Improvement of maternal vitamin D status with 25-hydroxycholecalciferol positively impacts porcine fetal skeletal muscle development and myoblast activity.

There is little information available regarding the influence of maternal vitamin D status on fetal skeletal muscle development. Therefore, we investigated the effect of improved vitamin D status resulting from 25-hydroxycholecalciferol (25OHD3) supplementation of dams on fetal skeletal muscle developmental characteristics and myoblast activity using Camborough 22 gilts (n = 40) randomly assigned to 1 of 2 corn-soybean meal-based diets. The control diet (CTL) contained 2,500 IU cholecalciferol (D3)/kg diet, whereas the experimental diet contained 500 IU D3/kg diet plus 50 µg 25OHD3/kg diet. Gilts were fed 2.7 kg of their assigned diet once daily beginning 43 d before breeding through d 90 of gestation. On gestational d 90 (± 1), fetal LM and semitendinosus muscle samples were collected for analysis of developmental characteristics and myoblast activity, respectively. No treatment difference was observed in fetal LM cross-sectional area (P = 0.25). Fetuses from 25OHD3-supplemented gilts had more LM fibers (P = 0.04) that tended to be smaller in cross-sectional area compared with CTL fetuses (P = 0.11). A numerical increase in the total number of Pax7+ myoblasts was also observed in fetuses from 25OHD3-supplemented gilts (P = 0.12). Myoblasts derived from the muscles of fetuses from 25OHD3-fed dams displayed an extended proliferative phase in culture compared with those from fetuses of dams fed only D3 (P < 0.0001). The combination of additional muscle fibers and Pax7+ myoblasts with prolonged proliferative capacity could enhance the postnatal skeletal muscle growth potential of fetuses from 25OHD3-supplemented gilts. These data highlight the importance of maternal vitamin D status on the development of fetal skeletal muscle.

Feeding 25-hydroxycholecalciferol improves gilt reproductive performance and fetal vitamin D status.

Little information is available regarding the effects of vitamin D and its metabolites on reproduction in swine. To investigate the effects of feeding the circulating metabolite of vitamin D, 25-hydroxycholecalciferol (25OHD3, ROVIMIX Hy • D, DSM Nutritional Products, Basel, Switzerland) on maternal and fetal circulating 25OHD3 concentration and gilt reproductive performance, a total of 40 PIC Camborough-22 gilts (BW on d -6 = 138 kg) in 4 replicates were randomly assigned to 1 of 2 corn-soybean meal-based diets. The control diet (CTL) was formulated to contain 2,500 IU D3/kg diet, and the experimental diet (25OHD3) was formulated to contain 500 IU D3/kg diet + 50 µg 25OHD3/kg diet. Gilts were fed 2.7 kg of their assigned diet once daily beginning 43 d before breeding. Gilt BW were measured on gestational d -6 and d 90. Gilts were artificially inseminated with PIC 337-G semen 12 h and 24 h after showing signs of estrus. Blood samples were collected from the jugular vein on gestational d -43, -13, 46, and 89 for analysis of circulating 25OHD3 plasma concentration and overall vitamin D status of the gilts. At gestational d 90 ± 1, gilts were harvested and reproductive tracts were removed. Fetal weight, sex, crown-to-rump length (CRL), as well as the number of mummified fetuses were recorded. As expected, circulating plasma concentrations of 25OHD3 were not different among treatment groups at d -43 (CTL = 53.8 ng/mL, 25OHD3 = 57.4 ng/mL; P = 0.66). However, gilts fed 25OHD3 had greater (P < 0.001) circulating plasma concentrations of 25OHD3 on d -13 (89.7 vs. 56.7 ng/mL), d 46 (95.8 vs. 55.7 ng/mL), and d 89 (92.8 vs. 58.2 ng/mL) of gestation compared with CTL-fed gilts. Circulating 25OHD3 was also greater in fetuses from 25OHD3-fed gilts on d 90 (P < 0.001). A 23% increase in pregnancy rate was observed in 25OHD3-fed gilts compared with CTL (78% vs. 55%, respectively; P = 0.21). Maternal BW gain (without conceptus), number of mummified fetuses, mean fetal weight, and mean fetal CRL were similar among treatments (P > 0.05). However, litter size was larger (CTL = 10.2; 25OHD3 = 12.7; P = 0.04) in 25OHD3-fed gilts compared with CTL-fed gilts. Notably, mean fetal weight was not decreased in 25OHD3-fed gilts as frequently occurs when litter size is increased. Overall, feeding 25OHD3 to first-service gilts before and during gestation improved both maternal and fetal vitamin D status and improved maternal reproductive performance.

