Raman excess spectroscopy vs. principal component analysis: probing the intermolecular interactions between chiral molecules and imidazolium-based ionic liquids.

Raman spectroscopy is a very sensitive and specific measurement tool for probing intermolecular interaction structures. As imidazolium-based ionic liquids can favorably be used for enantioseparation, in this work two approaches for Raman signal analysis, namely by means of excess spectra and principal component analysis (PCA), are compared to extract detailed information about the interaction structure of the enantiomers d- and l-glucose in an aqueous solution of the ionic liquid [EMIM][EtSO4]. In contrast to the excess calculations, the loadings obtained from PCA lead to significant results since the interactions are weighted by their strength and significance. Moreover, the analysis of the weighted vibrations in the loadings indicate that hydrogen bonds are particularly formed between the ethyl sulphate anion of [EMIM][EtSO4] and the hydrogen atom of the OH-group at the C6-atom of glucose.

Association of polymorphisms in solute carrier family 27, isoform A6 (SLC27A6) and fatty acid-binding protein-3 and fatty acid-binding protein-4 (FABP3 and FABP4) with fatty acid composition of bovine milk.

The main goal of this study was to develop tools for genetic selection of animals producing milk with a lower concentration of saturated fatty acids (SFA) and a higher concentration of unsaturated fatty acids (UFA). The reasons for changing milk fatty acid (FA) composition were to improve milk technological properties, such as for production of more spreadable butter, and milk nutritional value with respect to the potentially adverse effects of SFA on human health. We hypothesized that genetic polymorphisms in solute carrier family 27, isoform A6 (SLC27A6) fatty acid transport protein gene and fatty acid binding protein (FABP)-3 and FABP-4 (FABP3 and FABP4) would affect the selectivity of FA uptake into, and FA redistribution inside, mammary epithelial cells, resulting in altered FA composition of bovine milk. The objectives of our study were to discover genetic polymorphisms in SLC27A6, FABP3, and FABP4, and to test those polymorphisms for associations with milk FA composition. The results showed that after pairwise comparisons between SLC27A6 haplotypes for significantly associated traits, haplotype H3 was significantly associated with 1.37 weight percentage (wt%) lower SFA concentration, 0.091 lower SFA:UFA ratio, and 0.17 wt% lower lauric acid (12:0) concentration, but 1.37 wt% higher UFA and 1.24 wt% higher monounsaturated fatty acid (MUFA) concentrations compared with haplotype H1 during the first 3 mo of lactation. Pairwise comparisons between FABP4 haplotypes for significantly associated traits showed that haplotype H3 was significantly associated with 1.04 wt% lower SFA concentration, 0.079 lower SFA:UFA ratio, 0.15 wt% lower lauric acid (12:0), and 0.27 wt% lower myristic acid (14:0) concentrations, but 1.04 wt% higher UFA and 0.91 wt% higher MUFA concentrations compared with haplotype H1 during the first 3 mo of lactation. Percentages of genetic variance explained by H3 versus H1 haplotype substitutions for SLC27A6 and FABP4 ranged from 2.50 to 4.86% and from 4.91 to 7.22%, respectively. Tag single nucleotide polymorphisms were identified to distinguish haplotypes H3 of SLC27A6 and FABP4 from others encompassing each gene. We found no significant associations between FABP3 haplotypes and milk FA composition. In conclusion, polymorphisms in FABP4 and SLC27A6 can be used to select for cattle producing milk with lower concentrations of SFA and higher concentrations of UFA.

Sterol regulatory element binding transcription factor 1 (SREBF1) polymorphism and milk fatty acid composition.

