Synthesis of Indazoles via N-N Bond-Forming Oxidative Cyclization from 2-Aminomethyl-phenylamines.

Herein we report a method for the synthesis of indazoles from readily available 2-aminomethyl-phenylamines via N-N bond-forming oxidative cyclization. Inspired by indazole formation initially observed as a side product by N. Coskun et al. we developed a robust protocol to access indazoles in all three tautomeric forms. The method selectively gives access to various 2-substituted 2H-indazoles which are frequently used in drug design, and we also demonstrated its applicability to less studied 3H-indazoles.

Two hypotrichs (Ciliophora, Hypotricha) from China: morphology and SSU rDNA sequence of Holosticha aestuarina nov. spec. and H. muuiensis Kim et al., 2017.

The morphology, four ontogenetic stages, and the phylogenetic relationships based on the small subunit RNA-gene of Holosticha aestuarina nov. spec. from the Yangtze River Estuary (Shanghai) were investigated. The new species differs from the congeners by the following combination of features: body size 160-196 × 27-36 µm in vivo, a vesicular structure containing one or two bean-shaped opaque particles, 24-32 adoral membranelles, 16-34 midventral pairs, 17-26 left marginal cirri, 22-32 right marginal cirri, six or seven transverse cirri, four dorsal kineties, and usually two macronuclear nodules. The anlage for the left marginal row of the proter very likely originates de novo (anterior portion) and from parental cirri (posterior portion). Further, we provide a morphological description of a Chinese population of H. muuiensis Kim et al., 2017, originally discovered in South Korea, indicating that it is confined to the Eastern Asia region. Its SSU rDNA sequence is identical with that of the type population and forms a clade with the available Holosticha sequences. By contrast, H. aestuarina nov. spec. is sister to the Holosticha + Uncinata clade, indicating that Uncinata is a subgenus of Holosticha.

Fragment-based discovery of a chemical probe for the PWWP1 domain of NSD3.

Here, we report the fragment-based discovery of BI-9321, a potent, selective and cellular active antagonist of the NSD3-PWWP1 domain. The human NSD3 protein is encoded by the WHSC1L1 gene located in the 8p11-p12 amplicon, frequently amplified in breast and squamous lung cancer. Recently, it was demonstrated that the PWWP1 domain of NSD3 is required for the viability of acute myeloid leukemia cells. To further elucidate the relevance of NSD3 in cancer biology, we developed a chemical probe, BI-9321, targeting the methyl-lysine binding site of the PWWP1 domain with sub-micromolar in vitro activity and cellular target engagement at 1 µM. As a single agent, BI-9321 downregulates Myc messenger RNA expression and reduces proliferation in MOLM-13 cells. This first-in-class chemical probe BI-9321, together with the negative control BI-9466, will greatly facilitate the elucidation of the underexplored biological function of PWWP domains.

Monographic treatment of Paraholosticha muscicola (Ciliophora, Keronopsidae), including morphological and molecular biological characterization of a brackish water population from Korea.

Paraholosticha muscicola, type species of Paraholosticha Wenzel, inhabits mainly terrestrial habitats, but also freshwater. A brackish water population from Korea is described, the first record from such a habitat. Principal component analysis shows that this population is more similar to a terrestrial population from Denmark than to a population from Antarctic soil. Keronopsids have two strong morphological/ontogenetic apomorphies (frontal corona formed from anlagen I-III; division in cysts). However, the SSU rRNA sequence of the Korean population does not cluster with that of the Antarctic population in the phylogenetic tree, but both branch off consecutively and immediately before a mixture of other non-dorsomarginalian hypotrichs, including two further keronopsids. Furthermore, the keronopsids cluster in the phylogenetic network, providing phylogenetic conflicts, which cannot be exemplified in the conventional gene tree. To complete the picture of P. muscicola, we provide a detailed overview about nomenclature, history, taxonomy, and its geographic distribution. From the four synonyms proposed so far, we tentatively accept only P. lichenicola and P. ovata. Paraholosticha algivora is likewise very similar. Thus we propose to include these three taxa as members of the P. muscicola complex. Stylonethes sterkii and P. algivora are transferred to Paraholosticha Wenzel. A key to the Paraholosticha species is provided.

