Search Results (21)

Real-time PCR (qPCR) is today’s definitive quantitative technology in molecular biology and diagnostics. Until 30 years ago, PCR product analyses were generally performed after amplification using gel-based methods. Quantification typically relied on visual inspection or densitometry of end-point products and was therefore relatively unreliable and poorly suited to high-throughput automation. To celebrate real-time PCR’s 30-year anniversary of commercial availability, Professor Stephen Bustin, Guest Editor for the special edition, “Advancing Molecular Science Through Reproducible qPCR: MIQE Guidelines and Beyond,” asked Russell Higuchi to give a historical account on how his idea of real-time PCR was conceived and brought to fruition. Dr. Higuchi then asked his collaborator, Lincoln McBride, who drove the development of the ABI 7700—the high-throughput real-time PCR instrument that gave researchers access to this technology—to co-author this dual memoir. This story is told from the perspectives of the two scientists most directly responsible for making real-time PCR practical and widely accessible. Taking turns, Russell Higuchi describes the conceptual and experimental steps at Cetus and then Roche that led from homogeneous PCR detection to continuous fluorescence monitoring, whilst Lincoln McBride details ABI’s parallel efforts to commercialize Russ’s invention. Together, they trace how experimental insight, engineering constraints, product development, and commercial decision-making shaped the Applied Biosystems 7700 Sequence Detection System and established real-time PCR as a practical and reliable quantitative technology. Their team’s efforts persevered through technological uncertainty and within a complex corporate collaboration. They share key historical documents in their original form. Their accounts show how the 7700 system emerged as the convergence of chemistry, optics, software, and product development. The eventual global reliance on real-time PCR during the COVID-19 pandemic demonstrated, at unprecedented scale, the profound and enduring impact of these early technical and organizational choices. Full article

Objectives: The aim of this study was to evaluate changes in corneal topography, wavefront aberrations, and densitometry after superficial keratectomy (SK) for Salzmann nodular degeneration (SND). Methods: This was an observational retrospective study. Pre- and post-operative data, including best spectacle-corrected visual acuity (BCVA), refraction, and Pentacam^® topography scans, were analysed. Changes in keratometry (K), wavefront aberrations, and corneal optical densitometry (COD) were evaluated. Results: A total of 21 eyes of 17 patients who underwent SK for SND were included. At an average follow-up of 6.3 ± 4.4 months, there was a significant reduction in RMS (root mean square) values for total wavefront aberrations after surgery (mean reduction: −3.89 ± 4.08 μm, p = 0.001), lower-order aberrations (mean reduction: −3.47 ± 3.52 μm, p = 0.001), and higher-order aberrations (mean: −0.42 ± 0.58 μm, p < 0.001). There was a significant improvement in BCVA (mean change: 0.27 ± 0.31 logMAR, p < 0.001) and spherical equivalent (mean change: −2.09 ± 2.73 D, p = 0.002), and a reduction in refractive cylinder (mean reduction: −0.85 ± 1.14 D, p = 0.008). There was a significant reduction in corneal astigmatism (mean reduction 1.04 ± 2.88 D; p = 0.041), and an increase in flat keratometry (mean change 1.46 ± 3.10 D; p = 0.029). Lastly, there was a significant decrease in total COD values in all zones except for the central 2 mm (p < 0.05) and in the overall total 12 mm cornea (p = 0.035). Conclusions: SK for SND resulted in better visual acuity and potentially improved quality as suggested by the reduction in corneal aberrations and improvement in corneal regularity and transparency. Full article

