Small Brain Research Articles

Main causes of death of free-ranging bats in Turin province (North-Western Italy): gross and histological findings and emergent virus surveillance

BackgroundBats are recognized as reservoir species for multiple viruses. However, little is known on bats’ health and mortality. Thus, this study aimed to investigate the main causes of death of bats from Turin province (North-western Italy) and to describe gross and histopathological lesions potentially associated with the presence of selected bat viruses.ResultsA total of 71 bats belonging to 9 different species of the families Vespertilionidae and Molossidae were necropsied and samples of the main organs were submitted to histopathological examination. Also, aliquots of the small intestine, liver, spleen, lung, and brain were collected and submitted to biomolecular investigation for the identification of Coronaviridae, Poxviridae, Reoviridae (Mammalian orthoreovirus species), Rhabdoviridae (Vaprio ledantevirus and Lyssavirus species) and Kobuvirus. The majority of bats died from traumatic lesions due to unknown trauma or predation (n = 40/71, 56.3%), followed by emaciation (n = 13/71,18.3%). The main observed gross lesions were patagium and skin lesions (n = 23/71, 32.4%), forelimbs fractures (n = 15/71, 21.1%) and gastric distension (n = 10/71,14.1%). Histologically, the main lesions consisted of lymphoplasmacytic pneumonia (n = 24/71, 33.8%), skin/patagium dermatitis (n = 23/71, 32.4%), liver steatosis and hepatitis (n = 12, 16.9%), and white pulp depletion in the spleen (n = 7/71, 9.8%). Regarding emergent bat viruses, only poxvirus (n = 2, 2.8%) and orthoreovirus (n = 12/71, 16.9%) were detected in a low percentage of bats.ConclusionsTrauma is the main lesion observed in bats collected in Turin province (North-western Italy) associated with forelimb fractures and the detected viral positivity rate seems to suggest that they did not represent a threat for human health.

Intracranial arteriosclerosis and the risk of dementia: A population-based cohort study.

The impact of intracranial arteriosclerosis on dementia remains largely unclear. In 2339 stroke-free and dementia-free participants (52.2% women, mean age 69.5 years) from the general population, we assessed intracranial carotid artery calcification (ICAC) and vertebrobasilar artery calcification (VBAC) as proxy for arteriosclerosis. Associations with dementia were assessed using Cox models. In addition, indirect effects through cerebral small vessel disease (cSVD) and subcortical brain structure volumes were assessed using causal mediation analyses. During a median of 13.4 years (25th-75th percentiles 9.9-14.5) of follow-up, 282 participants developed dementia. Both ICAC presence (hazard ratio [HR]: 1.53, 95% confidence interval [CI]: 1.00-2.32]) and volume (HR per standard deviation: 1.19, 95% CI: 1.01-1.40) increased dementia risk. For VBAC, severe calcifications increased dementia risk (HR for third vs first volume tertile: 1.89, 95% CI: 1.00-3.59). These effects were mediated partly through increased cSVD (percentage mediated for ICAC: 13% and VBAC: 24%). Intracranial arteriosclerosis increases the risk of dementia.

Drosophila CASK regulates brain size and neuronal morphogenesis, providing a genetic model of postnatal microcephaly suitable for drug discovery

BackgroundCASK-related neurodevelopmental disorders are untreatable. Affected children show variable severity, with microcephaly, intellectual disability (ID), and short stature as common features. X-linked human CASK shows dosage sensitivity with haploinsufficiency in females. CASK protein has multiple domains, binding partners, and proposed functions at synapses and in the nucleus. Human and Drosophila CASK show high amino-acid-sequence similarity in all functional domains. Flies homozygous for a hypomorphic CASK mutation (∆18) have motor and cognitive deficits. A Drosophila genetic model of CASK-related disorders could have great scientific and translational value.MethodsWe assessed the effects of CASK loss of function on morphological phenotypes in Drosophila using established genetic, histological, and primary neuronal culture approaches. NeuronMetrics software was used to quantify neurite-arbor morphology. Standard nonparametric statistics methods were supplemented by linear mixed effects modeling in some cases. Microfluidic devices of varied dimensions were fabricated and numerous fluid-flow parameters were used to induce oscillatory stress fields on CNS tissue. Dissociation into viable neurons and neurite outgrowth in vitro were assessed.ResultsWe demonstrated that ∆18 homozygous flies have small brains, small heads, and short bodies. When neurons from developing CASK-mutant CNS were cultured in vitro, they grew small neurite arbors with a distinctive, quantifiable “bushy” morphology that was significantly rescued by transgenic CASK+. As in humans, the bushy phenotype showed dosage-sensitive severity. To overcome the limitations of manual tissue trituration for neuronal culture, we optimized the design and operation of a microfluidic system for standardized, automated dissociation of CNS tissue into individual viable neurons. Neurons from CASK-mutant CNS dissociated in the microfluidic system recapitulate the bushy morphology. Moreover, for any given genotype, device-dissociated neurons grew larger arbors than did manually dissociated neurons. This automated dissociation method is also effective for rodent CNS.ConclusionsThese biological and engineering advances set the stage for drug discovery using the Drosophila model of CASK-related disorders. The bushy phenotype provides a cell-based assay for compound screening. Nearly a dozen genes encoding CASK-binding proteins or transcriptional targets also have brain-development mutant phenotypes, including ID. Hence, drugs that improve CASK phenotypes might also benefit children with disorders due to mutant CASK partners.

