Sawadalepis prima n. g., n. sp. (Cestoda: Cyclophyllidea) from the Schreiber’s bent-winged bat Miniopterus schreibersii Kuhl (Chiroptera: Vespertilionidae) from China

Sawadalepis n. g. is erected for Sawadalepis prima n. sp. in Schreiber’s bent-winged bat Miniopterus schreibersii Kuhl (Chiroptera: Vespertilionidae) from southern China. Diagnostic features of the currently monotypic genus include attributes of the osmoregulatory system and female genital organs. The dorsal osmoregulatory canals are shifted bilaterally towards the margins of proglottides in relation to the ventral canals. The genital pores are unilateral and sinistral. Among female attributes, the copulatory part of the vagina is covered externally by a dense layer of intensely stained cells; the conductive part of the vagina is clearly distinguishable from the seminal receptacle; the uterus has ventral and dorsal diverticula, extending bilaterally beyond the longitudinal osmoregulatory canals; and the eggs are spherical with thick outer coat.     

Peptidoglycan fragments stimulate resuscitation of “non-culturable” mycobacteria

Resuscitation promoting factors (Rpfs), belonging to a family of secreted actinobacterial proteins with predicted peptidoglycan (PG) hydrolytic activities, participate in the reactivation of dormant cells. In the present study we demonstrate that a recombinant truncated form of Micrococcus luteus Rpf hydrolyzes isolated PG of Mycobacterium smegmatis and Mycobacterium tuberculosis liberating PG fragments of different size. These fragments possess stimulatory activity toward “non-culturable” dormant M. smegmatis and M. tuberculosis cells, similar to the activity of recombinant Rpf. Relatively large PG fragments (0.1–0.5 μm) obtained either by Rpf digestion or by PG ultrasonication revealed resuscitation activities when added in concentrations 0.1–0.2 μg/ml to the resuscitation medium. It is suggested that PG fragments could either directly activate the resuscitation pathway of dormant mycobacteria or serve as a substrate for endogenous Rpf, resulting in low molecular weight products with resuscitation activity. Whilst both suggestions are plausible, it was observed that PG-dependent resuscitation activity was suppressed by means of a specific Rpf inhibitor (4-benzoyl-2-nitrophenylthiocyanate), which provides additional support for the second of these possibilities.     

Mechanisms controling the timing of spring migration in birds

The question “Which factors govern the timing of migration in birds?” has fascinated researchers for a long time. It was initially assumed that avian migration is triggered by environmental factors, such as ambient temperature and food availability. Later laboratory experiments in various avian species convincingly showed that timing of spring migration is mainly governed by daylength (photoperiod) and is controlled by circannual rhythms. As a result, the concept that environmental factors (air temperature, precipitation, food availability) have no significant impact on timing of spring migration generally took hold. However, in recent decades more and more data has become available showing that the timing of spring migration in many bird species has significantly changed. These data allow the formulation of a novel concept of regulation mechanisms of timing of spring migration which accounts not only for photoperiodic and endogenous control, but also for the already mentioned extrinsic factors. Studies of endocrine control of spring migratory disposition showed that features of endocrine mechanisms governing the onset of spring migration depend on speciesspecific migratory strategies and the stability of environmental conditions in winter quarters and on migratory routes. It is becoming clear precisely which endocrine mechanisms are involved in adjusting migratory behaviour to variation of the local environment. In recent years, progress has also been made in finding genetic mechanisms controlling the timing of spring migration.     

Presenilins form ER Ca2+ leak channels, a function disrupted by familial Alzheimer's disease-linked mutations

Alzheimer's disease (AD) is a progressive and irreversible neurodegenerative disorder. Mutations in presenilins 1 and 2 (PS1 and PS2) account for approximately 40% of familial AD (FAD) cases. FAD mutations and genetic deletions of presenilins have been associated with calcium (Ca(2+)) signaling abnormalities. We demonstrate that wild-type presenilins, but not PS1-M146V and PS2-N141I FAD mutants, can form low-conductance divalent-cation-permeable ion channels in planar lipid bilayers. In experiments with PS1/2 double knockout (DKO) mouse embryonic fibroblasts (MEFs), we find that presenilins account for approximately 80% of passive Ca(2+) leak from the endoplasmic reticulum. Deficient Ca(2+) signaling in DKO MEFs can be rescued by expression of wild-type PS1 or PS2 but not by expression of PS1-M146V or PS2-N141I mutants. The ER Ca(2+) leak function of presenilins is independent of their gamma-secretase activity. Our data suggest a Ca(2+) signaling function for presenilins and provide support for the "Ca(2+) hypothesis of AD."     

