A randomized placebo-controlled trial on the effects of Menthacarin, a proprietary peppermint- and caraway-oil-preparation, on symptoms and quality of life in patients with functional dyspepsia.

BACKGROUND: Functional dyspepsia (FD) is a very common condition affecting more than 10% of the population. While there is no cure, a few drugs have been found to be effective for the relief of symptoms, although most are only effective in a subgroup of patients. We assess and compare the efficacy of a fixed peppermint/caraway-oil-combination (Menthacarin) on symptoms and quality of life (QoL) in patients with FD symptoms consistent with epigastric pain syndrome (EPS) and postprandial distress syndrome (PDS). METHODS: In a prospective, double-blind, multicenter trial, 114 outpatients with chronic or recurrent FD were randomized and treated for 4 weeks with the proprietary peppermint- and caraway-oil-preparation Menthacarin or placebo (2×1 capsule/day). Improvement of abdominal pain and discomfort were used as co-primary efficacy measures (scores measured with the validated Nepean Dyspepsia Index). KEY RESULTS: After 2 and 4 weeks, active treatment was superior to placebo in alleviating symptoms consistent with PDS and EPS (P all <.001). After 4 weeks of treatment, pain and discomfort scores improved by 7.6±4.8 and 3.6±2.5 points (full analysis set; mean±SD) for Menthacarin and by 3.4±4.3 and 1.3±2.1 points for placebo, respectively. All secondary efficacy measures showed advantages for Menthacarin. CONCLUSIONS & INFERENCES: Menthacarin is an effective therapy for the relief of pain and discomfort and improvement of disease-specific QoL in patients with FD and significantly improves symptoms consistent with EPS and PDS.

Effect of replacing soybean meal protein with protein from upland cottonseed, Pima cottonseed, or extruded Pima cottonseed on production of lactating dairy cows.

Pima cotton production is increasing in the United States, but Pima cottonseed generally contains higher concentrations of the antinutritive pigment gossypol than conventional upland cottonseed. Heating promotes the reaction of gossypol with protein, reducing gossypol absorption and toxicity. The objective of this study was to assess the nutritional value for dairy cattle of Pima cottonseed cake (PCSC) that was heated and oil largely removed by an experimental extrusion process, compared with upland cottonseed (UCS) and Pima cottonseed (PCS). The PCS had greater crude protein (CP) and ether extract, less neutral detergent fiber (NDF) and acid detergent fiber (ADF), similar total gossypol, but higher (-)-gossypol isomer compared with UCS. Extrusion reduced lipid content by 73%, increased concentrations of CP, NDF, and ADF, and reduced total gossypol, (+)-gossypol, and (-)-gossypol in PCSC versus PCS. Forty lactating Holsteins (8 with ruminal cannulas) were blocked by days in milk into 5 squares in a replicated, incomplete 8 × 8 Latin square, and were fed diets containing, on a dry matter (DM) basis, 30% alfalfa silage, 31% corn silage, 21 to 25% high-moisture corn, and about 15% CP. Diets were fed as total mixed rations for ad libitum intake. Supplemental CP was from (1) solvent soybean meal (SSBM) only or 50% from SSBM plus 50% from (2) UCS, (3) PCS, (4) PCSC, (5) UCS plus PCS, and (6) UCS plus PCSC, or (7) 50% from expeller soybean meal (ESBM) plus 50% from PCS, and (8) 50% from ESBM plus 50% from PCSC. Periods were 4 wk long (total of 16 wk); production data were collected over the last 2 wk and blood and ruminal samples were taken on d 28 of each period. Data were analyzed using Proc Mixed of SAS (SAS Institute Inc., Cary, NC). Diet affected dry matter intake, with greatest intake on diet 6 and lowest intake on diets 1 and 3. The highest milk fat content was observed on diet 5 and the greatest fat yield on diet 7; fat content and yield were lowest on diet 1 (soybean meal control). Milk fat secretion was proportional to dietary fat content, indicating that cottonseed oil was used effectively for milk fat synthesis. We observed a trend for an effect on milk protein yield with the greatest protein secretion occurring on diet 7. Milk urea was lowest on diets 3, 7, and 8. Ruminal concentrations of branched-chain volatile fatty acids were lower, or tended to be lower, when PCSC replaced either UCS or PCS in the diet, suggesting reduced degradation and increased escape of PCSC protein. Among cottonseed-containing diets, total gossypol intake was lowest on PCSC, intermediate on PCS, and highest on UCS. Total gossypol and both (+)- and (-)-isomers of gossypol were higher in blood plasma on PCS and lower on PCSC than on the corresponding diets containing UCS, indicating that the extrusion process reduced gossypol absorption. In this trial, production on diets supplemented with UCS, PCS, or PCSC was comparable to that on diets containing soybean meal.