2004P PD-L1 Inhibitors combined with whole brain radiotherapy in patients with small cell lung cancer brain metastases: Real-world evidence

2004P PD-L1 Inhibitors combined with whole brain radiotherapy in patients with small cell lung cancer brain metastases: Real-world evidence

Investigation of the Relationship between Aperture Use for Treating Small and Shallow Brain Lesions in Proton Therapy with Different Spot Size Variations in Three Proton Therapy Systems

To compare plan quality among standard vs. aperture-based Intensity-Modulated Proton Therapy (IMPT) using cyclotron-generated proton beams (CPB), linear accelerator proton beams (LPB), and linear accelerator proton minibeams (LPMB) for multiple brain metastases. Fifty-five brain lesions from twenty patients were planned with three different spot size ranges using CPBs (σ: 2.7-7.0 mm) and compared against LPBs (σ: 2.9-5.5 mm), and LPMBs (σ: 0.9-3.9 mm). Apertures with a diameter of 0.3 cm were applied to beams irradiating all tumors < 1 cm3 in volume and any tumor < 2.5 cm depth in the patient and compared against the same patient plans containing no apertures. All plans were optimized with the multi-field optimization (MFO) technique using the Monte Carlo algorithm. Dose coverage to each lesion for each proton plan was set to 99% of the GTV receiving the prescription (Rx) dose for all plans. Robustness with ±2 mm setup uncertainty and ±2% range uncertainty was included in robust evaluation using V100%Rx > 95% of the GTV. Conformity index (CI) and gradient index (GI) were used to analyze the effect of apertures vs. no apertures (standard) for each IMPT plan type. CI was defined as the volume of the 100% isodose line divided by the volume of the GTV + 2 mm expansion to account for robust planning. The Wilcoxon signed rank test was utilized to determine the statistical significance of dosimetric results compared between aperture-based and standard IMPT plans. When apertures were implemented in the treatment planning for all shallow and small brain lesions, shallow brain tumors showed the most prominent improvement in conformity and gradient index. A 6.7% difference in average conformity was calculated for standard vs. aperture-based plans for LPMBs, followed by a 5.3% improvement for CPBs. Improvement in gradient index for standard vs. aperture-based plans was significant for both shallow and deeper tumors for CPB and LPB plan types, as shown in Table 1 below. CPB and LPB plan gradient indices were statistically significant for comparing aperture-based vs. standard IMPT plans. We successfully quantified plan quality and evaluated results for aperture- vs. standard IMPT plans using CPBs, LPBs, and LPMBs for brain metastases. Plan quality improves the greatest with apertures applied to beams irradiating shallow tumors. Apertures may not be necessary for small, deeper tumors with IMPT.

