Influence of reaction-induced convection on quorum sensing in enzyme-loaded agarose beads.

In theory, groups of enzyme-loaded particles producing an acid or base may show complex behavior including dynamical quorum sensing, the appearance of synchronized oscillations above a critical number or density of particles. Here, experiments were performed with the enzyme urease loaded into mm-sized agarose beads and placed in a solution of urea, resulting in an increase in pH. This behavior was found to be dependent upon the number of beads present in the array; however, reaction-induced convection occurred and plumes of high pH developed that extended to the walls of the reactor. The convection resulted in the motion of the mm-sized particles and conversion of the solution to high pH. Simulations in a simple model of the beads demonstrated the suppression of dynamical quorum sensing in the presence of flow.

Glioma big potassium channel expression in human cancers and possible T cell epitopes for their immunotherapy.

Big potassium (BK) ion channels have several spliced variants. One spliced variant initially described within human glioma cells is the glioma BK (gBK) channel. This isoform consists of 34 aa inserted into the intracellular region of the basic BK ion channel. PCR primers specific for this inserted region confirmed that human glioma cell lines and freshly resected surgical tissues from glioblastoma multiforme patients strongly expressed gBK mRNA. Normal human brain tissue very weakly expressed this transcript. An Ab specific for this gBK isoform confirmed that human glioma cells displayed this protein in the cell membrane, mitochondria, Golgi, and endoplasmic reticulum. Within the gBK region, two putative epitopes (gBK1 and gBK2) are predicted to bind to the HLA-A*0201 molecule. HLA-A*0201-restricted human CTLs were generated in vitro using gBK peptide-pulsed dendritic cells. Both gBK1 and gBK2 peptide-specific CTLs killed HLA-A2⁺/gBK⁺ gliomas, but they failed to kill non-HLA-A2-expressing but gBK⁺ target cells in cytolytic assays. T2 cells loaded with exogenous gBK peptides, but not with the influenza M1 control peptide, were only killed by their respective CTLs. The gBK-specific CTLs also killed a variety of other HLA-A*0201⁺ cancer cells that possess gBK, as well as HLA-A2⁺ HEK cells transfected with the gBK gene. Of clinical relevance, we found that T cells derived from glioblastoma multiforme patients that were sensitized to the gBK peptide could also kill target cells expressing gBK. This study shows that peptides derived from cancer-associated ion channels maybe useful targets for T cell-mediated immunotherapy.

Iris atrophy after administration of Dexycu: Additional evidence and possible mechanism for a rare complication.

Purpose: To report the observation of iris atrophy with the use of Dexycu in cataract surgery and describe a possible mechanism for this complication. Observations: Between 2020 and 2021, 2 patients underwent cataract surgery procedures with the use of Dexycu and experienced iris atrophy. The first case of iris atrophy was discovered by 6-month follow-up and the second case by 2-month follow-up. The timing and nature of this complication possibly represent more than steroid-induced hypopigmentation. Conclusions and Importance: This publication adds further evidence of iris atrophy and a proposed mechanism associated with the use of Dexycu. Surgeons should be aware of this complication, especially when operating on eyes with dark irides.

The spectrum of COVID-19-associated chorioretinal vasculopathy.

Purpose: Although conjunctivitis represents the most common ocular manifestation of COVID-19 infection, sight-threatening retinal involvement has been reported. Herein, we report and characterize with multimodal retinal imaging 5 cases of acute vision loss secondary to presumed chorioretinal vasculopathy temporally associated with COVID-19 infection with varying severity, visual morbidity, and treatment response, and review the available literature on the association between COVID-19 infection and retinal microvascular changes. Design: Observational case series and literature review. Methods: Multicenter case series of 5 patients who presented to academic centers and private offices with acute vision loss temporally associated with COVID-19 infection. A review of the literature was conducted using online databases. Results: 10 eyes of 5 patients, 3 men and 2 women, with a mean age of 30.8 years (median 33, range 16-44) were described. All patients had a recently preceding episode of COVID-19, with symptomatology ranging from mild infection to life-threatening encephalopathy. Treatment for their retinal disease included topical, oral, intravitreal, and intravenous steroids, steroid-sparing immunosuppression, retinal photocoagulation, antivirals, and antiplatelet and anticoagulant agents. Treatment response and visual recovery ranged from complete recovery of baseline acuity to permanent vision loss and need for chronic immunosuppression. Conclusions and Importance: Clinicians should be mindful of the potential for vision-threatening retinal involvement after COVID-19 infection. If found, treatment with both anti-inflammatory therapy and anticoagulation should be considered, in addition to close monitoring, as some patients with this spectrum of disease may require chronic immune suppression and/or anti-VEGF therapy.

Lung cancer: a classic example of tumor escape and progression while providing opportunities for immunological intervention.

Lung cancers remain one of the most common and deadly cancers in the world today (12.5% of newly diagnosed cancers) despite current advances in chemo- and radiation therapies. Often, by the time these tumors are diagnosed, they have already metastasized. These tumors demonstrate the classic hallmarks of cancer in that they have advanced defensive strategies allowing them to escape various standard oncological treatments. Immunotherapy is making inroads towards effectively treating other fatal cancers, such as melanoma, glioblastoma multiforme, and castrate-resistant prostate cancers. This paper will cover the escape mechanisms of bronchogenic lung cancer that must be overcome before they can be successfully treated. We also review the history of immunotherapy directed towards lung cancers.

