Correction: 'Semen rheology and its relation to male infertility' (2022), by Tomaiuolo et al.

[This corrects the article DOI: 10.1098/rsfs.2022.0048.][This corrects the article DOI: 10.1098/rsfs.2022.0048.].

Organic Electrochemical Transistor Immuno-Sensors for Spike Protein Early Detection.

The global COVID-19 pandemic has had severe consequences from the social and economic perspectives, compelling the scientific community to focus on the development of effective diagnostics that can combine a fast response and accurate sensitivity/specificity performance. Presently available commercial antigen-detecting rapid diagnostic tests (Ag-RDTs) are very fast, but still face significant criticisms, mainly related to their inability to amplify the protein signal. This translates to a limited sensitive outcome and, hence, a reduced ability to hamper the spread of SARS-CoV-2 infection. To answer the urgent need for novel platforms for the early, specific and highly sensitive detection of the virus, this paper deals with the use of organic electrochemical transistors (OECTs) as very efficient ion-electron converters and amplifiers for the detection of spike proteins and their femtomolar concentration. The electrical response of the investigated OECTs was carefully analyzed, and the changes in the parameters associated with the transconductance (i.e., the slope of the transfer curves) in the gate voltage range between 0 and 0.3 V were found to be more clearly correlated with the spike protein concentration. Moreover, the functionalization of OECT-based biosensors with anti-spike and anti-nucleocapside proteins, the major proteins involved in the disease, demonstrated the specificity of these devices, whose potentialities should also be considered in light of the recent upsurge of the so-called "long COVID" syndrome.

Correction: Post-liquefaction normospermic human semen behaves as a weak-gel viscoelastic fluid.

Correction for 'Post-liquefaction normospermic human semen behaves as a weak-gel viscoelastic fluid' by Giovanna Tomaiuolo et al., Soft Matter, 2023, https://doi.org/10.1039/d3sm00443k.

Post-liquefaction normospermic human semen behaves as a weak-gel viscoelastic fluid.

The rheological behavior of human semen is overlooked and essentially unexplored in the scientific literature. Here, we provide the first quantitative experimental evidence that post-liquafaction normospermic human semen behaves as a viscoelastic fluid and the shear moduli can be scaled according to the weak-gel model.

Semen rheology and its relation to male infertility.

Infertility affects 15% of couples of reproductive age worldwide. In spite of many advances in understanding and treating male infertility, there is still a number of issues that need further investigation and translation to the clinic. Here, we review the current knowledge and practice concerning semen rheology and its relation with pathological states affecting male infertility. Although it is well recognized that altered rheological properties of semen can impair normal sperm movement in the female reproductive tract, routine semen analysis is mostly focused on number, motility and morphology of spermatozoa, and includes only an approximate, operator-dependent measure of semen viscosity. The latter is based on the possible formation of a liquid thread from a pipette where a semen sample has been aspirated, a method that is sensitive not only to viscosity but also to elongational properties and surface tension of semen. The formation of a liquid thread is usually associated with a gel-like consistency of the sample and changes in spermatozoa motility in such a complex medium are still to be fully elucidated. The aim of this review is to point out that a more quantitative and reliable characterization of semen rheology is in order to improve the current methods of semen analysis and to develop additional tools for the diagnosis and treatment of male infertility.

Organic electrochemical transistors as novel biosensing platforms to study the electrical response of whole blood and plasma.

In this paper, for the first time to the best of our knowledge, organic electrochemical transistors are employed to investigate the electrical response of human blood, plasma and alternative buffer solutions that inhibit red blood cell (RBC) aggregation. Our focus is on selecting a suitable electrolytic platform and the related operating conditions, where the RBC effect on the OECT response can be observed separately from the strong ionic environment of plasma in whole blood. The transient response of whole blood to pulse experiments is characterized by two time constants, which can be related to blood viscosity and to the capacitive coupling between the ionic and electronic components of the overall system. The role of capacitive effects, likely due to enhanced double-layer formation by negatively charged RBCs, is also confirmed by the increase of transconductance which was found in RBC suspensions as compared to the suspending buffer. Overall, the complex behavior found in these experiments provides new insights for the development of innovative blood-based sensing devices for biomedical applications.

Membrane Fouling Phenomena in Microfluidic Systems: From Technical Challenges to Scientific Opportunities.