Milk is known to contain high concentrations of saturated fatty acids-such as palmitic (16:0), myristic (14:0), and lauric (12:0) acids-that can raise plasma cholesterol in humans, making their presence in milk undesirable. The main objective of our candidate gene study was to develop genetic markers that can be used to improve the healthfulness of bovine milk. The sterol regulatory element binding transcription factor 1 (SREBF1) known to regulate the transcription of lipogenic genes together with SREBF chaperone and insulin induced gene 1 were the candidate genes. The results showed significant association of the overall SREBF1 haplotypes with milk production and variations in lauric (12:0) and myristic (14:0) acid concentrations in milk. Haplotype H1 of SREBF1 was the most desirable to improve milk healthfulness because it was significantly associated with lower lauric (12:0) and myristic (14:0) acid concentrations compared with haplotype H3 of SREBF1, and lower lauric acid (12:0) concentration compared with haplotype H2 of SREBF1. Haplotype H1 of SREBF1, however, was significantly associated with lower milk production compared with haplotype H3 of SREBF1. We did not detect any significant associations between genetic polymorphisms in insulin induced gene 1 (INSIG1) and SREBF chaperone and milk fatty acid composition. In conclusion, genetic polymorphisms in SREBF1 can be used to develop genetic tools for the selection of animals producing milk with healthier fatty acid composition.

The complete mitochondrial DNA sequence of the bichir (Polypterus ornatipinnis), a basal ray-finned fish: ancient establishment of the consensus vertebrate gene order.

The evolutionary position of bichirs is disputed, and they have been variously aligned with ray-finned fish (Actinopterygii) or lobe-finned fish (Sarcopterygii), which also include tetrapods. Alternatively, they have been placed into their own group, the Brachiopterygii. The phylogenetic position of bichirs as possibly the most primitive living bony fish (Osteichthyes) made knowledge about their mitochondrial genome of considerable evolutionary interest. We determined the complete nucleotide sequence (16,624 bp) of the mitochondrial genome of a bichir, Polypterus ornalipinnis. Its genome contains 13 proteincoding genes, 22 tRNAs, two rRNAs and one major noncoding region. The genome's structure and organization show that this is the most basal vertebrate that conforms to the consensus vertebrate mtDNA gene order. Bichir mitochondrial protein-coding and ribosomal RNA genes have greater sequence similarity to ray-finned fish than to either lamprey or lungfish. Phylogenetic analyses suggest the bichir's placement as the most basal living member of the ray-finned fish and rule out its classification as a lobefinned fish. Hence, its lobe-fins are probably not a shared-derived trait with those of lobe-finned fish (Sarcopterygii).

Shielding design for the PANDA spectrometer at the Munich high-flux reactor FRM-II.

The start-up of the Munich high-flux reactor FRM-II is in progress on. At the beam tube SR-2 the spectrometer PANDA has been installed. It is at three-axis neutron spectrometer looking onto a slightly under-moderated cold neutron source. For polarisation analysis, PANDA is equipped with a vertical cryomagnet producing fields up to 14.5 T for the sample. To get an appropriate shielding of the high-intensity instrument, one has to take into account the large cross section of the primary beam, several restrictions using magnetic materials, limitations in loading the site and finally, has to keep the lateral extent of the shielding small to allow for high-scattering angles. The shielding has been designed on the basis of the results, which were achieved by the combined use of both the Monte Carlo code MCNP-4B2 and an analytical method based on one-dimensional dose transmission functions.

The greater vulnerability of bile duct cells to reoxygenation injury than to anoxia. Implications for the pathogenesis of biliary strictures after liver transplantation.

The occurrence of biliary strictures in allografts following liver transplantation correlates with the duration of preservation time. The correlation between preservation time and biliary strictures suggests that anoxic or reperfusion injury of the bile duct epithelium causes stricture formation. However, the relative susceptibility of bile duct cells to anoxic or reoxygenation injury is unknown. Our aims were to determine the vulnerability of rat liver bile duct cells to anoxic and reoxygenation injury and to compare the results with hepatocytes. During anoxia, bile duct epithelial cells were significantly more resistant to cell killing than hepatocytes. Rates of cellular proteolysis were also 2.5-fold lower in bile duct cells than in hepatocytes during anoxia. In contrast to anoxia, reoxygenation of anoxic cells increased cell killing of bile duct cells but improved viability of hepatocytes. The rate of toxic oxygen species formation by bile duct cells was 5-fold greater than in hepatocytes during reoxygenation. In addition, basal levels of glutathione are lower in bile duct cells than in hepatocytes. These data suggest that bile duct cells are more susceptible to reoxygenation injury than to anoxia. These studies support the hypothesis that reoxygenation injury during liver preservation leads to bile duct injury during liver transplantation.