Stereotactic Radiosurgery for Small Intact Brain Metastasis: A Comparative Evaluation of 3 Different Single Fraction Prescription Doses

While single-fraction stereotactic radiosurgery (SRS) in the treatment of small brain metastases (SBM, ≤ 2 cm) is well established, prescription dosing varies considerably across institutions and clinical trials. The choice of prescription dose is a delicate balance between local failure (LF) and radiation necrosis (RN) risks. In the modern era, historically-established dosing thresholds may no longer be applicable. To evaluate the impact of prescription dose on outcomes, we performed a comparative analysis of patients with SBM treated with definitive SRS using three different prescriptions, at a single tertiary institution. Consecutive patients with intact SBM treated with SRS from January 2017 and December 2021 were analyzed. Baseline patient characteristics and dosing parameters were abstracted from the medical record. To limit the integral brain dose when treating multiple brain metastases, the institutional practice was to reduce prescription dose as the total number of lesions increased (i.e., 24 Gy for ≤10 lesions, 22 Gy for 11-20, and 20 Gy for >20). A per lesion analysis, where each lesion was followed from the date of SRS to the last follow-up, was conducted with primary endpoints of LF and RN. Gray's test was used to compare the cumulative incidence of the LF and RN, with death as a competing risk. Factors affecting LF were analyzed using Cox hazard regression analysis. A total of 1318 SBM in 250 patients received SRS and met the inclusion criteria. The median age was 62 years (range: 18-90), median KPS was 90 (range: 50-100) and 66% were female. The most common primary tumors were lung (55.5%) and breast cancers (26.4%). With a median follow-up of 12 months, 136 (11%) LF in 44 patients and 70 (5.7%) RN events in 46 patients occurred. The actuarial 1-year cumulative rate of LF was lower in lesions treated with 24 Gy (6.4%, 95% CI: 4.7-8.6%) or 22 Gy (5.8%, 95% CI: 3.7-8.7%) compared to 20 Gy (15.4%, 95% CI: 10.9-20.5%) (p<0.01). 22 Gy and 24 Gy prescription doses were associated with a 44% and 52% reduction in risk in LF compared to 20 Gy (HR: 0.56; 95% CI: 0.36-0.9; p = 0.01 and HR: 0.48; 95% CI: 0.31-0.74; p<0.01, respectively). In a subset analysis of radiosensitive tumors, 1-year LF rate was still lower with 24 Gy (7.4%, 95% CI: 5.3-9.9%) and 22 Gy (6.1%, 95% CI: 3.7-9.4%) than 20 Gy (15.7%, 95% CI: 11.2-21%) (p = 0.01). The cumulative 1-year RN rate numerically declined with dose, but was not statistically significantly different, with 3.6% (95% CI: 2.3-5.3%) for 24 Gy, 2.6% (95% CI: 1.3-4.8%) for 22 Gy and 1.4% (95% CI: 0.4-3.7%) for 20 Gy. Patients treated with single fraction SRS to intact SBM were at increased risk of LF with prescription doses of 20 Gy compared to 22-24 Gy, without an increased risk of RN. Even in patients with radiosensitive histologies, higher LF rates were still observed following 20 Gy compared to 22-24 Gy.

Feasibility of Using Pseudo-CT for Dosimetry, Radiomics, and Efficacy Assessment in IMRT/VMAT of Brain Tumors: A Multi-Omics Analysis

Pseudo-CT generated by convolutional neural networks (CNN) and planning MRI has facilitated the promotion of MRI-Only. The technology not only reduces the time and money spent on CT scans, but also eliminates the cumbersome CT-MR registration. The feasibility in Stereotactic Brain Radiotherapy has been analyzed in previous studies by our team. However, when the prescribed requirements are not met, IMRT/VMAT are still selected. The study aims to evaluate the feasibility of pseudo-CT in IMRT/VMAT for brain cancer via the following 5 aspects: (1) image difference, (2) dose accuracy, (3) radiomics feature, (4) efficacy assessment, and (5) correlation analysis. Brain tumor patients who had received radiotherapy at our institution and had planning MRI and CT were included in the study. Redesign of IMRT and VMAT radiotherapy plans according to 3 × 15Gy for each patient. Hounsfield unit (HU) values for PTV and OARs were used to assess image differences. And dose accuracy analysis contained a 2D dose volume histogram (DVH) metrics (Dmax, Dmean, D2%, D50%, D98%, HI, CI) and 3D gamma metrics (criteria: 1-3%/2mm, 1%/1mm, 10% threshold). Then 107 original image features of PTV and OARs were extracted for radiometry analysis. And tumor control probability (TCP) of PTV (Poisson model) and normal tissue complication probability (NTCP) of OARs (Lyman-Kutcher-Burman model) were used for the variance analysis of efficacy assessment. Wilcox-test was used for significance of differences test (0.05), and spearman correlation analysis was used to explore the key features of the dose bias. A total of 42 patients were included, with 42 planning CTs and pseudo-CTs (mDixon-T1), and 38 pseudo-CTs (mDixon-T1-CE). The median volume of PTV was 4.1 cc (range 0.5-27.3), with no significant differences in HU, DVH, 3D gamma, and NTCP/TCP metrics. The median local gamma passing rates (1%/1mm) between planning CTs and pseudo-CTs (mDixon-T1) were 93.1% (range 65.5%-99.7%, IMRT) and 93.3% (range 63.9%-99.6%, VMAT). And more than 85% original radiomics features have significant difference. Further, the feature HU-Min was found to be more correlated with dose metrics in the correlation analysis. We speculate that it may be caused by the smoothing of the low frequency signal before outputting image. And since Shape_MeshVolume, Shape_VoxelVolume and PTV volume difference are highly correlated with dose deviation, it indicates that dose deviation affected by CT-MR registration. This study has the potential to provide guidance for the clinical application of pseudo-CT in the MRI-Only workflow with IMRT/VMAT for brain tumors. These quantitative results strongly indicate pseudo-CT can be used as a substitute for the initial CT in IMRT/VMAT for small brain lesions (size <5 cm, numbers <5), but not for radiomics analysis. Additionally, the impact of inter-image differences on dose accuracy is less significant compared to the deviation caused by image registration.

