“Action potentials” in NEUROSPORA CRASSA, a mycelial fungus

Occasional spontaneous “action potentiais” are found in mature hyphae of the fungus Neurospora crassa. They can arise either from low-level sinusoidal oscillations of the membrane potential or from a linear slow depolarization which accelerates into a rapid upstroke at a voltage 5–20 mV depolarized from the normal resting potential (near − 180 mV). The “action potentiais” are long-lasting, 1–2 min and at the peak reach a membrane potential near −40 mV. A 2− to 8−fold increase of membrane conductance accompanies the main depolarization, but a slight decrease of membrane conductance occurs during the slow depolarization. Two plausible mechanisms for the phenomenon are (a) periodic increases of membrane permeability to inorganic ions, particularly H⁺ or Cl^- and (b) periodic decreases in activity of the major electrogenic pump (H⁺) of the Neurospora membrane, coupled with a nonlinear (inverse sigmoid) current-voltage relationship.Identification of action potential-like disturbances in fungi means that such behavior has now been found in all major biologic taxa which have been probed with suitable electrodes. As yet there is no obvious function for the events in fungi.     

A stable, “near-haploid” mammalian cell line (Dipodomys ordii)

An established SV40-transformed cell line of Dipodomys ordii was cloned for selective loss of chromosomal material. A clone is described which has a modal chromosome number of 50 (in the normal diploid 2n=72), and has about 66% of the DNA content of normal diploid cells. Karyotype analysis shows that, although some chromosome rearrangement has taken place, 23 chromosomes are completely unpaired and 7 chromosomes are partially unpaired. Buoyant density analysis in neutral CsCl and Cs₂SO₄+Ag gradients of the DNA of this clone shows that there has been no selective retention or loss of any of the satellite DNA components present in D. ordii during the elimination of 34% of the genetic material.     

Sphenophyllum miravallis Vetter and Bowmanites cupulatus sp.n. from the “Illinger Flözzone” (“Heusweiler Schichten”, Lower Stephanian, Saar Basin, German Federal Republic)

This paper deals with a detailed study of Sphenophyllum miravallis Vetter, a member of the “Sphenophyllum thonii group”. New material from the Reisbach colliery, working the “Illinger Flözzone” of the “Heusweiler Schichten” (Lower Stephanian, Saar Basin, German Federal Republic), is described morphologically and anatomically, and the species is discussed. The new material enlarges the known range of variability of the normal aspect of the foliage, i.e. the foliage of the thinner branches. Thicker stems with their aberrant polymorphous foliage, and cellular details, are described for the first time. An emended diagnosis is given. Comparisons with other species are made.

The new species Bowmanites cupulatus is introduced to accommodate fructufications most probably belonging to Sphenophyllum miravallis.

S. crenulatum Knight ex Wagner is considered to be a heterotypic synonym of S. miravallis, the latter name having priority.     

The Exceptional “Blind” Gut of Appendicularia sicula (Appendicularia, Tunicata)

In this work, we studied for the first time the histology and ultrastructure of the gut of Appendicularia sicula and demonstrated the absence of any trace of anus. Appendicularians are small holoplanktonic tunicates, characterised by very fast ingestion and quick food transfer along their gut. The high production of faecal pellets released in the aqueous environment, associated with a high filtration rate, highlights their important role in marine ecosystems. Due to the absence of an anus, in contrast with other appendicularians, A. sicula, one of the smallest species, accumulates undigested faecal material within its body, with consequent extreme enlargement of its rectum. The gut, the epithelium of which is generally extremely reduced, is formed of an oesophagus, a globular stomach, thin proximal and mid-intestine, and a huge rectum. The latter, when filled with faecal material, may occupy most of the volume of the trunk in fully grown specimens. Although profoundly altered, the gut of these animals does show several similarities to that of Fritillaria (a genus of the same family, Fritillariidae), with which it has in common many features such as specialised mitochondrial pump cells. In A. sicula, the structural simplifications of organs seem to reach their extreme condition in comparison to other appendicularians.     

The “erythromycin-resistance” methylated sequence of Staphylococcus aureus ribosomal RNA

We have determined the sequence of an oligonucleotide from the large ribosomal subunit RNA of Staphylococcus aureus whose methylation renders the organism resistant to erythromycin and other antibiotics (the “MLS” phenotype). Analysis of RNase A digests of [³H]methyl-, ³²P-labeled RNA yielded the sequence GG · m₂⁶A · AAGACp, where m₂⁶A is an N⁶-dimethylated adenosine residue that in sensitive cells is unmethylated. Comparison with homologous sequences recently reported for Saccharomyces cerevesiae mitochondria indicates that an A to G mutation in this latter system mimics dimethylation in St. aureus with regard to functional consequences.     