Highly Selective PTK2 Proteolysis Targeting Chimeras to Probe Focal Adhesion Kinase Scaffolding Functions.

Focal adhesion tyrosine kinase (PTK2) is often overexpressed in human hepatocellular carcinoma (HCC), and several reports have linked PTK2 depletion and/or pharmacological inhibition to reduced tumorigenicity. However, the clinical relevance of targeting PTK2 still remains to be proven. Here, we present two highly selective and functional PTK2 proteolysis-targeting chimeras utilizing von Hippel-Lindau and cereblon ligands to hijack E3 ligases for PTK2 degradation. BI-3663 (cereblon-based) degrades PTK2 with a median DC50 of 30 nM to >80% across a panel of 11 HCC cell lines. Despite effective PTK2 degradation, these compounds did not phenocopy the reported antiproliferative effects of PTK2 depletion in any of the cell lines tested. By disclosing these compounds, we hope to provide valuable tools for the study of PTK2 degradation across different biological systems.

Morphology, cell-division, and phylogeny of Schmidingerothrix elongata spec. nov. (Ciliophora, Hypotricha), and brief guide to hypotrichs with Gonostomum-like oral apparatus.

The hypotrich Schmidingerothrix elongata spec. nov., discovered in saline (20‰) soil of the Longfeng Wetland, Daqing, northern China, was studied using live observation and protargol impregnation. It is characterized, inter alia, by colorless cortical granules arranged in short rows, three frontoventral cirral rows with the rightmost extending far posteriorly, and 4-8, usually six macronuclear nodules. Cell division proceeds as in congeners and confirms the lack of dorsal ciliature. In phylogenetic analyses based on SSU rDNA, S. elongata is sister of S. salinarum+Paracladotricha salina. A re-investigation of the type slides of P. salina, type of Paracladotricha, revealed a misobservation in the original description. Since P. salina lacks, like Schmidingerothrix spp., a dorsal ciliature, Paracladotricha becomes a junior, subjective synonym of Schmidingerothrix with S. salina comb. nov. as fourth species. A review of the phylogenetic analyses dealing with Schmidingerothrix shows that its position is variable. However, together with the gonostomatid oral apparatus it can be hypothesized that Schmidingerothrix is a member of the Gonostomatidae or a close relative. A list of genera (14) and species (58) which have - like Schmidingerothrix - a gonostomatid oral apparatus, as well as a key to these genera are provided.

Chemically Induced Degradation of the Oncogenic Transcription Factor BCL6.

The transcription factor BCL6 is a known driver of oncogenesis in lymphoid malignancies, including diffuse large B cell lymphoma (DLBCL). Disruption of its interaction with transcriptional repressors interferes with the oncogenic effects of BCL6. We used a structure-based drug design to develop highly potent compounds that block this interaction. A subset of these inhibitors also causes rapid ubiquitylation and degradation of BCL6 in cells. These compounds display significantly stronger induction of expression of BCL6-repressed genes and anti-proliferative effects than compounds that merely inhibit co-repressor interactions. This work establishes the BTB domain as a highly druggable structure, paving the way for the use of other members of this protein family as drug targets. The magnitude of effects elicited by this class of BCL6-degrading compounds exceeds that of our equipotent non-degrading inhibitors, suggesting opportunities for the development of BCL6-based lymphoma therapeutics.

Redescription of Keronopsis helluo Penard, 1922 from Antarctica and Paraholosticha pannonica Gellért and Tamás, 1959 from Alaska (Ciliophora, Hypotricha).

