In vivo imaging of nanoparticle-labeled CAR T cells.

Metastatic osteosarcoma has a poor prognosis with a 2-y, event-free survival rate of ∼15 to 20%, highlighting the need for the advancement of efficacious therapeutics. Chimeric antigen receptor (CAR) T-cell therapy is a potent strategy for eliminating tumors by harnessing the immune system. However, clinical trials with CAR T cells in solid tumors have encountered significant challenges and have not yet demonstrated convincing evidence of efficacy for a large number of patients. A major bottleneck for the success of CAR T-cell therapy is our inability to monitor the accumulation of the CAR T cells in the tumor with clinical-imaging techniques. To address this, we developed a clinically translatable approach for labeling CAR T cells with iron oxide nanoparticles, which enabled the noninvasive detection of the iron-labeled T cells with magnetic resonance imaging (MRI), photoacoustic imaging (PAT), and magnetic particle imaging (MPI). Using a custom-made microfluidics device for T-cell labeling by mechanoporation, we achieved significant nanoparticle uptake in the CAR T cells, while preserving T-cell proliferation, viability, and function. Multimodal MRI, PAT, and MPI demonstrated homing of the T cells to osteosarcomas and off-target sites in animals administered with T cells labeled with the iron oxide nanoparticles, while T cells were not visualized in animals infused with unlabeled cells. This study details the successful labeling of CAR T cells with ferumoxytol, thereby paving the way for monitoring CAR T cells in solid tumors.

Advances in biomimetic collagen mineralisation and future approaches to bone tissue engineering.

With an ageing world population and ~20% of adults in Europe being affected by bone diseases, there is an urgent need to develop advanced regenerative approaches and biomaterials capable to facilitate tissue regeneration while providing an adequate microenvironment for cells to thrive. As the main components of bone are collagen and apatite mineral, scientists in the tissue engineering field have attempted in combining these materials by using different biomimetic approaches to favour bone repair. Still, an ideal bone analogue capable of mimicking the distinct properties (i.e., mechanical properties, degradation rate, porosity, etc.) of cancellous bone is to be developed. This review seeks to sum up the current understanding of bone tissue mineralisation and structure while providing a critical outlook on the existing biomimetic strategies of mineralising collagen for bone tissue engineering applications, highlighting where gaps in knowledge exist.

Rapid treatment of endophthalmitis with intravitreal antibiotics is associated with better vision outcomes.

BACKGROUND: To discover whether the timing of intravitreal antibiotics, from the time of presentation to injection, affects the visual outcomes of patients with endophthalmitis from all causes. METHODS: A retrospective study of consecutive patients managed at Auckland District Health Board between 1 January 2004 and 1 July 2021. The main outcome measure was final visual acuity at follow-up and the proportion of subjects with severe vision loss (≤6/60). RESULTS: Three hundred and seventy-four eyes were included in the study, with a median age of 69.7 years at presentation; 192 subjects (51.6%) were female. The median presenting visual acuity was hand movements and hypopyon was present in 194 subjects (51.9%). Cataract surgery was the most frequent aetiology in 115 subjects (30.7%), followed by intravitreal anti-vascular endothelial growth factor injections in 89 subjects (23.8%) and endogenous endophthalmitis in 54 subjects (14.4%). Median time to injection of intravitreal antibiotics was 3.5 h (Interquartile range 2-6). On multivariate analysis, early treatment with intravitreal antibiotics was associated with better visual outcome, particularly for those treated within 2 h, while poor presenting visual acuity and culture-positive endophthalmitis were associated with worse outcomes. CONCLUSION: Rapid intravitreal antibiotic administration is associated with better final visual acuity outcomes, particularly for those receiving treatment within 2 h of presentation. Patients with severe vision loss on presentation, benefit the most with improved final visual acuity following expedited treatment.

Diagnostic Uses of Common Eye Drops.

Medicated eye drops may have dual therapeutic and diagnostic uses that form part of the ophthalmic assessment paradigm. In this review article, commonly administered and prescribed eye drops were analyzed for their use as a diagnostic tool. It examines the common categories of eye drops-antimicrobial agents, topical anesthetics, mydriatics, and ocular anti-hypertensives, with respect to their therapeutic and diagnostic applications. Knowledge of the pharmacological effects of eye drops is an important aspect in performing clinical duties. Diagnostic tests by utilization of eye drops are safe, efficient, noninvasive, and informative to the eye care professional.

Dose-Related Visual Outcomes in the Treatment of Choroidal Melanoma with Stereotactic Radiotherapy.