Reproducible amplification of RAPD markers from vertebrate DNA.

Randomly amplified polymorphic DNA (RAPD) is a modification of PCR that uses short, randomly generated primers to amplify genomic DNA. Generally, many bands of mixed intensity (i.e., strong, faint, fuzzy or sharp) are generated with each primer. Mixed-intensity bands are inherent with the RAPD technique because (i) the target DNAs are undefined, (ii) one or more copies of the target DNA may exist per genome and (iii) the percentage of hybridization of primer to target may vary. The problem of mixed-intensity band exacerbates the well-known sensitivity of PCRs to reagent and template concentrations, pH and other reaction parameters. These complications have discouraged many investigators from using RAPD. Our goal was to optimize the RAPD amplification conditions for vertebrate DNA. We present the optimized protocol along with an experimental strategy for obtaining reproducible, interpretable RAPD banding patterns in vertebrates.

[The efficiency of cold staining methods at the light microscopical evidence of acid fast bacilli (author's transl)]. / Die Leistungsfähigkeit von Kaltfärbeverfharen beim lichtmikroskopischen Nachweis säurefester Bakterien.

Comparing studies has been demonstrated wether or not the cold staining of acid fast bacilli is qualitative and quantitative better than the Ziehl-Neelsen-method. The hot staining was superior to all cold staining methods and their modifications tested. If a cold staining is necessary two methods are recommended, the Kinyoun method and the staining with carbolfuchsin which chloroform is added to. Thin smear and homogeneous material provide better results. Additional investigations concerning decolorizations have shown, that 10% sulfuric acid is as effective as hydrochloric acid-ethanol mixture.

[Tracheal wall cyst. A rare cause of stridor in the adult]. / Trachealwandzyste. Eine seltene Ursache eines Stridors beim Erwachsenen.

A case of tracheal cyst is described in a 43-year-old man who was admitted with nonspecific symptoms of an obstruction in the upper respiratory tract. A CT scan of the thorax showed a solid tumor on the inner tracheal wall. Bronchoscopy revealed a tumor with a broad base and smooth surface that almost completely obstructed the trachea. Intraoperatively, the tumor was found to be a cyst and was successfully removed endoscopically. Tracheal cysts are quite rare and most of the reported cases have been confined to pediatric patients as a congenital disease. More frequently, congenital intraluminal cysts of the upper respiratory tract are found as epiglottic retention cysts and laryngoceles.

[Interferon-induced effects of lectins in vitro and following oral administration]. / Interferoninduzierende Effekte von Lektinen in vitro und nach oraler Applikation.

The interferon inducing effects of lectins such as ConA, PHA, LCA und WGA had been demonstrated. The studies were carried out in human-blood-cell cultures and in serum of mice after an oral uptake of substances subjected. In both experiments under determined conditions the lectins showed an interferogen effect. Especially efficacious was WGA. Most interesting was the systemic IFN-activation after oral stimulation using WGA.

Lectins and their role in a new polyvalent bacterial vaccine against ARI.

