Immunohistochemical demonstration of villin in the normal human pancreas and in chronic pancreatitis

Human pancreatic tissue was investigated by immunohistochemistry using a polyclonal antibody against the actin binding protein villin, which participates in the formation of actin filament bundles in the microvilli. In cells of the different parts of the pancreatic duct system as well as in the acinar cells villin immunoreactivity was located mainly at the apical cell surface. This was confirmed by the ultrastructural demonstration of microvilli on the surface of duct and acinar cells, which exhibited the typical actin bundles. In chronic pancreatitis the staining for villin in duct-like structures of degenerative pancreatic tissue was irregular or even absent. This correlated with the electron microscopic observation of duct-like structures known as tubular complexes composed of cells devoid of microvilli at the apical cell surface. At the light microscopical level degenerative structures without lumen and of unknown origin showed a strong staining for villin at their basal cell surface.     

Auxin-binding protein from coleoptile membranes of corn (Zea mays L.). II. Localization of a putative auxin receptor

With the aid of affinity chromatography on auxin-binding protein-Sepharose (ABP-Sepharose) monospecific IgGanti-ABP from rabbit antisera were isolated as judged by immuno-double diffusion test and sodium dodecyl sulfate-polyacrylamide gel electrophoresis. With this IgGanti-ABP the ABP is localized within the outer epidermal cells of coleoptiles using indirect immunofluorescence labeling. Auxin-induced growth of coleoptile segments can be inhibited by IgGanti-ABP, and the auxin response of split coleoptile sections is also strongly reduced by IgGanti-ABP. The ABP, therefore, is referred to as an auxin receptor. This auxin receptor is localized at the plasmalemma of the outer epidermal cells of the coleoptile.     

Beta-blocker therapy in unstable severe heart failure,evidence or experience?

Currently, no evidence exists on the effects of beta-receptor blocker (BRB) treatment in patients with unstable severe heart failure. When confronted with this specific patient category, clinical experience in our centre has consistently guided us to lower the dose or stop BRB therapy. To share this experience, we present three clinical case scenarios and discuss background literature motivating our approach in these patients.     

Purification and immunohistochemical localization of the auxin binding site i protein from maize coleoptiles

An auxin binding protein fraction prepared by means of affinity chromatography on 2-OH-3,5-diiodobenzoic acid-Sepharose and gel filtration was used as antigen. The obtained rabbit antisera contained antibodies against the auxin, binding protein (ABP) and several contaminating proteins (nonABP). The nonABP could be separated on an appropriate affinity matrix omitting the TIBA analogue. After their immobilization on Sepharose antibodies directed towards contaminating, the proteins were isolated and immobilized, too. This IgGanti nonABP-Sepharose retains almost all contaminating proteins present in the specific eluates of the auxin affinity matrix. In a final affinity chromatography step on IgG-Sepharose a highly purified ABP could be eluted. This ABP was immobilized on Sepharose for the separation of monospecific antibodies against ABP (IgGanti abp). Using these antibodies the ABP could be localized within the outer epidermal cells of the coleoptile by immunofluorescence microscopy. From the inhibition of auxin induced elongation of coleoptile tissue by IgGanti abp it is concluded that the ABP is localized at the plasmalemma of the epidermal cells and that the ABP is involved in auxin action as a true hormone receptor. Presented at the International Symposium “Plant Growth Regulators” held on June 18–22, 1984 at Liblice, Czechoslovakia.     

FIM Imaging and FIMtrack: Two New Tools Allowing High-throughput and Cost Effective Locomotion Analysis

The analysis of neuronal network function requires a reliable measurement of behavioral traits. Since the behavior of freely moving animals is variable to a certain degree, many animals have to be analyzed, to obtain statistically significant data. This in turn requires a computer assisted automated quantification of locomotion patterns. To obtain high contrast images of almost translucent and small moving objects, a novel imaging technique based on frustrated total internal reflection called FIM was developed. In this setup, animals are only illuminated with infrared light at the very specific position of contact with the underlying crawling surface. This methodology results in very high contrast images. Subsequently, these high contrast images are processed using established contour tracking algorithms. Based on this, we developed the FIMTrack software, which serves to extract a number of features needed to quantitatively describe a large variety of locomotion characteristics. During the development of this software package, we focused our efforts on an open source architecture allowing the easy addition of further modules. The program operates platform independent and is accompanied by an intuitive GUI guiding the user through data analysis. All locomotion parameter values are given in form of csv files allowing further data analyses. In addition, a Results Viewer integrated into the tracking software provides the opportunity to interactively review and adjust the output, as might be needed during stimulus integration. The power of FIM and FIMTrack is demonstrated by studying the locomotion of Drosophila larvae.     

The substrate specificity of trypsin. The interrelationship between the structure and reactivity for quasi-substrates, derivatives of O-alkylmethylphosphonic acid and carboxylic acids

The kinetics of trypsin phosphorylation by thioesters of O-n-alkylmethylphosphonic acids, and reactivation of corresponding phosphoryl enzymes as well as kinetics of trypsin-catalyzed hydrolysis of p-nitrophenylcarboxylates have been studied. The rate constants for phosphorylation and dephosphorylation of trypsin depend on hydrophobicity of non-polar fragments in both substrate series in the same degree. On the other hand, the deacylation rate constants for a series of acyl trypsins do not change significantly while the apparent Michaelis constants change consistently with variations of non-polar acyl substituent. The study of substrate specificity of trypsin in terms of the transition state theory has allowed to elucidate the basis for low reactivity of trypsin towards the quasisubstrates.     

Study of the growth and development of chlorella populations in the culture as a whole

Исследовалась цитоморфология двух видов Chlorella в условиях, при которых наблудается в боляшей или меняцей степени циссоциация процессов роста и размножения. В средах различного состава можно было наблуюдатявполне тиничный и воспроизводимый характер (trend) роста и развития кулятуры как целого. Chlorella в основном размножается так, как размножаются клеточные ядра, митохондрии и пластиды. Из них и из их плазмы строятся постепенно сначала безоболочечные и поцвижные проаутоспоры, которые, приобретая оболочку, преврацаются в непоцвижные аутоспоры. Экбалаты и экбластаты —это различные формы ранних фаз развитня проаутоспор. Недостаток магния и серы вызывает быстроепрекрацение способности к размножению и преобладания роста в величниу в сравнении со способностяю водорослей к делению. Чрезмерный рост в величину у Chlorella является не признаком физиологического благополучия, а признаком дегенерации. При зтом Chlorella подвергается вакуолизации и позднее погибает, обычно в результате осмотического растягивания.