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Superresolution, structured illumination microscopy (SIM) is an ideal modality for imaging live cells due to its relatively high speed and low photon-induced damage to the cells. The rate-limiting step in observing a superresolution image in SIM is often the reconstruction speed of the algorithm used to form a single image from as many as nine raw images. Reconstruction algorithms impose a significant computing burden due to an intricate workflow and a large number of often complex calculations to produce the final image. Further adding to the computing burden is that the code, even within the MATLAB environment, can be inefficiently written by microscopists who are noncomputer science researchers. In addition, they do not take into consideration the processing power of the graphics processing unit (GPU) of the computer. To address these issues, we present simple but efficient approaches to first revise MATLAB code, followed by conversion to GPU-optimized code. When combined with cost-effective, high-performance GPU-enabled computers, a 4- to 500-fold improvement in algorithm execution speed is observed as shown for the image denoising Hessian-SIM algorithm. Importantly, the improved algorithm produces images identical in quality to the original.

Super-resolution structured illumination microscopy (SR-SIM) is finding increasing application in biomedical research due to its superior ability to visualize subcellular dynamics in living cells. However, during image reconstruction artifacts can be introduced and when coupled with time-consuming postprocessing procedures, limits this technique from becoming a routine imaging tool for biologists. To address these issues, an accelerated, artifact-reduced reconstruction algorithm termed joint space frequency reconstruction-based artifact reduction algorithm (JSFR-AR-SIM) was developed by integrating a high-speed reconstruction framework with a high-fidelity optimization approach designed to suppress the sidelobe artifact. Consequently, JSFR-AR-SIM produces high-quality, super-resolution images with minimal artifacts, and the reconstruction speed is increased. We anticipate this algorithm to facilitate SR-SIM becoming a routine tool in biomedical laboratories.

Super-resolution structured illumination microscopy (SIM) routinely performs image reconstruction in the frequency domain using an approach termed frequency-domain reconstruction (FDR). Due to multiple Fourier transforms between the spatial and frequency domains, SIM suffers from low reconstruction speed, constraining its applications in real-time, dynamic imaging. To overcome this limitation, we developed a new method for SIM image reconstruction, termed spatial domain reconstruction (SDR). SDR is intrinsically simpler than FDR, does not require Fourier transforms and the theory predicts it to be a rapid image reconstruction method. Results show that SDR reconstructs a super-resolution image 7-fold faster than FDR, producing images that are equal to either FDR or the widely-used FairSIM. We provide a proof-of-principle using mobile fluorescent beads to demonstrate the utility of SDR in imaging moving objects. Consequently, replacement of the FDR approach with SDR significantly enhances SIM as the desired method for live-cell, instant super-resolution imaging. This means that SDR-SIM is a “What You See Is What You Get” approach to super-resolution imaging.

The maintenance of genome stability requires the coordinated actions of multiple proteins and protein complexes, that are collectively known as genome guardians. Within this broadly defined family is a subset of proteins that contain oligonucleotide/oligosaccharide-binding folds (OB-fold). While OB-folds are widely associated with binding to single-stranded DNA this view is no longer an accurate depiction of how these domains are utilized. Instead, the core of the OB-fold is modified and adapted to facilitate binding to a variety of DNA substrates (both single- and double-stranded), phospholipids, and proteins, as well as enabling catalytic function to a multi-subunit complex. The flexibility accompanied by distinctive oligomerization states and quaternary structures enables OB-fold genome guardians to maintain the integrity of the genome via a myriad of complex and dynamic, protein-protein; protein-DNA, and protein-lipid interactions in both prokaryotes and eukaryotes.

Super-resolution microscopy surpasses the diffraction limit to enable the observation of the fine details in sub-cellular structures and their dynamics in diverse biological processes within living cells. Structured illumination microscopy (SIM) uses a relatively low illumination light power compared with other super-resolution microscopies and has great potential to meet the demands of live-cell imaging. However, the imaging acquisition and reconstruction speeds limit its further applications. In this article, recent developments all targeted at improving the overall speed of SIM are reviewed. These comprise both hardware and software improvements, which include a reduction in the number of raw images, GPU acceleration, deep learning and the spatial domain reconstruction. We also discuss the application of these developments in live-cell imaging.