Physiological and ultrastructural changes in “green islands” on Avena sterilis leaves caused by (8R,16R)-(–)-pyrenophorin

The biochemical and ultrastructural changes in "green islands" (GIs) on detached Avena sterilis leaves caused by the macrodiolide (8R,16R)-(-)-pyrenophorin in the dark were examined. In the absence of light, leaf segments retained their photosynthetic pigments for 96 h after treatment with (8R,16R)-(-)-pyrenophorin (70 muM), whereas in the untreated leaves complete senescence, loss of photosynthetic pigments and cell disorganization were observed 72 h after detachment. Proteolytic enzyme activity in treated tissues with pyrenophorin remained at low levels for 96 h after treatment and protein dissipation was lower in the treated than in the untreated. Although tissues in "GIs" seem macroscopically healthy, electron microscopy observations revealed structurally disorganized cells filled with granular, electron-dense material. Chloroplasts were severely damaged and contained a large number of plastoglobuli. Similar ultrastructural changes were also observed in A. sterilis tissues treated with the phytotoxin under illumination, indicating a mechanism operating both under illumination and in the dark.     

A moss “canopy” – Small-scale differences in microclimate and physiological traits in Tortula ruralis

We studied vertical changes in light regime and water content (WC) in combination with vertical gradients in several physiological response variables, i.e. net CO₂ uptake, chlorophyll content and nitrogen content in the moss species, Tortula ruralis. The rate of light attenuation within the moss turf, which was determined with a custom-made optical microprobe system, was strongly dependent on plant WC. Curling movements of the upper leaves associated with the beginning desiccation of the uppermost parts of a turf allowed increased penetration of light to greater depth with decreasing WC. The capacity to fix carbon declined steeply with depth: below ca. 9 mm no net CO₂ uptake occurred, even when removing the shading parts above. The potential rates of photosynthesis in different depths were highly correlated with chlorophyll content, but not nitrogen content.     

“Candidatus Paraholospora nucleivisitans”, an intracellular bacterium in Paramecium sexaurelia shuttles between the cytoplasm and the nucleus of its host

An intracellular bacterium was discovered in two isolates of Paramecium sexaurelia from an aquarium with tropical fish in Münster (Germany) and from a pond in the Wilhelma zoological–botanical garden, Stuttgart (Germany). The bacteria were regularly observed in the cytoplasm of the host, but on some occasions they were found in the macronucleus of the host cell. In these cases, only a few, if any, bacteria were observed remaining in the cytoplasm. The bacterium was not infectious to P. sexaurelia or other species of Paramecium and appeared to be an obligate intracellular bacterium, while bacteria-free host cells were completely viable. The fluorescence in situ hybridisation (FISH) and comparative 16SrDNA sequence analyses showed that the bacterium belonged to a new genus, and was most closely, yet quite distantly, related to Holospora obtusa. In spite of this relationship, the new bacteria differed from Holospora by at least two biological features. Whereas all Holospora species reside exclusively in the nuclei of various species of Paramecium and show a life cycle with a morphologically distinct infectious form, for the new bacterium no infectious form and no life cycle have been observed. For the new bacterium, the name Candidatus Paraholospora nucleivisitans is suggested. The host P. sexaurelia is usually known from tropical and subtropical areas and is not a species typically found in Germany and central Europe. Possibly, it had been taken to Germany with fish or plants from tropical or subtropical waters. Candidatus Paraholospora nucleivisitans may therefore be regarded as an intracellular neobacterium for Germany.     

Antioxidant activities of Ptychopetalum olacoides (“muirapuama”) in mice brain

Ptychopetalum olacoides (PO) roots are used by Amazonian peoples to prepare traditional remedies for treating various central nervous system conditions in which free radicals are likely to be implicated. Following the identification of PO ethanol extract (POEE) free-radical scavenging properties in vitro, the aim of this study was to verify the in vivo antioxidant effect of POEE. Aging mice (14 months) were treated (i.p.) with saline, DMSO (20%) or POEE (100mg/kg body wt.), and the hippocampi, cerebral cortex, striata, hypothalamus and cerebellum dissected out 60 min later to measure antioxidant enzyme activities, free-radical production and damage to macromolecules. POEE administration reduced free-radical production in the hypothalamus, lead to significant decrease in lipid peroxidation in the cerebral cortex, striatum and hypothalamus, as well as in the carbonyl content in cerebellum and striatum. In terms of antioxidant enzymes, catalase activity was increased in the cortex, striatum, cerebellum and hippocampus, while glutathione peroxidase activity was increased in the hippocampus. This study suggests that POEE contains compounds able to improve the cellular antioxidant network efficacy in the brain, ultimately reducing the damage caused by oxidative stress.     