Pelargonium sidoides preparation (EPs 7630) in the treatment of acute bronchitis in adults and children.

Acute bronchitis, although mostly caused by viral infections, is commonly treated with antibiotics. As antibiotics should only be prescribed upon strict indication, treatment options like a liquid herbal drug preparation from the roots of Pelargonium sidoides (EPs 7630) gain more and more interest. To evaluate the efficacy and safety of treatment with EPs 7630 in patients with acute bronchitis, a multi-centre, prospective, open observational study was conducted in 440 study sites located in Germany. A total of 2099 patients aged 0-93 years with productive cough for less than six days without indication for treatment with antibiotics were given EPs 7630-solution in an age-dependent dosage for 14 days. The primary outcome criterion was the mean change of the Bronchitis Severity Score (BSS: cough, sputum, rales/rhonchi, chest pain at cough, dyspnoea) from baseline to patient's individual last observation. During treatment, the mean BSS of all patients decreased from 7.1+/-2.9 points at baseline to 1.0+/-1.9 points at patients' individual last visit. Subgroup analysis for children showed a decrease of mean BSS from 6.3+/-2.8 points to 0.9+/-1.8 points and analysis of children younger than three years showed a decrease of mean BSS from 5.2+/-2.5 points to 1.2+/-2.1 points. Adverse events occurred in 26/2099 (1.2%) patients. Serious adverse events were not reported. In conclusion, EPs 7630 is an effective and well tolerated treatment of acute bronchitis in adults, children and infants outside the strict indication for antibiotic treatment.

[Local tolerability of two preparations with eucalyptus oil and pine-needle oil]. / Lokale Verträglichkeit zweier eucalyptus- und kiefernnadelöl- haltiger Topika.

UNLABELLED: The objective of this open-label, single-centre study was to assess the tolerability of two locally applicable eucalyptus oil and pine-needle oil preparations using patch testing as an accepted and standardised method. METHOD: Two eucalyptus oil and pine-needle oil preparations were applied to the forearm of 46 subjects with healthy skin using standardised patches. The patches were removed after 48 hours and skin reactions were assessed immediately and after 24 and 48 hours. The major objective was the reaction of the patch test. RESULT: Neither preparation caused any positive skin reaction in any of the subjects. CONCLUSION: Skin tolerability to the two eucalyptus oil and pine-needle oil preparations is very good with regard to their active ingredients and other excipients.

Efficacy and Safety of the Platelet-Activating Factor Receptor Antagonist BN 52021 (Ginkgolide B) in Patients with Severe Sepsis : A Randomised, Double-Blind, Placebo-Controlled, Multicentre Trial.

