Engineering a Point-of-Care Paper-Microfluidic Electrochemical Device Applied to the Multiplexed Quantitative Detection of Biomarkers in Sputum.

Health initiatives worldwide demand affordable point-of-care devices to aid in the reduction of morbidity and mortality rates of high-incidence infectious and noncommunicable diseases. However, the production of robust and reliable easy-to-use diagnostic platforms showing the ability to quantitatively measure several biomarkers in physiological fluids and that could in turn be decentralized to reach any relevant environment remains a challenge. Here, we show the particular combination of paper-microfluidic technology, electrochemical transduction, and magnetic nanoparticle-based immunoassay approaches to produce a unique, compact, and easily deployable multiplex device to simultaneously measure interleukin-8, tumor necrosis factor-α, and myeloperoxidase biomarkers in sputum, developed with the aim of facilitating the timely detection of acute exacerbations of chronic obstructive pulmonary disease. The device incorporates an on-chip electrochemical cell array and a multichannel paper component, engineered to be easily aligned into a polymeric cartridge and exchanged if necessary. Calibration curves at clinically relevant biomarker concentration ranges are produced in buffer and artificial sputum. The analysis of sputum samples of healthy individuals and acutely exacerbated patients produces statistically significant biomarker concentration differences between the two studied groups. The device can be mass-produced at a low cost, being an easily adaptable platform for measuring other disease-related target biomarkers.

Validation of discriminant functions from the rib necks in two Portuguese adult identified populations.

Sexual dimorphism is the basis for sex estimation research in skeletal remains, helping the positive identification of individuals in forensic backgrounds. In this regard, it has been proved that the metrical analysis of the costal neck of the first four ribs in Spanish contemporary adult skeletons shows high sexual dimorphism, especially in the first rib. However, the validation of any method developed in identified skeletal collections must be compulsorily reproduced in other skeletonized sets to verify either its potential application in individuals from other locations, or the existence of possible biases associated with inter-population variation. Due to the geographical and socioeconomic proximity between Spain and Portugal, this paper aims to check the utility of the discriminant functions designed by Partido-Navadijo et al. (2021) in the two skeletal collections of the University of Coimbra (Portugal). Results show the utility of these discriminant functions in Portuguese population, with frequencies of cases correctly assigned reaching up to 93.6% in the Coimbra Identified Skeletal Collection (CISC); and up to 92.6% in the 21st Century Identified Skeletal Collection (CEI/XXI). However, two particularities between the Spanish and Portuguese osteological assemblages can be highlighted: the possible existence of secular changes in relation to the CISC collection (1904-1938); and a reversal in the degree of costal dimorphism, being the third and fourth ribs of the CEI/XXI collection more dimorphic than the first ones.

Methods for sex estimation of prepubertal individuals (< 12 years old): bibliographic review and future directions.

Sex estimation is essential in both bioarchaeological and forensic research. Based on bibliographical research, this study aims to systematize and compare the existing methods and glance at the future on this research topic. The survey in Web of Science and Google Scholar revealed 37 papers describing the generation of original procedures, and 25 validation studies. The results show that the pelvis, namely the ilium, is the most analyzed anatomical region, followed by the deciduous teeth, the cranium, the mandible, and the long bones. The morphometric approaches are addressed in greater proportion, both as original methods (n = 19) or validations (n = 24). Only one of the 19 approaches that require expensive equipment (conventional radiology, computer tomography, geometric morphometrics and 3D modelling) was tested. Most of them were generated during the 21st century; however, 24 have not been tested yet. Comparing all the methods tested, the percentages of correctly estimated cases are always lower in validation studies. It is suggested that the most promising ones be tested in documented samples by a group of researchers. Thus, the interobserver error would be evaluated and may also lead to new methodological proposals and refinements.

Compact Microfluidic Platform with LED Light-Actuated Valves for Enzyme-Linked Immunosorbent Assay Automation.

Lab-on-a-chip devices incorporating valves and pumps can perform complex assays involving multiple reagents. However, the instruments used to drive these chips are complex and bulky. In this article, a new wax valve design that uses light from a light emitting diode (LED) for both opening and closing is reported. The valves and a pumping chamber are integrated in lab-on-a-foil chips that can be fabricated at low cost using rapid prototyping techniques. A chip for the implementation of enzyme-linked immunosorbent assays (ELISA) is designed. A porous nitrocellulose material is used for the immobilization of capture antibodies in the microchannel. A compact generic instrument with an array of 64 LEDs, a linear actuator to drive the pumping chamber, and absorbance detection for a colorimetric readout of the assay is also presented. Characterization of all the components and functionalities of the platform and the designed chip demonstrate their potential for assay automation.

Elemental Composition in Female Dry Femora Using Portable X-Ray Fluorescence (pXRF): Association with Age and Osteoporosis.