Outcomes of 30 Gy/5 Fr Hypofractionated Stereotactic Radiation Therapy for Small Brain Metastases (≤2 cm).

Stereotactic radiosurgery (SRS)-used for brain metastases (BMs) with a tumor diameter of ≤2 cm-has a high local control rate, however, it can cause symptomatic radiation-induced brain necrosis. Hypofractionated stereotactic radiation therapy (HFSRT) is not commonly used for such lesions and its effectiveness remains unknown. Herein, the efficacy of 30 Gy 5-fraction HFSRT for treating BMs of <2 cm was retrospectively evaluated. Patients who received HFSRT and had a gross tumor volume (GTV) of ≤2 cm in maximum diameter were included in the study (49 patients; 179 BMs; median follow-up period, 11.9 months). The mean GTV Peripheral Dose (D95) was 36.2 Gy. The local control (LC) rates at 1 and 2 years were 93.0% and 81.5%, respectively, for all lesions. The 1-year LC rates were 93.6% and 92.0% for ≤1.0-cm and 1.0-2.0-cm lesions, respectively. Multivariate analysis revealed that the only significant difference was in GTV maximal tumor diameter (HR=1.961, p=0.0002). Notably, only one patient had asymptomatic radiation necrosis. Owing to the high toxicity of SRS, 5-fraction HFSRT can be an effective treatment strategy for BMs of <2 cm.

Stereotactic Radiosurgery (SRS) for Large Brain Metastases: Dosimetric and Clinical Predictors of Local Progression and Radionecrosis

Stereotactic radiosurgery (SRS) provides high rates of local control for small brain metastases with low rates of radionecrosis (RN). Larger targets are associated with increased risk of both local progression (LP) and RN. In this analysis, we hypothesized that dosimetric and clinical parameters predict for risk of LP and RN in SRS targets larger than two centimeters. We retrospectively reviewed patients with one or more targets with either an intact versus post-operative cavity larger than 2.0 cm treated with LINAC-based SRS between 2017 and 2022 at one institution. We assessed for association between patient, treatment, and disease variables with LP and RN. Variables assessed included tumor resection status, PDL1 positivity, target volume, maximum and minimum target dose, EQD2 and BED (a/b = 2 for necrosis and a/b = 10 for tumor control), as well as receipt of steroids, bevacizumab, or systemic therapy before or after SRS. Radionecrosis was determined by characteristic radiographic changes. Analyses were performed for the entire cohort and within subsets including by resection status and dose fractionation. A total of 178 lesions in 143 patients were included. Targets with volume diameters measuring at least 2 cm were used. Median follow-up was 2.3 years. Overall survival at 1 and 2 years was 56% and 32%, respectively. Most lesions (n = 119) were resected and treated with SRS post-operatively. The most common dose and fractionation schemes used were 30 Gy in 5 fractions (n = 89) and 27 Gy in 3 fractions (n = 63). For the entire cohort, the cumulative incidence of LP 1 and 2 years was 26% and 34%, respectively. The cumulative incidence of radiographic radionecrosis at 1 and 2 years was 12% and 17%, respectively. There was no difference in LP or RN between 27 Gy in 3 fractions versus 30 Gy in 5 fractions (p>0.5 for both). Median planning target volume (PTV) size was 18.5 cc for the 27 Gy in 3 fraction group compared to 21.9 cc in the 30 Gy in 5 fraction group. Minimum or maximum dose within the target was not associated with increased risk of LP or RN. Among patients receiving 27 Gy in 3 fractions, patients treated with resection followed by SRS had lower risk of LP compared to those treated with SRS alone (HR: 0.15, 95% CI: 0.03-0.64, p = 0.011). Among patients receiving 30 Gy in 5 fractions, patients who received corticosteroids prior to SRS had a lower risk of RN (HR: 0.14, 95% CI: 0.03-0.66, p = 0.013). For the entire cohort as well as within all subgroups, PD-L1≥1% was associated with increased risk of RN (p<0.001 for all). Selecting the optimal SRS dose fractionation and planning parameters to minimize both LP and RN remains a challenge for large targets. In this analysis, 27 Gy in 3 fractions appeared to provide equivalent LP and RN compared to 30 Gy in 5 fractions, and may be more convenient for patients. Patients with PD-L1≥1% with large brain targets treated with SRS may be at increased risk of RN; corticosteroid prophylaxis may be considered in this population.