OBJECTIVE: To evaluate the efficacy and safety of the natural platelet-activating factor receptor antagonist, BN 52021 (ginkgolide B) in the treatment of patients with severe sepsis related to Gram-negative and mixed bacterial infection. DESIGN AND SETTING: Prospective, randomised, double-blind, placebo-controlled, multicentre study carried out in 13 academic medical intensive care centres in Germany with up to 14 patients per centre. PATIENTS: 88 patients with severe sepsis under standard medical and surgical care: nine patients with pure Gram-positive infection, 79 patients with Gram-negative or mixed bacterial infections (subgroup for which efficacy was to be established). INTERVENTIONS: Patients were randomised to receive either placebo or BN 52021 1.25 mg/kg bodyweight intravenously every 12h over a 4-day period in addition to their standard medical and surgical care. MAIN OUTCOME MEASURES AND RESULTS: The primary efficacy variable was the 28-day all-cause mortality rate. The treatment groups were similar with respect to demographic data and prognostic factors influencing the outcome except for bodyweight and adequacy of antibiotic therapy. Analysis of patients with Gram-negative or mixed bacterial infection, for which efficacy was to be established, resulted in a 28-day all-cause mortality of 42.5% in the placebo group (n = 40; 17 deaths) versus 38.5% in the BN 52021 group (n = 39; 15 deaths). Among all randomised patients, the 28-day all-cause mortality rate was 40.9% in the placebo group (n = 44; 18 deaths) and 38.6% in the BN 52021 group (n = 44; 17 deaths). There were no differences in frequency and severity of adverse events between the two treatment groups. CONCLUSIONS: Four-day administration of BN 52021 failed to demonstrate a statistically significant reduction in mortality in patients with severe sepsis suspected or confirmed to be related to infections other than Gram-positive bacterial infection.

Miniature radio-frequency mobility analyzer as a gas chromatographic detector for oxygen-containing volatile organic compounds, pheromones and other insect attractants.

A high electric field, radio-frequency ion mobility spectrometry (RF-IMS) analyzer was used as a small detector in gas chromatographic separations of mixtures of volatile organic compounds including alcohols, aldehydes, esters, ethers, pheromones, and other chemical attractants for insects. The detector was equipped with a 2 mCi 63Ni ion source and the drift region for ion characterization was 5 mm wide, 15 mm long and 0.5 mm high. The rate of scanning for the compensation voltages was 60 V s(-1) and permitted four to six scans to be obtained across a capillary chromatographic elution profile for each component. The RF-IMS scans were characteristic of a compound and provided a second dimension of chemical identity to chromatographic retention adding specificity in instances of co-elution. Limits of detection were 1.6-55 x 10(-11) g with an average detection limit for all chemicals of 9.4 x 10(-11) g. Response to mass was linear from 2-50 x 10(-10) g with an average sensitivity of 4 pA ng(-1). Separations of pheromones and chemical attractants for insects illustrated the distinct patterns obtained from gas chromatography with RF-IMS scans in real time and suggest an analytical utility of the RF-IMS as a small, advanced detector for on-site gas chromatographs.

Regulation of avian fibroblast growth factor receptor 1 (FGFR-1) gene expression during skeletal muscle differentiation.

Myogenic cell proliferation and differentiation are regulated by a fibroblast growth factor (FGF) signal transduction cascade mediated by a high-affinity fibroblast growth factor receptor (FGFR). Exogenous FGF added to myogenic cultures has a mitogenic effect promoting myoblast proliferation while repressing differentiation. We have examined the regulation of the FGFR-1 gene (cek-1) in avian myogenic cultures by immunocytochemistry and Northern blot analysis. FGFR-1 protein was readily detected in undifferentiated myoblast cultures and was significantly reduced in differentiated muscle fiber cultures. Similarly, FGFR-1 mRNA was 2.5-fold more abundant in myoblast cultures than in differentiated cultures. To define the molecular mechanism regulating FGFR-1 gene expression in proliferating myoblasts and post-mitotic muscle fibers, we have isolated and partially characterized the avian FGFR-1 gene promoter. Transfection of FGFR-1 promoter-chloramphenicol acetyltransferase gene constructs into myogenic cultures identified two regions regulating expression of this gene in myoblasts. A distal region of 2226 bp conferred a high level of expression in myoblasts. This region functioned in an orientation-dependent manner and interacted with a promoter element(s) in a proximal 1058 bp promoter region to direct transcription. Deletion analysis revealed a 78 bp region that confers a high level of cek1 promoter activity in myoblasts. This DNA segment also contains Spl binding sites and interacts with a component in myoblast nuclear protein extracts. The proximal promoter region alone demonstrated no activity in directing transcription in either myoblasts or muscle fibers. Using the full-length promoter, gene expression was significantly decreased in differentiated muscle fibers relative to undifferentiated myoblasts indicating that the promoter-reporter gene constructs contain elements regulating expression of the endogenous FGFR-1 gene in both myoblasts and muscle fibers.

