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PubMed1994

Comparison of traditional and molecular methods of typing isolates of Staphylococcus aureus.

Fifty-nine Staphylococcus aureus isolates and 1 isolate of Staphylococcus intermedius were typed by investigators at eight institutions by using either antibiograms, bacteriophage typing, biotyping, immunoblotting, insertion sequence typing with IS257/431, multilocus enzyme electrophoresis, restriction analysis of plasmid DNA, pulsed-field or field inversion gel electrophoresis, restriction analysis of PCR-amplified coagulase gene sequences, restriction fragment length polymorphism typing by using four staphylococcal genes as probes, or ribotyping. Isolates from four well-characterized outbreaks (n = 29) and a collection of organisms from two nursing homes were mixed with epidemiologically unrelated stock strains from the Centers for Disease Control and Prevention. Several isolates were included multiple times either within or between the sets of isolates to analyze the reproducibilities of the typing systems. Overall, the DNA-based techniques and immunoblotting were most effective in grouping outbreak-related strains, recognizing 27 to 29 of the 29 outbreak-related strains; however, they also tended to include 3 to 8 epidemiologically unrelated isolates in the same strain type. Restriction fragment length polymorphism methods with mec gene-associated loci were less useful than other techniques for typing oxacillin-susceptible isolates. Phage typing, plasmid DNA restriction analysis, and antibiogram analysis, the techniques most readily available to clinical laboratories, identified 23 to 26 of 29 outbreak-related isolates and assigned 0 to 6 unrelated isolates to outbreak strain types. No single technique was clearly superior to the others; however, biotyping, because it produced so many subtypes, did not effectively group outbreak-related strains of S. aureus.

Bacterial Typing Techniques↗

PubMed2003

Real-time magnetic resonance imaging-guided coronary catheterization in swine.

BACKGROUND: We tested the hypothesis that real-time magnetic resonance imaging (MRI) can guide coronary artery catheterization in swine via a percutaneous femoral artery approach. METHODS AND RESULTS: In 12 pigs, we accessed femoral arteries percutaneously. We used 6- or 7-French coronary Judkins catheters filled with dilute 4% gadolinium (Gd) contrast agent and coaxially inserted 0.030-inch diameter active guidewires as endovascular devices. For catheter tracking, we used a 2-dimensional (2D) inversion recovery-prepared spoiled gradient echo sequence at a temporal resolution of 7 frames/s. For guidewire tracking, we used 2D steady-state free precession imaging at a temporal resolution of 9 frames/s. Coronary artery catheterization under MRI guidance was successful in 12/12 pigs. Successful coronary catheterization was verified by obtaining MR angiographic images after direct catheter-based injections of dilute Gd. CONCLUSIONS: Real-time MRI-guided catheterization of coronary arteries in swine is feasible via a percutaneous femoral artery approach. Selective coronary MR angiography can then be performed with dilute contrast agent injections.

Animals↗

PubMed1993

Insertion of a 5' truncated L1 element into the 3' end of exon 44 of the dystrophin gene resulted in skipping of the exon during splicing in a case of Duchenne muscular dystrophy.

We report here the second evidence of retrotransposition of L1, which was found inserted into the dystrophin gene of a patient, causing Duchenne muscular dystrophy (DMD). When the PCR was used to amplify a region of the dystrophin gene encompassing exon 44 from genomic DNA of two Japanese brothers with DMD, it was found to be approximately 600 bp larger than expected. Both the normal and the abnormally large products were amplified from the DNA of their mother. However, the maternal grandparents did not have the abnormal allele, and the mutation must therefore have occurred in the mother. Analysis of nucleotide sequence of the amplified product from a patient disclosed that the insertion was present zero to two bases upstream from the 3' end of exon 44 and that two to four bases of the exon sequence were deleted from the insertion site. The insertion sequence was found to be composed of 606-608 bp and to be almost identical to the inverse complement of 3' portion of the L1 retrotransposon consensus sequence. The dystrophin gene transcript from peripheral lymphocytes of one of the patients was analyzed by using reverse transcription/semi-nested PCR. The size of the amplified product encompassing exon 42 to 46 was smaller than expected. Sequencing of the amplified product disclosed that the sequence of exon 43 was directly joined to that of exon 45. Exon 44 of the transcript was thus shown to be skipped during splicing. This novel mutation of the dystrophin gene has important implications regarding retrotransposition of an active L1 element and provides a new insight into the origins of mutations in the dystrophin gene.