The genus Pavona includes massive to submassive hermatypic corals and represents one of the main reef builders of the coral reefs in the Eastern Tropical Pacific (ETP). However, its development and specific ecological role, particularly on offshore reefs (e.g., oceanic Atolls), remain poorly understood. This study aims to determine the sclerochronological characteristics of the four Pavona species (Pavona duerdeni, Pavona clavus, Pavona maldivensis, and Pavona varians) in Clipperton Atoll, and their contributions to reef maintenance. Using the optical densitometry technique, Pavona annual growth parameters were obtained, showing that skeletal density (1.26 ± 0.23 g cm⁻³), extension rate (0.94 ± 0.31 cm year⁻¹), and calcification rate (1.17 ± 0.36 g cm⁻² year⁻¹) were consistent with previous data from the ETP. However, differences at the species level showed that P. duerdeni, P. varians, and P. maldivensis invested their calcification resources into building denser skeletons, demonstrating the morphological plasticity of the genus, likely driven by local factors, such as strong hydrodynamics and depth, rather than regional conditions (e.g., El Niño-Southern Oscillation events). Pavona’s growth strategies contribute to the preservation potential on a geological timescale of Clipperton Atoll, highlighting their importance as one of the main reef builders at a massive coral-dominated reef. Full article

Background and Objectives: Descemet stripping automated endothelial keratoplasty (DSAEK) is a widely used surgical technique for treating corneal endothelial dysfunctions such as Fuchs endothelial corneal dystrophy (FECD) and pseudophakic bullous keratopathy (PBK). This study aimed to investigate the association between postoperative visual acuity and various corneal morphological and optical parameters, including corneal densitometry (CD) and higher-order aberrations (HOAs), measured using the Pentacam (OCULUS Optikgeräte GmbH, Wetzlar, Germany), as well as graft thickness, which was assessed by anterior segment optic coherence tomography (AS-OCT), (Optovue Inc., Fremont, CA, USA), and corneal thickness, assessed by both AS-OCT and Pentacam. Materials and Methods: This prospective, observational cohort study included 17 eyes from 13 patients who underwent DSAEK. Data on preoperative visual acuity were collected, while postoperative parameters were assessed during follow-up visits. Corneal measurements included the total corneal and corneal graft thickness, corneal densitometry in 20 defined subregions, and corneal higher-order aberrations. Associations between these parameters and postoperative visual acuity were evaluated using nonparametric statistical tests. Results: The postoperative visual acuity improved significantly (p < 0.001). Strong correlations were found between poorer visual acuity and higher CD values. The strongest correlations with visual acuity were found for CD 2–6 mm total (Rho = 0.795; p < 0.001), CD central 2–6 mm (Rho = 0.791; p < 0.001), and CD central 0–2 mm (Rho = 0.730; p < 0.001). Significant associations were also observed with anterior and posterior HOAs (Rho = 0.624, p = 0.01; and Rho = 0.556, p = 0.02, respectively). No correlation was found between visual outcomes and graft thickness measured by AS-OCT (Rho = 0.051; p = 0.85), nor with total corneal thickness measured by AS-OCT (Rho = −0.227; p = 0.38) or Pentacam (Rho = −0.369; p = 0.14). Conclusions: This study demonstrates that CD and HOAs are more strongly associated with postoperative visual acuity after DSAEK than traditionally monitored parameters such as graft or corneal thickness. The results highlight the value of detailed corneal imaging and support the use of advanced optical diagnostics in postoperative evaluation. Full article

(1) Background: A decrease in bone mineral density has been noted not only in at-risk patients (e.g., postmenopausal women) but also in young and middle-aged individuals due to changes in lifestyle. The aim of the study was to find a possible correlation for dual-energy X-ray absorptiometry (DXA) and quantitative computed tomography (QCT) with cone beam computed tomography (CBCT) of the jaws. (2) Methods: A total of 24 patients (14 women and 10 men aged 25 to 50 years) with partial secondary tooth loss and vitamin D insufficiency underwent cone beam computed tomography of the jaws and skeletal mineral density assessment using DXA (n = 12) and QCT (n = 12). (3) Results: When conducting CBCT of the jaws, a predominance of bone tissue type D3 (350–850 Hu) on the upper jaw (p > 0.05 (F = 0.68) and D2 (850–1350 Hu) on the lower jaw (p > 0.05 (F = 1) was revealed. According to the results of QCT densitometry of the skeleton, signs of osteopenia were found in four patients (with vitamin D3 deficiency) (33%) according to DXA; signs of osteopenia were found in six patients (with severe deficiency and deficiency of vitamin D3) (50%). The difference between QCT and DXA was not significant (p > 0.05) for each group. The significant strong correlation between CBCT and DXA or QCT was not found (p > 0.05). (4) Conclusions: Primary changes in bone density can be detected earlier in the dental system using cone beam computed tomography of the jaws. At the same time, the question of using a specific densitometry method—DXA or QCT—remains open, as their results correlating with CBCT optical density was not approved. Full article

