Analytical methods of antibody surface coverage and orientation on bio-functionalized magnetic beads: application to immunocapture of TNF-α.

The use of magnetic beads bio-functionalized by antibodies (Ab) is constantly increasing with a wide range of biomedical applications. However, despite an urgent need for current methods to monitor Ab's grafting process and orientation, existing methods are still either cumbersome and/or limited. In this work, we propose a new simple and rapid analytical approach to evaluate antibody orientation and density on magnetic beads. This approach relies on the cleavage by IdeS, a highly specific protease for human immunoglobulin G (hIgG), of immobilized antibodies. The F(ab)2 and Fc fragments could be then accurately quantified by size exclusion chromatography (SEC)-coupled to fluorescent detection (FLD), and the ratio of these fragments was used to give insight on the IgG orientation at the bead surface. Four different commercially available magnetic beads, bearing carboxyl groups, tosyl groups, streptavidin, or protein G on their surface have been used in this study. Results obtained showed that this approach ensures reliable information on hIgG orientation and bead surface coverage. Protein G magnetic beads demonstrated an optimal orientation of antibodies for antigen capture (75% of accessible F(ab)2 fragment) compared to tosylactivated, carboxylated, and streptavidin ones. Capture efficiency of the different functionalized beads towards human TNF-α immunocapture, a biomarker of inflammation, has been also compared. Protein G beads provided a more efficient capture compared to other beads. In the future, this approach could be applied to any type of surface and beads to assess hIgG coverage and orientation after any type of immobilization. A rapid and simple approach to evaluate orientation and density of antibodies immobilized on magnetic beads.

Surface Biochemical Modification of Poly(dimethylsiloxane) for Specific Immune Cytokine Response.

Recent evidence suggests that proinflammatory cytokines, such as tumor necrosis factor α (TNF-α), play a pivotal role in the development of inflammatory-related pathologies (covid-19, depressive disorders, sepsis, cancer, etc.,). More importantly, the development of TNF-α biosensors applied to biological fluids (e.g. sweat) could offer non-invasive solutions for the continuous monitoring of these disorders, in particular, polydimethylsiloxane (PDMS)-based biosensors. We have therefore investigated the biofunctionalization of PDMS surfaces using a silanization reaction with 3-aminopropyltriethoxysilane, for the development of a human TNF-α biosensor. The silanization conditions for 50 µm PDMS surfaces were extensively studied by using water contact angle measurements, electron dispersive X-ray and Fourier transform infrared spectroscopies, and fluorescamine detection. Evaluation of the wettability of the silanized surfaces and the Si/C ratio pointed out to the optimal silanization conditions supporting the formation of a stable and reproducible aminosilane layer, necessary for further bioconjugation. An ELISA-type immunoassay was then successfully performed for the detection and quantification of human TNF-α through fluorescent microscopy, reaching a limit of detection of 0.55 µg/mL (31.6 nM). Finally, this study reports for the first time a promising method for the development of PDMS-based biosensors for the detection of TNF-α, using a quick, stable, and simple biofunctionalization process.

Reusable Embedded Microcoils for Magnetic Nano-Beads Trapping in Microfluidics: Magnetic Simulation and Experiments.

In this study, a microfluidic chip with integrated coil was designed and fabricated for the aim of effectively trapping magnetic nanobeads (Adembeads®, 300 nm) and measuring the chip's temperature during the working time. In addition, a reversible technique of bonding Polydimethylsiloxane (PDMS) channels was presented. This bonding process used a coating layer of CYTOPproduct as a protection, insulation and low-adhesion layer. The reversible packaging technique allows the bottom substrate to be reused, possibly equipped with sensors, and to use a disposable microchannels network. The FE method was employed to calculate the magnetic field and power consumption by the ANSYS® version 12.1 software. Merit factors were defined in order to synthetically represent the ability of the simulated coil to trap beads for a unit power consumption, i.e. a given heat generation. The simulation results propose a new approach to optimize the design criteria in fabricating planar microcoils. The optimal microcoils were fabricated and then used to realize a magnetic immunoassay in a microfluidic chip. The aim was to integrate these microcoils into a lab-on-chip and obtain a fast and highly sensitive biological element detection.

Cadmium in Human Diseases: It's More than Just a Mere Metal.

Cadmium (Cd), poisoning has been reported from all around the World, causing many deaths annually. Cd is a toxic heavy metal, and is widely present in environment. It has been reported that chronic Cd exposure is associated with kidney disease, osteoporosis, cardiovascular diseases and cancer. Smoking causes exposure to significantly higher Cd levels in humans. Tobacco smoke transports Cd into the lungs. Blood then transport it to the rest of the body where it increases effects by potentiating Cd that is already present from Cd-rich food. Other high exposures of Cd can occur with people, who live near hazardous waste sites, or factories that release Cd into the air and people who work in the metal refinery industry. Breathing of Cd can severely damage the lungs and may even cause death. Multiple studies have shown an association between environmental exposure to hazardous chemicals including toxic metals and obesity, diabetes, and metabolic syndrome. At the same time, the existing data on the impact of Cd exposure on obesity and diabetes are contradictory. On the converse, results of epidemiologic studies linking Cd exposure and Osteoporosis, overweight or obesity are far less consistent and even conflicting, also depending on differences in exposure levels. In turn, laboratory studies demonstrated that Cd adversely affects adipose tissue physiopathology through several mechanisms, thus contributing to increased insulin resistance and enhancing diabetes. However, intimate biological mechanisms linking Cd exposure with human diseases are still to be adequately investigated. Therefore, the aim of the present review was to explore the impact of Cd exposure and status on the risk of Cd in human diseases.