The morphology of Keronopsis helluo Penard, 1922, type species of Keronopsis, and Paraholosticha pannonica Gellért and Tamás, 1959, two little-known members of the Keronopsidae Jankowski, 1979, was described using standard methods. In addition, we sequenced the SSU rRNA of both species. Keronopsis helluo was isolated from a mossy soil from Robert Island (Antarctica) while P. pannonica was found in terrestrial moss from Alaska. Our data correspond very well with the original descriptions. The frontal ciliature of K. helluo is identical with that of Paraholosticha spp., indicating that some Keronopsis species (K. tasmaniensis, K. dieckmanni) are misclassified in the keronopsids. The type species has distinctly more transverse cirri (8-13) than K. wetzeli (1-3), type species of Parakeronopsis, which is thus perhaps a valid genus or subgenus. The phylogenetic analyses confirm the position of the keronopsids outside the Dorsomarginalia. The species sequenced so far (K. helluo, Paraholosticha muscicola, P. pannonica) emerge from a soft polytomy, which also comprises Bistichella-like species and a large cluster composed of amphisiellids, trachelostylids, and gonostomatids, that is, the method failed to resolve the relationships within the keronopsids. The Keronopsidae and the two species studied are characterized based on previous studies and our data.

Morphology, morphogenesis, and molecular phylogeny of Uroleptus (Caudiholosticha) stueberi (Foissner, 1987) comb. nov. (Ciliophora, Hypotricha), and reclassification of the remaining Caudiholosticha species.

The morphology, morphogenesis, and SSU rDNA sequence of a Caudiholosticha stueberi population from Southwest China soil were analyzed. The studies confirm the assumption of previous workers that this species has dorsomarginal kineties and thus does not belong to the urostyloids, but to the non-oxytrichid dorsomarginalian genus Uroleptus whose members have, in contrast to C. stueberi, a distinct tail. On the basis of two morphological features we split Uroleptus into three subgenera: U. (Uroleptus) (tail present; more than five transverse cirri; habitat freshwater), U. (Paruroleptus) (present; five or less; freshwater or soil), and U. (Caudiholosticha) stat. nov. (lacking; five or less; soil). Since Uroleptus (Caudiholosticha) stueberi comb. nov. is the type of Caudiholosticha, the other 16 species so far assigned to Caudiholosticha have to be reclassified because they obviously lack dorsomarginal kineties. Based on published data, six new urostyloid genera are established: Extraholosticha gen. nov. (type: Holosticha sylvatica; monotypic); Adumbratosticha gen. nov. (type: H. tetracirrata; three species); Acuholosticha gen. nov. (type: U. paranotabilis; five species); Limnoholosticha gen. nov. (type: H. (Holosticha) navicularum; four species); Multiholosticha gen. nov. (type: H. multicaudicirrus; two species); and Caudikeronopsis gen. nov. (type: Caudiholosticha marina; monotypic). Urosomoida sejongensis is transferred to Oxytrichella: O. sejongensis comb. nov.

Morphology, Cell Division, and Phylogeny of Uroleptus longicaudatus (Ciliophora, Hypotricha), a Species of the Uroleptus limnetis Complex.

A Chinese population of the little-known freshwater hypotrich Uroleptus longicaudatus was investigated with emphasis on its living morphology and infraciliature. The characteristic, tripartite body consists of a narrowed (cephalized) anterior portion, a slender trunk, and a long, slender, and strongly contractile tail occupying up to 30% of body length. Contracted specimens with a tail length of about 12% closely resemble Uroleptus limnetis which has, like U. longicaudatus, its type locality on the East Coast of the United States so that it cannot be excluded that these two species are synonymous. Thus, we propose to subsume these and few other little-known species, which are not clearly distinguishable at the present state of knowledge, as U. limnetis complex. The morphogenesis of U. longicaudatus proceeds as in most congeners. The phylogenetic analyses reveal that Uroleptus is a monophyletic group, but due to the lack of detailed morphological data of the populations sequenced so far, the relationships within this taxon remain obscure. For the objective determination of the tail length of hypotrichs, we propose the "1/3-method", which says that the tail commences at that body width which corresponds one-third of the maximum width. Paruroleptus ophryoglena Gelei, 1954 is transferred to Uroleptus: Uroleptus ophryoglena (Gelei, 1954) comb. nov.

