Experimente mit Hefe

Why bread dough rises, although yeast cannot metabolize starch: Experiments with yeast Yeast is ideal for simple experiments to visualize metabolic processes, such as those known from baking bread, for example. Thus the metabolism of different sugars to carbon dioxide can be qualitatively demonstrated by inflating balloons or quantitatively by gas production in a fermentation tube (glass airlock for fermentation). The experiments presented can be used to answer the question of why yeast is added to bread dough even though it is not able to convert the starch present in the flour.     

Therapie mit Stammzellen

Therapeutic use of stem cells Here the hematopoetic system of blood‐ and immune cell renewal is reviewed. Curing of chronic leucemias and malignant lymphomas is the most successful stem cell based therapy up to date. However, mismatches of histocompatibility‐complexes (HLA‐types) between receiver and donor set narrow limits to such therapies. Whether other diseases such as Parkinson could be cured by infusion of stem cells is still in question.     

Experimente mit Verlagerung polnahen Eimaterials zur Analyse der Bedingungen für die metamere Gliederung des Embryos vonBruchidius (Coleoptera)

Ohne ZusammenfassungHerrn Prof. Dr. Dr. h. c.Hanns Langendorff zum 65. Geburtstag gewidmet.Wir danken der Deutschen Forschungsgemeinschaft für ihre Unterstützung.     

Identification and analysis of the amylase-binding protein B (AbpB) and gene (abpB) from Streptococcus gordonii

The binding of salivary amylase to Streptococcus gordonii has previously been shown to involve a 20-kDa amylase-binding protein (AbpA). S. gordonii also releases an 82-kDa protein into the supernatant that binds amylase. To study this 82-kDa component, proteins were precipitated from bacterial culture supernatants by the addition of acetone or purified amylase. Precipitated proteins were separated by SDS-PAGE and transferred to a sequencing membrane. The P2 kDa band was then sequenced, yielding a 25 N-terminal amino acid sequence, CGFIFGRQLTADGSTMFGPTEDYP. Primers derived from this sequence were used in an inverse PCR strategy to clone the full-length gene from S. gordonii chromosomal DNA. An open reading frame of 1959 bp was noted that encoded a 652 amino acid protein having a predicted molecular mass of 80 kDa. The first 24 amino acid residues were consistent with a hydrophobic signal peptide, followed by a 25 amino acid N-terminal sequence that shared identity (24 of 25 residues) with the amino acid sequence of purified AbpB. The abpB gene from strains of S. gordonii was interrupted by allelic exchange with a 420-bp fragment of the abpB gene linked to an erythromycin cassette. The 82-kDa protein was not detected in supernatants from these mutants. These abpB mutants retained the ability to bind soluble amylase. Thus, AbpA, but not AbpB, appears sufficient to be the major receptor for amylase binding to the streptococcal surface. The role of AbpB in bacterial colonization remains to be elucidated.     

Auge in Auge mit dem Wasserfloh

The Experimental‐Lab Biology FLOX The Experimental‐Lab Biology FLOX of the University of Kassel is a competence‐oriented School Laboratory that focuses on the autonomous experimentation of pupils. Currently water fleas (Daphnia magna) are the preferred experimental object. Student teachers learn to mentor those pupils in their process of knowledge acquisition offering support and advice. Simultaneously, the contradiction between the given and fixed experimental instructions and unstructured experiments is solved by work instructions in form of V‐diagrams. Feedback of pupils and teachers reveal that this is a concept worth pursuing and developing.     

Transport und Umsatz von Polyalkoholen bei der Hefe Rhodotorula gracilis (glutinis)

Zusammenfassung Die Polyalkohole Sorbitol, d-Mannitol, Ribitol, Xylitol, d-Arabitol, l-Arabitol und Erythritol werden von der obligat aeroben Hefe Rhodotorula gracilis über einen beweglichen Träger in der Zellmembran aufgenommen. Der Transportmechanismus ist aktiv, das erreichte Akkumulationsverhältnis ist jedoch bei allen Polyalkoholen erheblich geringer als bei Monosacchariden. Es nimmt, wie auch bei Monosacchariden, mit steigender Außenkonzentration ab, sogar auf Werte kleiner als 1.Kinetische Daten weisen darauf hin, daß das Trägersystem für Polyalkohole identisch ist mit dem für Monosaccharide, jedoch für Polyalkohole eine wesentlich geringere Affinität und maximale Geschwindigkeit aufweist. Aufgrund des hohen Affinitätsunterschiedes wird die Polyalkoholaufnahme in der Anwesenheit von Monosacchariden unterbunden.Die aufgenommenen Polyalkohole werden im Zellinneren nicht umgesetzt; eine Ausnahme stellen Ribitol und l-Arabitol dar, in deren Anwesenheit ein Abbausystem für Pentitole induziert wird.

