Matrix Metalloproteinases in Asthma-Associated Airway Remodeling – Dr. Jekyll or Mr. Hyde ?

Matrix metalloproteinases (MMPs) are Zn2+-dependent endoproteases, which digest extracellular matrix (ECM) components and various non-ECM molecules. Main physiological role of MMPs concerns regulation of tissue remodeling and regeneration. The production and activity of MMPs are tightly supervised by multistage control mechanisms. These mechanisms include regulation of gene expression, and various post-transcriptional/post-translational modifications. However, without proper control MMPs reveal dual nature, similarly to character from the novella by R.L. Stevenson, “Strange Case of Dr Jekyll and Mr Hyde”. They become dangerous molecules, involved in cancer metastasis, or cardiovascular diseases

Photometric flow system for the determination of serum lactate dehydrogenase activity

The routine method for LDH (Lactate dehydrogenase) activity determination is to monitor the increase of NADH concentration at 340 nm. There are some inconvenience in taking measurements in the near-UV region, especially in the case of serum samples analysis. In this work, two modifications of the routine LDH activity assay based on the use of reducing properties of NADH have been compared. Both methods involved the reduction of compounds that can be easily determined by well-known methods, ferric ion (with ferrozine) and nitrotetrazolium blue (NBT). A fully-mechanized Multicommutated Flow Analysis-Paired Emitter Detector Diode (MCFA-PEDD) system based on solenoid devices was developed and applied for both methods. The linear ranges obtained for Fe ferrozine and NBT methods are 6.0-200.0 U L -1 and 10.0-250.0 U L -1 with estimated detection limits at 0.2 U L -1 and 4.5 U L -1 , respectively. The low LOQ values enabled 10-fold sample dilutions, which is advantageous for samples with limited available volume. The Fe-ferrozine method is more selective for LDH activity in the presence of glucose, ascorbic acid, albumin, bilirubin, copper and calcium ions than NBT method. To confirm the analytical usefulness of the proposed flow system, the analysis of real human serum samples was carried out. The statistic tests showed satisfactory correlation between the results obtained for both developed methods and those received using the reference method.Polish National Science Centr

Label-free smartphone quantitation of bacteria by darkfield imaging of light scattering in fluoropolymer micro capillary film allows portable detection of bacteriophage lysis

Conventional methods for the detection and quantitation of bacteria are slow, laborious and require a laboratory. Microfluidic systems offer faster and portable testing, and smartphone cameras can record colorimetric or fluorometric bioassays, but this requires dye addition. Here, we demonstrate for the first time label-free smartphone detection of bacterial light scattering by darkfield microfluidic imaging to measure bacteria and bacteriophage lysis. A single LED and portable 3D printed imaging box allowed bacterial concentration and growth to be measured by direct imaging of bacterial light scattering. Bacteriophage lysis was detected within a 10-channel microfluidic device made from melt-extruded fluoropolymer micro capillary film, allowing rapid detection of host specificity. Elimination of unwanted reflections and optimising illumination angle are critical for successful darkfield bacterial imaging, with 15° giving maximal intensity. Bacterial sedimentation was directly observed within microfluidic devices, and detection sensitivity significantly increased by allowing bacteria to sediment for 30 min. With this simple, low-cost, 3D printed system bacterial concentrations down to an optical density of 0.1 could be measured corresponding to 8 × 104 colony forming units (CFU) per microdevice, approaching the sensitivity of conventional spectrophotometers. Bacteriophage lysis could be detected at a range of starting cell concentrations. With a low starting cell concentration, the increase in light scatter signal with incubation was prevented in the presence of bacteriophage. Conversely, with high starting cell concentration, the light scatter signal detected at the start was clearly eliminated when phage were added, indicating this simple system allows direct visualisation of bacteriophage eliminating light scattering by lysis

Mikrosolenoidowe przepływowe systemy bioanalityczne do oznaczania wybranych analitów istotnych w diagnostyce medycznej

Koncki, RobertBobilewicz, DagnaLink archiwalny https://depotuw.ceon.pl/handle/item/138

Diagnostic Value of IP-10 Level in Plasma and Bronchoalveolar Lavage Fluid in Children with Tuberculosis and Other Lung Diseases

Objectives: IP-10 has been proposed as a new diagnostic biomarker for Mycobacterium tuberculosis infection (MTBI). However, data on IP-10 concentration in bronchoalveolar lavage fluid (BALF) for pediatric tuberculosis are lacking. Aim: To determine IP-10 levels in unstimulated BALF and plasma in children with and without MTBI. Methods: IP-10 concentrations in BALF and plasma were measured in children hospitalized with suspected tuberculosis or other respiratory disease and scheduled for bronchoscopy. Thirty-five children were enrolled: 13 with suspected tuberculosis and 22 controls. The association between IP-10 and age was examined. Results: The IP-10 expression was increased in BALF compared to plasma (p = 0.008). We noticed higher BALF IP-10 levels in children with asthma, interstitial lung disease, and lung anomaly than in children with MTBI and other respiratory tract infections, but the differences were statistically insignificant. There was a moderate correlation between plasma and BALF IP-10 concentrations (rs = 0.46, p = 0.018). No correlation between IP-10 level and age was detected. Conclusions: IP-10 is detectable in unstimulated BALF in children with respiratory diseases, reaches higher concentrations in unstimulated BALF vs plasma, and does not correlate with age. However, it could not discriminate MTBI from other respiratory diseases