A pH-Based Single-Sensor Array for Discriminating Metal Ions in Water.

Human activities, such as mining and manufacturing, expose society and the natural environment to harmful levels of metal ions. Recently, optical sensor arrays for metal ion detection have become popular owing to their favourable features, such as facile sample preparation and the requirement of less expensive instrumentation compared to traditional, spectrometry-based analysis techniques. Sensor arrays usually consist of numerous optical probes that are used in combination to generate unique analyte responses. In contrast, here we present an array that comprises a single fluorescent sensor, Coum4-DPA, that produces unique responses to metal ions in different pH environments. With this simple sensing platform, we were able to classify 10 metal ions in different water sources and quantify Pb2+ in tap water using just one fluorescent sensor, a few pH buffers and two sets of spectral data. This novel approach significantly decreases time and costs associated with probe synthesis and data collection, making it highly transferrable to real-world metal sensing applications.

Reaction-diffusion hydrogels from urease enzyme particles for patterned coatings.

The reaction and diffusion of small molecules is used to initiate the formation of protective polymeric layers, or biofilms, that attach cells to surfaces. Here, inspired by biofilm formation, we present a general method for the growth of hydrogels from urease enzyme-particles by combining production of ammonia with a pH-regulated polymerization reaction in solution. We show through experiments and simulations how the propagating basic front and thiol-acrylate polymerization were continuously maintained by the localized urease reaction in the presence of urea, resulting in hydrogel layers around the enzyme particles at surfaces, interfaces or in motion. The hydrogels adhere the enzyme-particles to surfaces and have a tunable growth rate of the order of 10 µm min-1 that depends on the size and spatial distribution of particles. This approach can be exploited to create enzyme-hydrogels or chemically patterned coatings for applications in biocatalytic flow reactors.

Preserving Vision: Rethinking Burn Patient Monitoring to Prevent Orbital Compartment Syndrome.

Burn patients receiving aggressive fluid resuscitation are at risk of developing orbital compartment syndrome (OCS). This condition results in elevated orbital pressures and can lead to rapid permanent vision loss. Risk factors and monitoring frequency for OCS remain largely unknown. A retrospective review was therefore conducted of admitted burn patients evaluated by the ophthalmology service at an American Burn Association verified Burn Treatment Center. Demographic, burn, examination, and fluid resuscitation data were compared using two-sided t-tests, Fisher's exact tests, and linear regression. Risk factors for elevated intraocular pressures (IOPs; a surrogate for intraorbital pressure) in patients resuscitated via the Parkland formula were found to be total body surface area (% TBSA) burned, resuscitation above the Ivy Index (>250 ml/kg), and Parkland formula calculated volume. Maximum IOP and actual fluid resuscitation volume were linearly related. Analysis of all patients with elevated IOP found multiple patients with significant IOP increases after initial evaluation resulting in OCS within the first 24 hours postinjury. While %TBSA, Ivy Index, and resuscitation calculated volume are OCS risk factors in burn patients, two patients with facial burns developed OCS (25% of all patients with OCS) despite not requiring resuscitation. Orbital congestion can develop within the first 24 hours of admission when resuscitation volumes are the greatest. In addition to earlier and more frequent IOP checks in susceptible burn patients during the first day, the associated risk factors will help identify those most at risk for OCS and vision loss.

Acute Unilateral Proptosis After Blunt Orbital Trauma in an Adolescent Patient.

A male patient aged 16 years with a recent history of blunt trauma to the periorbita presented with blurred vision, pain, and proptosis of the right eye. MRI imaging revealed an intraconal mass. What would you do next?

Small cell lung cancer cells express the late stage gBK tumor antigen: a possible immunotarget for the terminal disease.

Big Potassium (BK) ion channels have several splice variants. One splice variant initially described within human glioma cells is called the glioma BK channel (gBK). Using a gBK-specific antibody, we detected gBK within three human small cell lung cancer (SCLC) lines. Electrophysiology revealed that functional membrane channels were found on the SCLC cells. Prolonged exposure to BK channel activators caused the SCLC cells to swell within 20 minutes and resulted in their death within five hours. Transduction of BK-negative HEK cells with gBK produced functional gBK channels. Quantitative RT-PCR analysis using primers specific for gBK, but not with a lung-specific marker, Sox11, confirmed that advanced, late-stage human SCLC tissues strongly expressed gBK mRNA. Normal human lung tissue and early, lower stage SCLC resected tissues very weakly expressed this transcript. Immunofluorescence using the anti-gBK antibody confirmed that SCLC cells taken at the time of the autopsy intensely displayed this protein. gBK may represent a late-stage marker for SCLC. HLA-A*0201 restricted human CTL were generated in vitro using gBK peptide pulsed dendritic cells. The exposure of SCLC cells to interferon-γ (IFN-γ) increased the expression of HLA; these treated cells were killed by the CTL better than non-IFN-γ treated cells even though the IFN-γ treated SCLC cells displayed diminished gBK protein expression. Prolonged incubation with recombinant IFN-γ slowed the in vitro growth and prevented transmigration of the SCLC cells, suggesting IFN-γ might inhibit tumor growth in vivo. Immunotherapy targeting gBK might impede advancement to the terminal stage of SCLC via two pathways.