Replacement of the essential Dictyostelium Arp2 gene by its Entamoeba homologue using parasexual genetics

Background: Cell motility is an essential feature of the pathogenesis and morbidity of amoebiasis caused by Entamoeba histolytica. As motility depends on cytoskeletal organisation and regulation, a study of the molecular components involved is key to a better understanding of amoebic pathogenesis. However, little is known about the physiological roles, interactions and regulation of the proteins of the Entamoeba cytoskeleton. Results: We have established a genetic strategy that uses parasexual genetics to allow essential Dictyostelium discoideum genes to be manipulated and replaced with modified or tagged homologues. Our results show that actin related protein 2 (Arp2) is essential for survival, but that the Dictyostelium protein can be complemented by E. histolytica Arp2, despite the presence of an insertion of 16 amino acids in an otherwise highly conserved protein. Replacement of endogenous Arp2 with myc-tagged Entamoeba or Dictyostelium Arp2 has no obvious effects on growth and the protein incorporates effectively into the Arp2/3 complex. Conclusion: We have established an effective two-step method for replacing genes that are required for survival. Our protocol will allow such genes to be studied far more easily, and also allows an unambiguous demonstration that particular genes are truly essential. In addition, cells in which the Dictyostelium Arp2 has been replaced by the Entamoeba protein are potential targets for drug screens

Evidence of volcanic ash at a K-T boundary section: Ocean drilling program hole 690 C, Maud Rise, Weddell Sea off East Antarctica

Rare vitric volcanogenic ash but more abundant clay minerals considered volcanogenic in origin are associated with an expanded and essentially complete K-T boundary sequence from Ocean Drilling Project (ODP) Hole 690 C on Maud Rise in the Weddell Sea off East Antarctica. Results at this writing are preliminary and are still based to some extent on shipboard descriptions. Further shore-based studies are in progress. It would appear, however, that the presence of volcanic ash and altered ash in the Danian section beginning at the biostratigraphically and paleomagnetically determined K-T boundary on Maud Rise can be cited as evidence of significant volcanic activity within the South Atlantic-Indian Ocean sector of the Southern Ocean coincident with the time of biotic crises at the end of the Maestrichtian. This is a postulated time of tectonic and volcanic activity within this Southern Hemisphere region, including possible initiation of the Reunion hot spot and a peak in explosive volcanism on Walvis Ridge (1) among other events. A causal relationship with the biotic crisis is possible and volcanism should be given serious consideration as a testable working hypothesis to explain these extinctions

Determinants of postnatal spleen tissue regeneration and organogenesis

Abstract The spleen is an organ that filters the blood and is responsible for generating blood-borne immune responses. It is also an organ with a remarkable capacity to regenerate. Techniques for splenic auto-transplantation have emerged to take advantage of this characteristic and rebuild spleen tissue in individuals undergoing splenectomy. While this procedure has been performed for decades, the underlying mechanisms controlling spleen regeneration have remained elusive. Insights into secondary lymphoid organogenesis and the roles of stromal organiser cells and lymphotoxin signalling in lymph node development have helped reveal similar requirements for spleen regeneration. These factors are now considered in the regulation of embryonic and postnatal spleen formation, and in the establishment of mature white pulp and marginal zone compartments which are essential for spleen-mediated immunity. A greater understanding of the cellular and molecular mechanisms which control spleen development will assist in the design of more precise and efficient tissue grafting methods for spleen regeneration on demand. Regeneration of organs which harbour functional white pulp tissue will also offer novel opportunities for effective immunotherapy against cancer as well as infectious diseases

Utilization of multiparametric prostate magnetic resonance imaging in clinical practice and focal therapy: report from a Delphi consensus project

To codify the use of multiparametric magnetic resonance imaging (mpMRI) for the interrogation of prostate neoplasia (PCa) in clinical practice and focal therapy (FT). An international collaborative consensus project was undertaken using the Delphi method among experts in the field of PCa. An online questionnaire was presented in three consecutive rounds and modified each round based on the comments provided by the experts. Subsequently, a face-to-face meeting was held to discuss and finalize the consensus results. mpMRI should be performed in patients with prior negative biopsies if clinical suspicion remains, but not instead of the PSA test, nor as a stand-alone diagnostic tool or mpMRI-targeted biopsies only. It is not recommended to use a 1.5 Tesla MRI scanner without an endorectal or pelvic phased-array coil. mpMRI should be performed following standard biopsy-based PCa diagnosis in both the planning and follow-up of FT. If a lesion is seen, MRI-TRUS fusion biopsies should be performed for FT planning. Systematic biopsies are still required for FT planning in biopsy-naïve patients and for patients with residual PCa after FT. Standard repeat biopsies should be taken during the follow-up of FT. The final decision to perform FT should be based on histopathology. However, these consensus statements may differ for expert centers versus non-expert centers. The mpMRI is an important tool for characterizing and targeting PCa in clinical practice and FT. Standardization of acquisition and reading should be the main priority to guarantee consistent mpMRI quality throughout the urological communit

Cryo-electron microscopy structure of the di-domain core of Mycobacterium tuberculosis polyketide synthase 13, essential for mycobacterial mycolic acid synthesis

