Pan-cortical cellular imaging in freely behaving mice using a miniaturized micro-camera array microscope (mini-MCAM).

Understanding how circuits in the brain simultaneously coordinate their activity to mediate complex ethnologically relevant behaviors requires recording neural activities from distributed populations of neurons in freely behaving animals. Current miniaturized imaging microscopes are typically limited to imaging a relatively small field of view, precluding the measurement of neural activities across multiple brain regions. Here we present a miniaturized micro-camera array microscope (mini-MCAM) that consists of four fluorescence imaging micro-cameras, each capable of capturing neural activity across a 4.5 mm x 2.55 mm field of view (FOV). Cumulatively, the mini-MCAM images over 30 mm2 area of sparsely expressed GCaMP6s neurons distributed throughout the dorsal cortex, in regions including the primary and secondary motor, somatosensory, visual, retrosplenial, and association cortices across both hemispheres. We demonstrate cortex-wide cellular resolution in vivo Calcium (Ca 2+ ) imaging using the mini-MCAM in both head-fixed and freely behaving mice.

Thermodynamic profiles for cotranslational trigger factor substrate recognition.

Molecular chaperones are central to the maintenance of proteostasis in living cells. A key member of this protein family is trigger factor (TF), which acts throughout the protein life cycle and has a ubiquitous role as the first chaperone encountered by proteins during synthesis. However, our understanding of how TF achieves favorable interactions with such a diverse substrate base remains limited. Here, we use microfluidics to reveal the thermodynamic determinants of this process. We find that TF binding to empty 70S ribosomes is enthalpy-driven, with micromolar affinity, while nanomolar affinity is achieved through a favorable entropic contribution for both intrinsically disordered and folding-competent nascent chains. These findings suggest a general mechanism for cotranslational TF function, which relies on occupation of the exposed TF-substrate binding groove rather than specific complementarity between chaperone and nascent chain. These insights add to our wider understanding of how proteins can achieve broad substrate specificity.

Polymeric nanomaterials-based theranostic platforms for triple-negative breast cancer (TNBC) treatment.

Breast cancer, the second leading global cause of death, affects 2.1 million women annually, with an alarming 15 percent mortality rate. Among its diverse forms, Triple-negative breast cancer (TNBC) emerges as the deadliest, characterized by the absence of hormone receptors. This article underscores the urgent need for innovative treatment approaches in tackling TNBC, emphasizing the transformative potential of polymeric nanomaterials (PNMs). Evolved through nanotechnology, PNMs offer versatile biomedical applications, particularly in addressing the intricate challenges of TNBC. The synthesis methods of PNMs, explored within the tumor microenvironment using cellular models, showcase their dynamic nature in cancer treatment. The article anticipates the future of TNBC therapeutics through the optimization of PNMs-based strategies, integrating them into photothermal (PT), photodynamic (PT), and hyperthermia therapy (HTT), drug delivery, and active tumor targeting strategies. Advancements in synthetic methods, coupled with a nuanced understanding of the tumor microenvironment, hold promise for personalized interventions. Comparative investigations of therapeutic models and a thorough exploration of polymeric nanoplatforms toxicological perspectives become imperative for ensuring efficacy and safety. We have explored the interdisciplinary collaboration between nanotechnology, oncology, and molecular biology as pivotal in translating PNMs innovations into tangible benefits for TNBC patients.

Emerging natural polymer-based architectured nanotherapeutics for the treatment of cancer.

The field of cancer therapy is advancing rapidly, placing a crucial emphasis on innovative drug delivery systems. The increasing global impact of cancer highlights the need for creative therapeutic strategies. Natural polymer-based nanotherapeutics have emerged as a captivating avenue in this pursuit, drawing substantial attention due to their inherent attributes. These attributes include biodegradability, biocompatibility, negligible toxicity, extended circulation time, and a wide range of therapeutic payloads. The unique size, shape, and morphological characteristics of these systems facilitate profound tissue penetration, complementing active and passive targeting strategies. Moreover, these nanotherapeutics exploit specific cellular and subcellular trafficking pathways, providing precise control over drug release kinetics. This comprehensive review emphasizes the utilization of naturally occurring polymers such as polysaccharides (e.g., chitosan, hyaluronic acid, alginates, dextran, and cyclodextrin) and protein-based polymers (e.g., ferritin, gelatin, albumin) as the foundation for nanoparticle development. The paper meticulously examines their in vitro characteristics alongside in vivo efficacy, particularly focusing on their pivotal role in ameliorating diverse types of solid tumors within cancer therapy. The amalgamation of material science ingenuity and biological insight has led to the formulation of these nanoparticles, showcasing their potential to reshape the landscape of cancer treatment.