Adult↗

PubMed1997

A biomechanical comparison of intramedullary nail and crossed lag screw fixation for tibiotalocalcaneal arthrodesis.

This study compared the mechanical bending and torsional properties of intramedullary nail fixation and lag screw fixation for tibiotalocalcaneal arthrodesis. Seven matched pairs of human cadaver lower extremities were studied, with one hindfoot in each pair stabilized with a 12 mm x 150 mm interlocked intramedullary nail inserted retrograde across the subtalar and ankle joints. The contralateral hindfoot was stabilized with two crossed 6.5 mm cannulated screws inserted across both the ankle and subtalar joints. Specimens were subjected to cantilever bending tests in plantarflexion, dorsiflexion, inversion, and eversion and to torsional tests in internal and external rotation. The intramedullary nail construct was significantly (P < 0.05) stiffer than the crossed lag screw construct in all four bending directions and both rotational directions: plantarflexion (nail, 42.8 N/mm; screws, 16.4 N/mm; P = 0.0003), dorsiflexion (nail, 43.0 N/mm; screws, 10.3 N/mm; P = 0.0005), inversion (nail, 37.7 N/mm; screws, 12.3 N/mm; P = 0.0024), eversion (nail, 35.4 N/mm; screws, 10.8 N/mm; P = 0.0004), internal rotation (nail, 1.29 N-m/degree; screws, 0.82 N-m/degree; P = 0.01), external rotation (nail, 1.35 N-m/degree; screws, 0.44 N-m/degree; P = 0.0001). Intramedullary fixation is biomechanically stiffer than crossed lag screws in all bending and torsional directions tested and therefore this construct may aid in maintaining alignment of the hindfoot during union and may help increase fusion rate through increased stability of the internal fixation.

Ankle Joint↗

PubMed2001

Chromosomal distribution of the transposable elements Osvaldo and blanco in original and colonizer populations of Drosophila buzzatii.

Chromosomal distribution of transposable elements (TEs) Osvaldo and blanco in D. buzzatii was studied in three original natural populations from Argentina (Berna, Puerto Tirol and La Nostalgia) and a colonizer population from the Iberian Peninsula (Carboneras). The Spanish population showed significant differences for Osvaldo and blanco copy numbers when we compared the X chromosome and the autosomes; but it is mainly the accumulation of copies in chromosome 2, where most sites with high insertion frequency were located, that causes the discrepancy with the negative selection model. We found no significant differences in TE frequency between chromosomal regions with different exchange rates, and no evident accumulation of TE was detected within chromosomal inversions where recombination rate is reduced. The Carboneras population shows euchromatic sites of Osvaldo and blanco with high occupancy and others with low copy number. On the contrary, Argentinian populations show only a generalized low occupancy per insertion site. Moreover, the mean copy number of both elements is higher in Spain than in Argentina. All these results suggest an important role of the colonization process in the distribution of TEs. The increase in the copy number of the TEs analysed and their elevated frequency in some chromosomal sites in Carboneras is, most probably, a sequel of the founder event and drift that took place at the time of the colonization of the Old World by D. buzzatii from the New World some 300 years ago.

Animals↗

PubMed1989

Regulated expression of muscle-specific genes introduced into mouse embryonal stem cells: inverse correlation with DNA methylation.