GABAergic transmission and chloride equilibrium potential are not modulated by pyruvate in the developing optic tectum of Xenopus laevis tadpoles

In the developing mammalian brain, gamma-aminobutyric acid (GABA) is thought to play an excitatory rather than an inhibitory role due to high levels of intracellular Cl(-) in immature neurons. This idea, however, has been questioned by recent studies which suggest that glucose-based artificial cerebrospinal fluid (ACSF) may be inadequate for experiments on immature and developing brains. These studies suggest that immature neurons may require alternative energy sources, such as lactate or pyruvate. Lack of these other energy sources is thought to result in artificially high intracellular Cl(-) concentrations, and therefore a more depolarized GABA receptor (GABAR) reversal potential. Since glucose metabolism can vary widely among different species, it is important to test the effects of these alternative energy sources on different experimental preparations. We tested whether pyruvate affects GABAergic transmission in isolated brains of developing wild type Xenopus tadpoles in vitro by recording the responsiveness of tectal neurons to optic nerve stimulation, and by measuring currents evoked by local GABA application in a gramicidin perforated patch configuration. We found that, in contrast with previously reported results, the reversal potential for GABAR-mediated currents does not change significantly between developmental stages 45 and 49. Partial substitution of glucose by pyruvate had only minor effects on both the GABA reversal potential, and the responsiveness of tectal neurons at stages 45 and 49. Total depletion of energy sources from the ACSF did not affect neural responsiveness. We also report a strong spatial gradient in GABA reversal potential, with immature cells adjacent to the lateral and caudal proliferative zones having more positive reversal potentials. We conclude that in this experimental preparation standard glucose-based ACSF is an appropriate extracellular media for in vitro experiments.     

Finding of the Low Molecular Weight Inhibitors of Resuscitation Promoting Factor Enzymatic and Resuscitation Activity

Background

Resuscitation promoting factors (RPF) are secreted proteins involved in reactivation of dormant actinobacteria, including Mycobacterium tuberculosis. They have been considered as prospective targets for the development of new anti-tuberculosis drugs preventing reactivation of dormant tubercle bacilli, generally associated with latent tuberculosis. However, no inhibitors of Rpf activity have been reported so far. The goal of this study was to find low molecular weight compounds inhibiting the enzymatic and biological activities of Rpfs.

Methodology/Principal Findings

Here we describe a novel class of 2-nitrophenylthiocyanates (NPT) compounds that inhibit muralytic activity of Rpfs with IC₅₀ 1–7 µg/ml. Fluorescence studies revealed interaction of active NPTs with the internal regions of the Rpf molecule. Candidate inhibitors of Rpf enzymatic activity showed a bacteriostatic effect on growth of Micrococcus luteus (in which Rpf is essential for growth protein) at concentrations close to IC₅₀. The candidate compounds suppressed resuscitation of dormant (“non-culturable”) cells of M. smegmatis at 1 µg/ml or delayed resuscitation of dormant M. tuberculosis obtained in laboratory conditions at 10 µg/ml. However, they did not inhibit growth of active mycobacteria under these concentrations.

Conclusions/Significance

NPT are the first example of low molecular weight compounds that inhibit the enzymatic and biological activities of Rpf proteins.     

Plasma corticosterone concentrations in European robins during spring and autumn migration

To estimate differences in hormonal mechanisms of regulation of spring and autumn migration in European robins Erithacus rubecula, the plasma corticosterone (CORT) concentrations were compared in birds caught during both migratory seasons. A total of 414 blood samples were analyzed. It was found that the baseline and stress-induced CORT concentrations in free-living robins during spring migration were practically twice as high as during autumn passage. Our results demonstrate that autumn and spring migrations are independent stages of the avian annual cycle. Probably, the increase in the CORT concentrations in spring can be considered to be physiological preparation for the breeding season.     