Mycobacteria are known for their complex cell wall, which comprises layers of peptidoglycan, polysaccharides and unusual fatty acids known as mycolic acids that form their unique outer membrane. Polyketide synthase 13 (Pks13) of Mycobacterium tuberculosis, the bacterial organism causing tuberculosis, catalyses the last step of mycolic acid synthesis prior to export to and assembly in the cell wall. Due to its essentiality, Pks13 is a target for several novel anti-tubercular inhibitors, but its 3D structure and catalytic reaction mechanism remain to be fully elucidated. Here, we report the molecular structure of the catalytic core domains of M. tuberculosis Pks13 (Mt-Pks13), determined by transmission cryo-electron microscopy (cryoEM) to a resolution of 3.4 Å. We observed a homodimeric assembly comprising the ketoacyl synthase (KS) domain at the centre, mediating dimerization, and the acyltransferase (AT) domains protruding in opposite directions from the central KS domain dimer. In addition to the KS-AT di-domains, the cryoEM map includes features not covered by the di-domain structural model that we predicted to contain a dimeric domain similar to dehydratases, yet likely lacking catalytic function. Analytical ultracentrifugation data indicate a pH-dependent equilibrium between monomeric and dimeric assembly states, while comparison with the previously determined structures of M. smegmatis Pks13 indicates architectural flexibility. Combining the experimentally determined structure with modelling in AlphaFold2 suggests a structural scaffold with a relatively stable dimeric core, which combines with considerable conformational flexibility to facilitate the successive steps of the Claisen-type condensation reaction catalysed by Pks13

Growth rates of icicles

Experimental and theoretical studies on the growth rate of an icicle were carried out as a function of temperature, water-supply rate and wind speed; the relative humidity was also taken into account. The length of an icicle increases by the downward growth of thin dendritic crystals into the supercooled pendant water drop at the tip, and thus the growth is in the crystallographic a-axis direction. The diameter, on the other hand, increases by the freezing of a water film flowing down along the icicle wall. The ratio of measured length-and diameter-growth rates was large, namely 8–32.Both growth rates increased with decreasing temperature and increasing wind speed. The increase in water-supply rate led to the decrease in the length-growth rate but no significant change in the diameter-growth rate. These results could be well described by a numerical model of icicle growth which takes account of the dendritic growth at the tip and the wall and the effective heat transfer within the turbulent boundary layer around the icicle. A formation mechanism of ribs and hollows is discussed in relation to the flowing and freezing process of water on an icicle wall

Additecture

Preliminary design work for the Solar Decathlon 2014 entry Techstyle Haus completed in a wintersession 2013 RISD design studio in Erfurt, Germany taught by Jonathan Knowles. The Solar Decathlon competition challenges twenty collegiate teams to design and build sustainable homes that are powered exclusively by solar energy and incorporate sustainable architecture and design. Techstyle Haus is an international Brown University, RISD and University of Applied Sciences Erfurt,Germany collaboration designing a solar passivehaus out of high performance textiles

Techstyle Haus

Preliminary design work for the Solar Decathlon 2014 entry Techstyle Haus completed in a wintersession 2013 RISD design studio in Erfurt, Germany taught by Jonathan Knowles. The Solar Decathlon competition challenges twenty collegiate teams to design and build sustainable homes that are powered exclusively by solar energy and incorporate sustainable architecture and design. Techstyle Haus is an international Brown University, RISD and University of Applied Sciences Erfurt,Germany collaboration designing a solar passivehaus out of high performance textiles

Haus House

Preliminary design work for the Solar Decathlon 2014 entry Techstyle Haus completed in a wintersession 2013 RISD design studio in Erfurt, Germany taught by Jonathan Knowles. The Solar Decathlon competition challenges twenty collegiate teams to design and build sustainable homes that are powered exclusively by solar energy and incorporate sustainable architecture and design. Techstyle Haus is an international Brown University, RISD and University of Applied Sciences Erfurt,Germany collaboration designing a solar passivehaus out of high performance textiles

Prostate Magnetic Resonance Imaging for Local Recurrence Reporting (PI-RR): International Consensus -based Guidelines on Multiparametric Magnetic Resonance Imaging for Prostate Cancer Recurrence after Radiation Therapy and Radical Prostatectomy

BACKGROUND: Imaging techniques are used to identify local recurrence of prostate cancer (PCa) for salvage therapy and to exclude metastases that should be addressed with systemic therapy. For magnetic resonance imaging (MRI), a reduction in the variability of acquisition, interpretation, and reporting is required to detect local PCa recurrence in men with biochemical relapse after local treatment with curative intent. OBJECTIVE: To propose a standardised method for image acquisition and assessment of PCa local recurrence using MRI after radiation therapy (RP) and radical prostatectomy (RT). EVIDENCE ACQUISITION: Prostate Imaging for Recurrence Reporting (PI-RR) was formulated using the existing literature. An international panel of experts conducted a nonsystematic review of the literature. The PI-RR system was created via consensus through a combination of face-to-face and online discussions. EVIDENCE SYNTHESIS: Similar to with PI-RADS, based on the best available evidence and expert opinion, the minimum acceptable MRI parameters for detection of recurrence after radiation therapy and radical prostatectomy are set. Also, a simplified and standardised terminology and content of the reports that use five assessment categories to summarise the suspicion of local recurrence (PI-RR) are designed. PI-RR scores of 1 and 2 are assigned to lesions with a very low and low likelihood of recurrence, respectively. PI-RR 3 is assigned if the presence of recurrence is uncertain. PI-RR 4 and 5 are assigned for a high and very high likelihood of recurrence, respectively. PI-RR is intended to be used in routine clinical practice and to facilitate data collection and outcome monitoring for research. CONCLUSIONS: This paper provides a structured reporting system (PI-RR) for MRI evaluation of local recurrence of PCa after RT and RP. PATIENT SUMMARY: A new method called PI-RR was developed to promote standardisation and reduce variations in the acquisition, interpretation, and reporting of magnetic resonance imaging for evaluating local recurrence of prostate cancer and guiding therapy