Monomorphic epitheliotropic intestinal T-cell lymphoma (MEITL) complicated with intestinal obstruction and perforation.

We report the case of a patient who comes to the emergency department with abdominal pain, oral intolerance and bloody stools, being diagnosed with intestinal obstruction secondary to a jejunal tumor with contained perforation. A laparotomy was performed with resection of the jejunal tumor and taking biopsies from lesions with similar characteristics in the rest of the small bowel compatible with monomorphic epitheliotropic intestinal T-cell lymphoma (MEITL). The monomorphic epitheliotropic intestinal T-cell lymphoma is a rare intestinal tumor with a poor prognosis that is characterized by the proliferation of intraepithelial lymphocytes. The abdominal symptoms are nonspecific and the known major complications are intestinal perforation and obstruction. There is no standard therapeutic approach, being a combination of surgical resection, chemotherapy and autologous stem cell transplant.

Europium(III)-Doped Gadolinium(III) Complex for High-Sensitivity Temperature Sensing in the Physiological Range.

A new Eu3+-doped Gd3+ complex of formula [Eu0.0135Gd0.9865(pta)3me-phen] was synthesized and structurally characterized (Hpta = benzoyltrifluoroacetone, me-phen = 5-methyl-1,10-phenanthroline). The photoluminescence study revealed that when the compound was excited at RT, under a 457 nm continuous laser, the material exhibited high luminescence due to the antenna effect of the ligands, as well as a good balance between the phosphorescence from the spin-forbidden triplet (from the organic ligands), and the characteristic lanthanide f-f transitions. The ratio between the previous emissions drastically changed when the sample was heated up to 62 °C inside a tubular furnace. This ratio was investigated using the luminescence intensity ratio method, to analyze the capabilities of the sample as a temperature sensor. The relative sensitivity reached a maximum of 11.4 °C-1 %, maintaining a detection limit below 0.15 °C for the whole temperature range.

Autochthonous transmission of the Indomalayan parasite, Transversotrema patialense, in the Caribbean: Molecular, morphological, and experimental evidence.

The Asian freshwater snail Melanoides tuberculata has been established since the 1960s in the Americas, where it transmits cercariae of a small number of digenetic trematode species from its native range. In 2021-2022, 24 M. tuberculata were discovered shedding transversotrematid cercariae in Puerto Rico, where parasites of this snail have not been previously studied. Adult transversotrematids (in some cases, gravid) were found on field-caught fish and on fish exposed to shedding snails, including on fish species native to Puerto Rico. Adults and cercariae were identified as Transversotrema patialense (Soparkar, 1924), a species native to the Indomalayan region. Morphological identification was supported with 28S rDNA sequences closely matching that from unidentified transversotrematid cercariae in Thailand. The absence of T. patialense in snails collected prior to 2021, increasing prevalence of infection in snails collected thereafter, and lack of variation in parasite DNA sequences (28S, internal transcribed spacer 2, cytochrome c oxidase I) from three isolates are consistent with a recently introduced and possibly expanding parasite population. Transversotrema patialense has been recorded outside its native range before, but most studies (including a prior record in the Americas) reported the parasite from captive hosts from commercial sources such as pet shops. The present results thus provide the first demonstration of natural transmission of T. patialense in the Americas. Phylogenetic analysis of 28S but not of ITS2 show the transversotrematid genus Transversotrema Witenberg, 1944 is paraphyletic, with Crusziella Cribb, Bray and Barker 1992 nested within it.

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.

Advances in quantum dots as diagnostic tools.

Quantum dots (QDs) are crystalline inorganic semiconductor nanoparticles a few nanometers in size that possess unique optical electronic properties vs those of larger materials. For example, QDs usually exhibit a strong and long-lived photoluminescence emission, a feature dependent on size, shape and composition. These special optoelectronic properties make them a promising alternative to conventional luminescent dyes as optical labels in biomedical applications including biomarker quantification, biomolecule targeting and molecular imaging. A key parameter for use of QDs is to functionalize their surface with suitable (bio)molecules to provide stability in aqueous solutions and efficient and selective tagging biomolecules of interest. Researchers have successfully developed biocompatible QDs and have linked them to various biomolecule recognition elements, i.e., antibodies, proteins, DNA, etc. In this chapter, QD synthesis and characterization strategies are reviewed as well as the development of nanoplatforms for luminescent biosensing and imaging-guided targeting. Relevant biomedical applications are highlighted with a particular focus on recent progress in ultrasensitive detection of clinical biomarkers. Finally, key future research goals to functionalize QDs as diagnostic tools are explored.