---

Transport and utilization of alditols in the yeast Rhodotorula gracilis (glutinis) I. Constitutive transport of alditols

The obligate aerobic yeast Rhodotorula gracilis was found to take up the alditols d-glucitol, d-mannitol, ribitol, xylitol, d-arabinitol, l-arabinitol and erythritol by means of a constitutive mobile membrane carrier. This uptake involved active transport, that is, it was dependent on the supply of metabolic energy, leading to the accumulation of alditols inside the cells. The accumulation ratio (intracellular concentration to extracellular concentration, S _i /S _o ) was much lower for alditols than for monosaccharides. As for monosaccharides, this ratio decreased with increasing extracellular concentration, even to values below 1.The kinetic data showed that the carrier system for alditols was identical to that for monosaccharides, though it had a much lower affinity and maximum velocity for alditols. Hence the uptake of alditols was blocked in the presence of monosaccharides.Only ribitol and l-arabinitol were catabolized following enzyme induction. The other alditols were not broken down.

     

Secretagogue activities in cod (Gadus morhua) and shrimp (Penaeus aztecus) extracts and alcalase hydrolysates determined in AR4-2J pancreatic tumour cells

Peptides with gastrin immunoreactivity were measured in cod muscle (Gadus morhua) and shrimp heads (Penaeus aztecus) extracts and alcalase hydrolysates and separated by two chromatographic steps. Secretagogue activities present in crude extracts fractions were examined with or without specific antagonists of CCK receptors in AR4-2J cells. Several sub-fractions significantly stimulate amylase release, up to 110%. These stimulatory effects could be completely inhibited by the presence of L 365, 260 specific antagonist of CCKB receptors. After hydrolysis of the raw material, the samples were partially fractionated by two chromatographic steps and potential active fractions detected by a gastrin-CCK radioimmunoassay. The molecular masses of the active fractions were lower than for the extracts. Stimulation of amylase release was higher than with extracts, and the inhibition by L 365, 260, less pronounced. These results show that some peptides remaining after hydrolysis or extraction still exert biological activities and have to be tested in nutritional studies.     

Experimente zur Bruth?hlenwahl der Kohlmeise (Parus major)

Zusammenfassung Obwohl die Kohlmeise in sehr verschiedenen Höhlen brütet, zeigt die Art eine signifikante Bevorzugung, wenn sie die Möglichkeit zur Auswahl hat. Versuche mit Holzbetonnisthöhlen ergaben eine eindeutige Präferenz tiefer Höhlen von 19 cm Tiefe (vom Flugloch ab) gegenüber Höhlen von 14 oder 9 cm Tiefe. Noch tiefere Höhlen wurden abgelehnt, ebenso horizontal verlaufende. Bruthöhlen mit großem Innenraum von 20 cm Durchmesser wurden gegenüber Modellen von 14 oder 11,5 cm bevorzugt. Der große Brutraum wurde gegenüber tieferen, aber engeren Höhlen bevorzugt; eine solche Höhle sollte 14 cm tief und nicht 19 cm tief sein. In der Diskussion wird ein möglicher Selektionsvorteil in der Sicherheit vor Nestfeinden sowie der besseren Isolation des Nistmaterials gegenüber engeren Höhlen vermutet. Der Bruterfolg ist in großen Höhlen infolge größerer Gelege am größten.