Some studies have used physical techniques for the assessment of bone structure and composition. However, very few studies applied multiple techniques, such as those described below, at the same time. The aim of our study was to determine the chemical and mineralogical/organic composition of bovine tibial bone samples using geophysical/geochemical reference techniques. X-ray diffraction (XRD), thermogravimetry (TG), Fourier-transform infrared spectrometry with attenuated total reflectance accessory (FTIR-ATR), inductively coupled mass spectrometry (ICP-MS) and inductively coupled optical emission spectrometry (ICP-OES) were applied to measure the organic and inorganic composition of 14 bovine bone samples. In addition, peripheral quantitative CT (QCT) was used to assess BMD in these bones. We were able to define the total composition of the studied bone samples. ICP-OES and ICP-MS techniques were used to determine the major and trace element composition. The X-ray analysis could detect inorganic crystalline compounds of bones, such as bioapatite, and its degree of ordering, indicating whether the bones belong to a younger or older individual. The total volatile content of the samples was calculated using TG and resulted in about 35 weight% (wt%). This, together with the 65 wt% total resulting from the chemical analysis (i.e., inorganic components), yielded a total approaching 100 wt%. As a large portion of the volatile content (H₂O, CO₂, etc.) was liberated from the organic components and, subordinately, from bioapatite, it could be concluded that the volatile-to-solid ratio of the examined bone samples was ~35:65. The FTIR-ATR analysis revealed that the organic portion consists of collagens containing amide groups, as their typical bands (OH, CH, CO, NC) were clearly identified in the infrared spectra. Numerous parameters of bone composition correlated with BMD as determined by QCT. In conclusion, we performed a complex evaluation of bovine bones to test multiple geophysical/geochemical techniques in bone research in association with QCT bone densitometry. From a medical point of view, the composition of the studied bones could be reliably examined by these methods. Full article

The aim of this study was to investigate whether PolyJet technology, which uses rubber-like materials for printing and is known for its high resolution and performance, could be suitable for producing flexographic printing plates. In our research, we designed test plates that were printed using PolyJet technology with TangoBlackPlus FLX9870-DM resin. These 3D-printed plates were evaluated for their resistance to various flexographic inks and solvents, and their contact angles were measured. Subsequently, the prints were made on a Flexiproof device using water-based ink with both the test plates and traditional photopolymer plates across six different substrates. The print quality was assessed using densitometry and spectrophotometry. Our findings indicate that the 3D-printed plates are suitable for printing solid areas and lines with water-based inks. However, the print quality of the 3D-printed plates is slightly lower than that of the photopolymer plates, with the optical density values for the high-quality prints on coated papers being approximately 10% lower. Additionally, the plates printed with TangoBlack Plus resin appear to be suitable for UV inks due to their high resistance, but they are not resistant to the solvents used in solvent-based inks. Full article

