On the absence of circular muscle elements in the body wall of Dysponetus pygmaeus (Chrysopetalidae, 'Polychaeta', Annelida)

The annelid body wall generally comprises an outer layer of circular muscle fibres and an inner layer of longitudinal muscle fibres as well as parapodial and chaetal muscles. An investigation of Dysponetuspygmaeus (Chrysopetalidae) with confocal laser scanning microscopy showed that circular muscles are entirely absent. Further studies indicate that this feature is characteristic for all Chrysopetalidae. A scrutiny of the literature showed a similar situation in many other polychaetes. This lack of circular muscle fibres may either be due to convergence or represent a plesiomorphic character. Since circular muscles are very likely important for burrowing forms but not necessary for animals which proceed by movements of their parapodial appendages or cilia, this problem is also related to the question of whether the ancestral polychaete was epi‐ or endobenthic.     

Molecular Cloning of a Preprohormone from Hydra magnipapillata Containing Multiple Copies of Hydra-LWamide (Leu-Trp-NH2) Neuropeptides: Evidence for Processing at Ser and Asn Residues

Abstract: The simple, freshwater polyp Hydra is often used as a model to study development in cnidarians. Recently, a neuropeptide, 2, has been isolated from sea anemones that induces metamorphosis in a hydroid planula larva to become a polyp. Here, we have cloned a preprohormone from Hydra magnipapillata containing 11 (eight different) immature neuropeptide sequences that are structurally related to the metamorphosis-inducing neuropeptide from sea anemones. During the final phase of our cloning experiments, another research team independently isolated and sequenced five of the neuropeptides originally found on the preprohormone. Comparison of these mature neuropeptide structures with the immature neuropeptide sequences on the preprohormone shows that most immature neuropeptide sequences are preceded by Ser or Asn residues, indicating that these residues must be novel processing sites. Thus, the structure of the Hydra prepro-hormone confirms our earlier findings that cnidarian pre-prohormones contain unusual or novel processing sites. Nearly all neuropeptide copies located on the Hydra preprohormone will give rise to mature neuropeptides with a C-terminal Gly-Leu-Trp-NH₂ sequence (the most frequent one being Gly-Pro-Pro-Pro-Gly-Leu-Trp-NH₂; Hydra-LWamide I; three copies). Based on their structural similarities with the metamorphosis-inducing neuropeptide from sea anemones, the mature peptides derived from the Hydra-LWamide preprohormone are potential candidates for being developmentally active neurohormones in Hydra .     

A Look into the Melting Pot: The mecC-Harboring Region Is a Recombination Hot Spot in Staphylococcus stepanovicii

Introduction

Horizontal gene transfer (HGT) is an important driver for resistance- and virulence factor accumulation in pathogenic bacteria such as Staphylococcus aureus.

Methods

Here, we have investigated the downstream region of the bacterial chromosomal attachment site (attB) for the staphylococcal cassette chromosome mec (SCCmec) element of a commensal mecC-positive Staphylococcus stepanovicii strain (IMT28705; ODD4) with respect to genetic composition and indications of HGT. S. stepanovicii IMT28705 was isolated from a fecal sample of a trapped wild bank vole (Myodes glareolus) during a screening study (National Network on “Rodent-Borne Pathogens”) in Germany. Whole genome sequencing (WGS) of IMT28705 together with the mecC-negative type strain CM7717 was conducted in order to comparatively investigate the genomic region downstream of attB (GenBank accession no. {"type":"entrez-nucleotide","attrs":{"text":"KR732654","term_id":"922979931"}}KR732654 and {"type":"entrez-nucleotide","attrs":{"text":"KR732653","term_id":"922979910"}}KR732653).

Results

The bank vole isolate (IMT28705) harbors a mecC gene which shares 99.2% nucleotide (and 98.5% amino acid) sequence identity with mecC of MRSA_LGA251. In addition, the mecC-encoding region harbors the typical blaZ-mecC-mecR1-mecI structure, corresponding with the class E mec complex. While the sequences downstream of attB in both S. stepanovicii isolates (IMT28705 and CM7717) are partitioned by 15 bp direct repeats, further comparison revealed a remarkable low concordance of gene content, indicating a chromosomal “hot spot” for foreign DNA integration and exchange.

Conclusion

Our data highlight the necessity for further research on transmission routes of resistance encoding factors from the environmental and wildlife resistome.     

Pulmonary vein isolation using new technologies to improve ablation lesion formation: Initial results comparing enhanced catheter tip irrigation (Surround Flow?) with contact force measurement (Smarttouch?)