The almost ubiquitous, though undesired, deposition and accumulation of suspended/dissolved matter on solid surfaces, known as fouling, represents a crucial issue strongly affecting the efficiency and sustainability of micro-scale reactors. Fouling becomes even more detrimental for all the applications that require the use of membrane separation units. As a matter of fact, membrane technology is a key route towards process intensification, having the potential to replace conventional separation procedures, with significant energy savings and reduced environmental impact, in a broad range of applications, from water purification to food and pharmaceutical industries. Despite all the research efforts so far, fouling still represents an unsolved problem. The complex interplay of physical and chemical mechanisms governing its evolution is indeed yet to be fully unraveled and the role played by foulants' properties or operating conditions is an area of active research where microfluidics can play a fundamental role. The aim of this review is to explore fouling through microfluidic systems, assessing the fundamental interactions involved and how microfluidics enables the comprehension of the mechanisms characterizing the process. The main mathematical models describing the fouling stages will also be reviewed and their limitations discussed. Finally, the principal dynamic investigation techniques in which microfluidics represents a key tool will be discussed, analyzing their employment to study fouling.

Complex Modes of Inheritance in Hereditary Red Blood Cell Disorders: A Case Series Study of 155 Patients.

Hereditary erythrocytes disorders include a large group of conditions with heterogeneous molecular bases and phenotypes. We analyzed here a case series of 155 consecutive patients with clinical suspicion of hereditary erythrocyte defects referred to the Medical Genetics Unit from 2018 to 2020. All of the cases followed a diagnostic workflow based on a targeted next-generation sequencing panel of 86 genes causative of hereditary red blood cell defects. We obtained an overall diagnostic yield of 84% of the tested patients. Monogenic inheritance was seen for 69% (107/155), and multi-locus inheritance for 15% (23/155). PIEZO1 and SPTA1 were the most mutated loci. Accordingly, 16/23 patients with multi-locus inheritance showed dual molecular diagnosis of dehydrated hereditary stomatocytosis/xerocytosis and hereditary spherocytosis. These dual inheritance cases were fully characterized and were clinically indistinguishable from patients with hereditary spherocytosis. Additionally, their ektacytometry curves highlighted alterations of dual inheritance patients compared to both dehydrated hereditary stomatocytosis and hereditary spherocytosis. Our findings expand the genotypic spectrum of red blood cell disorders and indicate that multi-locus inheritance should be considered for analysis and counseling of these patients. Of note, the genetic testing was crucial for diagnosis of patients with a complex mode of inheritance.

The microstructure of Carbopol in water under static and flow conditions and its effect on the yield stress.

In this work, experimental observations of the microstructure of neutralized polyacrylic acid (Carbopol) in water by confocal microscopy under both static and flow conditions are presented. In the former case, a Carbopol-rich phase made by swollen particles dispersed in a water-rich continuous phase is found, so that the system will be henceforth referred to as a suspension, as long as particles are observed. The swollen particles form dendritic-like aggregates, which span the entire solution volume above a critical concentration. In such conditions, a percolated network can be formed, leading to the onset of a yield stress behavior. By separating the dispersed and continuous phase through centrifugation, we provide evidence of a miscibility gap in the phase behavior of Carbopol in water. When the Carbopol suspensions flow in a microfluidic capillary, a particle-concentrated plug core can be distinguished from a less concentrated layer corresponding to a steep velocity decrease. Confocal imaging also shows that the apparent slip found in Carbopol suspensions is due to a particle-concentrated near-wall region, where no flow is observed. Such flow-induced microstructure is responsible for the different nature of the yield stress values measured by classical rheometry and by flow velocimetry. While the yield stress measured by the former can be here related to the presence of a percolated network, the yield stress obtained from the velocity profile is due to the heterogeneous particle distribution along the capillary radius. These results provide a novel insight on the mechanisms governing yield stress in complex fluids.

Confined flow behaviour of droplets in microcapillary flow.

The problem of droplets flowing in a micritions is relevant in several applications including flow in porous media. When the flow in the capillary is laminar with negligible gravity effects, droplet velocity and deformation depend upon three independent parameters: the droplet size relative to the capillary radius [Formula: see text][Formula: see text], which is a measure of confinement, the viscosity ratio [Formula: see text] between the droplet and the continuous phase and the capillary number Ca which measures the ratio of viscous to capillary forces. Although droplet microconfined flow behaviour in capillaries has been widely investigated by theoretical models, experimental results are still scarce. Here, an experimental campaign focused on the flow behaviour of axisymmetric confined droplets flowing in a microcapillary is carried out. Our experimental results were obtained by using a water in soybean oil emulsion with a low viscosity ratio and the effect of the aforementioned three parameters, [Formula: see text], [Formula: see text] and Ca, on droplet motion was investigated. Moreover, our experimental results are compared with numerical solutions available in the literature.

Measuring Interfacial Tension of Emulsions in Situ by Microfluidics.

