First-Year Common Nightingales (Luscinia megarhynchos) Have Smaller Song-Type Repertoire Sizes Than Older Males

Based on the assumptions that birdsong indicates male quality and that quality is related to age, one might expect older birds to signal their age. That is, in addition to actual body condition, at least some song features should vary with age, presumably towards more complexity. We investigated this issue by comparing repertoire sizes of free‐ranging common nightingale males in their first breeding season with those of older males. Nightingales are a good model species as they are open‐ended learners, where song acquisition is not confined to an early sensitive period of learning. Moreover, nightingales develop an extraordinarily large song‐type repertoire (approx. 180 different song types per male), and differences in repertoire size among males are pronounced. We analysed repertoire characteristics of the nocturnal song of nine nightingales in their first breeding season and compared them with the songs of nine older males. The repertoire size of older males was on average 53% larger than that of yearlings. When analysing two song categories of nightingales, whistle and non‐whistle songs separately, we found similar results. Our findings show marked differences in repertoire size between age categories, suggesting that this song feature may reflect a male's age. We discuss those mechanisms that may constrain the development of larger repertoires in first‐year males. Whether repertoire sizes are crucial for female mate choice or in vocal interactions among conspecific males remains open to further investigations.     

Produce Isolates of the Escherichia coli Ont:H52 Serotype That Carry both Shiga Toxin 1 and Stable Toxin Genes

Produce isolates of the Escherichia coli Ont:H52 serotype carried Shiga toxin 1 and stable toxin genes but only expressed Stx1. These strains had pulsed-field gel electrophoresis profiles that were 90% homologous to clinical Ont:H52 strains that had identical phenotypes and genotypes. All Ont:H52 strains had identical single nucleotide polymorphism profiles that are suggestive of a unique clonal group.     

Vaccine-Induced,Pseudorabies Virus-Specific,Extrathymic CD4+CD8+ Memory T-Helper Cells in Swine