P16.07.B PROSPECTIVE DOUBLE ARM PHASE 2 RANDOMIZED TRIAL FOR PATIENTS WITH MULTIPLE BRAIN METASTASIS AND/OR LEPTOMENINGEAL CARCINOMATOSIS: COMPARISON OF WHOLE BRAIN RADIATION THERAPY (WBRT) ALONE AND WBRT PLUS SILIBININ

Abstract BACKGROUND Occurrence of brain metastases (BMs) is increasing given availability of Magnetic Resonance Imaging (MRI) able to detect also small brain lesions, and most effective systemic therapy able to better control extra-cranial disease. Estimated 20% of patients with cancer will develop BMs. In case of limited brain metastases, radiosurgery or hypofractionated radiosurgery is treatment of choice. A subgroup of patients characterized by extensive brain disease and/or leptomeningeal carcinomatosis can require whole brain radiotherapy (WBRT). Since the 1950s, WBRT has been the most treatment used for patients with multiple BMs, given its effectiveness in palliation, widespread availability, and ease to deliver. However, the median overall survival (OS) recorded is restricted to 3 months, on the average. A better understanding of molecular and cellular mechanisms underlying brain metastasis might be expected to lead of improvements in overall survival. Recent studies demonstrated that active signal transducer and activator of transcription 3 (STAT3) in reactive astrocytes correlates with reduced survival from diagnosis of intracranial metastases. Blocking STAT3 signaling in reactive astrocytes reduces experimental brain metastasis, even at advanced stages of colonization. Silibinin is a natural polyphenolic flavonoid isolated from seed extracts of herb milk thistle. It has been shown to impair STAT3 activation, and in preclinical studies, show an anticancer effects in vitro and in vivo. Based on this background, we designed a double arm phase 2 randomized trial evaluating the benefit of Silibinin associated to WBRT respect to WBRT alone. Primary endpoint is OS, secondary endpoints are toxicity in term of corticosteroid therapy use, brain failure (BF) and progression free survival (PFS). MATERIAL AND METHODS Forty-four patients will be enrolled in two years. Patients with histological or cytological confirmation of solid tumor malignancy, clinical indication for WBRT and Karnofsky performance status (KPS) ≥60 will be enrolled. In arm A experimental, WBRT is concomitant to Silibinin, in arm B control, WBRT alone will be administered. In both arms, total dose of radiotherapy is 30 Gy in 10 fractions. Quality of life and use of corticosteroids will be evaluated first, last day of RT, and during follow-up. Outcome will be evaluated by MRI or CT at 2 months after RT, and every 3 months thereafter. RESULTS From March 2023, 6 patients were enrolled, 2 patients in Arm A and 4 in arm B. Primary histology was breast cancer in 3 patients and non small cell lung cancer in 3. In arm A, 1 patients dismissed RT for worsening of general conditions related to rapid lung progression. CONCLUSION WBRT with or without Silibinin was well tolerated, no acute toxicities or increase of corticosteroids were observed.