Intraoperatively diagnosed cerebellopontine angle facial nerve schwannoma: how to deal with it.

OBJECTIVE: This study aimed to report our experience in the management of patients with intraoperatively diagnosed intracranial facial nerve schwannomas (FNSs) and propose a decision-making strategy. METHODS: Twenty-three patients with FNS of the internal auditory canal and/or cerebellopontine angle operated on between 1992 and 2012 were identified. RESULTS: Preoperatively, all cases have been radiographically diagnosed as vestibular schwannomas. Operative procedures consisted of total tumor resection with grafting in 43.4% of patients, near-total resection leaving behind the tumor capsule overlying the facial nerve in 21.7%, total tumor resection with preservation of anatomic continuity of the facial nerve in 13%, and subtotal resection in 4.3%. Four patients (17.4%) underwent bony decompression with no tumor removal. CONCLUSION: Management of FNS diagnosed at surgery represents a significant clinical challenge. We considered total tumor resection with grafting when patients presented with preoperative facial nerve palsy (≥ grade III). Both subtotal and near-total tumor removal can be performed in patients with preoperative good facial function and/or large tumors with brainstem compression. Patients with small tumors who were selected for hearing preservation surgery can be considered for bony decompression. Fascicle preservation surgery may be an option when a clear cleavage plane between the tumor and the facial nerve is found.

Chemical engineering of self-assembled Alzheimer's peptide on a silanized silicon surface.

The aim of this work is to develop a sensitive and specific immune-sensing platform dedicated to the detection of potential biomarkers of Alzheimer's disease (AD) in biological fluids. Accordingly, a controlled and adaptive surface functionalization of a silicon wafer with 7-octenyltrichlorosilane has been performed. The surface has extensively been characterized by atomic force microscopy (AFM; morphology) and X-ray photoelectron spectroscopy (XPS; chemical composition) and contact angle measurements. The wettability of the grafted chemical groups demonstrated the gradual trend from hydrophilic to hydrophobic surface during functionalization. XPS evidenced the presence of silanes on the surface after silanization, and even carboxylic groups as products from the oxidation step of the functionalization process. The characterization results permitted us to define an optimal protocol to reach a high-quality grafting yield. The issue of the quality of controlled chemical preparation on bioreceiving surfaces was also investigated by the recognition of one AD biomarker, the amyloid peptide Aß 1-42. We have therefore evaluated the biological activity of the grafted anti Aß antibodies onto this silanized surface by fluorescent microscopy. In conclusion, we have shown, both qualitatively and quantitatively, the uniformity of the optimized functionalization on slightly oxidized silicon surfaces, providing a reliable and chemically stable procedure to determine specific biomarkers of Alzheimer disease. This work opens the route to the integration of controlled immune-sensing applications on lab-on-chip systems.

Spinal cord compression revealing an intraosseous schwannoma.

A 68-year-old female presented with inflammatory lumbalgia and cruralgia. Physical examination revealed a lumbar stiffness without neurological deficit. Secondarily, paraplegia and urinary retention appeared. Magnetic resonance imaging showed a vertebral compaction of L3 vertebra with medullar compression. Emergent surgery revealed an epidural tumor involving largely the L3 vertebral body. Histology found schwannoma with positive protein S100 on the immunohistochemical study. Metastasis screening revealed bilateral nodular lesions of the lungs and a trochanter high scintigraphic signal. It was a malignant schwannoma. The patient underwent radiotherapy in addition to the total tumor resection.

Surgical results and technical refinements in translabyrinthine excision of vestibular schwannomas: the Gruppo Otologico experience.

BACKGROUND: Vestibular schwannomas (VSs) are the most common cerebellopontine angle tumors, accounting for 75% of all lesions in this location. OBJECTIVE: To evaluate the results after removal of VS through the enlarged translabyrinthine approach, which is a widening of the classic translabyrinthine approach that gives larger access and provides more room to facilitate tumor removal and to minimize surgery-related morbidities. METHODS: This was a retrospective study of 1865 patients who underwent VS excision through the enlarged translabyrinthine approach between 1987 and 2009. Mean age was 50.39 years. Mean tumor size was 1.8 cm. Median follow-up was 5.7 years. RESULTS: Total removal was achieved in 92.33% of cases; 143 patients had incomplete resection with evidence of regrowth in 8. In the 1742 previously untreated patients, anatomic preservation of facial nerve was achieved in 1661 cases (95.35%), and House-Brackmann grade I or II was reached in 1047 patients (59.87%). Facial nerve outcome was significantly better in tumors ≤ 20 mm. Surgical complications included cerebrospinal fluid leakage in 0.85%, meningitis in 0.10%, intracranial bleeding in 0.80%, non--VII/VIII cranial nerve palsy in 0.96%, cerebellar ataxia in 0.69%, and death in 0.10%. The technical modifications that evolved with increasing experience are described. CONCLUSION: The enlarged translabyrinthine approach is a safe and effective approach for the removal of VS. In our experience, the complication rate is very low and tumor size is still the main factor influencing postoperative facial nerve function with a cutoff point at around 20 mm.