Pseudorabies virus (PRV; suid herpesvirus 1) infection causes heavy economic losses in the pig industry. Therefore, vaccination with live attenuated viruses is practiced in many countries. This vaccination was demonstrated to induce extrathymic virus-specific memory CD4⁺CD8⁺ T lymphocytes. Due to their major histocompatibility complex (MHC) class II-restricted proliferation, it is generally believed that these T lymphocytes function as memory T-helper cells. To directly prove this hypothesis, 15-amino-acid, overlapping peptides of the viral glycoprotein gC were used for screening in proliferation assays with peripheral blood mononuclear cells of vaccinated d/d haplotype inbred pigs. In these experiments, two naturally processed T-cell epitopes (T1 and T2) which are MHC class II restricted were identified. It was shown that extrathymic CD4⁺CD8⁺ T cells are the T-lymphocyte subpopulation that responds to epitope T2. In addition, we were able to show that cytokine secretion can be induced in these T cells through recall with inactivated PRV and demonstrated that activated PRV-primed CD4⁺CD8⁺ T cells are able to induce PRV-specific immunoglobulin synthesis by PRV-primed, resting B cells. Taken together, these results demonstrate that the glycoprotein gC takes part in the priming of humoral anti-PRV memory responses. The experiments identified the first T-cell epitopes so far known to induce the generation of virus-specific CD4⁺CD8⁺ memory T lymphocytes and showed that CD4⁺CD8⁺ T cells are memory T-helper cells. Therefore, this study describes the generation of virus-specific CD4⁺CD8⁺ T cells, which is observed during vaccination, as a part of the potent humoral anti-PRV memory response induced by the vaccine.Pseudorabies virus (PRV), a member of the Alphaherpesvirinae, is the causative agent of Aujeszky’s disease. This disease is lethal to young pigs and causes important economic losses (52). Therefore, vaccination of pigs is practiced in many countries.Several humoral immune system effector mechanisms are involved in the protection of pigs from PRV infection. Virus-neutralizing antibodies, antibodies mediating antibody-dependent cell-mediated cytotoxicity, and antibodies mediating complement-mediated lysis of PRV-infected target cells have been demonstrated (22, 23, 53, 54). The main targets of this humoral immune response were shown to be the viral glycoproteins (3, 45), and passive immunization with monoclonal antibodies (MAbs) against gB, gC, and gD protects pigs from a lethal challenge (20, 49).The protection conferred through cell-mediated immunity is poorly understood. An increase in major histocompatibility complex (MHC)-unrestricted cell-mediated cytotoxicity against uninfected and PRV-infected cells has been detected after infection or vaccination of pigs with PRV (16, 53, 54), and specific cellular immune responses to PRV infections could be demonstrated by stimulation of proliferation and lymphokine secretion of porcine PRV-immune lymphocytes (10, 17, 42, 43, 51) as well as by the detection of PRV-specific cytotoxic lymphocytes (21, 56).There are some difficulties in defining more precisely the impact of cell-mediated immune effector mechanisms to protection from PRV-infection and their interplay with the observed humoral immune response. Considerably fewer porcine than human or mouse differentiation markers are available (34). In addition, the immune system of swine differs considerably from that of humans and mice. The pig has a substantial number of CD4⁻CD8⁻ T lymphocytes in the peripheral blood (4, 6, 12, 36, 39). In young animals, this subpopulation of T lymphocytes comprises up to 60% of the T lymphocytes and contains mainly γδ T lymphocytes. The pig is also the only species so far known to contain a substantial number of resting extrathymic CD4⁺CD8⁺ T lymphocytes (28, 36, 39). This T-lymphocyte population shows morphologically the phenotype of mature T lymphocytes (40) and increases with age to up to 60% of peripheral T lymphocytes (29, 35, 39, 55). Further, it was demonstrated that CD4⁺CD8⁺ T lymphocytes comprise memory T cells which proliferate upon stimulation with recall antigen (43, 55). Since the observed proliferative response was shown to be MHC class II-restricted, it was speculated that the porcine CD4⁺CD8⁺ T-cell subset contains memory T-helper lymphocytes (43). However, the ability of these T lymphocytes to secrete cytokines or to provide help to B cells has so far not been demonstrated.To gain a better understanding of immune effector mechanisms conferring protection from PRV infection, the function of these unusual extrathymic T-lymphocyte subsets has to be elucidated. In the present study, we identified two T-cell epitopes on glycoprotein gC which are primed during vaccination of d/d haplotype inbred pigs (41) against PRV and demonstrated that MHC class II-restricted, peripheral CD4⁺CD8⁺ memory T lymphocytes are the responding T lymphocytes. We were further able to show that PRV-specific, extrathymic CD4⁺CD8⁺ T lymphocytes are able to secrete cytokines and have the capacity to stimulate the secretion of PRV-specific immunoglobulins (Ig) by PRV-primed B cells. These results demonstrate that porcine CD4⁺CD8⁺ T lymphocytes can function as memory T-helper cells and can direct humoral anti-PRV memory responses.     

Storage behavior of Typha latifolia pollen at low water contents: Interpretation on the basis of water activity and glass concepts

Germplasm must be stored under optimal conditions to maximize longevity and efficiently maintain genetic resources. In order to identify optimal storage conditions, we investigated the effects of temperature (−5 to 45°C) and water content (<0.17 g H₂O g⁻¹ dry weight) on longevity of Typha latifolia L. pollen. Longevity was highest at water contents corresponding to storage relative humidity (RH) of 11‐15% which corresponded to the shoulder of water sorption isotherms. Also coinciding with this shoulder were abrupt changes in heat capacity of water present in the pollen. Consistent with changes in isotherms with temperature and the concept of critical RH for storage, optimum water contents increased with decreasing temperature. An attempt was made to explain the aging behavior according to the glass concept. The water content‐temperature combinations of optimal storage were found to be below the glass transition curve, indicating that optimum storage conditions are achieved when intracellular glasses are present. We also found a change in activation energy of aging in Arrhenius plots around T_g, demonstrating a change in aging kinetics when the glassy state is lost. We concluL that T_g curves cannot be used solely to predict precise conditions of optimum storage, but might be useful for predictions of storage longevity above optimum water contents. The data imply that too much drying reduces longevity and should be avoided, particularly when cryogenic storage is considered.     