Generation of the iPSC line FINi002-A from a male Parkinson's disease patient carrying compound heterozygous mutations in the PRKN gene.

The most common cause of autosomal recessive familial Parkinson's disease (PD) are mutations in the PRKN/PARK2 gene encoding an E3 ubiquitin protein-ligase PARKIN. We report the generation of an iPSC cell line from the fibroblasts of a male PD patient carrying a common missense variant in exon 7 (p.Arg275Trp), and a 133 kb deletion encompassing exon 8, using transiently-present Sendai virus. The established line displays typical human primed iPSC morphology and expression of pluripotency-associated markers, normal karyotype without SNP array-detectable copy number variations and can give rise to derivatives of all three embryonic germ layers. We envisage the usefulness of this iPSC line, carrying a common and well-studied missense mutation in the RING1 domain of the PARKIN protein, for the elucidation of PARKIN-dependent mechanisms of PD using in vitro and in vivo models.

Parallelized computational 3D video microscopy of freely moving organisms at multiple gigapixels per second.

Wide field of view microscopy that can resolve 3D information at high speed and spatial resolution is highly desirable for studying the behaviour of freely moving model organisms. However, it is challenging to design an optical instrument that optimises all these properties simultaneously. Existing techniques typically require the acquisition of sequential image snapshots to observe large areas or measure 3D information, thus compromising on speed and throughput. Here, we present 3D-RAPID, a computational microscope based on a synchronized array of 54 cameras that can capture high-speed 3D topographic videos over an area of 135 cm2, achieving up to 230 frames per second at spatiotemporal throughputs exceeding 5 gigapixels per second. 3D-RAPID employs a 3D reconstruction algorithm that, for each synchronized snapshot, fuses all 54 images into a composite that includes a co-registered 3D height map. The self-supervised 3D reconstruction algorithm trains a neural network to map raw photometric images to 3D topography using stereo overlap redundancy and ray-propagation physics as the only supervision mechanism. The resulting reconstruction process is thus robust to generalization errors and scales to arbitrarily long videos from arbitrarily sized camera arrays. We demonstrate the broad applicability of 3D-RAPID with collections of several freely behaving organisms, including ants, fruit flies, and zebrafish larvae.

CONVIQT: Contrastive Video Quality Estimator.

Perceptual video quality assessment (VQA) is an integral component of many streaming and video sharing platforms. Here we consider the problem of learning perceptually relevant video quality representations in a self-supervised manner. Distortion type identification and degradation level determination is employed as an auxiliary task to train a deep learning model containing a deep Convolutional Neural Network (CNN) that extracts spatial features, as well as a recurrent unit that captures temporal information. The model is trained using a contrastive loss and we therefore refer to this training framework and resulting model as CONtrastive VIdeo Quality EstimaTor (CONVIQT). During testing, the weights of the trained model are frozen, and a linear regressor maps the learned features to quality scores in a no-reference (NR) setting. We conduct comprehensive evaluations of the proposed model against leading algorithms on multiple VQA databases containing wide ranges of spatial and temporal distortions. We analyze the correlations between model predictions and ground-truth quality ratings, and show that CONVIQT achieves competitive performance when compared to state-of-the-art NR-VQA models, even though it is not trained on those databases. Our ablation experiments demonstrate that the learned representations are highly robust and generalize well across synthetic and realistic distortions. Our results indicate that compelling representations with perceptual bearing can be obtained using self-supervised learning.

Parallelized computational 3D video microscopy of freely moving organisms at multiple gigapixels per second.

