagnosed on the basis of pulmonary function and radiographic data. It should be considered in the differential diagnosis of new diffuse interstitial infiltrates or hilar adenopathy in a patient with cystic fibrosis; biopsy of lung, lymph node, or skin lesions and interleukin-2 receptor levels may help to obtain a diagnosis.

Constitutive and IL 1-regulated murine complement gene expression is strain and tissue specific.

To study the molecular mechanisms accounting for strain- and tissue-specific variations in the production of complement proteins, complementary DNA probes were used to assess qualitative and quantitative differences in specific mRNA content of complement proteins C2, factor B, and C3 in extracts of tissues (liver, lung, spleen, kidney, and peritoneal macrophages) isolated from various mouse strains. Northern blot analysis of total hepatic RNA revealed differences in C2, factor B, and C3 mRNA levels in strains that share B10 background but differ in the H-2 region (e.g., H-2k, H-2u, H-2d, H-2f). In each instance, hepatic mRNA specific for the individual gene product corresponded in amount to the serum levels. By contrast, specific mRNA content of C2 and factor B in macrophages differed significantly from those observed in liver for each strain. Modulation of C2, factor B, and C3 expression was studied after in vivo administration of recombinant IL 1 or endotoxin to H-2k (B10.AKM) or H-2u (B10.PL) strain mice. As assessed by Northern blot analysis, neither endotoxin nor IL 1 affected liver C2-specific mRNA but increased specific C2 mRNA levels in kidney and lung. For both strains, IL 1 increased specific factor B mRNA in all tissues examined except for the H-2u strain liver factor B mRNA content, which was not affected by IL 1, whereas that of H-2k mice was increased. The lack of factor B modulation by IL 1 in the H-2u lines was specific to that gene and not a reflection of a generalized IL 1 unresponsiveness. Differences in tissue and strain specific constitutive and IL 1-regulated expression of the C3 gene were also observed in the H-2u and H-2k strains.

Macrophage maturation: differences in complement secretion by marrow, monocyte, and tissue macrophages detected with an improved hemolytic plaque assay.

In order to examine one function of mononuclear phagocytes during maturation from bone marrow precursors to tissue macrophages, an improved hemolytic plaque assay for the detection of synthesis of the second (C2) and fourth (C4) components of C by single cells was developed. With this method, production of C2 and C4 was assessed in cell populations derived from bone marrow, blood, lung, peritoneum, and spleen. The proportion of cells producing C2 and C4 in each population varied. Approximately 10% of bone marrow cells produced C4, but not detectable C2 plaque-forming cells (PFC) were detected. Circulating monocytes yielded about 10% PFC each for C2 and C4. The proportion of C2-producing cells in tissue macrophages varied from approximately 2% in bronchoalveolar macrophages to about 45% in peritoneal and splenic macrophage populations, whereas C4 production by macrophages from lung, peritoneum, and spleen were all approximately 45%. These data suggest that differences in C biosynthesis characterize mononuclear phagocytes at different stages of maturation.

Isolation of guinea pig macrophages bearing surface C4 by fluorescence-activated cell sorting: correlation between surface C4 antigen and C4 protein secretion.

Studies of complement (C) secretion by single cells indicate that only a subset of a guinea pig macrophage population is capable of secreting the second (C2) and fourth (C4) components of C. Moreover, cell-surface bound C4 antigen is also found on a proportion of guinea pig peritoneal macrophages. The availability of methods for the isolation of macrophages bearing surface membrane C4 antigen and for the detection of the secretion of C by single cells made it possible to ascertain the relationship between these two subsets. Approximately 25% of the freshly isolated peritoneal macrophages were surface C4 antigen positive. When incubated in conditioned medium containing C4 or incubated in small volumes to increase the concentration of fluid phase C4, the proportion of macrophages bearing surface C4 increased to approximately 80%. The surface C4 antigen was adsorbed from the medium and was predominantly native C4. The proportion of peritoneal macrophages secreting functionally active C4 or C2 was approximately 45% as measured by a hemolytic plaque assay technique. The proportion of C4-secreting cells decreased to 5% after incubation in conditioned medium containing preformed C4, whereas the proportion of C2-producing macrophages was unchanged, i.e., it remained at about 45%. The removal of secreted C4 with F(ab')2 anti-C4 or effectively decreasing C4 concentration by increasing the volume of the culture medium abrogated the decrease in the proportion of C4-secreting cells. Conditioned medium derived from cells genetically deficient in C4 had no effect on the proportion of C4- or C2-producing macrophages. The macrophage population bearing surface membrane C4, isolated by fluorescence-activated cell sorting, contained more than 90% of the C4-producing cells. Cells producing C2 were distributed equally in both subpopulations. Maintenance of the surface C4-positive, C4-producing cells in culture for 12 hr resulted in a decrease in the proportion of C4-secreting cells. Conversely, isolated macrophages initially surface C4 negative and not producing C4, developed the capacity to produce C4 in culture. C4 production in the isolated surface C4-negative population was inhibited by incubation in medium containing preformed C4. These results suggest the presence of a negative feedback effect on C4 secretion that is mediated by extracellular C4.(ABSTRACT TRUNCATED AT 400 WORDS)

Alternate-day prednisone reduces morbidity and improves pulmonary function in cystic fibrosis.

A randomised, double-blind, placebo-controlled study examined the effects of alternate-day prednisone therapy on morbidity and progression of lung disease in cystic fibrosis (CF). At baseline the patients (aged 1-12 years) had mild to moderate lung disease, and the prednisone group did not differ significantly from the placebo group for any values measured. After 4 years, the prednisone-treated group had significant advantages over the placebo group for height, weight, vital capacity, forced expiratory volume in 1 s, peak flow rate, erythrocyte sedimentation rate, and serum IgG. The prednisone-treated group required 9 admissions to hospital for CF-related pulmonary disease compared with 35 for the placebo group. There were no steroid-induced side-effects. To rule out bias in case selection, 69 CF clinic patients comparable in age and clinical status but not included in the study were compared with the placebo group at 4 years; no significant differences between the groups were found.

Tissue-specific pretranslational regulation of complement production in human mononuclear phagocytes.

The net production of the complement protein C2, C4, and factor B differ among mononuclear phagocytes from peripheral blood and different tissues in experimental animals and in humans. To examine the mechanisms that regulate these differences in humans, the proportions of C2-producing cells, the average single-cell production rate of C2, the posttranslational glycosylation and kinetics of secretion of C2 and factor B, and the amounts of C2 and factor B mRNA were examined in freshly isolated peripheral blood monocytes, monocytes maintained up to 2 wk in culture, and freshly isolated tissue macrophages from breast milk and bronchoalveolar lavage. In addition, the biosynthesis of two other proteins synthesized and secreted by mononuclear phagocytes, C3 and lysozyme, were examined. We report that despite comparable rates of C3 and lysozyme synthesis and similar processing and kinetics of secretion of C2 and factor B, the freshly isolated tissue macrophage differs from the monocyte-derived macrophage in the proportion of C2-producing cells, in the average single-cell production rate of complement, and in the amounts of specific C2 and factor B mRNA. These differences are tissue specific, because C2-specific mRNA content in bronchoalveolar macrophages is considerably greater than in breast milk macrophages, although the amounts of factor B mRNA are comparable. These data suggest that tissue-specific regulation of complement production in human mononuclear phagocytes occurs at a pretranslational level. These studies now provide a basis for investigation of the molecular effects of agents that modulate the biologic functions of monocytes and macrophages.