Background: To compare corneal densitometry (CD) patterns at the graft–host interface between Descemet Membrane Endothelial Keratoplasty (DMEK) and Descemet Stripping Automated Endothelial Keratoplasty (DSAEK). Corneal densitometry is a quantitative assessment that objectively evaluates corneal clarity and optical quality by measuring the light backscatter from the cornea. Methods: Fifty-one eyes that received DMEK or DSAEK surgery for corneal endothelium dysfunction were evaluated. The primary endpoint included CD patterns at the graft–host interface, which were assessed by the Pentacam HR device at the center point of the corneal horizontal meridian (CD_central), and at six points on the central circumference of the cornea (with a total diameter of 4 mm) (CD_{I,II,III,IV,V,VI}). Secondary endpoints included the best-corrected distance visual acuity (BCDVA), central corneal thickness (CCT), and graft thickness (GT). All of the evaluations were performed at follow-up appointments one, three, six and twelve months after the procedure. Results: DMEK showed a significant overall CD reduction of −7.9 ± 8.5 grayscale unit (GSU) compared to DSAEK (p < 0.001). In addition, the DMEK group showed significantly lower CD_{Central,I,II,III,IV,V,VI} values at follow-up appointments one, three, six and twelve months after the procedure compared to the DSAEK group (p < 0.001). BCDVA, CCT and GT were in favor of the DMEK group with a mean value of 0.39 ± 0.35 LogMar, 552.2 ± 71.1 µm and 11.03 ± 1.4 µm, respectively (p < 0.001). Conclusions: CD patterns at the graft–host interface seem to be different depending on the endothelial keratoplasty procedure. This provides specific insight into CD changes in this critical region of surgery, which may provide a better understanding of the postoperative evolution of these patients. Full article

During operation, cables of overhead power lines (OPLs) are exposed to the impact that differs in separate parts of the OPL span due to the different responses of cables near the clamps and far from them. This paper presents the results of a study of aluminum and steel wires cut from such separate parts of ACSR cables before and after exploitation. Structural, microstructural, and elastic–microplastic properties of wires and their changes during operation were studied through optical microscopy, energy-dispersive X-ray microanalysis, electron backscattering diffraction, X-ray diffraction, densitometry, and acoustic measurements. The characteristics of the properties of the wires along the span were found to change in a coordinated manner. Numerical estimates of the influence of the steel core on aging the ACSR cable were obtained. Changes in the properties of the wires, as well as oxidization and corrosion of their near-surface layers, were studied in detail. Quantitative values of the characteristics of properties, the most distant from those observed in the new wires, were revealed for samples of aluminum and steel wires cut from the cable at 1/4 span and near clamps. It is assumed that these cable parts should be the most crucial for cable durability. Full article

A pressing health problem, both in clinical and socio-economic terms, is the increase in the number of patients with liver damage caused by viral diseases (hepatitis), cancer, toxicological damage, or metabolic disorders. Liver function assessment is a complex task, for which various existing diagnostic methods are used. Unfortunately, they all have several limitations which frequently make prompt and accurate diagnosis impossible. The high level of disability and mortality caused by liver diseases makes the development of new liver diagnostic methods very urgent. In this paper, we describe a new joint methodology for studying liver function based on optical densitometry and dynamic light scattering. This will help to diagnose and predict the dynamics of liver function during treatment with greater efficiency, due to including in consideration the individual characteristics of the cardiovascular system and tissue metabolism. In this paper, we present a laboratory model of a combined sensor for optical densitometry and dynamic light scattering. We also developed special software for controlling the sensor and processing the recorded data. Modeling experiments and physical medical studies were carried out to adjust and calibrate the sensor and software. We also assessed the sensor resolution when registering the concentration of dye in the human body and the minimum measured flow rate. Full article