Protein kinase C activity regulates slow myosin heavy chain 2 gene expression in slow lineage skeletal muscle fibers.

Expression of the slow myosin heavy chain (MyHC) 2 gene defines slow versus fast avian skeletal muscle fiber types. Fetal, or secondary, skeletal muscle fibers express slow MyHC isoform genes in developmentally regulated patterns within the embryo, and this patterning is at least partly dependent on innervation in vivo. We have previously shown that slow MyHC 2 gene expression in vitro is regulated by a combination of innervation and cell lineage. This pattern of gene expression was indistinguishable from the pattern observed in vivo in that it was restricted to innervated muscle fibers of slow muscle origin. We show here that slow MyHC 2 gene expression in the slow muscle fiber lineage is regulated by protein kinase C (PKC) activity. Inhibition of PKC activity induced slow MyHC 2 gene expression, and the capacity to express the slow MyHC 2 gene was restricted to muscle fibers of slow muscle (medial adductor) origin. Fast muscle fibers derived from the pectoralis major did not express significant levels of slow MyHC 2 with or without inhibitors of PKC activity. This differential expression pattern coincided with different inherent PKC activities in fast versus slow muscle fiber types. Furthermore, over-expression of an unregulated PKCalpha mutant suppressed slow MyHC 2 gene expression in muscle fibers of the slow lineage. Lastly, denervation of skeletal muscles caused an increase in PKC activity, particularly in the slow medial adductor muscle. This increase in PKC activity was associated with lack of slow MyHC 2 gene expression in vivo. These results provide a mechanistic link between innervation, an intracellular signaling pathway mediated by PKC, and expression of a muscle fiber type-specific contractile protein gene. Dev Dyn 1999;216:177-189.

Identification of a lectin that induces cell death in developing chicken B cells.

The bursa of Fabricius is required for the development of a diverse B cell repertoire in chickens. Bursal B cells are dependent on survival signals within the bursa and their removal from the bursa results in death by apoptosis. To find molecules that regulate B cell survival, a panel of mAb and lectins was screened for the ability to either accelerate or prevent B cell death in culture. The fucose-specific lectin Aleuria aurantia agglutinin (AAA) rapidly rendered B cells permeable to propidium iodide. Incubation with the lectin also accelerated the appearance of internucleosomal DNA fragmentation and nuclear condensation, characteristics of apoptotic cell death. On Western blots the lectin detects a single protein band of approximately 48-50 kDa molecular weight. AAA detects fucose in an alpha 1-6 linkage and the restriction of this fucose linkage to a single protein suggests that it may be functionally important in the regulation of cell survival.

The avian chB6 (Bu-1) alloantigen can mediate rapid cell death.

The control of cell death is critical in the immune system. T and B lymphocytes must be censored during their development to remove nonfunctional or self-reactive lymphocytes. However, the molecules controlling cell deletion during lymphopoiesis have not been defined. B cells removed from the avian bursa of Fabricius rapidly undergo cell death in culture. We screened bursal B cells with a panel of Abs and lectins to identify molecules affecting their viability. Abs to the chB6 alloantigen caused a rapid loss of cell viability as measured by staining with propidium iodide. ChB6 Abs also cause adhesion between B cells. Transfection of cDNA encoding chB6 reconstituted the allele-specific cell death and adhesion effects in avian cell lines. These effects can be separated by binding cells onto Ab-coated plastic dishes. In these experiments, cells were killed in the absence of cell:cell contact. The ability of chB6 cross-linking to evoke cell aggregation and cell death is also observed when chB6 is expressed in growth factor-dependent mammalian cells. In these cells growth factor can almost completely prevent cell death but not cell aggregation. This suggests that known cell survival stimuli can suppress the cell death brought about by chB6 cross-linking. These results show that chB6 may have an important role in controlling cell survival and/or adhesion during avian B cell development.

Avian B cell development.