Introduction

Pulmonary vein reconnection after pulmonary vein isolation (PVI) is a significant problem in the treatment of paroxysmal atrial fibrillation (AF). We report about patients who underwent contact force (CF) guided PVI using CF catheter and compared them to patients with PVI using an ablation catheter with enhanced tip irrigation.

Methods

A total of 59 patients were included in the analysis. In 30 patients circumferential PVI was performed using the Thermocool Smarttouch^® ablation catheter (ST) whereas in 29 patients circumferential PVI using the Thermocool Surround Flow SF^® ablation catheter (SF) was performed. Patients were compared in regard to procedure time, fluoroscopy time/dose as well as RF-application duration and completeness of PVI. Adverse events (pericardial effusion, PV stenosis, stroke, death) were evaluated. The presence of sinus rhythm off antiarrhythmic medication was assessed during 6 months follow-up using multiple 7 day Holter-ECGs.

Results

In both groups, all PVs were isolated without serious adverse events. Procedure time was 2.15 ± 0.5 h (ST) vs. 2.37 ± 1.13 h (SF) (p = 0.19). Duration of RF-applications was 46.6 ± 18 min (ST) and 49.8 ± 19 min (SF) (p = 0.52). Fluoroscopy time was 25.2 ± 13 min (ST) vs. 29 ± 18 min (SF), fluoroscopy dose 2675.6 ± 1658 versus 3038.3 ± 1997 cGym² (p = 0.36 and 0.46 respectively). Sinus rhythm off antiarrhythmic medication validated with 7 day Holter ECGs was present in both groups in 72% of patients after 6 months of follow up.

Conclusion

PVI using the new contact force catheter is safe and effective in patients with paroxysmal AF.     

A Theoretical Study of the Molecular Coupled Structures of Aristolochic Acids and Humic Acid,Potential Environmental Contaminants

An understanding of the fate of organic compounds originating from plants in soil is crucial for determining their persistence and concentrations in the environment. Aristolochic acids are believed to be the causal agents that induce Balkan endemic nephropathy by food contamination through soil adsorption of humic acids, major components of soil. Aristolochic acids are active chemicals in Aristolochia plant species found in endemic villages. In this article, molecular structure interactions between 18 structures of aristolochic acids with an inserted humic acid structure were studied. These structures were optimized in vacuo and by periodic box simulation with water solvate using the computational molecular mechanics MM+ method with HyperChem software. The QSPR models were used for correlation of the relationship between the hydrophobicity values of 18 AA structures coupled with a HA structure by MM+ and QSAR+ properties. Computational hydrophobicity values were considered dependent variables and were related to the structural features obtained by molecular and quantum mechanics calculations by multiple linear regression approaches. The obtained model was validated, and the results indicated differing hydrophobicity between the MM+ and QSAR+ properties.     

High resolution multicolor fluorescence in situ hybridization using cyanine and fluorescein dyes: Rapid chromosome identification by directly fluorescently labeled alphoid DNA probes

We tested DNA probes directly labeled by fluorescently labeled nucleotides (Cy3-dCTP, Cy5-dCTP, FluorX-dCTP) for high resolution uni- and multicolor detection of human chromosomes and analysis of centromeric DNA organization by in situ hybridization. Alpha-satellite DNA probes specific to chromosomes 1, 2, 3, 4 + 9, 5 + 19, 6, 7, 8, 10, 11, 13 + 21, 14 + 22, 15, 16, 17, 18, 20, 22, X and Y were suitable for the accurate identification of human chromosomes in metaphase and interphase cells. Cy3-labeled probes had several advantages: (1) a high level of fluorescence (5–10 times more compared with fluorescein-labeled probes); (2) a low level of fluorescence in solution, allowing the detection of target chromosomes in situ during hybridization without the washing of slides; and (3) high resistance to photobleaching during prolonged (1-2 h) exposure to strong light, thus allowing the use of a high energy mercury lamp or a long integration time during image acquisition in digital imaging microscopy for the determination of weak signals. For di- and multicolor fluorescence in situ hybridization (FISH), we successfully used different combinations of directly fluorophorated probes with preservation of images by conventional microscopy or by digital imaging microscopy. FluorX and Cy3 dyes allowed the use of cosmid probes for mapping in a one-step hybridization experiment. Cyanine-labeled fluorophorated DNA probes offer additional possibilities for rapid chromosome detection during a simple 15-min FISH procedure, and can be recommended for basic research and clinical studies, utilizing FISH.