Molecular phylogeny and ontogeny of a new ciliate genus, Paracladotricha salina n. g., n. sp. (Ciliophora, Hypotrichia).

A hypotrichous ciliate, Paracladotricha salina n. g., n. sp., was discovered in hypersaline waters (salinity about 80‰) from Qingdao, China. Its morphology and some major ontogenetic stages were studied and the phylogenetic position was estimated using standard methods. Paracladotricha salina is characterized by a flexible, more or less slender body (size 50-120 × 20-35 µm), a gonostomatid oral apparatus, one short and two long frontoventral rows, four macronuclear nodules, almost completely reduced dorsal kineties 1-3, and a loss of several parts of the ciliature, namely, the slightly shortened ciliary row of the adoral membranelles, the paroral, and the buccal, the postoral and pretransverse ventral, the transverse, and the caudal cirri. The ontogenesis is rather simple: anlage II of both filial products and anlage III of the opisthe originate de novo, while anlagen IV and V are formed within the parental rows. This combination of features requires the establishment of a new genus, Paracladotricha, which is, according to the morphological data, closely related to Schmidingerothrix and Cladotricha. The small-subunit rRNA gene was sequenced, indicating that P. salina is, as also demonstrated by the oral apparatus, a member of the gonostomatids. We provide a first, vague hypothesis about the phylogenetic relationships of the Gonostomatidae, Cladotrichidae, and Schmidingerotrichidae. However, since molecular data of the type species of these higher taxa are lacking, their validity and relationships remain obscure.

Typification of the genus Dileptus Dujardin, 1841 (Ciliophora, Rhynchostomatia).

In their monograph of the dileptids, Vdacný and Foissner (2012) could not clarify the type species of the genus DileptusDujardin, 1841. Thus, they suggested that the problem be referred to the International Commission on Zoological Nomenclature. However, recently we discovered that Dujardin (1841) has originally typified Dileptus with Amphileptus anser sensu Ehrenberg (1838) which is in fact a misidentified Amphileptus margaritiferEhrenberg, 1833, a common species also originally classified in Dileptus. Under Article 70.3.2 of the Code, Dileptus margaritifer (Ehrenberg, 1833) Dujardin, 1841, thoroughly redescribed by Foissner et al. (1995), is now the type of Dileptus. This has the great advantages of historical continuity and that new combinations (names) are not required.

Three-gene based phylogeny of the Urostyloidea (Protista, Ciliophora, Hypotricha), with notes on classification of some core taxa.

Classifications of the Urostyloidea were mainly based on morphology and morphogenesis. Since molecular phylogeny largely focused on limited sampling using mostly the one-gene information, the incongruence between morphological data and gene sequences have risen. In this work, the three-gene data (SSU-rDNA, ITS1-5.8S-ITS2 and LSU-rDNA) comprising 12 genera in the "core urostyloids" are sequenced, and the phylogenies based on these different markers are compared using maximum-likelihood and Bayesian algorithms and tested by unconstrained and constrained analyses. The molecular phylogeny supports the following conclusions: (1) the monophyly of the core group of Urostyloidea is well supported while the whole Urostyloidea is not monophyletic; (2) Thigmokeronopsis and Apokeronopsis are clearly separated from the pseudokeronopsids in analyses of all three gene markers, supporting their exclusion from the Pseudokeronopsidae and the inclusion in the Urostylidae; (3) Diaxonella and Apobakuella should be assigned to the Urostylidae; (4) Bergeriella, Monocoronella and Neourostylopsis flavicana share a most recent common ancestor; (5) all molecular trees support the transfer of Metaurostylopsis flavicana to the recently proposed genus Neourostylopsis; (6) all molecular phylogenies fail to separate the morphologically well-defined genera Uroleptopsis and Pseudokeronopsis; and (7) Arcuseries gen. nov. containing three distinctly deviating Anteholosticha species is established.