---

Experiments on nest-site selection in Great Tit,Parus major

Summary Although the Great Tit nests in a wide variety of holes, it shows a significant preference when a choice is available. Experiments with cement-sawdust nest-boxes indicated a clear preference for deep boxes of 19-cm depth (measured from entrance hole) over boxes 14 or 9 cm deep. Still deeper boxes were rejected as were those horizontally mounted. Nest-boxes with large internal space of 20-cm diameter were preferred to models of diameter 14 or 11.5 cm. The large box was preferred to deeper boxes with smaller internal diameter; such a box should be 14 and not 19 cm deep. It is suggested in the discussion that the larger-diameter box offers a possible selective advantage over smaller models in security against nest-predators as well as better insulation in the form of more extensive nest material. Breeding success is highest in large-diameter boxes due to larger clutches.

     

The taphonomic fate of isorenieratene in Lower Jurassic shales—controlled by iron?

Fossil derivatives of isorenieratene, an accessory pigment in brown‐colored green sulfur bacteria, are often used as tracers for photic zone anoxia through Earth's history, but their diagenetic behavior is still incompletely understood. Here, we assess the preservation of isorenieratene derivatives in organic‐rich shales (1.5–8.4 wt.% TOC) from two Lower Jurassic anoxic systems (Bächental oil shale, Tyrol, Austria; Posidonia Shale, Baden‐Württemberg, Germany). Bitumens and kerogens were investigated using catalytic hydropyrolysis (HyPy), closed‐system hydrous pyrolysis (in gold capsules), gas chromatography–mass spectrometry (GC–MS) and gas chromatography combustion isotope ratio‐mass spectrometry (GC‐C‐IRMS). Petrography and biomarkers indicate a syngenetic relationship between bitumens and kerogens. All bitumens contain abundant isorenieratane, diverse complex aromatized isorenieratene derivatives, and a pseudohomologous series of 2,3,6‐trimethyl aryl isoprenoids. In contrast, HyPy and mild closed‐system hydrous pyrolysis of the kerogens yielded only minor amounts of these compounds. Given the overall low maturity of the organic matter (below oil window), it appears that isorenieratene and its abundant derivatives from the bitumen had not been incorporated into the kerogens. Accordingly, sulfur cross‐linking, the key mechanism for sequestration of functionalized lipids into kerogens in anoxic systems, was not effective in the Jurassic environments studied. We explain this by (i) early cyclization/aromatization and (ii) hydrogenation reactions that have prevented effective sulfurization. In addition, (iii) sulfide was locally removed via anoxygenic photosynthesis and efficiently trapped by the reaction with sedimentary iron, as further indicated by elevated iron contents (4.0–8.7 wt.%) and the presence of abundant pyrite aggregates in the rock matrix. Although the combined processes have hampered the kerogen incorporation of isorenieratene and its derivatives, they may have promoted the long‐term preservation of these biomarkers in the bitumen fraction via early defunctionalization. This particular taphonomy of aromatic carotenoids has to be considered in studies of anoxic iron‐rich environments (e.g., the Proterozoic ocean).     

Utilization of horticultural waste (Apple Pomace) for multiple carbohydrase production from Rhizopus delemar F2 under solid state fermentation

The brown rot fungus Rhizopus delemar F₂ was shown to produce extracellular thermostable and multiple carbohydrase enzymes. The potential of Rhizopus delemar F2 in utilizing apple pomace under solid state fermentation (SSF) is the purpose of the study. Solid state fermentation (SSF) is a very effective technique opposed to submerged fermentation in various aspects. Enhanced production of multiple carbohydrases 18.20?U?g^?1 of cellulose, 158.30?U?g^?1 of xylanase, 61.50?U?g^?1 of pectinase and amylase 21.03?U?g^?1 was released by microwave pretreatment of apple pomace at 450?W for 1?min and then by incubation the culture thus obtained at 30?°C for 6 days with moisture content of 1:4.5. Apple pomace can serve as a potential source of raw material for the production of multiple carbohydrases. Besides, it can find great commercial significance in production of bioethanol and various industries like textile, fruit juice, paper and pulp industry.     

Warum Spinnen (nur) acht Beine haben

Why spiders have (only) eight legs The difference in leg number between insects and spiders serves as a model for the study of the genetics and evolution of bodyplan differences. In this context the Hox genes are important factors: especially the Hox gene Antennapedia has apparently changed its function and interactions with so far unidentified factors to result in the eight‐legged spider bodyplan.     