Development of B cells in chickens proceeds via a series of discrete developmental stages that includes the maturation of committed B cell progenitors in the specialized microenvironment of the bursa of Fabricius. The bursa has been shown to be required for the amplification of the B cell pool and selects for cells with productive immunoglobulin rearrangement events. Other events regulating chicken B cell development such as lymphocyte trafficking and apoptosis are just beginning to be elucidated. Within the bursa, the variable regions of immunoglobulin genes of B cell progenitors are diversified by a process of intrachromosomal gene conversion, where blocks of sequence information are transferred from pseudo-V regions to the recombined variable regions of the immunoglobulin genes. Recently gene conversion has been determined to play a role in the diversification of the immune repertoire in other species. In this review we focus on the current understanding and recent advances of B cell development in the chicken.

Identification and partial characterization of an Actinomyces pyogenes hemolysin.

Twenty-two Actinomyces pyogenes isolates were recovered from hepatic abscesses in cattle and evaluated for hemolysin production. Hemolysin was collected from supernatant of cultures grown in 6% CO2 in brain heart infusion (BHI) broth. The effect of oxidizing and reducing agents, enzymes, temperatures and pH on hemolytic activity were studied using sheep erythrocytes as the target cells. Our study showed that A. pyogenes hemolysin is oxygen stable; sensitive to treatment by protease, trypsin, and amylase; and destroyed by treatment at extreme temperatures (56 and 100 degrees C) and pH (pH 3 and 11). Production of hemolysin was studied in BHI, RPMI-1640, and a defined serum-free A. pyogenes medium under aerobic and anaerobic conditions. Maximum hemolysin was produced in BHI incubated aerobically in 6% CO2 and to a lesser degree anaerobically in RPMI-1640. No hemolysin was produced in the defined A. pyogenes medium. Differential filtration, isoelectric focusing and sodium dodecyl sulfate polyacrylamide gel electrophoresis identified two hemolysin proteins with pI values of 3.40 and 9.45 and estimated molecular masses of 62 and 58 kDa, respectively. Cell-free supernatant samples positive for hemolysin activity also were screened for leukotoxin activity. Significant levels of leukotoxin were detected in all samples screened.

Current concepts in chicken B cell development.

The chicken has provided fundamental insights into the workings of vertebrate immunity. In particular, the development of B cells in a unique organ, the bursa of Fabricius, has provided a novel opportunity to study B cell development. Although chickens generate their Ig repertoire in a different way than mice and humans, there are many striking similarities in the developmental process. In particular, the control of lymphocyte migration and survival is key to the development of an immune system. The evolutionary distance of chickens and mammals underscore how common the problems are as well as how the solutions are often similar. Such commonalities serve to maintain the chicken as a compelling animal in which to study B cell development.

Vascular cell adhesion molecule 1-positive reticular cells express interleukin-7 and stem cell factor in the bone marrow.

In vitro studies have defined an essential role for stromal cells in supporting B-cell development, including production of lymphopoietic cytokines. It has been suggested that stromal cells are equivalent to adventitial reticular cells in the marrow; however, evidence of reticular cells producing cytokines has been difficult to obtain. Staining of bone marrow (BM) sections with antibodies to interleukin-7 (IL-7) showed a reticular pattern, mimicking that obtained using antibodies to vascular cell adhesion molecule 1 (VCAM-1), a molecule present on both stromal cells in vitro and reticular cells. To more closely examine cytokine production within normal marrow, an immunomagnetic separation scheme was devised to directly enrich VCAM-1+ stromal cells. Twenty to thirty percent of cells isolated in the VCAM-1+ fraction shared characteristics with stromal cells from long term BM cultures, including cellular morphology and expression of alkaline phosphatase and alpha actin. These were termed "reticular stromal" cells. Immunohistochemical staining showed that virtually all of the latter cells possessed cytoplasmic IL-7 protein, and about half expressed stem cell factor. In contrast with cultured stromal cells, very few had detectable macrophage-colony-stimulating factor. These data constitute the first report of cytokine expression by marrow reticular cells in vivo. The implications of this data with respect to the existence of stromal cell subsets and their regulation of lymphopoiesis is discussed.

Native associations of early hematopoietic stem cells and stromal cells isolated in bone marrow cell aggregates.