Introduction: Stereotactic radiotherapy (SRT) in the treatment of choroidal melanoma (CM) may be indicated if the tumour is located close to the optic nerve or is unsuitable for a radiotherapeutic plaque. It is thought that the rate of visual decline and ocular sequelae with SRT is influenced by dose and location of radiation in relation to important visual structures. This study therefore aimed to look at these prognoses with respect to localisation and dose of radiation when treatment of CM with SRT occurs. Methods: A retrospective data analysis was conducted on all patients at Dunedin Hospital (DH) from August 2001 to May 2017 who were followed up for 4 years. SRT consisted of 50 Gy divided into five fractions over 5 days to tumours, with 2-mm treatment margins. The primary outcome measure was retention of functional vision - better than hand movements (HMs) within the treated eye. Secondary outcome measures included time to non-functional vision (HM or less) in relation to location, dose and tumour thickness, the presence of radiation retinopathy, local and metastatic tumour progression, enucleation, and disease-specific mortality. Results: Seventy-five patients were identified in this study. Follow-up was incomplete in 10 patients, and 4 patients became deceased within the 4-year study period. Twenty-nine patients (48%) retained visual acuity (VA) better than HMs in the treated eye at 4 years, and thirty-two (52%) of patients did not. Calculated dose to the optic nerve and macula and proximity of the tumour to the optic nerve and macula were not statistically determinative of vision outcomes, although presenting VA was. Fifty-six per cent of patients developed radiation retinopathy involving the macula. The local progression, metastatic progression and enucleation rates were 4.6%, 6%, and 12.3%, representing 3, 4, and 8 patients, respectively. Conclusion: This study demonstrates that approximately half of patients treated with SRT can expect to maintain functional vision better than HM at 4 years. The rate of visual decline and final vision outcome are independent of location of the tumour in relation to the optic nerve and macula. While it affirms that SRT achieves high rates of local tumour control and eye retention, preservation of functional VA remains an unpredictable endpoint for individual cases and highlights the therapeutic challenge of this treatment modality.

Vincristine, irinotecan, and temozolomide for treatment of relapsed alveolar rhabdomyosarcoma.

Event-free survival for recurrent alveolar rhabdomyosarcoma (ARMS) is poor, and a consensus approach to treatment in the relapse setting has not been established. Recent studies suggest that a combination regimen of vincristine, irinotecan, and temozolomide (VITA) is active against recurrent sarcomas. We present our single-institution experience with this regimen for relapsed ARMS in heavily pretreated patients, including those with prior exposure to a combination regimen of vincristine and irinotecan. We observed a complete radiographic response in 1 of 4 patients who received VITA as a fifth attempted salvage regimen. Radiographic remission for the responsive patient was sustained for 27 weeks before disease recurrence. All therapies were administered in the outpatient setting and no grade III or grade IV toxicities were observed. These findings suggest that for patients with refractory ARMS, VITA in combination should be among the treatment options considered. They also reinforce the need for biological correlates to prospectively identify patients who may benefit from this treatment.

Comparative Outcomes of Primary Vitrectomy Versus Tap and Inject for Endophthalmitis Following Phacoemulsification Cataract Surgery.

PURPOSE: To compare functional and anatomical outcomes, rates of culture positivity and number of procedures in eyes with endophthalmitis following phacoemulsification surgery, treated with either primary vitrectomy and intravitreal antibiotics or vitreous tap and antibiotic injection (T&I). METHODS: Patients developing endophthalmitis after phacoemulsification surgery between 2007 and 2016 were identified, and outcomes were compared between the two treatment groups. RESULTS: 19 patients underwent a primary vitrectomy and 22 underwent a T&I. There was a significant improvement in visual acuity after T&I (p=.003) and primary vitrectomy (p=.00005). The median improvement in visual acuity was significantly greater for the primary vitrectomy group than the T&I group (p=.024). 64% of eyes were culture positive with the initial T&I, and 63% with primary vitrectomy (p=1.00). Two eyes initially culture negative with a T&I, and three eyes that were culture positive with a T&I were subsequently culture positive with a vitrectomy 24-72 hours later. 68% of patients who underwent a T&I required an additional procedure, compared to 26% of the vitrectomy group (p=.01). The T&I group underwent a mean of 2.3 procedures each, and the primary vitrectomy group underwent 1.5 (p=.03). CONCLUSIONS: Eyes with endophthalmitis treated with a primary vitrectomy demonstrated greater visual improvement and needed fewer procedures than those initially treated with a T&I. Viable bacteria were only seen in subsequent procedures in the T&I group, indicating that primary vitrectomy was superior at sterilizing the eye.

Nanoscale morphology, tribology and electrical properties of polyaniline/graphene oxide/LAPONITE® composites investigated using atomic force microscopy.

Polyaniline composites with graphene and graphene oxide have received broad interest for applications in charge separation and storage in electrical devices. Syntheses via in situ polymerization of aniline in colloidal dispersions of graphene oxide have afforded nanocomposites of polyaniline intercalated within graphene oxide nanosheets. The simultaneous inclusion of LAPONITE® nanoparticles has improved aqueous phase processability and thin film formation. Mechanical, morphological and conductivity studies including atomic force microscopy and scanning electron microscopy were employed to study the host-guest interactions and heterointerfaces that govern self-assembly. Adhesion and conductivity values within graphene oxide/polyaniline/LAPONITE® (GOPL) nanocomposites were tuned by varying the weight ratios of polyaniline : graphene oxide. These studies inform ongoing work to use GOPL nanomaterials for clean energy harvesting and storage applications.