Pluripotent embryonal stem cell lines (ES) were isolated from cultured normal mouse blastocysts. These cells retained their capacity to differentiate into a great variety of cell types in cell cultures or in tumors formed after subcutaneous injection of the cells into nude mice. A chimeric actin/globin gene containing about two-thirds of the rat skeletal muscle actin gene and 730 bp of its 5' flanking region fused to the 3' end of the human embryonic epsilon-globin gene, was inserted into a plasmid containing a neomycin resistance gene (neor) whose transcription is regulated by the SV40 early control elements. The prokaryotic vector DNA sequences of this plasmid (pAG-Neo) were deleted and the two linked genes were introduced into the ES cells by electroporation. G418-resistant clones were isolated, amplified and injected subcutaneously into nude mice. From the teratocarcinoma-like tumors which developed we isolated myogenic as well as nonmyogenic cell lines. In cell lines derived from three independent transfected ES clones, expression of the actin/globin gene was developmentally regulated in myogenic cells. In contrast, in a number of experiments in which the actin/globin gene or other muscle-specific genes were introduced into the ES cells without the removal of the pBR sequences, no expression could be detected at any stage. Moreover, in the differentiated lines derived from these clones, G418 resistance was lost, and no neor transcripts could be detected. Southern-blot analysis of MSPI- or HpaII-digested DNA revealed extensive methylation in the clones that did not express the foreign DNA, whereas no significant methylation of the inserted DNA was observed in clones which expressed the transfected genes. Examination of the DNA extracted from transgenic mice carrying the same actin/globin gene revealed an inverse correlation between methylation of the exogenous gene and its potential to be expressed in the transgenic strain. However, no tissue-specific differences in methylation, related to the tissue specificity of expression of the exogenous gene, could be detected in these experiments. These results suggest that the process of methylation reported here is causally related to constitutive inactivation of the exogenous genes.

Actins↗

PubMed2000

The mechanical waveform of the basilar membrane. II. From data to models--and back.

Mechanical responses in the basal turn of the guinea-pig cochlea are measured with low-level broad-band noise as the acoustical stimulus [for details see de Boer and Nuttall, J. Acoust. Soc. Am. 101, 3583-3592 (1997)]. Results are interpreted within the framework of a classical three-dimensional model of the cochlea that belongs to a very wide class of nonlinear models. The use of linear-systems analysis for this class of nonlinear models has been justified earlier [de Boer, Audit. Neurosci. 3, 377-388 (1997)]. The data are subjected to inverse analysis with the aim to recover the "effective basilar-membrane impedance." This is a parameter function that, when inserted into the model, produces a model response, the "resynthesized" response, that is similar to the measured response. With present-day solution methods, resynthesis leads back to an almost perfect replica of the original response in the spatial domain. It is demonstrated in this paper that this also applies to the response in the frequency domain and in the time domain. This paper further reports details with regard to geometrical properties of the model employed. Two three-dimensional models are studied; one has its dimensions close to that of the real cochlea, the other is a stylized model which has homogeneous geometry over its length. In spite of the geometric differences the recovered impedance functions are very similar. An impedance function computed for one model can be used in resynthesis of the response in the other one, and this leads to global amplitude deviations between original and resynthesized response functions not exceeding 8 dB. Discrepancies are much larger (particularly in the phase) when a two-dimensional model is compared with a three-dimensional model. It is concluded that a stylized three-dimensional model with homogeneous geometric parameters will give sufficient information in further work on unraveling cochlear function via inverse analysis. In all cases of a sensitive cochlea stimulated by a signal with a stimulus level of 50 dB SPL per octave or less, the resulting basilar-membrane impedance is found to be locally active, that is, the impedance function shows a region where the basilar membrane is able to amplify acoustic power or to reduce dissipation of power by the organ of Corti. Finally, the influence of deliberate errors added to the data is discussed in order to judge the accuracy of the results.

Animals↗

PubMed1986

Inversions and recombinations in mitochondrial DNA of the (SG-1) cytoplasmic mutant in two Neurospora species.

The mitochondrial DNAs of [SG-1] cytoplasmically-mutant and wild-type strains of Neurospora crassa and Neurospora sitophila were examined by comparative restriction endonuclease analyses. The mtDNA of N. sitophila wild type of Whitehouse differs from type II mtDNA of N. crassa by insertions of 3.3 kb in EcoRI-9, and 1.2 kb in EcoRI-3, and a deletion of 1.1 kb in EcoRI-5. These DNA heteromorphisms provided convenient markers for tracing N. crassa [SG-1] mtDNA during and after its transfer into N. sitophila. The [SG-1] cytoplasmic mutant in both N. crassa and N. sitophila has a distinctive inversion that connects the fragment EcoRI-4 with HindIII-10a. The [SG-1] mtDNA from N. crassa remained essentially intact after it was transferred by crosses into N. sitophila. In each species, a unique second inversion occurred in the [SG-1] mtDNA after the transfer was made. In N. sitophila, polar recombination in heteroplasmons between [SG-1] and wild-type preferentially yields strains with mtDNAs that contain the maximum possible number of insertions in the cob and co-1 loci of the EcoRI-3 region of the mitochondrial chromosome.