Strip modification and alternative architectures for signal amplification in nanoparticle-based lateral flow assays.

Nanoparticle (NP)-based lateral flow assay (LFA) technology has outstanding characteristics that make it ideal for point-of-care bioanalytical applications. However, LFAs still have important limitations, especially related to sensitivity, which is in general worse than that of other well-established bioassays such as ELISA or PCR. Many efforts have been made for enhancing the sensitivity of LFAs, mainly actuating on the nanoparticle labels and on alternative optical detection modes. However, strip pads modification for such a purpose is an incipient vast field of research. This article gives a brief overview on the recent advances proposed for signal amplification actuating on different pads and the general architecture of the LFA strips. Such strategies offer universal tools that can be adapted to any LFA, independently of the kind of sample, analyte, and label. The principles of the different strategies developed to achieve novel signal amplification and sensitive detection are discussed, and some examples of relevant approaches are highlighted, together with future prospects and challenges.

Quantum Dot Bioconjugates for Diagnostic Applications.

Quantum dots (QDs) are a special type of engineered nanomaterials with outstanding optoelectronic properties that make them as a very promising alternative to conventional luminescent dyes in biomedical applications, including biomolecule (BM) targeting, luminescence imaging and drug delivery. A key parameter to ensure successful biomedical applications of QDs is the appropriate surface modification, i.e. the surface of the nanomaterials should be modified with the appropriate functional groups to ensure stability in aqueous solutions and it should be conjugated with recognition elements capable of ensuring an efficient tagging of the BMs of interest. In this review we summarize the most relevant strategies used for surface modification of QDs and for their conjugation to BMs in preparation of their application in nanoplatforms for luminescent BM sensing and imaging-guided targeting. The applications of conjugations of photoluminescent QDs with different BMs in both in vitro and in vivo chemical sensing, immunoassays or luminescence imaging are reviewed. Recent progress in the application of functionalized QDs in ultrasensitive detection in bioanalysis, diagnostics and imaging strategies are reported. Finally, some key future research goals in the progress of bioconjugation of QDs for diagnosis are identified, including novel synthetic approaches, the need for exhaustive characterization of bioconjugates and the design of signal amplification schemes.

Bioelectrochromic hydrogel for fast antibiotic-susceptibility testing.

Materials science offers new perspectives in the clinical analysis of antimicrobial sensitivity. However, a biomaterial with the capacity to respond to living bacteria has not been developed to date. We present an electrochromic iron(III)-complexed alginate hydrogel sensitive to bacterial metabolism, here applied to fast antibiotic-susceptibility determination. Bacteria under evaluation are entrapped -and pre-concentrated- in the hydrogel matrix by oxidation of iron (II) ions to iron (III) and in situ formation of the alginate hydrogel in less than 2min and in soft experimental conditions (i.e. room temperature, pH 7, aqueous solution). After incubation with the antibiotic (10min), ferricyanide is added to the biomaterial. Bacteria resistant to the antibiotic dose remain alive and reduce ferricyanide to ferrocyanide, which reacts with the iron (III) ions in the hydrogel to produce Prussian Blue molecules. For a bacterial concentration above 107 colony forming units per mL colour development is detectable with the bare eye in less than 20min. The simplicity, sensitivity, low-cost and short response time of the biomaterial and the assay envisages a high impact of these approaches on sensitive sectors such as public health system, food and beverage industries or environmental monitoring.

Aqueous Exfoliation of Transition Metal Dichalcogenides Assisted by DNA/RNA Nucleotides: Catalytically Active and Biocompatible Nanosheets Stabilized by Acid-Base Interactions.