Interfacial tension is a key parameter affecting industrially relevant properties of emulsions, such as morphology and stability. Although several methods are available to measure interfacial tension, they are based on generation of droplets starting from separate emulsion components and cannot directly probe the interfacial tension of an emulsion as such. Here, a novel microfluidic tensiometry device to measure interfacial tension of a water-in-oil emulsion in situ as a function of surfactant concentration is presented. In our approach, interfacial tension is obtained from a quantitative analysis of the deformation of individual emulsion droplets under steady state shear flow in microfluidic channels. The technique is validated by comparing the results with experimental data obtained by the pendant drop method in a broad range of interfacial tension values. A very good agreement is found, and an estimate of the surfactant critical micellar concentration (CMC) is also obtained. The proposed microfluidic setup can be used even at high surfactant concentrations, where the measurement is made more challenging by sample viscoelasticity, thus providing a powerful tool to determine the interfacial tension of complex systems in an extended concentration range. The technique could be also used for in-line monitoring of emulsion processing.

Emulsions in porous media: From single droplet behavior to applications for oil recovery.

Emulsions are suspensions of droplets ubiquitous in oil recovery from underground reservoirs. Oil is typically trapped in geological porous media where emulsions are either formed in situ or injected to elicit oil mobilization and thus enhance the amount of oil recovered. Here, we briefly review basic concepts on geometrical and wetting features of porous media, including thin film stability and fluids penetration modes, which are more relevant for oil recovery and oil-contaminated aquifers. Then, we focus on the description of emulsion flow in porous media spanning from the behaviour of single droplets to the collective flow of a suspension of droplets, including the effect of bulk and interfacial rheology, hydrodynamic and physico-chemical interactions. Finally, we describe the particular case of emulsions used in underground porous media for enhanced oil recovery, thereby discussing some perspectives of future work. Although focused on oil recovery related topics, most of the insights we provide are useful towards remediation of oil-contaminated aquifers and for a basic understanding of emulsion flow in any kind of porous media, such as biological tissues.

The effect of flow on viscoelastic emulsion microstructure.

Emulsions made of oil, water and surfactants are widespread soft materials with complex structures depending on composition and temperature. Emulsion phase behavior at rest has been widely investigated but flow-induced effects, which are very relevant in many applications, can still be further explored towards improved emulsion microstructural design. In this work, we use low energy emulsification processing to create small-sized emulsions. In a previous report, we showed the emulsion morphology development and the effect of flow on the microstructure of a highly viscoelastic attractive emulsion which result in a concentrated nanoemulsion after viscoelastic droplet filaments are disrupted. Here, we show that upon stopping the flow, the filaments slowly buckle, recoil and finally form clusters of randomly flocculated droplets. We thus obtain two completely different emulsion morphologies simply induced by the action of flow, where in both cases attractive interactions play a key role. The emulsion high interfacial area represents a valuable feature for several applications such as upstream operations, microreaction media and drug delivery.

Multi-gene panel testing improves diagnosis and management of patients with hereditary anemias.

Mutations in more than 70 genes cause hereditary anemias (HA), a highly heterogeneous group of rare/low frequency disorders in which we included: hyporegenerative anemias, as congenital dyserythropoietic anemia (CDA) and Diamond-Blackfan anemia; hemolytic anemias due to erythrocyte membrane defects, as hereditary spherocytosis and stomatocytosis; hemolytic anemias due to enzymatic defects. The study describes the diagnostic workflow for HA, based on the development of two consecutive versions of a targeted-NGS panel, including 34 and 71 genes, respectively. Seventy-four probands from 62 unrelated families were investigated. Our study includes the most comprehensive gene set for these anemias and the largest cohort of patients described so far. We obtained an overall diagnostic yield of 64.9%. Despite 54.2% of cases showed conclusive diagnosis fitting well to the clinical suspicion, the multi-gene analysis modified the original clinical diagnosis in 45.8% of patients (nonmatched phenotype-genotype). Of note, 81.8% of nonmatched patients were clinically suspected to suffer from CDA. Particularly, 45.5% of the probands originally classified as CDA exhibited a conclusive diagnosis of chronic anemia due to enzymatic defects, mainly due to mutations in PKLR gene. Interestingly, we also identified a syndromic CDA patient with mild anemia and epilepsy, showing a homozygous mutation in CAD gene, recently associated to early infantile epileptic encephalopathy-50 and CDA-like anemia. Finally, we described a patient showing marked iron overload due to the coinheritance of PIEZO1 and SEC23B mutations, demonstrating that the multi-gene approach is valuable not only for achieving a correct and definitive diagnosis, but also for guiding treatment.

Endothelial glycocalyx regulates cytoadherence in Plasmodium falciparum malaria.