GAMMA GLUTAMYL TRANSFERASE LEVELS IN PATIENTS WITH ACUTE STROKE: AN ANALYTICAL STUDY

Objectives: Stroke is a significant global health issue with a major socioeconomic burden, ranking as the second leading cause of death worldwide. While stroke rates have declined in high-income countries, they have increased in low- to middle-income countries, including India. The World Health Organization defines stroke as a disturbance of cerebral function caused by vascular factors, classified as ischemic or hemorrhagic. Methods: Ischemic stroke is primarily caused by atherosclerosis in the brain’s arteries, including the proximal aorta, leading to embolus formation. Other factors contributing to stroke include arterial stenosis, coexisting thrombosis, artery-to-artery embolism, micro atheroma, lipohyalinosis, and occlusive diseases of small brain arteries. Cardiogenic embolism, often associated with atrial fibrillation, contributes to a significant proportion of ischemic strokes. Gamma-glutamyl transferase (GGT), commonly used as a marker for alcohol consumption and liver diseases, is also linked to oxidative stress, inflammation, and atherosclerosis. Elevated GGT levels are associated with various stroke risk factors. Results: GGT plays a crucial role in cellular intake of glutathione, an important antioxidant. Paradoxically, it can generate reactive oxygen species in the presence of certain metals. Studies have found a correlation between high serum GGT levels and stroke risk, potentially due to oxidative stress and atherosclerosis progression. Conclusion: Despite its primary use as an alcohol consumption marker, GGT has emerged as a potential independent biomarker for vascular diseases, including stroke. However, limited research exists on the association between GGT and acute stroke. This study aims to evaluate GGT levels in patients with acute stroke, exploring potential differences among stroke types to better understand the impact of GGT on acute stroke.

Iron oxide nanoparticles induce ferroptosis via the autophagic pathway by synergistic bundling with paclitaxel.

In recent years, inhibiting tumor cell activity by triggering cell ferroptosis has become a research hotspot. The development of generic targeted nanotherapeutics might bring new ideas for non‑invasive applications. Currently, the potential mechanism underlying the universal application of paclitaxel (PTX)‑loaded iron oxide nanoparticles (IONP@PTX) to different types of tumors is unclear. The present study aimed to prepare IONP@PTX for targeted cancer therapy and further explore the potential mechanisms underlying the inhibitory effects of this material on the NCI‑H446 human small cell lung cancer and brain M059K malignant glioblastoma cell lines. First, a CCK‑8 assay was performed to determine cell viability, and then the combination index for evaluating drug combination interaction effect was evaluated. Intracellular reactive oxygen species (ROS) and lipid peroxidation levels were monitored using a DCFH‑DA fluorescent probe and a C11‑BODIPY™ fluorescent probe, respectively. Furthermore, western blotting assay was performed to determine the expression of autophagy‑ and iron death‑related proteins. The experimental results showed that, compared with either IONP monotherapy, PTX monotherapy, or IONP + PTX, IONP@PTX exerted a synergistic effect on the viability of both cell types, with significantly increased total iron ion concentration, ROS levels and lipid peroxidation levels. IONP@PTX significantly increased the expression of autophagy‑related proteins Beclin 1 and histone deacetylase 6 (HDAC6) in both cell lines (P<0.05), increased the expression of light chain 3 (LC3)‑II/I in NCI‑H446 cells (P<0.05) and decreased that of sequestosome1 (p62) in M059K cells (P<0.05). Moreover, the addition of rapamycin enhanced the IONP@PTX‑induced the upregulation of Beclin 1, LC3‑II/I and HDAC6 and the downregulation of mTORC1 protein in both cell lines (P<0.05). Moreover, rapamycin enhanced the IONP@PTX‑induced downregulation of p62 protein in NCI‑H446 cells (P<0.05), suggesting that IONP@PTX induces ferroptosis, most likely through autophagy. Collectively, the present findings show that IONP works synergistically with PTX to induce ferroptosis via the autophagic pathway.

Measurements of slow tissue dynamics with short-separation speckle contrast optical spectroscopy.