In suspensions of murine bone marrow, many stromal cells are tightly entwined with hematopoietic cells. These cellular aggregations appear to exist normally within the marrow. Previous studies showed that lymphocytes and stem cells adhered to stromal cells via vascular cell adhesion molecule 1 (VCAM1). Injection of anti-VCAM1 antibody into mice disrupts the aggregates, showing the importance of VCAM1 in the adhesion between stromal cells and hematopoietic cells in vivo. Early hematopoietic stem cells were shown to be enriched in aggregates by using a limiting-dilution culture assay. Myeloid progenitors responsive to WEHI-3CM in combination with stem cell factor (c-kit ligand) and B220- B-cell progenitors responsive to insulin-like growth factor-1 in combination with interleukin-7 are not enriched. We propose a scheme of stromal cell-hematopoietic cell interactions based on the cell types selectively retained within the aggregates. The existence of these aggregates as native elements of bone marrow organization presents a novel means to study in vivo stem cell-stromal cell interaction.

Activity of stem cell factor and IL-7 in combination on normal bone marrow B lineage cells.

The production of B cells is regulated by soluble and cell contact signals presumably provided by bone marrow stromal cells. Among these is IL-7, a well characterized proliferative stimulus for a subset of pre-B cells. Stem cell factor (SCF), a stromal cell-derived cytokine with broad hemopoietic effects, has been reported to synergize with IL-7 to drive the proliferation and differentiation of B220- bone marrow cells into B220+ B cell precursors in long term culture. A subsequent report has cast doubt on this result by showing that SCF and IL-7 were incapable of producing mu+ pre-B cells after short term culture. Here, using the cell sorter to assure discrete separation of B220+ and B220- cells followed by soft agar culture to prevent interaction with accessory cells, we demonstrate that the combination of SCF and IL-7 does not stimulate the expansion or differentiation of B220- lymphoid precursors but can act synergistically in the clonal proliferation of B220+ cells.

Enrichment of primary lymphocyte-supporting stromal cells and characterization of associated B lymphocyte progenitors.

Studies of Whitlock/Witte long-term bone marrow cultures have revealed the necessity of two cell types for B lymphopoiesis, a stem cell and the stromal cell. While a number of stromal cell lines exist they have been found to be heterogeneous with respect to cell surface marker expression and growth factor production. Separation and analysis of fresh bone marrow stromal cells is, therefore, necessary to understand the regulation of lymphopoiesis in vivo. Here we report the early stages of such studies. We demonstrate that stromal cells, as assessed by morphology and alkaline phosphatase reactivity after short-term culture, are enriched in cellular aggregates that can be separated from bone marrow suspensions. Stromal cells are present in aggregates at a frequency of one per thousand cells, whereas marrow from which the aggregates have been removed contains only one stromal cell per fifty-thousand cells. These aggregates are able to form Whitlock cultures from greatly reduced numbers of initiating cells, indicating that they contain culturable B lineage precursors as well as stromal cells capable of supporting B lymphopoiesis. The aggregates appear to be naturally formed and provide a means to examine native B cell precursor-stromal cell contacts. We find little evidence for sequestering of late-stage B cell precursors within the aggregates. Terminal deoxynucleotidyl transferase-positive cells, on the other hand, are approximately three times more frequent in bone marrow aggregates, suggesting close contact between very early B cell progenitors and stromal cells within the aggregates. The finding that stromal cells are enriched in cellular aggregates is an important first step in the ultimate isolation of these cells from marrow suspensions, which is vital to understanding stromal cell function in vivo.

Professional attitudes of Ohio pharmacists working in three practice settings.

The attitudes of pharmacists in three practice settings toward a set of attributes associated with professionalism were studied. A questionnaire that included the 40-item Shack and Hepler professionalism instrument was mailed to 1999 pharmacists in Ohio. Responses from 617 pharmacists who worked in hospital, independent community, or chain community pharmacy practice were used. There were significant differences between the responses of pharmacists who worked in the different practice settings (multivariate analysis of variance, p less than 0.01). However, the practice setting accounted for less than 10% of the variance in the responses. Statistically significant differences were also observed when the pharmacists were grouped according to sex, years since graduation, degree, and job position; however, less than 8% of the variance in the responses could be explained by the demographic variables. There did not appear to be any practical differences in beliefs about professional attributes among pharmacists working in hospital, independent community, and chain community practice settings.