Txp40, a ubiquitous insecticidal toxin protein from Xenorhabdus and Photorhabdus bacteria.

Xenorhabdus and Photorhabdus are gram-negative bacteria that produce a range of proteins that are toxic to insects. We recently identified a novel 42-kDa protein from Xenorhabdus nematophila that was lethal to the larvae of insects such as Galleria mellonella and Helicoverpa armigera when it was injected at doses of 30 to 40 ng/g larvae. In the present work, the toxin gene txp40 was identified in another 59 strains of Xenorhabdus and Photorhabdus, indicating that it is both highly conserved and widespread among these bacteria. Recombinant toxin protein was shown to be active against a variety of insect species by direct injection into the larvae of the lepidopteran species G. mellonella, H. armigera, and Plodia interpunctella and the dipteran species Lucilia cuprina. The protein exhibited significant cytotoxicity against two dipteran cell lines and two lepidopteran cell lines but not against a mammalian cell line. Histological data from H. armigera larvae into which the toxin was injected suggested that the primary site of action of the toxin is the midgut, although some damage to the fat body was also observed.

Clinical evaluation of the electronic nose in the diagnosis of ear, nose and throat infection: a preliminary study.

The term electronic nose describes an electronic system that is able to mimic the human sense of smell. Electronic noses have been developed over the last 10 or more years to perform a variety of identification tasks in various industries. More recently electronic noses have attracted new interest in their application in the field of medical diagnosis. The aim of this study is to explore the use of an electronic nose to identify and classify pathogens associated with ear, nose and throat (ENT) infections. In this study 90 bacterial swab samples were collected from 90 patients with ENT infections. Some of these samples were analysed immediately with a commercial electronic nose (Cyranose C320). Similar numbers of swabs were also taken from the same site of infection and were sent for microbiology culture and sensitivity. The electronic nose diagnosis was compared with the microbiology diagnosis and it was found that the electronic nose diagnosis was correct in 88.2 per cent of the cases, which is an encouraging result.

[Raman scattering study of water up to 600 MPa at 290 K].

Raman scattering studies of the stretching band from liquid water have been conducted up to 600 MPa at 290 K. It is shown that (r1)(max) decreases with increasing pressure initially and reaches the minimum at about 200 MPa, and increases at higher pressure up to about 400 MPa, then decreases with increasing pressure up to 600 MPa. And this is also in accordance with the behavior of r(oo) under pressure. The authors conclude that, just like ice phase transition ice ( I h) --> ice(II) --> ice (V), there also probably exists water phase transition water ( I h) --> water (III) --> water (V).

Electronic nose based tea quality standardization.

In this paper we have used a metal oxide sensor (MOS) based electronic nose (EN) to analyze five tea samples with different qualities, namely, drier month, drier month again over-fired, well fermented normal fired in oven, well fermented overfired in oven, and under fermented normal fired in oven. The flavour of tea is determined mainly by its taste and smell, which is generated by hundreds of Volatile Organic Compounds (VOCs) and Non-Volatile Organic Compounds present in tea. These VOCs are present in different ratios and determine the quality of the tea. For example Assamica (Sri Lanka and Assam Tea) and Assamica Sinesis (Darjeeling and Japanese Tea) are two different species of tea giving different flavour notes. Tea flavour is traditionally measured through the use of a combination of conventional analytical instrumentation and human or ganoleptic profiling panels. These methods are expensive in terms of time and labour and also inaccurate because of a lack of either sensitivity or quantitative information. In this paper an investigation has been made to determine the flavours of different tea samples using an EN and to explore the possibility of replacing existing analytical and profiling panel methods. The technique uses an array of 4 MOSs, each of, which has an electrical resistance that has partial sensitivity to the headspace of tea. The signals from the sensor array are then conditioned by suitable interface circuitry. The data were processed using Principal Components Analysis (PCA), Fuzzy C Means algorithm (FCM). We also explored the use of a Self-Organizing Map (SOM) method along with a Radial Basis Function network (RBF) and a Probabilistic Neural Network classifier. Using FCM and SOM feature extraction techniques along with RBF neural network we achieved 100% correct classification for the five different tea samples with different qualities. These results prove that our EN is capable of discriminating between the flavours of teas manufactured under different processing conditions, viz. over-fermented, over-fired, under fermented, etc.