The exfoliation and colloidal stabilization of layered transition metal dichalcogenides (TMDs) in an aqueous medium using functional biomolecules as dispersing agents have a number of potential benefits toward the production and practical use of the corresponding two-dimensional materials, but such a strategy has so far remained underexplored. Here, we report that DNA and RNA nucleotides are highly efficient dispersants in the preparation of stable aqueous suspensions of MoS2 and other TMD nanosheets at significant concentrations (up to 5-10 mg mL-1). Unlike the case of common surfactants, for which adsorption on 2D materials is generally based on weak dispersive forces, the exceptional colloidal stability of the TMD flakes was shown to rely on the presence of relatively strong, specific interactions of Lewis acid-base type between the DNA/RNA nucleotide molecules and the flakes. Moreover, the nucleotide-stabilized MoS2 nanosheets were shown to be efficient catalysts in the reduction of nitroarenes (4-nitrophenol and 4-nitroaniline), thus constituting an attractive alternative to the use of expensive heterogeneous catalysts based on noble metals, and exhibited an electrocatalytic activity toward the hydrogen evolution reaction that was not impaired by the possible presence of nucleotide molecules adsorbed on their active sites. The biocompatibility of these materials was also demonstrated on the basis of cell proliferation and viability assays. Overall, the present work opens new vistas on the colloidal stabilization of 2D materials based on specific interactions that could be useful toward different practical applications.

Multiple actuation microvalves in wax microfluidics.

Microvalves are an essential component of microfluidic devices. In this work, a low-consumption (<35 mJ), fast-response (<0.3 s), small footprint (<0.5 mm2) wax microvalve capable of multiple actuation is described. This phase-change microvalve is electrically controlled, simple to operate and can be easily fabricated as a fully integrated element of wax microfluidic devices through a special decal-transfer microlithographic process. The valve is inherently latched and leak-proof to at least 100 kPa. A minimum pressure of 3 kPa is required for valve opening. Maximum pressures for a successful closing in air and liquid are 90 and 40 kPa, respectively. The wax valve exhibits reversible open-close behaviour without failure for up to 10 actuation cycles in air (60 kPa) and 5 in water (30 kPa). To the best of our knowledge, this microvalve has the lowest energy consumption (two orders of magnitude lower) reported so far for a plug-type phase-change valve. Furthermore, its size, actuation mechanism and fabrication technology make it suitable for large-scale integration in microfluidic devices. Detailed characteristics in fabrication and actuation of the wax microfluidic valve as well as a test example of its performance for liquid dispensing are reported.

Development and validation of an HPLC-MS method for a simultaneous measurement of oxalate and citrate in urine / Desarrollo y validación de un método de HPLC-MS para la medida simultánea de oxalato y citrato en orina

Objectives. An increased urinary oxalate and reduced urinary citrate are considered major risk factors in the formation of calcium oxalate kidney stones. In this work, an HPLC-MS method is presented for the simultaneous measurement of oxalate and citrate in urine. Methods. Sample preparation was carried out using a liquid-liquid extraction with ethyl acetate. Chromatographic separation was performed in a C18 column by gradient elution with methanol and 1 M formate buffer at 35 °C. Citrate and oxalate were monitored on a single-quadrupole MS system. Results. The method was linear in the concentration range of 0.5 mg/L to 450 mg/L for oxalate and from 2.5 mg/L to 950 mg/L for citrate. The Lower Limit of Measurement was 0.56 mg/L for oxalate and 2.5 mg/L for citrate. The within-day imprecision was 6% for oxalate and 3% for citrate, and the between day imprecision was lower than 15% for both analytes. LC-MS method was compared with capillary electrophoresis and it was shown that both methods were interchangeable to measure oxalate, but not citrate. Conclusions. HPLC-MS method is a good approach to measure oxalate and citrate in 24-hour urine, and it is applicable in clinical routine for patients with recurrent stone formation (AU)