Use of Pichia pastoris for the Expression, Purification, and Characterization of Rat Calretinin “EF-Hand” Domains

Calretinin (CR) is a calcium-binding, neuronal protein of undefined function. Related proteins either buffer intracellular calcium concentrations or are involved in calcium-signaling pathways. We transformed three CR gene fragment sequences, corresponding to its three complementary domains (I–II, III–IV, and V–VI), into Pichia pastoris. High yields of extracellular expression, of more than 200 mg/liter, were achieved. Simple purification protocols provide high yields of homogenous proteins: dialysis and DEAE–cellulose chromatography for domains I–II and III–IV or ammonium sulfate precipitation and octyl–Sepharose chromatography for domain V–VI. To our knowledge, this is the first report of the expression of an EF-hand protein using P. pastoris. Direct comparison of the purified yields of domain I–II indicates a 20-fold improvement over Escherichia coli. N-terminal amino acid sequencing confirmed our gene products and two anti-calretinin antibodies recognized the appropriate domains. All three CR domains bind ⁴⁵Ca and the domain containing EF-hands V and VI seems to have a lower calcium capacity than the other domains. Circular dichroism indicates a high helix content for each of the domains. Calcium-induced structural changes in the first two domains, followed by tryptophan fluorescence, correspond with previous studies, while tyrosine emission fluorescence indicates calcium-induced structural changes also occur in domain V–VI. The methods and expression levels achieved are suitable for future NMR labeling of the proteins, with ¹⁵N and ¹³C, and structure–function studies that will help to further understand CR function.     

Heterogeneous Single Atom Electrocatalysis,Where “Singles” Are “Married”

There is an intensive search for heterogeneous single atom catalysts (SACs) of high activity, efficiency, durability, and selectivity for a wide variety of electrocatalytic conversion and chemical reactions, such as the hydrogen evolution reaction (HER), oxygen evolution/reduction reaction (OER and ORR), CO₂ reduction reaction (CO₂ RR), and nitrogen reduction reaction (NRR). With the downsizing from nanoparticles and clusters to single atoms, there are steady changes in the bond and coordination environment for each and every atom involved. Indeed, the single atoms in these electrocatalysts are not “singles”; they are “married” to the supporting surfaces, and their performance is controlled by the bonding and coordination with the substrate surfaces. Herein, an overview is presented on the brief history leading to the rapid development of SACs and their current status, by focusing on their synthesis, control of composition, strategies to realize single atoms with the desired bonds and coordination, and targeted performance in selected reactions. Their applications in the selected spectrum of energy conversion and chemical reactions are discussed, in relation to their structures at varying length scales down to the atomic level. A particular emphasis is placed on on‐going research activities, together with the future perspectives and particular challenges for SACs.     

Novel antiplatelet and antithrombotic activities of essential oil from Lavandula hybrida Reverchon “grosso”

Lavender extracts are known to produce several mild effects at central and peripheral level. However, no studies are so far available about the potential effects of lavender essential oil on the hemostatic system. In this work, we demonstrated antiplatelet properties of lavender oil towards platelet aggregation induced by arachidonic acid, U46619, collagen and ADP (IC₅₀=51, 84, 191 and 640 μg/ml, respectively) on guinea-pig platelet rich plasma (PRP) and its ability to destabilize clot retraction (IC₅₀=149 μg/ml) induced by thrombin on rat PRP.

Furthermore, antithrombotic properties were studied in an in vivo model of pulmonary thromboembolism induced by intravenous injection of a collagen–epinephrine mixture in mice subacutely treated with lavender oil. In this model, lavender oil (100 mg/kg/day os for 5 days) significantly reduced thrombotic events without inducing prohemorrhagic complications at variance with acetylsalicylic acid used as reference drug. Finally, main components of the oil were studied in vitro in order to assess their antiplatelet effects, but none of them possessed an activity comparable to the oil itself. These results provide the first experimental evidence of lavender oil's antiplatelet/antithrombotic properties which could be due to a synergistic effect of its components.     

Reinvestigations of Three “Well-known” Marine Scuticociliates: Uronemella filificum (Kahl, 1931) nov. gen., nov. comb., Pseud cohnilembus hargisi Evans & Thompson, 1964 and Cyclidium citrullus Cohn 1865, with Description of the New Genus Uronemella (Protozoa, Ciliophora, Scuticociliatida)

The living morphology, infraciliature and silverline system of 3 “well-known” marine scuticociliates, Uronemella filificum (Kahl, 1931) nov. gen., nov. comb. (formerly Uronema filificum Kahl, 1931), Pseudocohnilembus hargisi Evans & Thompson, 1964 and Cyclidium citrullus Cohn 1865 are reinvestigated and redescribed.