Chromosome Inversion↗

PubMed2001

Biased distribution of inverted and direct Alus in the human genome: implications for insertion, exclusion, and genome stability.

Alu sequences, the most abundant class of large dispersed DNA repeats in human chromosomes, contribute to human genome dynamics. Recently we reported that long inverted repeats, including human Alus, can be strong initiators of genetic change in yeast. We proposed that the potential for interactions between adjacent, closely related Alus would influence their stability and this would be reflected in their distribution. We have undertaken an extensive computational analysis of all Alus (the database is at http://dir.niehs.nih.gov/ALU) to better understand their distribution and circumstances under which Alu sequences might affect genome stability. Alus separated by <650 bp were categorized according to orientation, length of regions sharing high sequence identity, distance between highly identical regions, and extent of sequence identity. Nearly 50% of all Alu pairs have long alignable regions (>275 bp), corresponding to nearly full-length Alus, regardless of orientation. There are dramatic differences in the distributions and character of Alu pairs with closely spaced, nearly identical regions. For Alu pairs that are directly repetitive, approximately 30% have highly identical regions separated by <20 bp, but only when the alignments correspond to near full-size or half-size Alus. The opposite is found for the distribution of inverted repeats: Alu pairs with aligned regions separated by <20 bp are rare. Furthermore, closely spaced direct and inverted Alus differ in their truncation patterns, suggesting differences in the mechanisms of insertion. At larger distances, the direct and inverted Alu pairs have similar distributions. We propose that sequence identity, orientation, and distance are important factors determining insertion of adjacent Alus, the frequency and spectrum of Alu-associated changes in the genome, and the contribution of Alu pairs to genome instability. Based on results in model systems and the present analysis, closely spaced inverted Alu pairs with long regions of alignment are likely at-risk motifs (ARMs) for genome instability.

Alu Elements↗

PubMed1985

Novel rearrangements of herpes simplex virus DNA sequences resulting from duplication of a sequence within the unique region of the L component.

We constructed insertion mutants of herpes simplex virus type 1 that contained a duplication of DNA sequences from the BamHI-L fragment (map units 0.706 to 0.744), which is located in the unique region of the L component (UL) of the herpes simplex virus type 1 genome. The second copy of the BamHI-L sequence was inserted in inverted orientation into the viral thymidine kinase gene (map units 0.30 to 0.32), also located within UL. A significant fraction of the progeny produced by these insertion mutants had genomes with rearranged DNA sequences, presumably resulting from intramolecular or intermolecular recombination between the BamHI-L sequences at the two different genomic locations. The rearranged genomes either had an inversion of the DNA sequence flanked by the duplication or were recombinant molecules in which different regions of the genome had been duplicated and deleted. Genomic rearrangements similar to those described here have been reported previously but only for herpes simplex virus insertion mutants containing an extra copy of the repetitive a sequence. Such rearrangements have not been reported for insertion mutants that contain duplications of herpes simplex virus DNA sequences from largely unique regions of the genome. The implications of these results are discussed.

Base Sequence↗

PubMed1999

Multiple DNA binding activities of the novel site-specific recombinase, Piv, from Moraxella lacunata.

The recombinase, Piv, is essential for site-specific DNA inversion of the type IV pilin DNA segment in Moraxella lacunata and Moraxella bovis. Piv shows significant homology with the transposases of the IS110/IS492 family of insertion elements, but, surprisingly, Piv contains none of the conserved amino acid motifs of the lambda Int or Hin/Res families of site-specific recombinases. Therefore, Piv may mediate site-specific recombination by a novel mechanism. To begin to determine how Piv may assemble a synaptic nucleoprotein structure for DNA cleavage and strand exchange, we have characterized the interaction of Piv with the DNA inversion region of M. lacunata. Gel shift and nuclease/chemical protection assays, competition and dissociation rate analyses, and cooperativity studies indicate that Piv binds two distinct recognition sequences. One recognition sequence, found at multiple sites within and outside of the invertible segment, is bound by Piv protomers with high affinity. The second recognition sequence is located at the recombination cross-over sites at the ends of the invertible element; Piv interacts with this sequence as an oligomer with apparent low affinity. A model is proposed for the role of the different Piv binding sites of the M. lacunata inversion region in the formation of an active synaptosome.