A new baurusuchid crocodyliform (Archosauria) from the Late Cretaceous of Patagonia (Argentina)

A new baurusuchid, Wargosuchus australis gen. et sp. nov., coming from the Bajo de La Carpa Formation, Neuquén Province (Argentina), is described. This new taxon is based on a fragment of snout and a portion of the cranial roof. Wargosuchus differs from other crocodyliforms by possessing a deep median groove on the frontals, a contact between nasals and frontals extremely reduced, a large depression for the olfactory bulbs, three large foramina surrounding the large, smooth perinarial depression, and a hypertrophied, conical last premaxillary tooth followed by a large paracanine fossa. The finding of Wargosuchus in Patagonia (Argentina), a taxon with a strong resemblance to Brazilian baurusuchids, reinforces the hypothesis of a similar biota between both regions by the Late Cretaceous. Wargosuchus and Cynodontosuchus represent the only Argentinian mesoeucrocodylians to be included within Baurusuchidae. This finding extends the number of crocodyliforms from the Bajo de la Carpa Formation, which, in turn, corresponds to the most taxonomically diverse one in Argentina.     

Recharacterization of the mammalian cytosolic type 2 (R)-β-hydroxybutyrate dehydrogenase as 4-oxo-l-proline reductase (EC 1.1.1.104)

Early studies revealed that chicken embryos incubated with a rare analog of l-proline, 4-oxo-l-proline, showed increased levels of the metabolite 4-hydroxy-l-proline. In 1962, 4-oxo-l-proline reductase, an enzyme responsible for the reduction of 4-oxo-l-proline, was partially purified from rabbit kidneys and characterized biochemically. However, only recently was the molecular identity of this enzyme solved. Here, we report the purification from rat kidneys, identification, and biochemical characterization of 4-oxo-l-proline reductase. Following mass spectrometry analysis of the purified protein preparation, the previously annotated mammalian cytosolic type 2 (R)-β-hydroxybutyrate dehydrogenase (BDH2) emerged as the only candidate for the reductase. We subsequently expressed rat and human BDH2 in Escherichia coli, then purified it, and showed that it catalyzed the reversible reduction of 4-oxo-l-proline to cis-4-hydroxy-l-proline via chromatographic and tandem mass spectrometry analysis. Specificity studies with an array of compounds carried out on both enzymes showed that 4-oxo-l-proline was the best substrate, and the human enzyme acted with 12,500-fold higher catalytic efficiency on 4-oxo-l-proline than on (R)-β-hydroxybutyrate. In addition, human embryonic kidney 293T (HEK293T) cells efficiently metabolized 4-oxo-l-proline to cis-4-hydroxy-l-proline, whereas HEK293T BDH2 KO cells were incapable of producing cis-4-hydroxy-l-proline. Both WT and KO HEK293T cells also produced trans-4-hydroxy-l-proline in the presence of 4-oxo-l-proline, suggesting that the latter compound might interfere with the trans-4-hydroxy-l-proline breakdown in human cells. We conclude that BDH2 is a mammalian 4-oxo-l-proline reductase that converts 4-oxo-l-proline to cis-4-hydroxy-l-proline and not to trans-4-hydroxy-l-proline, as originally thought. We also hypothesize that this enzyme may be a potential source of cis-4-hydroxy-l-proline in mammalian tissues.     

Der Anionenmetabolismus in stärkehaltigen und stärkefreien Schliefbzellprotoplasten

Mit isolierten und gereinigten stärkehaltigen (Vicia faba) und stärkefreien (Allium cepa) Schließzellprotoplasten wurde der Anionenmetabolismus in Abhängigkeit von der Volumenregulation der Protoplasten untersucht. Es wurden die Korrelation von Malatsynthese und dem Anschwellen in stärkehaltigen sowie der Blockierung dieser Korrelation in stärkefreien Schließzellprotoplasten gezeigt. Die Untersuchung der CO₂-Fixierung und des Stärkemetabolismus ergab, daß die Licht-unabhängige und schnelle Stärkeakkumulation in den Protoplasten von Vicia auf eine CO₂-Fixierung durch die PEP-Carboxylase zurückzuführen ist, wobei das entstandene Malat auf dem Weg der Gluconeogenese zur Stärke umgewandelt werden dürfte. Die schnelle, K+-induzierte Volumenzunahme der Schließzellprotoplasten, die von einer Malatsynthese und einem Stärkeabbau begleitet ist, weist auf ein enges Stärke-Malat-Gleichgewicht hin. Umgekehrt werden nach Reduzierung der Kationenkonzentration ein Anschwellen der Protoplasten (Vicia) und somit eine Stärkemobilisierung zum Malat verhindert. Ein Modell wurde aufgestellt, um einen Beitrag zur Klärung der Rolle des Malats und den Ort seiner Synthese in den stärkehaltigen und stärkefreien Schließzellprotoplasten zu liefern.     