Quantitative measurement of small forces and small displacement using holographic optical tweezers (HOTs) is finding increasing applications due to the features of non-contact and high accuracy manipulation. Although hologram optimization algorithms have been widely reported, the holographic optical trapping performance relying on the algorithms has not been studied systematically. In this paper, we investigated the force measuring the performance of various types of HOTs generated with six different hologram algorithms (GSW, GAA, GS, SR, S, and RM). To do this, we built up a HOT instrument and compared the light fields’ intensity distribution, trap stiffness, efficiency, and calculation time of multi-point trap arrays generated by six hologram algorithms with this setup. Our work will provide a better understanding of the performance of different hologram algorithms in HOTs.

Exosomes are endosome-derived small membrane vesicles that are secreted by most cell types including tumor cells. Tumor-derived exosomes usually contain tumor antigens and have been used as a source of tumor antigens to stimulate anti-tumor immune responses. However, many reports also suggest that tumor-derived exosomes can facilitate tumor immune evasion through different mechanisms, most of which are antigen-independent. In the present study we used a mouse model of delayed-type hypersensitivity (DTH) and demonstrated that local administration of tumor-derived exosomes carrying the model antigen chicken ovalbumin (OVA) resulted in the suppression of DTH response in an antigen-specific manner. Analysis of exosome trafficking demonstrated that following local injection, tumor-derived exosomes were internalized by CD11c+ cells and transported to the draining LN. Exosome-mediated DTH suppression is associated with increased mRNA levels of TGF-β1 and IL-4 in the draining LN. The tumor-derived exosomes examined were also found to inhibit DC maturation. Taken together, our results suggest a role for tumor-derived exosomes in inducing tumor antigen-specific immunosuppression, possibly by modulating the function of APCs.

Cardiogenic shock refractory to conventional therapy has very high mortality and limited therapeutical options. Aim of the study was to evaluate the impact of the use of ECMO as a life-saving measure when optimal conventional treatment has been reached and mechanical circulatory support is the only option for survival. Material and methods: Between January 2009 and May 2011, 32 patients in cardiogenic shock refractory to optimal conventional therapy (inotropes and intra-aortic-balloonpump) were treated with the extracorporeal life support implantation. Veno-arterial extracorporeal membrane oxygenation has been implanted either at bedside under local anesthesia or in operating room. Results: The mean age of the population (24 male and 8 female) was 49 ± 16 years, all patients presented with cardiogenic shock refractory to medical therapy due to various etiology. Veno-arterial extracorporeal membrane oxygenation was implanted at bedside under local anesthesia in 20 awake patients (63%) and in the operating room in the remaining 12 (37%). Average duration of ECMO support was 12.3 ± 10.2 days (range 1–46). Twenty-six patients (81%) were weaned from veno-arterial extracorporeal membrane oxygenation or bridged to either a ventricular assist device or heart transplantation. ECMO was used as bridge to transplantation in 7 patients (22%), bridge to recovery in 10 patients (31%) and bridge to bridge in 9 patients (28%). Six patients (18.7%) died during ECMO support, whereas 30-day overall survival after ECMO removal was 80.7% (21/26 pts). Sixteen patients (50%) were discharged from the hospital, with a 100% survival at sixmonths follow-up. Conclusions: In our experience the use of ECMO as a "bridge to decision" significantly improved the outcome of cardiogenic shock patients, greatly reducing the expected mortality.

Limitations of conventional treatment and dosage forms prompted the investigation of novel approaches that combine efficacy, selectivity and fewer adverse effects. These are the main promises of nanomedicine, generally defined as the application of nanotechnology to the biomedical field. Despite the considerable advances over the years and the large number of publications resulting from the growth of nanomedicine, there are still many hurdles and unknown factors limiting its successful translation from promise to reality. These factors range from the lack of standardization in the terminology and regulations, to the lack of understanding of interactions between nanocarriers and the biological system, and their influence on drug pharmacodynamics. In this manuscript, we aim at presenting and discussing concepts and terminologies used in the nanomedicine field and examining the main types of nanocarriers and their effects to modulate biological barriers and aid drug transport, bioavailability and targeting.