Pathophysiological conditions can modify the skeletal chemical concentration. This study analyzes the elemental composition in two anatomical regions from dry femoral bone using a portable X-Ray Fluorescence (pXRF) and evaluates its impact in the bone mineral density (BMD). The left femora of 97 female skeletons (21-95 years old individuals) from the Coimbra Identified Skeletal Collection were studied. Diagenetic biases were discarded at the outset and BMD was determined with Dual-energy X-ray absorptiometry. Chemical measurements were performed at the midpoint of the femoral neck and at the midshaft using a pXRF device, and comparisons were made considering the age and the BMD values. Only elements with a Technical Measurement Error ≤ 5% were selected: P, S, Ca, Fe, Zn, As, Sr, Pb and the Ca/P ratio. Statistically significant differences were found between regions, with higher concentrations of P, Ca, Zn and S at the midshaft, and the Ca/P ratio at the femoral neck. The concentration of P is higher in individuals < 50 years, while S and Ca/P ratio increase in individuals ≥ 50 years. The decrease of P with age can be simultaneously related to the decline of its concentration in osteoporosis. Decreased BMD is also associated with higher levels of S and Pb. Osteoporosis enhances the absorption of osteolytic elements in specific locations. This fast and non-destructive technique has proved effective for the comprehension of chemical changes related to bone mass loss. This study highlights the potential of identified skeletal collections to improve the knowledge about bone fragility.

Test of the auricular surface sex estimation method in fetuses and non-adults under 5 years old from the Lisbon and Granada Reference Collections.

Non-adult sex estimation is an active field of forensic inquiry as morphological variations between males and females are subtle, but observable, even from intrauterine development. The objectives of this study are threefold: to test the validity of the auricular surface method for sex estimation (Int J Osteoarchaeol 27:898-911, 2017) in fetuses and children under the age of 5 years old; to evaluate if health conditions, reported as the cause of death, influence its accuracy; and to detect possible secular trends in sexual dimorphism. One-hundred and ninety-seven skeletal individuals from the Lisbon and Granada Identified Collections were studied. Individuals were divided according to the hormonal peaks (< 0, 0-2, < 2, and 2.1-5 years old), cause, and year of death (before and after 1960). As in previous studies, two ratios (FI/CF and DE/AD) and two qualitative variables (OM and MRS) showed the highest frequencies of correct estimation (0.81-0.86). The correct sex allocations increased when the discriminant function (0.85) and logistic regression (0.86) were applied. Males of the age groups < 0 and 2.1-5 years were all correctly sexed by both formulae, and the same was observed for the female probabilities of adequate allocation. The cause and year of death were identified as variables without statistical significance. It is proposed that this method can be incorporated with confidence into the multifactorial laboratory protocols for non-adult sex estimation from skeletal remains.

Evaluation of the auricular surface method for non-adult sex estimation on the Lisbon documented collection.

OBJECTIVES: Sex is usually not estimated in skeletonized non-adult individuals because sexual dimorphism is considered minimal before puberty. In 2017, a new approach based on the shape of the auricular surface was proposed, showing that this anatomic area of the ilium is dimorphic. This study tests the reproducibility and evaluates the accuracy of the method in a bigger sample. MATERIALS AND METHODS: Thirty males and 31 females from the Lisbon Collection (1 day-18 years old) were analyzed by two researchers, one experienced, and the other without previous knowledge of the method. The sample was divided into three groups according to prepubertal and pubertal hormonal peaks (<2, 0-12, and 13-18 years old). Two metric variables (DE/AD and FI/CF ratios), two morphological features (Overall Morphology, OM and Morphology of the Retroauricular End of the Superior Demiface, MRS), and two formulae were tested based on the adequate results obtained in the original article. RESULTS: Data shows a low interobserver error (ICC > 0.92; K > 0.74). Morphological features provide better results than the metrics (DE/AD = 77.05%; FI/CF = 73.77%; OM = 80.33%; MRS = 85.24%). The discriminant function correctly classified 86.66% of the males and the logistic regression, 83.33%. The method was equally reliable in 0-12 and 13-18 years-old age groups and more accurate in males. The accuracy of most of the variables reached 100% for males under 2 years-old, and the probabilities were higher than for older individuals. The percentages of correct estimations are not influenced by the age, year, and cause of death. DISCUSSION: Despite the relatively small sample size, this study confirms the usefulness of the auricular surface for non-adult sexual estimation, especially for those younger than 2 years old. Additional validation tests in documented individuals from other geographic regions are suggested.

A possible Echinococcus granulosus calcified cyst found in a medieval adult female from the churchyard of Santo Domingo de Silos (Prádena del Rincón, Madrid, Spain).

Calcification, or mineralisation, can occur as part of a natural process, or by pathological processes. The purpose of this work is to examine an unidentified semi-spherical and perforate hollow mass, found near the pelvis of an adult female, dated 12th-13th century AD, exhumed of the Church of Santo Domingo de Silos (Prádena del Ricón, Madrid, Spain). The mass was examined by SEM and Energy Dispersive X-Ray Spectroscopy. These procedures revealed a heterogeneous inner surface with both smooth and irregular areas. A larger spherical and several smaller crescent-shaped perforations were noticed. X-ray microanalysis revealed the presence of the elements C, K, P, Ca, Al, Si, Fe, and Mg. The co-localisation of Ca and P suggests that they may be combined in a mineral matrix, likely formed in vivo. Other minerals probably came from the soil, although Fe could be related to the presence of blood. The macroscopic and microscopic appearances, chemical composition, and location of the calcified mass point to a possible hydatid cyst from Echinococcus granulosus, common in agricultural populations. This study used a suite of analytical techniques that are useful in the diagnosis of unknown calcified masses and can, therefore, be recommended for use in future analytical work.