Intensives Wachstum von Ricciocarpus natans (Lebermoos) in Durchlüftungskultur

Das Lebermoos Ricciocarpus natans wurde nichtaxenisch in Durchlüftungskulturen (etwa 2% CO₂ in Luft) im Lichtthermostaten angezogen und seine morphologischen Merkmale beschrieben. Als optimal für eine hohe Produktivität erwiesen sich bei der verwendeten Nährlösung 29 C, ein Licht-Dunkel-Wechsel von 14:10 Stunden und eine Beleuchtungsstärke von maximal 10000 Lux. Unter diesen Bedingungen ist das Wachstum bei relativ kleinen Schwankungen sehr gleichmäß. Die Verdopplungszeit für Masse und Individuen beträgt 48 Stunden. In eine 300 ml-Kulturröhre wurde 1 g Frischgewicht in 200 ml Nährlösung eingeimpft (Trockengewichtsanteil etwa 4 %). NH₄ (bei Konstanthalten des pH-Wertes) und NO₃ sind als Stickstoffquelle gleich gut geeignet. Bei Mangel an Stickstoff nimmt die Thallusgröße stark ab, die Ventralschuppen werden auffällig lang und violett gefärbt. Das Lebermoos eignet sich gut für eine standardisierte, effektive Laborkultur zum Zwecke physiologischer und biochemischer Untersuchungen.     

Untersuchungen zur Synthese der Brenztraubensäuredecarboxylase (E.C.4.1.1)1) bei Hefe

Zusammenfassung Bei der Vergärung eines synthetischen Substrates und von sterilisiertem Traubenmost ist die. Pyruvatbildung wesentlich größer als die Acetaldehydbildung.Mit Thiamin nivelliert sich die Bildung der beiden aufeinanderfolgenden Metaboliten, der Brenztraubensäureanstau ist nicht mehr nachweisbar, Acetaldehyd fällt in etwa gleicher Menge an wie Brenztraubensäure. Das beweist, daß die Synthese des Coenzyms TPP für die Synthese der Brenztraubensäuredecarboxylase geschwindigkeitsbestimmend ist.Bei Zugabe der beiden Thiaminbestandteile Pyrimidin und Thiazol erfolgt ebenfalls eine Nivellierung des Pyruvat/Acetaldehyd-Verhältnisses. Bei Zusatz von nur einem der beiden Teilstücke allein ist dies a ber nicht mehr der Fall. Pyrimidin oder Thiazol allein zeigt die gleichen Verhältnisse wie ohne Thiamin. Die Hefe ist also nicht fähig, die jeweils fehlende andere Hälfte des Thiamins entsprechend schnell zu synthetisieren.Die Synthese der Brenztraubensäuredecarboxylase wird also limitiert von der Synthesegeschwindigkeit des Coenzymes. Diese Ergebnisse beweisen erneut die Induktion der Synthese der Brenztraubensäuredecarboxylase durch das Coenzym, die von anderer Seite mit anderen Methoden gefunden worden war. Diese Coenzym-Induktion kann ebenfalls erfolgen durch Pyrimidin und Thiazol, aber nicht durch einen der beiden Thiaminbestandteile allein.