Objetivos. La hiperoxaluria e hipocitraturia están consideradas el principal factor de riesgo en la formación de cálculos renales de oxalato cálcico. En este trabajo se ha desarrollado un método de HPLC-MS para la medida simultánea de oxalato y citrato en orina. Métodos. La preparación de muestras se realizó por una extracción líquido-líquido con etilacetato. La separación cromatográfica se llevó a cabo en una columna C-18 a 35 °C con un gradiente de elución con metanol y ácido fórmico 1 M. El citrato y el oxalato se monitorizaron mediante espectrometría de masas con un cuadrupolo simple. Resultados. El método fue lineal para el oxalato en el rango de concentración de 0,5 a 450 mg/l y para el citrato de 2,5 a 950 mg/l. El límite menor de intervalo de medida fue de 0,56 mg/l para el oxalato y de 2,5 mg/l para el citrato. La imprecisión intradía fue del 6% para el oxalato y del 3% para el citrato, y la interdía fue inferior al 15% para ambos analitos. El método de LC-MS desarrollado se comparó con un método de electroforesis capilar y se demostró que ambos eran intercambiables para el oxalato, pero no para el citrato. Conclusiones. El método de HPLC-MS desarrollado constituye una buena aproximación para medir oxalato y citrato en orina de 24 horas y es aplicable en la rutina clínica para pacientes con riesgo de formación de cálculos renales (AU)

A microfluidic device for the automated electrical readout of low-density glass-slide microarrays.

Microarrays are a powerful platform for rapid and multiplexed analysis in a wide range of research fields. Electrical readout systems have emerged as an alternative to conventional optical methods for microarray analysis thanks to its potential advantages like low-cost, low-power and easy miniaturization of the required instrumentation. In this work an automated electrical readout system for low-cost glass-slide microarrays is described. The system enables the simultaneous conductimetric detection of up to 36 biorecognition events by incorporating an array of interdigitated electrode transducers. A polydimethylsiloxane microfluidic structure has been designed that creates microwells over the transducers and incorporates the microfluidic channels required for filling and draining them with readout and cleaning solutions, thus making the readout process fully automated. Since the capture biomolecules are not immobilized on the transducer surface this readout system is reusable, in contrast to previously reported electrochemical microarrays. A low-density microarray based on a competitive enzymatic immunoassay for atrazine detection was used to test the performance of the readout system. The electrical assay shows a detection limit of 0.22±0.03 µg L(-1) similar to that obtained with fluorescent detection and allows the direct determination of the pesticide in polluted water samples. These results proved that an electrical readout system such as the one presented in this work is a reliable and cost-effective alternative to fluorescence scanners for the analysis of low-density microarrays.

Microbial trench-based optofluidic system for reagentless determination of phenolic compounds.

Phenolic compounds are one of the main contaminants of soil and water due to their toxicity and persistence in the natural environment. Their presence is commonly determined with bulky and expensive instrumentation (e.g. chromatography systems), requiring sample collection and transport to the laboratory. Sample transport delays data acquisition, postponing potential actions to prevent environmental catastrophes. This article presents a portable, miniaturized, robust and low-cost microbial trench-based optofluidic system for reagentless determination of phenols in water. The optofluidic system is composed of a poly(methyl methacrylate) structure, incorporating polymeric optical elements and miniaturized discrete auxiliary components for optical transduction. An electronic circuit, adapted from a lock-in amplifier, is used for system control and interfering ambient light subtraction. In the trench, genetically modified bacteria are stably entrapped in an alginate hydrogel for quantitative determination of model phenol catechol. Alginate is also acting as a diffusion barrier for compounds present in the sample. Additionally, the superior refractive index of the gel (compared to water) confines the light in the lower level of the chip. Hence, the optical readout of the device is only altered by changes in the trench. Catechol molecules (colorless) in the sample diffuse through the alginate matrix and reach bacteria, which degrade them to a colored compound. The absorbance increase at 450 nm reports the presence of catechol simply, quickly (~10 min) and quantitatively without addition of chemical reagents. This miniaturized, portable and robust optofluidic system opens the possibility for quick and reliable determination of environmental contamination in situ, thus mitigating the effects of accidental spills.

Reusable conductimetric array of interdigitated microelectrodes for the readout of low-density microarrays.