Base Sequence↗

PubMed1993

The immunoglobulin kappa locus-or-what has been learned from looking closely at one-tenth of a percent of the human genome.

The immunoglobulin kappa locus and its immediate surroundings, which are described in the present report, comprise 3 Mb of DNA, i.e., 0.1% or one per mill of the 3000 Mb of the human genome. Based on the work of our group during the past 12 years, we can now (1) depict in much detail the structure of the kappa locus with its 76 V kappa genes and pseudo genes, five J kappa elements and one C kappa gene; (2) specify the size of the germ-line repertoire of kappa light chains, which is one of the sources of the practically unlimited antibody diversity; (3) assign the known transcription products (studied as cDNAs) and kappa proteins to certain germ-line V kappa genes and attribute the differences in sequences to hypermutation and, to a lesser extent, to allelic variation; (4) analyze the hypermutation patterns which may contribute to the understanding of this enigmatic process; (5) describe the V kappa-J kappa rearrangements for half of the V kappa genes by a deletion mechanism and for the other half by a mechanism involving inversions of Mb-sized (i.e., 0.5 mm long) DNA fragments; (6) define various regulatory and other conserved sequence elements; (7) get clues as to the variation of the structure of the kappa locus in different individuals and populations, including a haplotype with only half the number of V kappa genes; (8) interpret many aspects of the evolution of the kappa locus in terms of duplications, insertions, deletions and gene conversions; (9) attribute the formation of the 24 V kappa orphons (i.e., genes outside the locus), whose sequences were determined, to pericentric inversions and other transposition processes; (10) answer a series of questions of biomedical interest; and (11) contribute 12.5 Mb of restriction maps, 1.8 Mb of clones and 250 kb of sequences to the elucidation of the human genome.

Animals↗

PubMed2006

Studies of potential inversion in the electrochemical reduction of 11,11,12,12-tetracyano-9,10-anthraquinodimethane and 2,3,5,6-tetramethyl-7,7,8,8-tetracyano-1,4-benzoquinodimethane.

The electrochemical reduction of the title compounds, 2a and 3a, and 7,7,8,8-tetracyanoquinodimethane, 1a, was studied in acetonitrile. The reduction of 1a shows normal ordering of potentials, i.e., the potential for insertion of the first electron, E degrees1, is more positive than the potential for the second step of reduction, E degrees2. Thus, E degrees1 - E degrees2 > 0. By contrast, 2a and 3a show inversion of potentials where introduction of the second electron occurs with greater ease than the first (E degrees1 - E degrees2 < 0). The extent of inversion has been determined by simulation of the cyclic voltammograms obtained at 298 and 257 K. Electron paramagnetic resonance measurements at room temperature of solutions containing equimolar mixtures of the neutral and dianion allow determination of the concentration of anion radicals from which the disproportionation equilibrium constant and E degrees1 - E degrees2 can be calculated. The results were in good agreement with the voltammetric determinations. Calculations were conducted to characterize the structural changes accompanying reduction to the anion radical and dianion forms. Fast scan experiments at low temperatures (up to 10 000 V/s at 257 K; 500 V/s at 233 K) were conducted in an attempt to detect intermediates in the reduction, but none was found. Thus, it is not possible to state whether structural change and electron transfer are concerted or occur in discrete steps.

Journal Article↗

PubMed1975

Patient package inserts. II. Toward a rational patient package insert.

Analysis of the response to a questionnaire on various aspects of drug information showed several significant trends. The importance of the physician as an information source varied directly with age and inversely with educational level of patients. The doctor's importance as a source of information was also greater for those who visited physicians' offices or clinics more and if they were on long-term treatment. Younger persons, those with higher levels of education, and those who were not in regular contact with a physician placed greater importance on non-physician sources. Persons consulting physicians regularly desired less information and knew less about drugs than those who did not regularly see a physician. These findings suggest that the educational role of the physician could be improved and should be directed particularly at the low-income, older patient. In the design of the package insert, particular attention should be given to self-prescribed medications where no other information sources will be operating.

Adult↗

PubMed1997

The highly rearranged chloroplast genome of Trachelium caeruleum (Campanulaceae): multiple inversions, inverted repeat expansion and contraction, transposition, insertions/deletions, and several repeat families.