To study the behavior of freely moving model organisms such as zebrafish (Danio rerio) and fruit flies (Drosophila) across multiple spatial scales, it would be ideal to use a light microscope that can resolve 3D information over a wide field of view (FOV) at high speed and high spatial resolution. However, it is challenging to design an optical instrument to achieve all of these properties simultaneously. Existing techniques for large-FOV microscopic imaging and for 3D image measurement typically require many sequential image snapshots, thus compromising speed and throughput. Here, we present 3D-RAPID, a computational microscope based on a synchronized array of 54 cameras that can capture high-speed 3D topographic videos over a 135-cm^2 area, achieving up to 230 frames per second at throughputs exceeding 5 gigapixels (GPs) per second. 3D-RAPID features a 3D reconstruction algorithm that, for each synchronized temporal snapshot, simultaneously fuses all 54 images seamlessly into a globally-consistent composite that includes a coregistered 3D height map. The self-supervised 3D reconstruction algorithm itself trains a spatiotemporally-compressed convolutional neural network (CNN) that maps raw photometric images to 3D topography, using stereo overlap redundancy and ray-propagation physics as the only supervision mechanism. As a result, our end-to-end 3D reconstruction algorithm is robust to generalization errors and scales to arbitrarily long videos from arbitrarily sized camera arrays. The scalable hardware and software design of 3D-RAPID addresses a longstanding problem in the field of behavioral imaging, enabling parallelized 3D observation of large collections of freely moving organisms at high spatiotemporal throughputs, which we demonstrate in ants (Pogonomyrmex barbatus), fruit flies, and zebrafish larvae.

Gigapixel imaging with a novel multi-camera array microscope.

The dynamics of living organisms are organized across many spatial scales. However, current cost-effective imaging systems can measure only a subset of these scales at once. We have created a scalable multi-camera array microscope (MCAM) that enables comprehensive high-resolution recording from multiple spatial scales simultaneously, ranging from structures that approach the cellular scale to large-group behavioral dynamics. By collecting data from up to 96 cameras, we computationally generate gigapixel-scale images and movies with a field of view over hundreds of square centimeters at an optical resolution of 18 µm. This allows us to observe the behavior and fine anatomical features of numerous freely moving model organisms on multiple spatial scales, including larval zebrafish, fruit flies, nematodes, carpenter ants, and slime mold. Further, the MCAM architecture allows stereoscopic tracking of the z-position of organisms using the overlapping field of view from adjacent cameras. Overall, by removing the bottlenecks imposed by single-camera image acquisition systems, the MCAM provides a powerful platform for investigating detailed biological features and behavioral processes of small model organisms across a wide range of spatial scales.

High-speed multi-objective Fourier ptychographic microscopy.

The ability of a microscope to rapidly acquire wide-field, high-resolution images is limited by both the optical performance of the microscope objective and the bandwidth of the detector. The use of multiple detectors can increase electronic-acquisition bandwidth, but the use of multiple parallel objectives is problematic since phase coherence is required across the multiple apertures. We report a new synthetic-aperture microscopy technique based on Fourier ptychography, where both the illumination and image-space numerical apertures are synthesized, using a spherical array of low-power microscope objectives that focus images onto mutually incoherent detectors. Phase coherence across apertures is achieved by capturing diffracted fields during angular illumination and using ptychographic reconstruction to synthesize wide-field, high-resolution, amplitude and phase images. Compared to conventional Fourier ptychography, the use of multiple objectives reduces image acquisition times by increasing the area for sampling the diffracted field. We demonstrate the proposed scaleable architecture with a nine-objective microscope that generates an 89-megapixel, 1.1 µm resolution image nine-times faster than can be achieved with a single-objective Fourier-ptychographic microscope. New calibration procedures and reconstruction algorithms enable the use of low-cost 3D-printed components for longitudinal biological sample imaging. Our technique offers a route to high-speed, gigapixel microscopy, for example, imaging the dynamics of large numbers of cells at scales ranging from sub-micron to centimetre, with an enhanced possibility to capture rare phenomena.