Low-density protein microarrays are emerging tools in diagnostics whose deployment could be primarily limited by the cost of fluorescence detection schemes. This paper describes an electrical readout system of microarrays comprising an array of gold interdigitated microelectrodes and an array of polydimethylsiloxane microwells, which enabled multiplexed detection of up to thirty six biological events on the same substrate. Similarly to fluorescent readout counterparts, the microarray can be developed on disposable glass slide substrates. However, unlike them, the presented approach is compact and requires a simple and inexpensive instrumentation. The system makes use of urease labeled affinity reagents for developing the microarrays and is based on detection of conductivity changes taking place when ionic species are generated in solution due to the catalytic hydrolysis of urea. The use of a polydimethylsiloxane microwell array facilitates the positioning of the measurement solution on every spot of the microarray. Also, it ensures the liquid tightness and isolation from the surrounding ones during the microarray readout process, thereby avoiding evaporation and chemical cross-talk effects that were shown to affect the sensitivity and reliability of the system. The performance of the system is demonstrated by carrying out the readout of a microarray for boldenone anabolic androgenic steroid hormone. Analytical results are comparable to those obtained by fluorescent scanner detection approaches. The estimated detection limit is 4.0 ng mL(-1), this being below the threshold value set by the World Anti-Doping Agency and the European Community.

Fabrication of biofunctionalized microfluidic structures by low-temperature wax bonding.

In this work, a new fabrication technology for microfluidics based on the use of wax is described. Microfluidic structures are assembled using wax as both a thermoplastic adhesive layer between two glass substrates and a spacer layer defining the microchannels. Wax patterns with dimensions down to 25 µm are easily produced on glass substrates using specially developed decal-transfer microlithography. A complete microfluidic system is created by bonding the wax patterned layer with an additional glass substrate. On the basis of the special melting behavior of waxes, an effective glass-wax bonding is achieved at 40 °C by applying a soft pressure and without the requirement of any glass pretreatment. Wax bonding provides an effective sealing of the fluidic networks even on nonflat glass substrates (i.e., containing metal electrodes). The mild conditions required for the bonding process enables the fabrication of lab-on-a-chip devices incorporating biomolecules, as is demonstrated with the implementation of a simple heterogeneous immunoassay in a microfluidic device with amperometric detection.

Phenolic compounds as enhancers in enzymatic and electrochemical oxidation of veratryl alcohol and lignins.

Sixteen phenolic compounds, 14 of which naturally occurring, were compared to the synthetic 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt (ABTS) and violuric acid (VA) in terms of their ability to act as mediators/enhancers in: (1) laccase oxidation of veratryl alcohol as a lignin model compound, and (2) electrochemical oxidation of kraft and flax lignins. HPLC analysis revealed that the syringyl-type phenols methyl syringate and acetosyringone were the most efficient natural enhancers in the laccase oxidation of veratryl alcohol. Both compounds, though far from the performance of ABTS were able to generate veratraldehyde in amount similar to that obtained with VA. By contrast, the best performing phenolic enhancers for the electrochemical oxidation of lignins were sinapinaldehyde, vanillin, acetovanillone, and syringic acid. Catalytic efficiencies close to those achieved with ABTS and VA were calculated for these phenolic compounds.

DNA hybridization biosensors using polylysine modified SPCEs.

Two electrochemical DNA hybridization biosensors (genosensors) for the detection of a 30-mer sequence unique to severe acute respiratory syndrome (SARS) virus are described in this work. Both genosensors rely on the hybridization of the oligonucleotide target with its complementary probe, which is immobilized on positively charged polylysine modified screen-printed carbon electrodes (SPCEs), through electrostatic interactions. In one design, a biotinylated target is used and the detection of the hybridization reaction is monitored using alkaline phosphatase labeled streptavidin (S-AP). This enzyme catalyzes the hydrolysis of the substrate 3-indoxyl phosphate (3-IP) to indigo, which is then solubilized to indigo carmine and detected by means of cyclic voltammetry (CV). In the other design, the target is labeled using an Au(I) complex, sodium aurothiomalate, and the duplex formation is detected by measuring, for first time, the current generated by the hydrogen evolution catalyzed by the gold label. Using 30 min of hybridization time, a detection limit of 8 pM is calculated for the enzymatic genosensor. Although this good sensitivity cannot be reached with the metal label (0.5 nM), the use of this label allows a considerable decrease of the analysis time. Both genosensors do not require the modification of the oligonucleotide probe and using stringent experimental conditions (60 min of hybridization time and 50% formamide in the hybridization buffer) can discriminate between a complementary oligonucleotide and an oligonucleotide with a three-base mismatch.