This study aimed to assess clinical application of the Scheimpflug corneal tomography for objective evaluation of corneal optical density in eyes undergoing Descemet’s stripping endothelial keratoplasty (DSEK). In this prospective study, 39 pseudophakic eyes with bullous keratopathy were enrolled. All eyes underwent primary DSEK. Ophthalmic examination included best corrected visual acuity (BCVA) measurement, biomicroscopy, Scheimpflug tomography, pachymetry, and endothelial cell count. All measurements were taken preoperatively and within a 2-year follow-up period. Gradual BCVA improvement was observed in all patients. After two years, the mean and median BCVA values were 0.18 logMAR. A decrease in central corneal thickness was noted only during the first 3 months postoperatively and was followed by a gradual increase. Corneal densitometry decreased constantly and most significantly in the first 3 months postoperatively. The consecutive decrease in endothelial cell count of the transplanted cornea was most significant during the first 6 months postoperatively. Six months postoperatively, the strongest correlation (Spearman’s r = −0.41) with BCVA was found for densitometry. This tendency was maintained throughout the entire follow-up period. Corneal densitometry is applicable for objective monitoring of early and late outcomes of endothelial keratoplasty, showing a higher correlation with visual acuity than pachymetry and endothelial cell density. Full article

The diverse application potential of metal-organic framework (MOF) materials are currently limited by their challenging and complicated preparation processes. In this study, we successfully developed a novel strategy for the rapid synthesis of a sustainable MOF composite membrane under neutral conditions with improved physicochemical and antibacterial properties. Our reaction pipeline comprised visible light that induced the production of reactive oxygen species (ROS) from ZIF-8 particles, which facilitated the rapid oxidative polymerization of dopamine to polydopamine. The physicochemical properties of the composite membrane were assessed using imaging methods, including scanning and transmission electron microscopy, X-ray photoelectron spectrometry, and nitrogen adsorption/desorption; its antibacterial effects against Staphylococcus aureus, Escherichia coli, and Pseudomonas aeruginosa were measured using optical densitometry. The bactericidal potency of the synthesized membrane was >99% against all tested strains under the conditions of simulated sunlight. Moreover, the composite membrane retained its structural integrity and antibacterial effect after multiple cycles of use and recovery, showcasing remarkable stability. Overall, this study displays a ROS-mediated method for the rapid preparation of sustainable MOF composite membranes under neutral conditions with optimal physicochemical characteristics, antibacterial properties, and performance. Our study provides insights into the use of membrane materials as design platforms for a range of diverse practical applications. Full article

Purpose: To evaluate the radiation effect of fractionated robotic radiotherapy of benign tumors located in the parasellar region on the anterior and posterior segments of the eye. Methods: A prospective observational study based on the expanded ophthalmological examination. The pre-treatment baseline was used as a control for the post-radiotherapy follow-up examinations. The study group consists of 34 patients (68 eyes) irradiated using the CyberKnife system. There were ten patients with cavernous sinus meningioma, nine with pituitary adenoma, five with meningioma of the anterior and middle cranial fossa, five with meningioma in the region close to optic chiasm, three with craniopharyngioma, and two with meningioma of the orbit. All patients were treated using three fractions of 600–800 cGy. We assessed the impact of radiation on the eye based on changes in anatomical and functional features. The condition of the eye surface, central corneal thickness (CCT), endothelial cell density (ECD), lens densitometry, central macular thickness (CMT), and retinal nerve fiber layer (RNFL) were the anatomical features assessed. The functional tests were best-corrected visual acuity (BCVA), intraocular pressure (IOP), visual field (VF) and visual-evoked potentials (VEP). An ophthalmologic examination was performed before and 6, 12, 18, and 24 months after radiotherapy. Results: We did not observe any significant changes in BCVA, IOP, CCT, CMT, VF, and VEP, nor in the slit-lamp examination during the two-years observation. We found a significant decrease in ECD at all follow-up measurements. The drop in ECD exceeded approximated age-related physiological loss. The reduction in ECD was not large enough to disrupt corneal function and thus affect vision. We also observed a statistically significant reduction of RNFL in all observation time points. However, there was no correlation between the dose delivered to the optic pathway and the decrease in RNFL thickness. The thinning of the RNFL was not significant enough to impair visual function. Conclusion: Fractionated robotic radiotherapy of the tumors located close to the optical pathway is safe and does not impair patient’s vision. Minor changes found in optic nerve anatomy (RNFL thinning) might be related to radiation effect or tumor compression. The causal relation between low doses of radiation delivered to the cornea and the observed significant but slight decrease in ECD is uncertain. The observed changes did not cause visual disturbances perceivable by the patients. Full article