Mikrosolenoidowe przepływowe systemy bioanalityczne do oznaczania wybranych analitów istotnych w diagnostyce medycznej

Diagnostyka medyczna jest procesem rozpoznania chorób na podstawie stwierdzonych objawów przy pomocy wielu rodzajów badań. Jednym z obszarów diagnostyki medycznej jest analiza kliniczna. Wynika to ze znaczenia i wkładu wyników otrzymanych podczas badań laboratoryjnych w toku identyfikacji zespołów chorobowych. Znaczna większość podejmowanych decyzji medycznych jest oparta właśnie o jakościowe lub/i ilościowe dane analityczne. Od technik pomiarowych wprowadzanych na potrzeby analityki klinicznej wymagany jest szereg parametrów, które przyczynią się do coraz efektywniejszego oznaczania wielu, różnorodnych analitów w złożonych matrycach materiałów biologicznych. Dlatego też wysoka precyzja, powtarzalność i odtwarzalność oraz zmniejszenie kosztów i czasu przeprowadzanych analiz to główne parametry charakteryzujące nowoczesne układy pomiarowe. W dużym stopniu zostało to osiągnięte poprzez wprowadzenie zmechanizowania lub zautomatyzowania procedur analitycznych i zaimplementowania ich w systemach klinicznych. Jednym z trzech głównych typów analizatorów dostępnych w laboratoriach klinicznych, poza analizatorami wirówkowymi i dyskretnymi, są analizatory przepływowe. Koncepcja analiza przepływowej powstała w późnych latach pięćdziesiątych jako odpowiedź na problemy analizy klinicznej (do których głównie należy zaliczyć niewielką ilość przeprowadzanych oznaczeń oraz niską precyzję pomiarów) i od samego początku była prężnie rozwijana. Dowodem tego są powstałe na jej podstawie liczne techniki z wykorzystaniem coraz to wydajniejszych urządzeń do sterowania przepływem. Zalicza się do nich między innymi technika z zastosowaniem urządzeń mikrosolenoidowych (określana w literaturze anglojęzycznej jako MCFA – MultiCommutated Flow Analysis). Celem prezentowanych badań była konstrukcja zmechanizowanych układów analitycznych typu MCFA z detekcją optoelektroniczną typu PEDD (Paired Emitter Detector Diode) do oznaczania wybranych biomarkerów w płynach ustrojowych, istotnych z punktu widzenia diagnostyki medycznej, w tym: enzymów (fosfataza kwaśna i zasadowa), metabolitów (kreatynina), białka całkowitego oraz frakcji albuminowej. Dodatkowo, elastyczność i uniwersalność mikrosolenoidowych układów multikomutacyjnych pozwoliła na sprawne i efektywne analizowanie próbek o bardzo złożonej matrycy (mocz, surowica, płyn mózgowo-rdzeniowy). W pracy została przedstawiona budowa, zasada działania oraz charakterystyka opisywanych systemów wraz z potwierdzeniem eksperymentalnym ich przydatności analitycznej do zastosowania w analizie klinicznej.Medical diagnostics is the process of disease recognition based on the symptoms observed with many types of research. One of the areas of medical diagnostics is clinical analysis. The contribution and importance of the results obtained in clinical laboratories are significant in identification process of syndromes. The vast majority of medical decisions are based on the qualitative and / or quantitative analytical data. From analytical techniques, which are introduced for the clinical analysis purpose, a number of parameters is required to efficient determination of many analytes in complex biological matrices. Therefore, high precision, repeatability, reproducibility and reduction of costs and time of the analysis are the main parameters that characterize modern measuring systems. It has been achieved by the introduction of mechanization or automation of analytical procedures and implementation them in clinical systems. One of the three main types of analyzers available in clinical laboratories, besides centrifugal and discrete analyzers are flow analyzers. The concept of flow analysis was introduced in the late fifties as an answer to the problems of clinical analysis (which mainly include a small amount of performed determinations and low precision of measurements). From the very beginning, flow analysis has been rapidly developed. A number of techniques with the use of increasingly efficient flow control devices have been invented on the basis of flow analysis concept. One of them is MultiCommutated Flow Analysis (MCFA) - the technique using solenoid flow control devices. The aim of this study was to construct mechanized analytical MCFA systems with optoelectronic PEDD (Paired Emitter Detector Diode) detection for the determination of selected biomarkers in body fluids, significant from medical diagnostics point of view, including enzymes (acid phosphatase and alkaline), metabolites (creatinine ), total protein and albumin. In addition, the flexibility and versatility solenoid multicommutation systems allowed for efficient and effective sample analysis of very complex matrices (urine, serum, cerebrospinal fluid). In presented dissertation, design, principle of operation and characterization of described systems is presented with experimental confirmation of their analytical usefulness in clinical analysis

A remote-controlled immunochemical system for nephelometric detection of human serum transferrin

127Nie dotycz