Comprehensive gene mapping reveals that the chloroplast genome of Trachelium caeruleum is highly rearranged relative to those of other land plants. Evolutionary scenarios that consist of seven to ten inversions, one or two transpositions, both expansion and contraction of the typically size-conserved inverted repeat, a presumed gene loss, deletions within two large open reading frames and several insertions, are sufficient to derive the Trachelium arrangement from the ancestral angiosperm chloroplast DNA arrangement. Two of the rearrangements disrupt transcriptional units that are otherwise conserved among land plants. At least five families of small dispersed repeats exist in the Trachelium chloroplast genome. Most of the repeats are associated with inversion endpoints and may have facilitated inversions through recombination across homologous repeats.

Biological Evolution↗

PubMed2001

Microsatellite instability, mitochondrial DNA large deletions, and mitochondrial DNA mutations in gastric carcinoma.

Mitochondrial DNA (mtDNA) large deletions and mtDNA mutations have been demonstrated in various types of human cancer. The relationship between the occurrence of such alterations and the nuclear microsatellite instability (MSI) status of the neoplastic cells remains controversial. In an attempt to clarify the situation in gastric carcinoma, we studied, by PCR/SSCP and sequencing, five mitochondrial genes and two D-loop regions in 32 gastric carcinomas that had been previously screened for MSI and mitochondrial common deletion. MtDNA alterations were detected in 26 carcinomas (81%). All the mtDNA mutations, which occurred mainly in the D-loop and ND1 and ND5 genes, were transitions. D-loop alterations (insertions and/or deletions) were not significantly associated with mutations in the coding regions. There was a trend towards an inverse relationship between the occurrence of mitochondrial common deletion and mtDNA mutations. No significant relationship was observed between MSI status and mtDNA mutations, whereas the mitochondrial common deletion appeared to be almost exclusively restricted to MSI-negative tumors. The latter finding--almost no gastric carcinoma with MSI-positive phenotype has large deletions of mtDNA--needs to be confirmed in a larger series and in tumors from other organs.

Carcinoma↗

PubMed1993

Sec dependent and sec independent assembly of E. coli inner membrane proteins: the topological rules depend on chain length.

Translocation of proteins across the inner membrane of Escherichia coli normally requires the participation of the sec machinery. A number of proteins are known, however, where translocation can proceed unhindered even when the function of either SecA or SecY, central components of the sec machinery, is blocked. We now show that there is a linear correlation between the length of a translocated region and its degree of dependence on SecA and SecY for lengths between 25 and 55 residues. We also find that positively charged residues have distinctly different topological effects during SecA dependent and SecA independent membrane protein insertion, and that a short cytoplasmic segment in Lep can be converted to a translocated segment (with a concomitant inversion of the original topology of the whole molecule) by increasing its length into the SecA/Y dependent realm.

Adenosine Triphosphatases↗

PubMed2004

The elusive alpha(1D)-adrenoceptor: molecular and cellular characteristics and integrative roles.

alpha(1)-Adrenoceptors seem to play key roles in cardiovascular, genitourinary, and central nervous system functions. This review will be focused on alpha(1D)-adrenoceptors. These receptors have intrinsic activity, and many of the more commonly used antagonists are in reality inverse agonists. alpha(1D)-Adrenoceptors are phosphorylated in the basal state, and the natural agonists, adrenaline and noradrenaline, increase their phosphorylation; similar effects are induced by direct activation of protein kinase C and through activation of nonadrenergic receptors. Interestingly, a large proportion of alpha(1D)-adrenoceptors are located in intracellular vesicles. Such intracellular location can be changed to surface expression through the use of inverse agonists and coexpression of alpha(1B)-adrenoceptors, which seem to act as pharmacological chaperons for proper plasma membrane insertion. The alpha(1D)-adrenoceptor amino terminus seems to contain a signal that keeps the receptor intracellularly, but interaction with other proteins may also contribute. The precise relationship between the intrinsic activity, phosphorylation, and intracellular location is currently unknown. alpha(1D)-Adrenoceptor activation induces contraction in a variety of vessels, and a role in the control of blood pressure has been suggested. Studies using young prehypertensive and adult spontaneously hypertensive rats as well as knockout mice suggest that vascular alpha(1D)-adrenoceptors are involved in the genesis/maintenance of hypertension.

Adrenergic alpha-1 Receptor Antagonists↗