Ion channels in microglia (brain macrophages)

Microglia are immunocompetent cells in the brain that have manysimilarities with macrophages of peripheral tissues. In normal adultbrain, microglial cells are in a resting state, but they becomeactivated during inflammation of the central nervous system, afterneuronal injury, and in several neurological diseases. Patch-clamp studies of microglial cells in cell culture and in tissue slices demonstrate that microglia express a wide variety of ion channels. Sixdifferent types of K⁺ channelshave been identified in microglia, namely, inward rectifier, delayedrectifier, HERG-like, G protein-activated, as well as voltage-dependentand voltage-independentCa²⁺-activatedK⁺ channels. Moreover, microgliaexpress H⁺ channels,Na⁺ channels, voltage-gatedCa²⁺ channels,Ca²⁺-release activatedCa²⁺ channels, andvoltage-dependent and voltage-independentCl channels. With respectto their kinetic and pharmacological properties, most microglial ionchannels closely resemble ion channels characterized in othermacrophage preparations. Expression patterns of ion channels inmicroglia depend on the functional state of the cells. Microglial ionchannels can be modulated by exposure to lipopolysaccharide or variouscytokines, by activation of protein kinase C or G proteins, by factorsreleased from astrocytes, by changes in the concentration of internalfree Ca²⁺, and by variations ofthe internal or external pH. There is evidence suggesting that ionchannels in microglia are involved in maintaining the membranepotential and are also involved in proliferation, ramification, and therespiratory burst. Further possible functional roles of microglial ionchannels are discussed.

     

Effect of radiation on the biosynthesis of L-Tryptophan from 14C-Anthranilic acid in kohlrabi (Brassica oleracea L., var.gongylodes L.)

The metabolism of¹⁴C-anthranilic acid (¹⁴C-AA) in kohlrabi (Brassica oleracea L. var.gongylodes L.) and the effect of radiation gamma⁶⁰Co on this metabolism was investigated. In hypocotylar segmnents of seven days old etiolated seedlings¹⁴C-AA was metabolised par, tially to its detoxication product¹⁴C-β-glucoside of AA. Simultaneously L-tryptophan was also formed, which in these plants is a precursor of indolic glucosinolates glucobrassicin and neoglucobrassicin. The metabolism of¹⁴C-AA was followed for 97 h. Radiation, applied both to seeds and to seven days old plants did not affect the metabolism of¹⁴C-AA substantially. The intermediary reaction AA → L-tryptophan in the biosynthesis of L-tryptophan is not a radiosensitive part of the synthesis of this amino acid. A not too high radiation sensitivity (max. 45%) was observed in the metabolic pathway leading from L-tryptophan to glucobrassicin.     

Die sog. Asphalt-Flecke der kultivierten Herzmuskelzellen — eine im Phasenkontrastmikroskop sichtbare Formation des Glykogens

Zusammenfassung Bei einer korrespondierenden Betrachtung dreier Asphaltflecke von lebenden Herzmuskelzellen in der Kultur und den identischen Stellen im Elektronenmikroskop erweist sich der Fleckeninhalt als eine Anhäufung locker beieinanderliegender -Teilchen des Glykogens.Präparate für die elektronenmikroskopische Untersuchung konnten hergestellt werden, nachdem der schweren Fixierbarkeit des Fleckeninhaltes mit der Verwendung des Glutaraldehyds in Kakodylatpuffer und der Spülung mit 50%igem Alkohol nach der Reynold'schen Kontrastierung Rechnung getragen war.Mit dieser Diagnose ist zugleich bewiesen, daß Glykogen im Phasenkontrastbild der lebenden Zelle sichtbar werden kann.

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The asphalt coloured spots of living heart muscle cells — glycogen formations visible under phase contrast

Summary The contents of three asphalt coloured spots previously examined in the living heart muscle cells in a culture by a phase contrast microscope and subsequently identified in the electron microscope by the method of Gross and Riedel proved to be accumulations of -particles of glycogen loosely lying together.Suitable sections could be manufactured for electron microscope after complying with the difficulty of the spot contents to be fixated — revealed during the histochemical investigations. One had to use glutaraldehyde in cacodylate buffer as the first fixative and 50% alcohol instead of distilled water for rinsing after the Reynold's staining.By this diagnosis at the same time, it is proven that certain glycogen formations in living cells are visible under phase contrast.

Mit Unterstützung durch die Deutsche Forschungsgemeinschaft.