Systematics and species-specific response to pH of Oxytricha acidotolerans sp. nov. and Urosomoida sp. (Ciliophora, Hypotricha) from acid mining lakes.

We investigated the morphology, phylogeny of the 18S rDNA, and pH response of Oxytricha acidotolerans sp. nov. and Urosomoida sp. (Ciliophora, Hypotricha) isolated from two chemically similar acid mining lakes (pH~2.6) located at Langau, Austria, and in Lusatia, Germany. Oxytricha acidotolerans sp. nov. from Langau has 18 frontal-ventral-transverse cirri but a very indistinct kinety 3 fragmentation so that the assignment to Oxytricha is uncertain. The somewhat smaller species from Lusatia has a highly variable cirral pattern and the dorsal kineties arranged in the Urosomoida pattern and is, therefore, preliminary designated as Urosomoida sp. The pH response was measured as ciliate growth rates in laboratory experiments at pH ranging from 2.5 to 7.0. Our hypothesis was that the shape of the pH reaction norm would not differ between these closely related (3% difference in their SSU rDNA) species. Results revealed a broad pH niche for O. acidotolerans, with growth rates peaking at moderately acidic conditions (pH 5.2). Cyst formation was positively and linearly related to pH. Urosomoida sp. was more sensitive to pH and did not survive at circumneutral pH. Accordingly, we reject our hypothesis that similar habitats would harbour ciliate species with virtually identical pH reaction norm.

Angle-resolved photoemission spectroscopy at ultra-low temperatures.

The physical properties of a material are defined by its electronic structure. Electrons in solids are characterized by energy (ω) and momentum (k) and the probability to find them in a particular state with given ω and k is described by the spectral function A(k, ω). This function can be directly measured in an experiment based on the well-known photoelectric effect, for the explanation of which Albert Einstein received the Nobel Prize back in 1921. In the photoelectric effect the light shone on a surface ejects electrons from the material. According to Einstein, energy conservation allows one to determine the energy of an electron inside the sample, provided the energy of the light photon and kinetic energy of the outgoing photoelectron are known. Momentum conservation makes it also possible to estimate k relating it to the momentum of the photoelectron by measuring the angle at which the photoelectron left the surface. The modern version of this technique is called Angle-Resolved Photoemission Spectroscopy (ARPES) and exploits both conservation laws in order to determine the electronic structure, i.e. energy and momentum of electrons inside the solid. In order to resolve the details crucial for understanding the topical problems of condensed matter physics, three quantities need to be minimized: uncertainty* in photon energy, uncertainty in kinetic energy of photoelectrons and temperature of the sample. In our approach we combine three recent achievements in the field of synchrotron radiation, surface science and cryogenics. We use synchrotron radiation with tunable photon energy contributing an uncertainty of the order of 1 meV, an electron energy analyzer which detects the kinetic energies with a precision of the order of 1 meV and a He(3) cryostat which allows us to keep the temperature of the sample below 1 K. We discuss the exemplary results obtained on single crystals of Sr2RuO4 and some other materials. The electronic structure of this material can be determined with an unprecedented clarity.

Comparison of four specific dynamic office chairs with a conventional office chair: impact upon muscle activation, physical activity and posture.