The potency of the polyvalent bacterial vaccine (Infectvac) to prevent lethal infections with S. pneumoniae ATCC 6301 was examined. NMRI-mice were protected 2-5 times better than untreated controls. The protection is based on activation of resistance-mechanisms, e.g. interferon production. Most interesting is a strong activation of the phagocytosis-killing-system of alveolar macrophages after oral application of antigen (information: gut mucosa to lung mucosa). Using the same infection model the important role of bacterial lectins for infectious diseases was demonstrated. Blocking the combining site of the bacterial lectin of S. pneumoniae by intranasal application of N-acetylglucosamine (the specific carbohydrate for the lectin) was able to prevent a lethal infection with S. pneumoniae 3-times better than PBS or using not lectin relevant carbohydrates. Therefore, blocking the lectin receptor with specific carbohydrates might also be of clinical relevance to prevent acute respiratory infections (ARI).

Investigation of the patient with abnormal liver function tests.

About one-half of patients with ulcerative colitis develop abnormal liver function tests at some time during the course of the illness. This should prompt an investigation for primary sclerosing cholangitis and other common hepatobiliary diseases. Primary sclerosing cholangitis occurs in 2-10% of patients with ulcerative colitis. The diagnosis of primary sclerosing cholangitis is most often made by endoscopic retrograde cholangiography. Liver histopathology is often inconclusive but magnetic resonance cholangiography shows promise as a useful non-invasive diagnostic tool. Cholangiocarcinoma complicates 20-40% of patients with end-stage primary sclerosing cholangitis and is now one of the most common causes of death in patients with ulcerative colitis. Distinction between benign and malignant strictures can be difficult and is best done with a combination of clinical suspicion, repeated imaging for mass lesions, cholangiography, and endoscopic brushings and/or biopsies. Dominant lesions of the common bile duct or common hepatic duct produce progressive jaundice and liver damage. Early treatment may improve prognosis. Single strictures can be dilated endoscopically. If the stricture is more complicated and extends into the intrahepatic ducts or there is suspicion of cholangiocarcinoma, surgical resection may be more appropriate. Liver transplantation should be considered in end-stage disease.

Lung mechanics in orally immunized mice after aerolized exposure to influenza virus.

A method is described which allows the investigation of lung mechanics during artificial ventilation in live mice. Inhalation of nebulized influenza virus (A/PR/8/34, H1N1) decreased in NMRI mice compliance (means = 0.012 ml/cm H2O, n = 18) due to severe virus pneumonia and edema on days 5-7 after infection. In comparison to noninfected controls (means = 0.029 ml/cm H2O, n = 17) compliance was unchanged in infected, but orally immunized mice (means = 0.030 ml/cm H2O, n = 21). The results demonstrate that decreased compliance due to influenza virus pneumonia can be prevented by oral immunization with influenza viruses.

[Interferon-inducing effect of influenza viruses following aerogenic and oral administration in NMRI mice]. / Die interferoninduzierende Wirkung von Influenzaviren nach aerogener und oraler Applikation bei NMRI-Mäusen.

The aim of our study was to investigate the interferoninducing effect of aerogenic or oral uptake of influenza viruses in animals. NMRI-mice were infected with mouse-adapted influenza viruses (A/PR/8/34, H1 N1) by aerogen and oral route. In one-day-intervals interferon titers were determined in lung lavage fluids and in lung tissues within nine days after infection. The aerogenic infection led to an increase in interferon followed by a plateau and followed by a slope. The same course of interferon given in lower values was seen in animals after oral virus uptake. The results support the idea of a stimulation of local defense mechanisms in the lung after oral antigen uptake.

Influence of trivalent chromium on the beta-cell function.

Application of trivalent chromium (daily 1 mg, p.o.) to pregnant rats in the time of gestation and thereafter--beginning with the first day of the gestation and lasting for 50 days--did not change the i.p. glucose tolerance of the adult animals. The insulin concentration in serum 30 min after glucose loading was decreased significantly. The basal concentration of serum insulin of neonates was diminished, too. The insulin content of the whole pancreas was decreased in mother animals and increased in neonates. Secretion of insulin by isolated islets of the neonatal animals was unchanged. Insulin biosynthesis of the islets of these animals was increased in presence of 6 and 15 mM glucose, respectively. These data suggest an effect of trivalent chromium on endocrine pancreas.