Identifying the optimal developmental age of human pluripotent stem cell-derived midbrain dopaminergic progenitors for transplantation in a rodent model of Parkinson's disease.

Donor cell age can have a significant impact on transplantation outcomes. Despite the rapid advancement of human pluripotent stem cell (hPSC)-derived dopaminergic (DA) progenitors to the clinic for transplantation into Parkinson's Disease (PD), surprisingly limited data exists regarding the influence of cellular age on neural graft survival, composition, and integration. Here we examined the impact of transplanting ventral midbrain (VM) progenitors at varying days of differentiation (from day 13-30) into a rodent PD model, comparing two hPSC lines (an embryonic and an induced pluripotent cell line, hESC and hiPSC, respectively). Both hPSC lines expressed GFP under the promoter PITX3 enabling specific tracking of graft-derived DA neurons. Post-mortem analysis at 6 months revealed larger grafts from Day19 (D19), D22 and D25 progenitors, yet contained a higher proportion of non-DA and poorly specified (FOXA2-) cells. While D13 and D30 progenitors yielded smaller grafts. D13-derived grafts had the highest DA neuron proportion and proportionally more GIRK2+ DA neurons, the subpopulation critical for motor function. These younger progenitor grafts maintained their capacity to innervate developmentally relevant DA targets, with increased innervation capacity per DA neuron, collectively resulting in restoration of motor deficits with equal or greater proficiency than older donor cells. While donor age effects were reproducible for a given hPSC line and trends were similar between the two hPSC lines, grafts of D13 hiPSC-derived progenitors showed a 6-fold greater density of DA neurons compared to D13 hESC-derived grafts, highlighting between-line variability. These findings show that hPSC-derived VM donor age has a direct impact on graft survival, composition and maturation, and that careful assessment, on a line-to-line basis is required prior to translation.

Image Quality Assessment Using Contrastive Learning.

We consider the problem of obtaining image quality representations in a self-supervised manner. We use prediction of distortion type and degree as an auxiliary task to learn features from an unlabeled image dataset containing a mixture of synthetic and realistic distortions. We then train a deep Convolutional Neural Network (CNN) using a contrastive pairwise objective to solve the auxiliary problem. We refer to the proposed training framework and resulting deep IQA model as the CONTRastive Image QUality Evaluator (CONTRIQUE). During evaluation, the CNN weights are frozen and a linear regressor maps the learned representations to quality scores in a No-Reference (NR) setting. We show through extensive experiments that CONTRIQUE achieves competitive performance when compared to state-of-the-art NR image quality models, even without any additional fine-tuning of the CNN backbone. The learned representations are highly robust and generalize well across images afflicted by either synthetic or authentic distortions. Our results suggest that powerful quality representations with perceptual relevance can be obtained without requiring large labeled subjective image quality datasets. The implementations used in this paper are available at https://github.com/pavancm/CONTRIQUE.

ST-GREED: Space-Time Generalized Entropic Differences for Frame Rate Dependent Video Quality Prediction.

We consider the problem of conducting frame rate dependent video quality assessment (VQA) on videos of diverse frame rates, including high frame rate (HFR) videos. More generally, we study how perceptual quality is affected by frame rate, and how frame rate and compression combine to affect perceived quality. We devise an objective VQA model called Space-Time GeneRalized Entropic Difference (GREED) which analyzes the statistics of spatial and temporal band-pass video coefficients. A generalized Gaussian distribution (GGD) is used to model band-pass responses, while entropy variations between reference and distorted videos under the GGD model are used to capture video quality variations arising from frame rate changes. The entropic differences are calculated across multiple temporal and spatial subbands, and merged using a learned regressor. We show through extensive experiments that GREED achieves state-of-the-art performance on the LIVE-YT-HFR Database when compared with existing VQA models. The features used in GREED are highly generalizable and obtain competitive performance even on standard, non-HFR VQA databases. The implementation of GREED has been made available online: https://github.com/pavancm/GREED.

Low-cost, sub-micron resolution, wide-field computational microscopy using opensource hardware.