Classifying coronary dysfunction using neural networks through cardiovascular auscultation.

The paper applies artificial neural networks (ANNs) to the analysis of heart sound abnormalities through auscultation. Audio auscultation samples of 16 different coronary abnormalities were collected. Data pre-processing included down-sampling of the auscultated data and use of the fast Fourier transform (FFT) and the Levinson-Durbin autoregression algorithms for feature extraction and efficient data encoding. These data were used in the training of a multi-layer perceptron (MLP) and radial basis function (RBF) neural network to develop a classification mechanism capable of distinguishing between different heart sound abnormalities. The MLP and RBF networks attained classification accuracies of 84% and 88%, respectively. The application of ANNs to the analysis of respiratory auscultation and consequently the development of a combined cardio-respiratory analysis system using auscultated data could lead to faster and more efficient treatment.

Regulation of the rat NHE3 gene promoter by sodium butyrate.

Short-chain fatty acids, and especially butyrate (NaB), stimulate sodium and water absorption by inducing colonic Na+/H+ exchange (NHE). NaB induces NHE3 activity and protein and mRNA expression both in vivo and in vitro. NaB, as a histone deacetylase (HDAC) inhibitor, regulates gene transcription. We therefore studied whether NaB regulates transcription of the rat NHE3 promoter in transiently transfected Caco-2 cells. NaB (5 mM) strongly stimulated reporter gene activity, and this stimulation was prevented with actinomycin D, indicating transcriptional activation. NaB effects on the NHE3 promoter depended on the activity of Ser/Thr kinases, in particular, protein kinase A (PKA). However, PKA stimulation alone did not have an effect on promoter activity, and it did not act synergistically with NaB. Another HDAC inhibitor, Trichostatin A (TSA), stimulated NHE3 promoter in a Ser/Thr kinase-independent fashion. The putative NaB-responsive elements were localized within -320/-34 bp of the NHE3 promoter. These findings suggest that PKA mediates NaB effects on NHE3 gene transcription and that the mechanism of NaB action is different from that of TSA.

Intrinsic AHR in IL-5 transgenic mice is dependent on CD4(+) cells and CD49d-mediated signaling.

Overexpression of interleukin (IL)-5 by the airway epithelium in mice using the rat CC10 promoter (NJ.1726 line) leads to several histopathologies characteristic of human asthma, including airway hyperreactivity (AHR). We investigated the contribution of B and T cells, as well as CD4 expression, to the development of AHR in IL-5 transgenic mice. NJ.1726 mice on a T cell or CD4 knockout background, but not on a B cell knockout background, lost intrinsic AHR. These effects occurred without decreases in IL-5 or eosinophils. We further investigated the contribution of alpha(4)-integrin signaling to the development of AHR in IL-5 transgenic mice through the administration of anti-CD49d (alpha(4)-integrin) antibody (PS/2). Administration of PS/2 resulted in immediate (16-h) inhibition of AHR. The inhibition of AHR was not associated with a decrease in airway eosinophils. These studies demonstrate that, despite the presence of increased levels of IL-5 and eosinophils in the lungs of NJ.1726 mice, CD4(+) cells and alpha(4)-integrin signaling are necessary for the intrinsic AHR that develops in IL-5 transgenic mice.

Extensive eosinophil degranulation and peroxidase-mediated oxidation of airway proteins do not occur in a mouse ovalbumin-challenge model of pulmonary inflammation.

Paradigms of eosinophil effector function in the lungs of asthma patients invariably depend on activities mediated by cationic proteins released from secondary granules during a process collectively referred to as degranulation. In this study, we generated knockout mice deficient for eosinophil peroxidase (EPO) to assess the role(s) of this abundant secondary granule protein in an OVA-challenge model. The loss of EPO had no effect on the development of OVA-induced pathologies in the mouse. The absence of phenotypic consequences in these knockout animals extended beyond pulmonary histopathologies and airway changes, as EPO-deficient animals also displayed OVA-induced airway hyperresponsiveness after provocation with methacholine. In addition, EPO-mediated oxidative damage of proteins (e.g., bromination of tyrosine residues) recovered in bronchoalveolar lavage from OVA-treated wild-type mice was <10% of the levels observed in bronchoalveolar lavage recovered from asthma patients. These data demonstrate that EPO activities are inconsequential to the development of allergic pulmonary pathologies in the mouse and suggest that degranulation of eosinophils recruited to the lung in this model does not occur at levels comparable to those observed in humans with asthma.