[The appearance of specific IgG and IgA antibodies to Streptococcus pneumoniae in serum and in lung lavage fluid of mice after the administration of different antigens. 1. Development of an enzyme immunoassay (EIA) for the detection of specific IgG and IgA antibodies to Streptococcus pneumoniae]. / Auftreten von spezifischen IgG- und IgA-Antikörpern gegen Streptococcus pneumoniae im Serum und in der Lungenspülflüssigkeit von Mäusen nach unterschiedlicher Antigenapplikation. 1. Mitteilung: Aufbau eines Enzymimmunoassay (EIA) zum Nachweis von spezifischen IgG- and IgA-Antikörpern gegen Streptococcus pneumoniae.

Immunization against streptococcus pneumoniae is for the prophylaxis of pneumococcus diseases in an increased number of risk patients and because of an increasing presence of antibiotica resistance in present days necessary. In order to get better possibilities to test the immunization effect, improved methods for detection of antibodies are applied. In this publication a double sandwich-EIA and a protein-coupled EIA are presented and compared. The double sandwich-EIA has proved to be superior with regard to its day by day variability of 12.5% and its series variability of 3.5% and to be of the same value compared with international developments. The production of most of antigens and antisera is described and the application of gamma-irradiated micro-plates is proposed.

[The appearance of specific IgG and IgA antibodies to Streptococcus pneumoniae in serum and lung lavage fluid of mice after the administration of different antigens. 2. IgG and IgA antibody detection after intraperitoneal, aerosol and oral immunization with Streptococcus pneumoniae]. / Auftreten von spezifischen IgG- und IgA-Antikörpern gegen Streptococcus pneumoniae im Serum und in der Lungenspülflüssigkeit von Mäusen nach unterschiedlicher Antigenapplikation. 2. Mitteilung: IgG- und IgA-Antikörpernachweis nach intraperitonealer, aerogener und oraler Immunisierung mit Streptococcus pneumoniae.

The stimulation of secretory and systemic IgG- and IgA-antibodies in the lung lavage fluid was examined in an animal test. It was also examined in the serum of mice after once to four times oral, aerosol or intraperitoneal immunization with total formalin destroyed bacteria of the species Streptococcus pneumoniae type I. Not only in the lung lavage fluid, but also in the sera specific antibodies could be found with the enzyme immunoassay. An intraperitoneal antigen application mainly resulted in increase of the specific IgG in the serum, whereas an oral and especially aerosol antigen application mainly developed a stimulation of IgA in the lung lavage fluid.

[Detection of specific antibodies to influenza viruses of the genital tract of mice following oral immunization]. / Nachweis spezifischer Antikörper gegen Influenzaviren im Genitaltrakt von Müsen nach oraler Immunisierung.

Balb c-mice were immunized orally twice within 10 days with a live influenza vaccine (A PR 8/34, H1 N1). Specific antibodies were determined in sera, uterus homogenates and uterus lavage fluids. In all compartments specific antibodies of IgG class were estimated by means of an enzymimmunoassay. Regarding estrogen cyclus of mice the antibody titers were divided into diestrus, proestrus, estrus and metestrus. We have not seen any cyclic changes of antibody titers. The results support the concept of a common immune mucosal system. The continuous antibody titers during genital cyclus are evident for using oral vaccination aiming to induce secretory antibodies in genital tract.

Influenza specific antibodies in the female genital tract of mice after oral administration of live influenza vaccine.

In order to investigate the possible stimulation of antibodies in the genital tract by immunization female NMRI-mice were given orally a live influenza vaccine (A/PR/8/34, H1N1) on two occasions which were 10 days apart. Subsequently, virus specific IgA antibodies measured by an enzyme immunoassay in homogenates of urinary bladder, uterus and vagina and also in uterine washings. Specific IgA antibodies were not detectable in the sera of immunized mice. The high IgA titer in uterine washings, and in the homogenates suggests enhancement by vaccine of IgA antibody production in the genital tract.