The revolution in low-cost consumer photography and computation provides fertile opportunity for a disruptive reduction in the cost of biomedical imaging. Conventional approaches to low-cost microscopy are fundamentally restricted, however, to modest field of view (FOV) and/or resolution. We report a low-cost microscopy technique, implemented with a Raspberry Pi single-board computer and color camera combined with Fourier ptychography (FP), to computationally construct 25-megapixel images with sub-micron resolution. New image-construction techniques were developed to enable the use of the low-cost Bayer color sensor, to compensate for the highly aberrated re-used camera lens and to compensate for misalignments associated with the 3D-printed microscope structure. This high ratio of performance to cost is of particular interest to high-throughput microscopy applications, ranging from drug discovery and digital pathology to health screening in low-income countries. 3D models and assembly instructions of our microscope are made available for open source use.

Sixth hour transcutaneous bilirubin predicting significant hyperbilirubinemia in ABO incompatible neonates.

BACKGROUND: Neonates with ABO hemolytic disease are at greater risk for developing significant hyperbilirubinemia. We aimed to determine whether sixth hour transcutaneous bilirubin (TcB) could predict such a risk. METHODS: TcB measurements were obtained at the 6th hour of life in blood group A or B neonates born to blood group O, rhesus factor compatible mothers. Subsequent hyperbilirubinemia was monitored and considered significant if a neonate required phototherapy/exchange transfusion. The predictive role of sixth hour TcB was estimated. RESULTS: Of 144 ABO incompatible neonates, 41(OA, 24; O-B, 17) had significant hyperbilirubinemia. Mean sixth hour TcB was significantly higher among neonates who developed significant hyperbilirubinemia than those who did not (5.83±1.35 mg/dL vs. 3.65±0.96 mg/dL, P<0.001). Sixth hour TcB value >4 mg/dL had the highest sensitivity of 93.5% and >6 mg/dL had the highest specificity of 99%. Area under receiver operating characteristic curve was 0.898. CONCLUSION: Sixth hour TcB predicts subsequent significant hyperbilirubinemia in ABO incompatible neonates.

Validation of visual analogue scale for anxiety (VAS-A) in preanesthesia evaluation.

BACKGROUND: Anxiety is a relevant but still underscored perioperative problem. The Visual Analogue Scale for Anxiety (VAS-A) seems to be effective, fast and manageable, but has not been fully validated yet. The aim of this study is to validate VAS-A comparing it to, Corah's Dental Anxiety Scale (CDAS) Spielberger's State Trait Anxiety Inventory (STAI) and Beck Depression Inventory (BDI). METHODS: One hundred consecutive patients (38 males and 62 females, median age 49 years) submitted to oral surgery filled out the VAS-A, CDAS, STAI forms Y1 and Y2, and BDI at preoperative examination; the order of administration of tests was randomized. RESULTS: VAS-A score was significantly correlated to CDAS (P<0.0001), STAI-Y1 (P<0.0001), STAI-Y2 (P<0.002) but not to BDI (P=0.18). ROC curve analysis suggested VAS-A equal to 46 mm as threshold for anxiety when using STAI Y1 equal to 40 as reference cutoff. CONCLUSION: Our study confirms that VAS-A is a reliable indicator of preoperative anxiety and may detect patients with depressive symptoms also. Values of VAS-A around 50 mm are a reliable threshold for a clinically meaningful level of preoperative anxiety.

Identification of bacterial infections and clinical manifestation associated with cytomegalovirus in liver transplantation patients.

INTRODUCTION: Liver transplantation has become the most effective therapy for the treatment of patients with end-stage liver disease. With new immunosuppressive agents the incidence of acute rejection has been significantly reduced, but infection has become a serious problem. OBJECTIVE: Our objective was to correlate cytomegalovirus (CMV) positivity of antigenemia and polymerase chain reaction (PCR) with clinical manifestations and bacterial infections among patients undergoing liver transplantation. METHODS: This prospective study included patients monitored for 6 months for early detection of CMV infection. Sample collections were performed at the time of surgery and weekly until the second month followed by fortnightly in the third month, and monthly in the fourth to sixth month. CMV infection was defined by positive antigenemia (>3 cells) or 2 positive PCR tests associated or not with clinical symptoms. The methodology for the diagnosis of bacterial infection was through biochemical tests and the automated VITEK/bioMérieux (identification and antibiogram) using samples of urine and blood cultures. Chi-square test was used for dicotomic variables with significant differences when P < .05. RESULTS: Sixteen patients (32%) had CMV infections, including 13 (81%) with concomitant infections. Thirty-four patients (68%) did not have CMV infections and 8 of these (24%) had bacterial infection. There was a high correlation with bacterial infections among CMV-positive patients. CONCLUSION: Bacterial infections after liver transplantation were associated with CMV infection.