---

Investigations on the synthesis of pyruvate decarboxylase (E.C.4.1.1.1) by yeast

Summary During fermentation of a synthetic substrate as well as sterilized grape juice the formation of pyruvate is substantially higher than that of acetaldehyde.The presence of thiamine levels the formation of both successive metabolites, the accumulation of pyruvic acid does not occur and more or less equal amounts of acetaldehyde and pyruvic acid are formed. This proves that the rate of pyruvate decarboxylase synthesis is determined by the synthesis of the coenzyme TPP.The addition of the constituents of thiamine, pyrimidine and thiazole also effects a levelling of the pyruvate/acetaldehyde ratio. However, with the addition of only one of the constituents this phenomenon is not observed and the same conditions as in the absence of thiamine are obtained. Thus the yeast is not able to synthesize the respective missing constituent of thiamine.The synthesis of pyruvate decarboxylase is thus limited by the rate of coenzyme synthesis. These results demonstrate anew the induction of the synthesis of pyruvate decarboxylase by the coenzyme. Other investigators obtained these results by a different method. This coenzyme induction can also be obtained by pyrimidine+thiazole but not by one of these thiamine constituents alone.

     

Citizen Science

Citizen science Citizen science is performed on a honorary basis. Citizen scientists (”Citizen Science proper“) have no professional employment in the relevant field of research and differ very much in their educational background, specific knowledge and amount of time dedicated to the subject of research. Today, citizen science has become especially important in some fields neglected by professional science, e.g. regional research. Recently, another form of citizen science has gained much attention, in which usually many citizen scientist are active (”Citizen Science light“). Internet, smartphone and georeferencing by GPS are important tools for collecting, documenting and communicating the observation data. Besides the scientific results, social relevance through participation and information of citizens (e.g. for conservation issues) play a very important role in these projects. The recently strongly increasing interest in and contribution to citizen science plays an important role to strengthen the link and communication between science and society.     

Experimente am ungefurchten Ei vonPimpla turionellae L. (Hymenoptera) zur Funktionsanalyse des Oosombereichs

Summary In endoplasm close to the posterior pole of the egg ofPimpla one finds conglomerated oosome material, rich in RNA. Investigations after various operations in the oosome region (10% of egg length), before cleavage were intended to show whether pole cells develop, how many segments form and if gonads contain primordial germ cells.Oosome material was squashed with a blunt glass needle. The uninjured part of the egg in front of the oosome region develops blastoderm but no pole cells. It gives rise to a fully segmentated larva with germ cells in the gonads.After ligation of up to 15% of egg length complete embryos with germ cells can develop. The smaller the anterior isolates, the more abdominal segments are missing.By ligation and invagination of the hindpole of eggs with a blunt glass needle, anteriorly material from the oosome region is combined with ooplasm situated more. Translocation of only a small amount of ooplasm results in the same number of abdominal segments in the anterior isolate as in ordinary ligated eggs. Translocation of much ooplasm yields a significantly greater number of abdominal segments. It is immaterial for the metameric segmentation of the embryo whether the oosome is situated before or behind the ligature or is destroyed. But the depth of the invagination and how many segments result do not seem to be correlated.A completely segmentated embryo can develop also after extirpation of the oosome provided care is taken not to injure the hindpole-plasm. No pole-cells result when the complete oosome is missing and the hindpole-plasm is present; loss of part of the oosome results in the development of only a few pole-cells. Thus oosome material is a necessary and quantitative condition for pole-cell differentiation. In one favourable case pole-cells developed in the extraovate because the oosome was followed after some hours by endoplasm and cleavage nuclei.Functions of the oosome are discussed: together with cleavage nuclei it is responsible for pole-cell development. As pole-cells are not invariable precursors of germ-cells, the oosome cannot contain determinants for them. Possibly it includes postembryonic growth modifiers or it could be active in gametogenesis later on. As an egg without oosome-region is able to develop an embryo, this region does not or exclusively contain an activation-center (e. g.Platycnemis), or special hind-pole factors (e. g.Euscelis). In any case the oosome itself does not include these factors. A greater number of segments in the anterior isolate after translocation of ooplasm could be due to its special quality, as inEuscelis andBruchidius whose metameric organisations originate from a bipolar ooplasmic reaction system. Also it could depend only on the increase of ooplasm competent for differentiation-factors in the middle and anterior egg parts.

---

---

Durchgeführt mit Leihgaben der Deutschen Forschungsgemeinschaft und mit Hilfe von Euratom (Verträge Nr. 041-65-10 BIOD und Nr. 077-69-I BIOC mit dem Heiligenberg-Institut).