Anaphase transport of akinetochoric fragments in tipulid spermatocytes

Akinetochoric chromosomal fragments in spermatocytes of mutant Pales ferruginae are transported polewards in anaphase. During migration their surfaces form radial lamellar projections between which non-kinetochoric spindle microtubules become arranged in an orderly fashion. The same morphological features had been observed earlier in intact chromosomes in late anaphase. It is assumed that the fragments are transported by some kind of poleward directed "streaming" force of the anaphase spindle, which is applied to the fragment's surface. Non-kinetochoric microtubules are thought to be engaged in the generation or, at least, in the transmission of this spindle force. Due to the morphological similarities with akinetochoric fragments, extra-kinetochoral application sites for anaphase spindle forces can be also suggested for chromosomes possessing kinetochores.     

Akustische Differenzierung verwandtschaftlicher Beziehungen in derParus (Periparus)-Gruppe nach Untersuchungen im Nepal-Himalaya

Zusammenfassung Die fünf Formen der UntergattungParus (Periparus) des südlichen Zentral-Asien(rufonuchalis, rubidiventris, beavani, melanolophus, aemodius) werden morphologisch, biologisch-ökologisch und vor allem akustisch differenziert. Die Spezifität der Gesänge gilt als wesentliches Art-Kriterium; sie wird mit Attrappen-Versuchen getestet.1.Rufonuchalis, rubidiventris undbeavani (Fichtenmeisen-Gruppe) bilden im Nepal-Himalaya Arealgrenzen aus.Rufonuchalis undrubidiventris leben dort sympatrisch,rubidiventris undbeavani allopatrisch ohne Introgressionszone. Rubidiventris undbeavani haben übereinstimmende ökologische Ansprüche;rufonuchalis als größere und trockenadaptierte Form weicht merklich ab. Die Arealgrenzen vonrufonuchalis undrubidiventris sind im westlichen Nepal-Himalaya klimatisch bedingt (Monsun!), nicht durch Konkurrenz mit anderenPeriparus-Formen.Reviergesänge der farblich fast ununterscheidbarenrufonuchalis undbeavani divergieren sehr, die vonrubidiventris undbeavani sind gleich. Rufonuchalis verfügt über 2 Gesangstypen: Trillergesang dient der Reviermarkierung, Pfiffgesang der Revierverteidigung. Beide Gesangsformen entsprechen nicht demParus-Gesangsschema, wohl aber der Klappergesang vonrubidiventris undbeavani. Beavani reagiert im Attrappen-Test nur auf denrufonuchalis-Pfiffgesang und besitzt vergleichbare Lautäußerungen.Rufonuchalis-Trillergesang findet beibeavani ebenfalls Entsprechungen, jedoch nicht beirubidiventris.Spezialisierte Trillergesang-Elemente vonrufonuchalis undbeavani werden als homolog betrachtet. Bei diesen allopatrischen Formen konnten sie erhalten bleiben, der mitrufonuchalis sympatrischerubidiventris hat sie reduziert.Die gleichfarbigenrufonuchalis undbeavani haben das Himalaya-System nie kontinuierlich besiedelt; ihre Areale standen ursprünglich über die Waldzonen Zentralasiens miteinander in Verbindung.Akustische Analyse und Verbreitungsmuster weisenrufonuchalis undrubidiventris (mit den Subspeziesrubidiventris undbeavani) als eigene Spezies aus; sie werden zu einer Superspezies verklammert.2. Die Areale vonP. ater aemodius undP. melanolophus stehen im zentralen Himalaya in sekundärem Kontakt; sie bilden dort eine Hybrid-(Introgressions-)Zone von 70 km Breite.An keiner Stelle desaemodius- bzw.melanolophus-Areals kommen die beiden Ausgangsformen bzw. Hybriden und Ausgangsformen nebeneinander vor. Merkliche Zuwanderung in das Hybridgebiet aus östlichen oder westlichen Arealteilen findet nicht statt.Es existieren Hybriden, die nicht intermediär zwischen den Ausgangsformen stehen, sondern von ihnen in Farbmuster und Färbung markant abweichen (rostbäuchige Hybriden).Vom Gebiet reineraemodius-Phäne zu dem reinermelanolophus-Phäne verläuft eine Merkmalsprogression:aemodius-Beige der Unterseite zu Rostrot;aemodius-Beige der Flanken zumelanolophus-Grau. Rostrot der Unterseite schlägt fast ohne Übergang zumelanolophus-Grau um (bisher nur für eine Population belegt).Alle Populationen des Hybridgürtels sind voll fortpflanzungsfähig (Reviergesang, Revierabgrenzung, -Brutfleck, -Gonadenentwicklung). Die Ausgangsformen haben (noch) keine verschiedenen adaptiven Niveaus erreicht; die Hybriden sind bei der Paarbildung (offensichtlich) nicht unterlegen.Isolationsmechanismen zwischen Ausgangsformen und Hybriden sind (heute) nicht (mehr) entwickelt; füraemodius undmelanolophus werden gering entwickelte akustische (und optische) postuliert (Hinweise durch Hybridisation in Gefangenschaft).Rostfarbigkeit wird als altes Merkmal derPeriparus-Gruppe angesehen, sein Auftreten bei den Hybriden als Atavismus.Reviergesänge aus derater/melanolophus-Gruppe sind einander sehr ähnlich. Der Gesang mitteleuropäischera. ater und der vonmelanolophus zeichnet sich durch ansteigende Elemente aus, diea. aemodius fehlen. Auf solche Gesangstypen reagiertaemodius nur mangelhaft.Die Gesänge in den Hybridpopulationen gleichen jenen vonaemodius; das wird auf Lernen desaemodius-Repertoires der Hybriden während des ersten Kontaktes vonmelanolophus undaemodius zurückgeführt.Süddeutschea. ater reagieren nur zur Zeit höchster Revierverteidigungs-Bereitschaft auf einen bestimmtenaemodius-Strophentyp. Auf diesen Strophentyp reagierten auch syntop lebendeP. major; er enthält Merkmale der Gesänge beider Arten.Die genetische Divergenz vonaemodius undmelanolophus — kenntlich an den rostbäuchigen Hybriden — legt nahe, den Artstatus beider bedingt aufrechtzuerhalten: SuperspeziesP. ater mit den SemispeziesP. ater undP. melanolophus.