Psychometric features of temporomandibular disorders patients in relation to pain diffusion, location, intensity and duration.

The aim of the present investigation was to assess the psychological profile of a sample of patients with temporomandibular disorders (TMD) and to compare the psychometric scores between patients with pain of different diffusion, location, intensity and duration. One hundred and ten (N = 110) patients with painful TMD fulfilled three psychometric instruments. Pain features were assessed as categorical variables as concerns its diffusion, viz., diffuse or localised, duration, viz., more or <6 months, and location, viz., joint and/or muscles. Pain intensity was scored on a 0-100 Visual Analog Scale (VAS) rating. Patients with diffuse pain showed higher psychometric scores than patients with localised pain. No significant differences were detected between patients with pain lasting from more or equal than 6 months and those with pain lasting from <6 months as well as between patients with pain localised in the jaw muscles, joints or both, even if a trend for lower scores for patients with joint pain alone was observed. Pain intensity was significantly related with anxiety (ANX), depression (DEP) and somatisation(SOM) scores. In conclusion, pain diffusion and intensity were strongly related with high levels of SOM, ANX and DEP, while no differences in psychometric scores were detected between patients with pain of different duration and location.

Monitoring and detection of cytomegalovirus in liver transplant recipients.

Cytomegalovirus (CMV) is a ß-herpesvirus. CMV infections are a common complication contributing to morbidity and mortality after liver transplantation. Among organ transplant recipients, CMV can reactivate from latency during the first 6 months. This prospective study performed from February 2008 to December 2009 examined liver transplant recipients during the first 6 months. Two methods were performed to detect CMV infections: antigenemia (AGM) and nested (PCR). Ninety-four patients, including 72 men (76.6%) and 22 women (23.4%) underwent liver transplantation during this period. We analyzed 575 samples including 465 for AGM and PCR. Forty-three (9.25%) showed positive AGM as detected 2 to 179 days posttransplantation with a mean of 50 days and a median of 35 days, and 93/465 (20%) showed positive PCR at 0 to 186 days posttransplantation with a mean of 31 days and a median of 38 days. Among the 43 antigenemia patients, 38 samples were positive for up to 5 cells 18 of which were PCR-positive. Five samples were positive with more than 5 cells, including 3 that were PCR-positive. Only 4.51% had AGM and were PCR-positive in the same sample. Despite only 9.25% (43/465) showing AGM, the current study suggested the utility of routine monitoring to detect early CMV infection among liver transplantation patients seeking to reduce morbidity and mortality.

Weight gain related to treatment with atypical antipsychotics is due to activation of PKC-ß.

Atypical antipsychotics (APDs) are currently used in clinical practice for a variety of mental disorders such as schizophrenia, bipolar disorder and severe behavioral disturbances. A well-known disadvantage of using these compounds is a propensity for weight gain, resulting frequently in obesity. The mechanisms underlying pharmacologically induced weight gain are still controversial. The objective of this study was to evaluate in vitro the effects of different APDs on adipogenic events in cultured human pre-adipocytes and in rat muscle-derived stem cells (MDSCs), aiming to identify a common intracellular event contributable to these drugs. Culture behavior was evaluated in terms of cell proliferation, lipid accumulation, gene expression and morphological features. Results indicate that APDs influence adipogenic events through changes in the differentiation and proliferation of pre-adipocytes and MDSCs that are brought on by protein kinase C-ß (PKC-ß) activation. These data identify a signaling route that could be a potential target of pharmacological approaches for preventing the weight gain associated with APD treatment.