Intravenous lidocaine for the treatment of acute pain in the emergency department.

OBJECTIVE: To evaluate intravenous lidocaine's safety and efficacy as an analgesic agent in the treatment of a variety of painful conditions presenting to the emergency department. METHODS: This case series identified seventeen patients who received lidocaine over a six month period and recorded demographic data, amount of lidocaine administered, the amount of opioid medication administered before and after lidocaine, pre- and post-lidocaine pain scores, and any qualitative descriptors of the patient's pain recorded in the record. Side effects and adverse events were also recorded. RESULTS: Of the seven patients who had a pre- and post-lidocaine pain score recorded, the mean reduction was 3 points on a 10 point scale. Patients who received lidocaine used less opioid medication. One patient received an improperly high dose of lidocaine and suffered a brief seizure and cardiac arrest, but was quickly resuscitated. CONCLUSION: This series suggests that lidocaine may be a useful adjunct in the treatment of acutely painful conditions in the emergency department.

Adaptations to iron deficiency: cardiac functional responsiveness to norepinephrine, arterial remodeling, and the effect of beta-blockade on cardiac hypertrophy.

BACKGROUND: Iron deficiency (ID) results in ventricular hypertrophy, believed to involve sympathetic stimulation. We hypothesized that with ID 1) intravenous norepinephrine would alter heart rate (HR) and contractility, 2) abdominal aorta would be larger and more distensible, and 3) the beta-blocker propanolol would reduce hypertrophy. METHODS: 1) 30 CD rats were fed an ID or replete diet for 1 week or 1 month. Norepinephrine was infused via jugular vein; pressure was monitored at carotid artery. Saline infusions were used as a control. The pressure trace was analyzed for HR, contractility, systolic and diastolic pressures. 2) Abdominal aorta catheters inflated the aorta, while digital microscopic images were recorded at stepwise pressures to measure arterial diameter and distensibility. 3) An additional 10 rats (5 ID, 5 control) were given a daily injection of propanolol or saline. After 1 month, the hearts were excised and weighed. RESULTS: Enhanced contractility, but not HR, was associated with ID hypertrophic hearts. Systolic and diastolic blood pressures were consistent with an increase in arterial diameter associated with ID. Aortic diameter at 100 mmHg and distensibility were increased with ID. Propanolol was associated with an increase in heart to body mass ratio. CONCLUSIONS: ID cardiac hypertrophy results in an increased inotropic, but not chronotropic response to the sympathetic neurotransmitter, norepinephrine. Increased aortic diameter is consistent with a flow-dependent vascular remodeling; increased distensibility may reflect decreased vascular collagen content. The failure of propanolol to prevent hypertrophy suggests that ID hypertrophy is not mediated via beta-adrenergic neurotransmission.

Applied spectrophotometry: analysis of a biochemical mixture.

Spectrophotometric analysis is essential for determining biomolecule concentration of a solution and is employed ubiquitously in biochemistry and molecular biology. The application of the Beer-Lambert-Bouguer Lawis routinely used to determine the concentration of DNA, RNA or protein. There is however a significant difference in determining the concentration of a given species (RNA, DNA, protein) in isolation (a contrived circumstance) as opposed to determining that concentration in the presence of other species (a more realistic situation). To present the student with a more realistic laboratory experience and also to fill a hole that we believe exists in student experience prior to reaching a biochemistry course, we have devised a three week laboratory experience designed so that students learn to: connect laboratory practice with theory, apply the Beer-Lambert-Bougert Law to biochemical analyses, demonstrate the utility and limitations of example quantitative colorimetric assays, demonstrate the utility and limitations of UV analyses for biomolecules, develop strategies for analysis of a solution of unknown biomolecular composition, use digital micropipettors to make accurate and precise measurements, and apply graphing software.

Diet restriction and life history trade-offs in short- and long-lived species of Daphnia.

The life-extending effects of diet restriction are well documented. One evolutionary model that accounts for this widespread conservation is the resource allocation model, where the selected individuals are those that can delay reproduction during periods of resource limitation. In this study, we use closely related species of a model organism, Daphnia, with widely divergent lifespans to address the relationship between diet restriction and longevity and assess whether the relationships are owing to trade-offs between reproductive and somatic investment. Specifically, we conducted a common garden experiment and constructed reaction norms for lifespan, fecundity, and body size as a function of food concentration. Our study provides evidence that the short-lived species in our study, D. pulex, shows the classically observed relationship of enhanced lifespan in response to reduced diet intake, but does not divert resources to somatic maintenance at the expense of reproduction during chronic diet restriction. In contrast, we find no evidence that the long-lived species in our study, D. pulicaria, gains any life-extending effects through diet restriction. Combined, our results provide evidence that the resource allocation model is not sufficient to explain the evolution of diet-mediated lifespan plasticity.