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Acoustic differentiation of phylogenetic relations in theParus (Periparus) group from investigations in the Nepal Himalayas

Summary In the Nepal Himalayas 5 taxa of the subgenusParus (Periparus) exist — the greatest concentration of this subgenus in the whole Palearctic region. The taxa belong to 2 groups: 1.rufonuchalis, rubidiventris, beavani; 2. aemodius, melanolophus (fig. 1). All forms settle in the same altitude belt from about 2800 to 4200 m — three of them even in the same locality.Rufonuchalis is adapted to dry habitats mainly of West Central Asia and is thus restricted in Nepal to the Western parts (fig. 3, 4).The up to now unclarified status of all forms is determined as follows: Distinct species areP. rufonuchalis andP. rubidiventris (with highly different subspeciesP. r. rubidiventris andP. r. beavani) and they are regarded as a superspecies.P. rufonuchalis andP. r. rubidiventris live syntopically in West Nepal and display different territorial songs.Both differ remarkably in body size and bill size. Certain acoustic similarities indicating close relationship could be shown betweenrufonuchalis and the allopatric (fig. 3, 4)rubidiventris beavani; r. beavani reacts to the playback of whistle song (Pfiffgesang, fig. 10) ofP. rufonuchalis (see also fig. 14).The areas ofP. r. rubidiventris andP. r. beavani are separated apparently only by the Bhote Kosi river in East Nepal (fig. 4). Introgression is not (yet) known; the songs are identical (fig. 8). P. melanolophus andP. ater aemodius hybridize in West Nepal in a zone of secondary contact. Their areas are linked by a zone of introgression (fig. 17, 18). In that zone cinnamomeous bellied hybrids locally occur differing thus strikingly in colour from both parental species (fig. 1). Sympatric occurrence ofmelanolophus andater aemodius is not known and is not expected either.Territorial songs ofa. aemodius (fig. 21) andmelanolophus (fig. 25) are very similar and both species obviously do not differ in their ecology. Despite the small introgression belt, similar songs and apparently very similar or even identical adaptive peaks,P. ater andP. melanolophus are still treated as distinct species but reduced to semispecies level. In doing so the genetic divergence betweenater aemodius andmelanolophus indicated by the cinnamomeous bellied hybrids is emphasized.

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Mit einem Jahresstipendium des Deutschen Akademischen Austauschdienstes und einer Sachbeihilfe der Deutschen Forschungsgemeinschaft. — Ergebnisse der Nepal-Reisen 1969/70, 1973 und 1974, Nr. 36. — Nr. 35: Senckenbergiana biol., 56 (4/6): 247–256, 1975.     

Pitfalls in the sample preparation and analysis of N-acylethanolamines

N-acylethanolamines (NAEs) are a group of lipid mediators synthesized in response to a number of physiological and pathological stimuli. Because of the low tissue concentrations of NAEs, analyses often include liquid extraction followed by solid-phase extraction and subsequent quantitation by LC/MS or GC/MS. Reported levels of NAEs vary considerably, however, and often no explanation is given for these discrepancies. Brought on by difficulties encountered during method development, the effects of using four different brands of silica-containing solid phase extraction (SPE) columns and five different brands of chloroform for sample preparation were investigated. Considerable variation in the retention and recoveries of seven NAEs and 2-arachidonoylglycerol existed between the SPE columns. Furthermore, it was found that some chloroforms contained quantifiable amounts of N-palmitoylethanolamine and N-stearoylethanolamine. Finally, it was found that use of one of the chloroforms resulted in a loss of N-oleoylethanolamine from solution due to addition of chlorine to the ω-9 bond. The identity of this reaction product was confirmed by LC-MS/MS and NMR. It is recommended that these aspects of sample preparation and analysis should be thoroughly validated during method development and the relevant information on specific brands used be reported in future communications in order to better estimate the validity of reported quantitative data.     