Malaria is associated with significant microcirculation disorders, especially when the infection reaches its severe stage. This can lead to a range of fatal conditions, from cerebral malaria to multiple organ failure, of not fully understood pathogenesis. It has recently been proposed that a breakdown of the glycocalyx, the carbohydrate-rich layer lining the vascular endothelium, plays a key role in severe malaria, but direct evidence supporting this hypothesis is still lacking. Here, the interactions between Plasmodium falciparum infected red blood cells ( PfRBCs) and endothelial glycocalyx are investigated by developing an in vitro, physiologically relevant model of human microcirculation based on microfluidics. Impairment of the glycocalyx is obtained by enzymatic removal of sialic acid residues, which, due to their terminal location and net negative charge, are implicated in the initial interactions with contacting cells. We show a more than twofold increase of PfRBC adhesion to endothelial cells upon enzymatic treatment, relative to untreated endothelial cells. As a control, no effect of enzymatic treatment on healthy red blood cell adhesion is found. The increased adhesion of PfRBCs is also associated with cell flipping and reduced velocity as compared to the untreated endothelium. Altogether, these results provide a compelling evidence of the increased cytoadherence of PfRBCs to glycocalyx-impaired vascular endothelium, thus supporting the advocated role of glycocalyx disruption in the pathogenesis of this disease.

Margination of Fluorescent Polylactic Acid-Polyaspartamide based Nanoparticles in Microcapillaries In Vitro: the Effect of Hematocrit and Pressure.

The last decade has seen the emergence of vascular-targeted drug delivery systems as a promising approach for the treatment of many diseases, such as cardiovascular diseases and cancer. In this field, one of the major challenges is carrier margination propensity (i.e., particle migration from blood flow to vessel walls); indeed, binding of these particles to targeted cells and tissues is only possible if there is direct carrier-wall interaction. Here, a microfluidic system mimicking the hydrodynamic conditions of human microcirculation in vitro is used to investigate the effect of red blood cells (RBCs) on a carrier margination in relation to RBC concentration (hematocrit) and pressure drop. As model drug carriers, fluorescent polymeric nanoparticles (FNPs) were chosen, which were obtained by using as starting material a pegylated polylactic acid-polyaspartamide copolymer. The latter was synthesized by derivatization of α,ß-poly(N-2-hydroxyethyl)-d,l-aspartamide (PHEA) with Rhodamine (RhB), polylactic acid (PLA) and then poly(ethyleneglycol) (PEG) chains. It was found that the carrier concentration near the wall increases with increasing pressure drop, independently of RBC concentration, and that the tendency for FNP margination decreases with increasing hematocrit. This work highlights the importance of taking into account RBC-drug carrier interactions and physiological conditions in microcirculation when planning a drug delivery strategy based on systemically administered carriers.

Flow-induced nanostructuring of gelled emulsions.

Although the phase behavior of emulsions has been thoroughly investigated, the effect of flow on emulsion morphology, which is relevant for many applications, is far from being fully elucidated. Here, we investigate an emulsion based on two common nonionic surfactants in a range of water concentration where complex and diverse microstructures are found at rest, such as multilamellar and bicontinuous phases. In spite of such complexity, once subjected to shear flow, all the emulsions investigated are characterized by thinning filaments which eventually break up into a concentrated suspension of micro-sized water-based droplets dispersed in a continuous oil phase. The so-formed droplets tend to align in string-like structures. The emulsions exhibit a yield stress, whose value can be estimated by the plug-core velocity profiles in pressure-driven capillary flow, thus providing evidence of weakly attractive interdroplet interactions. The latter are consistent with droplet clustering and percolation observed at rest. These results can also be relevant to the flow behavior of other liquid-liquid systems, such as polymer blends, where the flow-induced microstructure is under debate as well.

Engineering approaches in siRNA delivery.

siRNAs are very potent drug molecules, able to silence genes involved in pathologies development. siRNAs have virtually an unlimited therapeutic potential, particularly for the treatment of inflammatory diseases. However, their use in clinical practice is limited because of their unfavorable properties to interact and not to degrade in physiological environments. In particular they are large macromolecules, negatively charged, which undergo rapid degradation by plasmatic enzymes, are subject to fast renal clearance/hepatic sequestration, and can hardly cross cellular membranes. These aspects seriously impair siRNAs as therapeutics. As in all the other fields of science, siRNAs management can be advantaged by physical-mathematical descriptions (modeling) in order to clarify the involved phenomena from the preparative step of dosage systems to the description of drug-body interactions, which allows improving the design of delivery systems/processes/therapies. This review analyzes a few mathematical modeling approaches currently adopted to describe the siRNAs delivery, the main procedures in siRNAs vectors' production processes and siRNAs vectors' release from hydrogels, and the modeling of pharmacokinetics of siRNAs vectors. Furthermore, the use of physical models to study the siRNAs vectors' fate in blood stream and in the tissues is presented. The general view depicts a framework maybe not yet usable in therapeutics, but with promising possibilities for forthcoming applications.