Pregledni članek predstavi protokol obravnave novorojenčkov z anorektalno malformacijo od kliničnega pregleda, diagnostičnih metod do kirurškega zdravljenja, ki novorojenčku omogoča odvajanje blata. ...Zgodnja prepoznava in ustrezno zdravljenje omogočata optimalne možnosti za dober funkcionalni izid. Pregledali smo sodobno literaturo o obravnavi anorektalnih malformacij in zbrali okviren pregled naših izkušenj v Univerzitetnem kliničnem centru Ljubljana. Naši rezultati so posledica uspešnega sodelovanja multidisciplinarnega tima in spoštovanja protokola obravnave.
Detection of bacteria in bloodstream infections and their antibiotic susceptibility patterns is critical to guide therapeutic decision-making for optimal patient care. Current culture-based assays ...are too slow (>48 h), leading to excessive up-front use of broad-spectrum antibiotics and/or incorrect antibiotic choices due to resistant bacteria, each with deleterious consequences for patient care and public health. To approach this problem, we describe a method to rapidly isolate bacteria from whole blood using inertial microfluidics and directly determine pathogen identity and antibiotic susceptibility with hybridization-based RNA detection. Using the principle of Dean flow fractionation, bacteria are separated from host blood cells in a label-free separation method with efficient recovery of even low abundance bacteria. Ribosomal RNA detection can then be applied for direct identification of low abundance pathogens (~100 per mL) from blood without culturing or enzymatic amplification. Messenger RNA detection of antibiotic-responsive transcripts after brief drug exposure permits rapid susceptibility determination from bacteria with minimal culturing (~10(5) per mL). This unique coupling of microfluidic cell separation with RNA-based molecular detection techniques represents significant progress towards faster diagnostics (~8 hours) to guide antibiotic therapy.
Highlights • Engineered protein switches are increasingly used in molecular diagnostics. • Engineered protein switches can be used to sense and actuate cellular functions. • Empirical design rules to ...construct tailor-engineered protein switches are emerging. • Future synthetic signaling circuits will see networks of engineered protein switches.
Highlights • Nanoparticle designs and interactions in the body determine their individual behavior. • The collective behavior of nanoparticles can be exploited to improve their performance. • ...Bio-inspired mechanisms of cooperation include self-assembly and communication. • Systems approaches build on new nanomaterials, biological insight, and computation.
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Cancer cells show an abnormal balance of reduced/oxidised species and the detection of cancer redox balance can be exploited for diagnostic purposes. In particular, it is known that ...the expression of oncogenic Ras changes the intracellular oxidised/reduced glutathione balance. Electrochemical imaging of living cells is currently feasible by scanning electrochemical microscopy and was used here, using ferrocenemethanol as redox mediator, to measure the oxidised/reduced glutathione balance in human breast epithelial MCF10A cells expressing constitutively active Ha-Ras Val12 mutant compared to normal MCF10A cells. Oxidized ferrocenemethanol is reduced by glutathione and the resulting maps of current over cell cultures were different for transformed cells compared to normal cells. Furthermore, scanning electrochemical microscopy using ferrocenemethanol redox mediator was able to distinguish human lung carcinoma cells from the surrounding normal epithelium of clinical specimens. We propose that scanning electrochemical microscopy analysis using specific redox mediator such as ferrocenemethanol may become a useful tool in cancer diagnostics.
Lung cancer shows the highest mortality rate worldwide (1.3 million/year) with respect to other common cancers as prostate, breast and colon cancer. It is the second most common cancer in men and the ...third in women (22% of all cancer cases). The treatment of the disease is long and difficult with survival scarcely attains to 5 years. The most critical point for best prognosis is to diagnose the disease at its early stage. For this aim, divers’ methods have been used including chest X-ray, computerized tomography, magnetic resonance imaging, positron emission tomography, sputum cytology and biopsy. Although some of these methods are not suitable for all people due to the other pathologies that the patient may have. Also since the current diagnostic tools are also expensive and time consuming, hence, a new sensitive and rapid method for screening is necessary for lung cancer detection. With this approach, biosensors as diagnostics tools have provided a promising technology for the detection of many important disease biomarkers through rapid, sensitive and easy applications. In this paper, lung cancer, state-of-the-art with current diagnostics tools, genetic and protein biomarkers of the disease and emerging biosensor technology for rapid detection are reviewed.
•A sulfite reductase (MW; 116kDa) was purified to homogeneity from Escherichia coli.•Purified enzyme was successfully used for synthesis of gold nanoparticles.•Gold nanoparticles showed strong ...antifungal activity against human pathogenic fungi.•Gold nanoparticles had no obvious cytotoxicity for Vero and Hep-2 cell lines in vitro.•Bioactive non-toxic gold nanoparticles have potential to use in biology and medicine.
Biological synthesis of nanomaterials using microorganisms is a growing field of industrial microbiology due to the numerous beneficial properties while lower toxicity for biological systems. We have developed a cell-free viable approach for synthesis of gold nanoparticles using α-NADPH-dependent sulfite reductase purified from Escherichia coli. The enzyme was purified by ion exchange chromatography on DEAE Sephadex A-50 and its molecular weight was determined by gel filtration on Sephacryl S-300. The purified enzyme with molecular weight of 116kDa composed of two subunits of 75 and 41kDa was successfully used for cell-free synthesis of gold nanoparticles which were spherical with an average size of 10nm and a zeta potential of −30±0.2. Gold nanoparticles showed strong antifungal activity toward a wide range of human pathogenic fungi. Evaluation of the in vitro cytotoxicity of gold nanoparticles showed no toxicity for two cell lines, i.e., Vero and Hep-2 at the concentrations ranged from 0.31 to 10%. Taken together, biologically active non-toxic gold nanoparticles green synthesized by a sulfite reductase of E. coli in the present work are potential candidates for application in different disciplines of medicine and biology from diagnostics to drug delivery systems, combating infectious diseases and anticancer therapies.
In nature there are various specific reactions for which highly selective detection or support is required to preserve their bio-specificity or/and functionality. In this respect, mimics of cell ...membranes and bio-compartments are essential for developing tailored applications in therapeutic diagnostics. Being inspired by nature, we present here biomimetic nanocompartments with ion-selective membrane permeability engineered by insertion of ionomycin into polymersomes with sizes less than 250 nm. As a marker to assess the proper insertion and functionality of ionomycin inside the synthetic membrane, we used a Ca(2+)-sensitive dye encapsulated inside the polymersome cavity prior to inserting the biopore. The calcium sensitive dye, ionomycin, and Ca(2+) did not influence the architecture and the size of polymersomes. Successful ionomycin functionality inside the synthetic membrane with a thickness of 10.7 nm was established by a combination of fluorescence spectroscopy and stopped-flow spectroscopy. Polymersomes equipped with ion selective membranes are ideal candidates for the development of medical applications, such as cellular ion nanosensors or nanoreactors in which ion exchange is required to support in situ reactions.
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