Based on the data obtained, we suggest an establishment of a new genus Uronemella. The diagnosis for the new taxon: thigmotactic Uronematidae with generally pear-shaped body and subequatorially positioned cytostome; apical plate dominant; oral apparatus Uronema-like, one-rowed membranelle 1 about as long as membranelle 2; paroral membrane extending anteriorly to about mid-level of membrane 2; one caudal cilium; in vivo exhibiting typical rotatory movement with help of a caudal-cilium-associated sticky thread; generally in marine habitats. According to this definition, three nominal species as new combinations are transferred into this genus: Uronemella binucleata (Song, 1993) nov. comb. (= Homalogastra binucleata Song, 1993), Uronemella filificum (Kahl, 1931) nov. comb. (= Uronema filificum Kahl, 1931) and Uronemella cymruensis (Pérez-Uz & Hope, 1997) nov. comb. (=Urocyclon cymruensis Pérez-Uz & Hope, 1997). For comparison with Uronemella, some other closely-related taxa are also briefly outlined in the present paper.     

Tri-locus sequence data reject a “Gondwanan origin hypothesis” for the African/South Pacific crab genus Hymenosoma

Crabs of the family Hymenosomatidae are common in coastal and shelf regions throughout much of the southern hemisphere. One of the genera in the family, Hymenosoma, is represented in Africa and the South Pacific (Australia and New Zealand). This distribution can be explained either by vicariance (presence of the genus on the Gondwanan supercontinent and divergence following its break-up) or more recent transoceanic dispersal from one region to the other. We tested these hypotheses by reconstructing phylogenetic relationships among the seven presently-accepted species in the genus, as well as examining their placement among other hymenosomatid crabs, using sequence data from two nuclear markers (Adenine Nucleotide Transporter [ANT] exon 2 and 18S rDNA) and three mitochondrial markers (COI, 12S and 16S rDNA). The five southern African representatives of the genus were recovered as a monophyletic lineage, and another southern African species, Neorhynchoplax bovis, was identified as their sister taxon. The two species of Hymenosoma from the South Pacific neither clustered with their African congeners, nor with each other, and should therefore both be placed into different genera. Molecular dating supports a post-Gondwanan origin of the Hymenosomatidae. While long-distance dispersal cannot be ruled out to explain the presence of the family Hymenosomatidae on the former Gondwanan land-masses and beyond, the evolutionary history of the African species of Hymenosoma indicates that a third means of speciation may be important in this group: gradual along-coast dispersal from tropical towards temperate regions, with range expansions into formerly inhospitable habitat during warm climatic phases, followed by adaptation and speciation during subsequent cooler phases.     

The seeds of Loasaceae subfam. Loasoideae (Cornales) II: Seed morphology of “South Andean Loasas” (Loasa, Caiophora, Scyphanthus and Blumenbachia)

South Andean Loasas (Blumenbachia, Caiophora, Loasa, Scyphanthus) are a monophyletic group of taxa within Loasaceae subfam. Loasoideae, comprising some 100 species, 49 of which are investigated here. They retain a many-layered testa in the mature seeds and usually have passive transfer testas with complex, spongiose wall outgrowths. Additional modifications concern the undulations of the testa epidermis, presence or absence of the outer periclinal wall, secondary sculpturing, the presence or absence of spines, warts and finally spongiose structures on the anticlinal walls of the testa epidermis and the inner periclinal wall. Seeds of the widespread “deeply pitted” type are plesiomorphic, while various subclades of South Andean Loasas have derivations underscoring their relationships and confirming the relationships found with molecular markers and other morphological characters. The genus Blumenbachia has either seeds with a many-layered testa forming longitudinal lamellae (sect. Angulatae), or balloon seeds with a loose outer testa layer and spongiose wall outgrowths on the inner periclinal walls (sect. Blumenbachia and sect. Gripidea) and is clearly monophyletic. Loasa s.str. (ser. Loasa, ser. Macrospermae, ser. Floribundae, ser. Deserticolae) is characterized by the presence of a subterminal hilum or hilar scar and one subgroup (ser. Loasa, ser. Macrospermae) by very large and heavy seeds with a collapsed testa. L. ser. Pinnatae, ser. Acaules, ser. Volubiles, Scyphanthus and Caiophora share more or less one seed types with minor modifications. Within Caiophora various derivations are observed, of which the gradual loss of the secondary sculpture of the inner periclinal wall is the most striking one. Anemochoria is the most widespread dispersal mechanism in South Andean Loasas and is achieved in at least five structurally different ways.