Different Glycosylation in Acetylcholinesterases from Mammalian Brain and Erythrocytes

Acetylcholinesterases (EC 3.1.1.7, AChE) have varying amounts of carbohydrates attached to the core protein. Sequence analysis of the known primary structures gives evidence for several asparagine-linked carbohydrates. From the differences in molecular mass determined on sodium dodecyl sulfate-polyacrylamide gel before and after deglycosylation with N-glycosidase F (EC 3.2.2.18), it is seen that dimeric AChE from red cell membranes is more heavily glycosylated than the tetrameric brain enzyme. Furthermore, dimeric and tetrameric forms of bovine AChE are more heavily glycosylated than the corresponding human enzymes. Monoclonal antibodies 2E6, 1H11, and 2G8 raised against detergent-soluble AChE from electric organs of Torpedo nacline timilei as well as Elec-39 raised against AChE from Electrophorus electricus cross-reacted with AChE from bovine and human brain but not with AChE from erythrocytes. Treatment of the enzyme with N-glycosidase F abolished binding of monoclonal antibodies, suggesting that the epitope, or part of it, consists of N-linked carbohydrates. Analysis of N-acetylglucosamine sugars revealed the presence of N-acetylglucosamine in all forms of cholinesterases investigated, giving evidence for N-linked glycosylation. On the other hand, N-acetylgalactosamine was not found in AChE from human and bovine brain or in butyrylcholinesterase (EC 3.1.1.8) from human serum, indicating that these forms of cholinesterase did not contain O-linked carbohydrates. Despite the notion that within one species, the different forms of AChE arise from one gene by different splicing, our present results show that dimeric erythrocyte and tetrameric brain AChE must undergo different postsynthetic modifications leading to differences in their glycosylation patterns.     

Bovine-associated mucoprotein: I. Distribution among adult and fetal bovine tissues and body fluids

Bovine-associated mucoprotein (BAMP), solubilized with water from the delipidated membranes of bovine milk fat globules, is not restricted to fat globules or to the alveolar epithelial cells from which they are formed. BAMP also has a widespread distribution on other bovine glandular epithelial cells and on undifferentiated cells in lymphoid germinal centers and in several fetal tissues. Free BAMP is present in bovine colostrum, milk, other secretory fluids, and in fetal serum but is absent from adult and colostrum-deprived calf sera. In bronchoalveolar fluids, BAMP is preferentially found in the mucus-rich fraction. BAMP is antigenically distinct from all adult serum proteins, free secretory component, beta 2-microglobulin, lactoferrin, alpha-lactalbumin, beta-lactoglobulin, and five different caseins. BAMP as a free protein constitutes one-sixth of the total amount of BAMP present in milk. The BAMP-related component of fetal serum lacks antigenic determinants present on the BAMP of milk as demonstrated by immunoprecipitation and partial blocking of immunofluorescence. The fetal component is not fetuin or alpha 1-fetoprotein. These data suggest that BAMP may be useful in studies of the membranes of proliferating or differentiating epithelial cells.     

Comparative phytochemistry and systematics of Anacyclus

Leaf flavonoids of 13 Anacyclus taxa have been identified and compared. The most common compounds are 3-, 7- or 5-glycosylated flavonols which, together with the accumulation of 2 diosmetin 7-glycosides, help to delimitate species groups according to recent morphological and cytological findings. In addition to quercetagetin, quercetagetin 3'-methyl ether, patuletin and spinacetin have been isolated as 7-glucosides from the yellow disc and ray flowers of Anacyclus radiatus. The distribution patterns of polyacetylenes and particularly related amides, characterize different Anacyclus species and apparently contribute to a more natural interpretation of relationships with other genera, which may also be underlined by the distribution of cyanogenic glycosides.     

Freeze-drying and analysis of plant and other biological material

The freeze-drying of biological material, which is to be quantitatively analyzed (micro-amount level) for compounds of low or intermediate molecular weight, should be either omitted or handled under strict control. This is because compounds such as amino acids, sugars, flavonoids, glycosides, coenzymes, peptides, etc., might be removed from concentrates and (or) the ground biological material by the high vacuum.     

Thermal stability of fatty acid-serum albumin complexes studied by differential scanning calorimetry.

Differential scanning calorimetry has been used to study the thermal stability of bovine serum albumin as affected by binding of fatty acids (lauric acid and stearic acid) and sodium dodecyl sulfate (SDS). All the ligands stabilized the protein molecules in a similar manner, but to different levels. A maximum increase in denaturation temperature of 30 degrees C was obtained with lauric acid. The thermograms indicate the presence of several ligand-albumin complexes having different heat stabilities. Variations in pH in 0.9% NaCl affected the heat stability of both ligand-poor and ligand-rich albumin, the former being more sensitive to variations in pH within the physiological range. Variations in NaCl concentration affected the thermal stabilities at neutral pH, expecially at low salt concentrations. While ligand-rich albumin was somewhat destabilized by increasing NaCl concentrations, ligand-poor albumin was strongly stabilized. The potential use of differential scanning calorimetry in ligand-albumin research is discussed.