Prolonged and static sitting postures provoke physical inactivity at VDU workplaces and are therefore discussed as risk factors for the musculoskeletal system. Manufacturers have designed specific dynamic office chairs featuring structural elements which promote dynamic sitting and therefore physical activity. The aim of the present study was to evaluate the effects of four specific dynamic chairs on erector spinae and trapezius EMG, postures/joint angles and physical activity intensity (PAI) compared to those of a conventional standard office chair. All chairs were fitted with sensors for measurement of the chair parameters (backrest inclination, forward and sideward seat pan inclination), and tested in the laboratory by 10 subjects performing 7 standardized office tasks and by another 12 subjects in the field during their normal office work. Muscle activation revealed no significant differences between the specific dynamic chairs and the reference chair. Analysis of postures/joint angles and PAI revealed only a few differences between the chairs, whereas the tasks performed strongly affected the measured muscle activation, postures and kinematics. The characteristic dynamic elements of each specific chair yielded significant differences in the measured chair parameters, but these characteristics did not appear to affect the sitting dynamics of the subjects performing their office tasks.

Office task effects on comfort and body dynamics in five dynamic office chairs.

In the present study, we investigated the effect of office tasks on posture and movements in field settings, and the comfort rating for chair characteristics and correlation with type of task. The tasks studied were: computer work, telephoning, desk work and conversation. Postures, movements, chair part inclinations and comfort rating data were collected from 12 subjects. Computer work showed the lowest physical activity, together with upright trunk and head position and low backrest inclination. Conversation shows the highest activity of head legs and low back together with the highest cervical spine extension. In contrast, desk work provoked the most cervical spine flexion and showed the second lowest activity. The telephoning tasks showed medium activity and the highest kyphosis. Conversation showed the highest backrest inclination. Positive comfort relations were found for computer work and a "swing system" chair, for telephoning and an active longitudinal seat rotation, and for desk work and a chair with a three-dimensionally moveable seat.

Morphology and small subunit (SSU) rRNA gene sequence of the new brackish water ciliate Neobakuella flava n. g., n. sp. (Ciliophora, Spirotricha, Bakuellidae) and SSU rRNA gene sequences of six additional hypotrichs from Korea.

The morphology and the small subunit (SSU) rRNA gene sequence of the hypotrich Neobakuella flava n. g., n. sp. from the estuary of the Taehwagang River (Ulsan, South Korea) were investigated. The three frontal cirri, the composition of the midventral complex of cirral pairs and rows, and the simple dorsal kinety pattern of three bipolar kineties assign it to the urostyloid taxon Bakuellidae. The increased number of buccal and parabuccal cirri, the presence of transverse cirri, and more than one left marginal row, as well as the lack of caudal cirri separate Neobakuella n. g. from the other bakuellids. Neobakuella flava n. sp. has many 0.3 µm sized green and/or yellow usually dark-green cortical granules and some sparsely distributed, 2 × 1 µm sized grass green with yellowish shimmer granules. The gene sequence data indicate a close relationship with Diaxonella and a distinct separation from the bakuellid Metaurostylopsis and parabirojimid Parabirojimia. The SSU rRNA gene sequences of four further urostyloids (i.e. Diaxonella pseudorubra, Anteholosticha monilata, Metaurostylopsis struederkypkeae, Pseudourostyla cristata) and two stylonychines (i.e. Sterkiella cavicola, Sterkiella histriomuscorum) from Korea were analyzed. Anteholosticha monilata, type of the genus, is clearly separated from the Holosticha clade, supporting the morphological separation from Holosticha. Sterkiella cavicola, type of Sterkiella, clusters within the stylonychines and is obviously closely related with S. histriomuscorum.

Microwave-assisted parallel synthesis of fused heterocycles in a novel parallel multimode reactor.

New rotor types using disposable glass vials for small-scale parallel synthesis in multimode microwave reactors are introduced. One rotor comprises 16 groups of four vials, whereas the second uses four silicon carbide plates with a 6 x 4 matrix to process the vials. Both rotors achieve utmost temperature homogeneity upon microwave irradiation and can be used for microwave-mediated reactions at temperatures of up to 200 degrees C and pressures of 20 bar. The generation of three different heterocycle libraries furnishing thiophenes, oxindoles, and benzimidazoles using the new rotor types is described.