Molecular cloning of murine sodium-phosphate cotransporter type IIb (Na/P(i)-IIb) gene promoter and characterization of gene structure.

We report the cloning of the murine Na/P(i)-IIb cotransporter gene, which spans more than 18 kilobases and consists of 12 introns and 13 exons. Three promoter/reporter gene constructs, -159/+73, -429/+73 and -954/+73, showed significant luciferase activity (22-82-fold over background) when transfected into in rat intestinal epithelial (RIE-1) cells.

Molecular cloning of the murine PHEX gene promoter.

We report the novel cloning of the murine PHEX promoter, the gene that is mutated in X-linked hypophosphatemic rickets (XLH). Four promoter/reporter gene constructs, -133/+104, -542/+104, -1061/+104, and -2866/+104, showed significant luciferase activity (4.9-13.2-fold over background) when transfected into rat osteogenic sarcoma (UMR-106) cells.

Neural network correction of ultrasonic C-scan images.

A neural network-based approach to the correction of C-scan images is presented. This allows the effects of a finite ultrasonic beam diameter in an immersion experiment to be considered, by training the network on a series of defects of known characteristics. The result is an image which is a better representation of the actual defect.

The use of a neural network for studying the relationship between air pollution and asthma-related emergency room visits.

To establish the relationship between air pollution levels and bronchial asthma-associated emergency room (ER) visits, we adapted artificial network technology to conduct this study which focused on three different pollutants, sulphur dioxide, nitrogen oxide and ozone. The study population was comprised of adults presenting to the emergency room of a large metropolitan hospital in Israel during a 3-month period with acute exacerbation of bronchial asthma and who had a past history of intermittent airway disease compatible with bronchial asthma. The range of mean daily pollutants levels for the whole period were: O3 = 15-26 micrograms m-3, NOx = 36-108 micrograms m-3, NO = 16-70 micrograms m-3, and SO2 = 11-32 micrograms m-3. The data sets were composed of input air pollution levels and output ER visits. The first 126 data sets used for the training phase showed that maximal ER visits were mainly associated with the highest cumulative values of air pollution and mostly with nitrogen oxide. In phase two, an attempt was made to predict ER visits based on air pollution level in 49 data sets. The study findings demonstrated that ordinary network technology can be used for learning the effect of air pollution ER visits and, although limited in accuracy, to also predict future ER visits.

Neural network based on adaptive resonance theory as compared to experts in suggesting treatment for schizophrenic and unipolar depressed in-patients.

A modified neural network based on adaptive resonance theory (ART) was trained with the records of 211 psychiatric inpatients (74 schizophrenic, 50 unipolar depressed, 34 bipolar depressed, 20 bipolar manic, 33 other) who improved by at least 40 points on the GAFS during 8 weeks of treatment. Thereafter, a comparison was made between the clinical response of another 26 schizophrenic patients and 28 unipolar depressed inpatients, to treatment suggested by the trained ART (N = 21) and by the consensus of two senior psychiatrists (N = 33). The patients were allocated blindly and randomly to the two treatment groups. The BPRS (for the schizophrenic patients) or the HDRS (for the unipolar depressed patients) was completed weekly for 5 weeks. Results showed no difference between decisions regarding treatment by the ART network and by the experts. Length of hospital stay was also similar. All ART suggestions included supportive psychotherapy. High potency antipsychotics were suggested for 7 schizophrenic inpatients, clozapine for one and the addition of community therapy for another. Depressed patients got a variety of treatment suggestions. No contraindicated treatment was suggested by ART; however, two incomplete treatment suggestions were dropped from the study. In conclusion, in a prospective study ART was successful in learning treatment strategies and performed under supervision similar to experts.

The sulfakinins of the blowfly Calliphora vomitoria. Peptide isolation, gene cloning and expression studies.