Phylogenetic relationships of the genus <Emphasis Type="Italic">Armadolepis</Emphasis> Spassky, 1954 (Eucestoda,Hymenolepididae), with descriptions of two new species from Palaearctic dormice (Rodentia,Gliridae)

Two new species of hymenolepidid cestodes belonging to the genus Armadolepis Spassky, 1954 are described from dormice (Gliridae) from the southern East European Plain and the northwestern Caucasus, Russia. Armadolepis (Bremserilepis) longisoma n. sp., with a rudimentary, unarmed rostellar apparatus is described from the fat dormouse Glis glis (Linnaeus) from the Republic of Adygeya, Russia. Additionally, A. (Armadolepis) dryomi n. sp., characterised by a well-developed rostellar apparatus and armed rhynchus is described from the forest dormouse Dryomys nitedula Pallas from Rostov Oblast’, Russia. Armadolepis (Bremserilepis) longisoma n. sp. differs from A. (Bremserilepis) myoxi (Rudolphi, 1819) in having a substantially longer strobila and cirrus-sac, wider scolex and ovary and larger rostellar pouch and testes. Armadolepis (Armadolepis) dryomi n. sp. is distinguishable from A. (Armadolepis) spasskii Tenora & Baru?, 1958, A. (Armadolepis) jeanbaeri Makarikov, 2017 and A. (Armadolepis) tenorai Makarikov, 2017 in having a substantially longer and wider strobila, and larger rostellar pouch and cirrus-sac. Furthermore, A. dryomi n. sp. can be distinguished from its congeners by the number and size of rostellar hooks and the arrangement of the testes. Phylogenetic affinities of Armadolepis were studied for the first time using partial sequences of the nuclear ribosomal 28S DNA gene. Phylogenetic analysis strongly supported the status of Armadolepis as a separate genus belonging to the “Rodentolepis clade”.     

Timing of nocturnal autumn migratory departures in juvenile European robins (Erithacus rubecula) and endogenous and external factors

Many passerine medium distance nocturnal migrants take off from stopover sites not only at the beginning of the night, but also in the middle and at the end of the night. In this paper, we tested two explanations for this phenomenon: (1) that departure time is governed by fuel stores, and (2) that departure time is influenced by the weather. The relationship of temporal distribution of migratory nocturnal departures with body condition and weather factors was studied in juvenile European robins (Erithacus rubecula) during autumn migration. The study was done on the Courish Spit on the Baltic Sea in 1997–2003 by retrapping 74 ringed birds in high mist nets during nocturnal migratory departure. Departure time was not related to fuel stores at arrival and departure, stopover duration, fuel deposition rate or progress of the season. Nor did the local weather at departure influence departure time. A possible reason was a large variation in the behaviour of the birds. European robins which made 1-day stopovers arrived and departed during better weather conditions than birds that stopped over for longer periods. In the former cohort, a significant model with four predictors explained 55% of variation in departure time. It is assumed that weather at the night of departure and during the previous night influenced the time of take-offs in these birds. In robins which made long stopovers, departure time is probably governed by their individual endogenous circadian rhythms of activity, which are related to the environment in a complex way.     

Muralytic activity of Micrococcus luteus Rpf and its relationship to physiological activity in promoting bacterial growth and resuscitation

The culturability of several actinobacteria is controlled by resuscitation-promoting factors (Rpfs). These are proteins containing a c. 70-residue domain that adopts a lysozyme-like fold. The invariant catalytic glutamate residue found in lysozyme and various bacterial lytic transglycosylases is also conserved in the Rpf proteins. Rpf from Micrococcus luteus, the founder member of this protein family, is indeed a muralytic enzyme, as revealed by its activity in zymograms containing M. luteus cell walls and its ability to (i) cause lysis of Escherichia coli when expressed and secreted into the periplasm; (ii) release fluorescent material from fluorescamine-labelled cell walls of M. luteus; and (iii) hydrolyse the artificial lysozyme substrate, 4-methylumbelliferyl-beta-D-N,N',N'-triacetylchitotrioside. Rpf activity was reduced but not completely abolished when the invariant glutamate residue was altered. Moreover, none of the other acidic residues in the Rpf domain was absolutely required for muralytic activity. Replacement of one or both of the cysteine residues that probably form a disulphide bridge within Rpf impaired but did not completely abolish muralytic activity. The muralytic activities of the Rpf mutants were correlated with their abilities to stimulate bacterial culturability and resuscitation, consistent with the view that the biological activity of Rpf results directly or indirectly from its ability to cleave bonds in bacterial peptidoglycan.