Background: Angiogenesis is a crucial process in physiological maintenance and tissue regeneration. To understand the contribution of angiogenesis, it is essential to replicate this process in an environment that reproduces the biochemical and physical properties which are largely governed by the extracellular matrix (ECM). We investigated vascularization in cardiac left ventricular ECM hydrogels to mimic post-myocardial repair. We set out to assess and compare different destructive and non-destructive methods, optical as well as non-optical, to visualize angiogenesis and associated matrix remodeling in myocardial ECM hydrogels. Methods: A total of 100,000, 300,000, and 600,000 Human Pulmonary Microvascular Endothelial Cells (HPMEC) were seeded in left ventricular cardiac ECM hydrogel in 48-well plates. After 1, 7, and 14 days of culture, the HPMEC were imaged by inverted fluorescence microscopy and 3D confocal laser scanning microscopy (Zeiss Cell Discoverer 7). In addition, cell-seeded ECM hydrogels were scanned by optical coherence tomography (OCT). Fixed and paraffin-embedded gels were thin-sectioned and assessed for ECM components via H&E, picrosirius red histochemical staining, and immunostaining for collagen type I. ImageJ-based densitometry was used to quantify vascular-like networks and GraphPad was used for statistical analyses. Results: Qualitative analyses were realized through fluoromicrographs obtained by the confocal laser scanning microscope which allowed us to visualize the extensive vascular-like networks that readily appeared at all seeding densities. Quantification of networks was only possible using fluoromicrographs from inverted microscopy. These showed that, after three days, the number of master junctions was seeding density-dependent. The resolution of optical coherence tomography was too low to distinguish between signals caused by the ECM and cells or networks, yet it did show that gels, irrespective of cells, were heterogeneous. Interestingly, (immuno)histochemistry could clearly distinguish between the cast cardiac-derived matrix and newly deposited ECM in the hydrogels. The H&E staining corroborated the presence of vascular-like network structures, albeit that sectioning inevitably led to the loss of 3D structure. Conclusions: Except for OCT, all methods had complementary merit and generated qualitative and quantitative data that allowed us to understand vascular network formation in organ-derived ECM hydrogels. Full article

The purpose of this study is to quantitatively assess lens opacity, using a swept-source optical coherence tomography (SS-OCT) device for anterior segment assessment, and establish the correlation with Scheimpflug imaging, corrected distance visual acuity (CDVA) and cumulative dissipated energy (CDE). This prospective cross-sectional single-center study enrolled 51 patients (51 eyes) with crystalline lens opacity. Patients with previous ocular surgery, pathologies or general disorders affecting vision were excluded. Eyes were scanned with an SS-OCT device, and lens densitometry was automatically analyzed using a custom MATLAB script which examined lens density, nuclear density and linear density. The same analyses were performed on Scheimpflug images. Preoperative CDVA and CDE during phacoemulsification were recorded. Spearman’s (ρ) and Pearson’s (r) correlation coefficients were assessed according to data normality. Statistically significant correlations were established between SS-OCT and Scheimpflug imaging using lens analysis (ρ = 0.47, p < 0.001), nuclear analysis (ρ = 0.73, p < 0.001) and linear analysis (r = 0.44, p < 0.001). A significant correlation with CDE was found with all the SS-OCT methods (r = 0.57, p < 0.001). Only the nuclear analysis of the SS-OCT scans (T_b = −0.33, p < 0.01) and Pentacam Nucleus Staging (T_b = −0.26, p < 0.05) showed a statistically significant correlation with CDVA. Good inter-device agreement in lens densitometry was found. However, SS-OCT yielded improved lens imaging compared with the Scheimpflug device and a higher correlation with clinical parameters. Thus, high-resolution SS-OCT has the potential to become a preferable option for automatic cataract grading and preoperative planning. Full article