The nonapeptide, Phe-Asp-Asp-Tyr(SO3)-Gly-His-Met-Arg-Phe-NH2 was isolated from heads of the blowfly Calliphora vomitoria. Designated callisulfakinin I, the peptide is identical to the earlier known drosulfakinin I of Drosophila melanogaster and to neosulfakinin I of Neobellieria bullata. It belongs to the sulfakinin family, all known members of which (from flies, cockroaches and locusts) have the C-terminal heptapeptide sequence Asp-Tyr(SO3)-Gly-His-Met-Arg-Phe-NH2. The callisulfakinin gene of C. vomitoria was cloned and sequenced. In addition to callisulfakinin I, the DNA revealed a coding sequence for the putative tetradecapeptide. Gly-Gly-Glu-Glu-Gln-Phe-Asp-Asp-Tyr-Gly-His- Met-Arg-Phe-NH2, callisulfakinin II. However, this peptide was not identified in the fly head extracts. Confocal laser scanning immunocytochemical studies with antisera raised against the synthetic undecapeptide C-terminal fragment of drosulfakinin II from D. melanogaster, Asp-Gln-Phe-Asp-Asp-Tyr(SO3)- Gly-His-Met-Arg-Phe-NH2, revealed only four pairs of sulfakinin neurones in the brain of C. vomitoria and no others anywhere else in the neural, endocrine or gut tissues. In situ hybridisation studies with a digoxigenin-labelled sulfakinin gene probe (from the blowfly Lucilia cuprina) also revealed only four pairs of neurones in the brain. The perikarya of two pairs of cells are situated medially in the caudo-dorsal region, close to the roots of the ocellar nerve. The other perikarya are slightly more posterior and lateral. Although it has been suggested by several authors that the insect sulfakinins are homologous to the vertebrate peptides gastrin and cholecystokinin, such arguments (based essentially on C-terminal structural similarities) do not take account of important differences in the C-terminal tetrapeptide. His-Met-Arg-Phe-NH2 in the sulfakinins, compared with Trp-Met-Asp-Phe-NH2 in gastrin and cholecystokinin. Furthermore, whereas the sulfakinin neurons of C. vomitoria are small in number and have a very specialised location, a greater number of cells throughout the nervous system react positively to gastrin/cholecystokinin antisera. Chromatographic profiles of the present study also revealed peaks of gastrin/cholecystokinin-immunoreactive material separate from the sulfakinin peptides. This evidence suggests that the insect and vertebrate peptides may not necessarily be homologous.

Pathophysiology of failure to wean from mechanical ventilation.

Weaning patients from mechanical ventilation constitutes a major portion of the workload in an intensive care unit, as over 40% of total ventilator time is consumed by the weaning process. Several pathophysiological mechanisms may be responsible for weaning failure, but the precise role of each is incompletely understood. Patients who fail a weaning trial commonly develop hypercapnia, which appears to be due to decreased tidal volume rather than a primary decrease in respiratory drive. Respiratory muscle performance is impaired as a result of dynamic hyperinflation and paradoxic motion of the rib cage and abdomen. Worsening of pulmonary mechanics will cause further embarrassment of the respiratory muscles. However, the clinical importance of respiratory muscle fatigue remains unclear. Afferent stimuli arising in the lung parenchyma, respiratory muscles, or as a consequence of impaired gas exchange will be transmitted to the respiratory control centers and result in severe dyspnea in patients who fail a weaning trial.

Characterization of a juvenile hormone binding lipophorin from the blowfly Lucilia cuprina.

The larval haemolymph of the sheep blowfly Lucilia cuprina (Weidemann) contains a juvenile hormone binding protein with a Kd for racemic JH III of 33 +/- 6 nM. The density of the binding sites is 212 +/- 33 pmol/mg haemolymph protein. The binding protein is equally specific for JH III and methyl farnesoate. Some natural juvenoids were ranked for their ability to displace [3H]JH III with JH III > JH II > JH I > JH III acid > JH III diol > JHB3 = no detectable displacement. These data, together with displacement studies for 14 synthetic juvenoids, indicate some characteristics of the JH binding cleft. The binding protein is a high density lipophorin (density = 1.15 g/ml) and has subunit molecular weights of 228 kDa (apolipophorin I) and 70 kDa (apolipophorin II). The N-terminal amino acid sequences of the subunits have no discernible homology to any previously sequenced protein. Lipophorin-specific immunocytochemical staining occurs in a subset of fat body cells.

Association of exogenous DNA with cattle and insect spermatozoa in vitro.

Spermatozoa isolated from domestic cattle (Bos taurus), the Australian sheep blowfly (Lucilia cuprina), and the honeybee (Apis mellifera) are capable of binding exogenous radiolabeled linear DNA. Both motile and nonmotile bovine sperm exhibit four distinct patterns of DNA association. Following treatment with DNase I, the relative proportion of one of these patterns increases specifically in living sperm, suggesting that a small proportion of DNA that associates with bovine sperm may be sequestered within the sperm head.