Laser speckle contrast imaging (LSCI) measures 2D maps of cerebral blood flow (CBF) in small animal brains such as mice. The contrast measured in LSCI also includes the static and slow-varying components that contain information about brain tissue dynamics. But these components are less studied as compared to the fast dynamics of CBF. In traditional wide-field LSCI, the contrast measured in the tissue is largely contaminated by neighboring blood vessels, which reduces the sensitivity to these static and slow components. Our goal is to enhance the sensitivity of the contrast to static and slow tissue dynamics and test models to quantify the characteristics of these components. To achieve this, we have developed a short-separation speckle contrast optical spectroscopy (ss-SCOS) system by implementing point illumination and point detection using multi-mode fiber arrays to enhance the static and slow components in speckle contrast measurements as compared to traditional wide-field LSCI (WF-LSCI). We observed larger fractions of the static and slow components when measured in the tissue using ss-SCOS than in traditional LSCI for the same animal and region of interest. We have also established models to obtain the fractions of the static and slow components and quantify the decorrelation time constants of the intensity auto-correlation function for both fast blood flow and slower tissue dynamics. Using ss-SCOS, we demonstrate the variations of fast and slow brain dynamics in animals before and post-stroke, as well as within an hour post-euthanasia. This technique establishes the foundation to measure brain tissue dynamics other than CBF, such as intracellular motility.

Brain organoid data synthesis and evaluation.

Datasets containing only few images are common in the biomedical field. This poses a global challenge for the development of robust deep-learning analysis tools, which require a large number of images. Generative Adversarial Networks (GANs) are an increasingly used solution to expand small datasets, specifically in the biomedical domain. However, the validation of synthetic images by metrics is still controversial and psychovisual evaluations are time consuming. We augment a small brain organoid bright-field database of 40 images using several GAN optimizations. We compare these synthetic images to the original dataset using similitude metrcis and we perform an psychovisual evaluation of the 240 images generated. Eight biological experts labeled the full dataset (280 images) as syntetic or natural using a custom-built software. We calculate the error rate per loss optimization as well as the hesitation time. We then compare these results to those provided by the similarity metrics. We test the psychovalidated images in a training step of a segmentation task. Generated images are considered as natural as the original dataset, with no increase of the hesitation time by experts. Experts are particularly misled by perceptual and Wasserstein loss optimization. These optimizations render the most qualitative and similar images according to metrics to the original dataset. We do not observe a strong correlation but links between some metrics and psychovisual decision according to the kind of generation. Particular Blur metric combinations could maybe replace the psychovisual evaluation. Segmentation task which use the most psychovalidated images are the most accurate.

The neurological and non-neurological roles of the primary microcephaly-associated protein ASPM.

Primary microcephaly (MCPH), is a neurological disorder characterized by small brain size that results in numerous developmental problems, including intellectual disability, motor and speech delays, and seizures. Hitherto, over 30 MCPH causing genes (MCPHs) have been identified. Among these MCPHs, MCPH5, which encodes abnormal spindle-like microcephaly-associated protein (ASPM), is the most frequently mutated gene. ASPM regulates mitotic events, cell proliferation, replication stress response, DNA repair, and tumorigenesis. Moreover, using a data mining approach, we have confirmed that high levels of expression of ASPM correlate with poor prognosis in several types of tumors. Here, we summarize the neurological and non-neurological functions of ASPM and provide insight into its implications for the diagnosis and treatment of MCPH and cancer.

Amplifying the Effects of Contrast Agents on Magnetic Resonance Images Using a Deep Learning Method Trained on Synthetic Data.

Artificial intelligence (AI) methods can be applied to enhance contrast in diagnostic images beyond that attainable with the standard doses of contrast agents (CAs) normally used in the clinic, thus potentially increasing diagnostic power and sensitivity. Deep learning-based AI relies on training data sets, which should be sufficiently large and diverse to effectively adjust network parameters, avoid biases, and enable generalization of the outcome. However, large sets of diagnostic images acquired at doses of CA outside the standard-of-care are not commonly available. Here, we propose a method to generate synthetic data sets to train an "AI agent" designed to amplify the effects of CAs in magnetic resonance (MR) images. The method was fine-tuned and validated in a preclinical study in a murine model of brain glioma, and extended to a large, retrospective clinical human data set. A physical model was applied to simulate different levels of MR contrast from a gadolinium-based CA. The simulated data were used to train a neural network that predicts image contrast at higher doses. A preclinical MR study at multiple CA doses in a rat model of glioma was performed to tune model parameters and to assess fidelity of the virtual contrast images against ground-truth MR and histological data. Two different scanners (3 T and 7 T, respectively) were used to assess the effects of field strength. The approach was then applied to a retrospective clinical study comprising 1990 examinations in patients affected by a variety of brain diseases, including glioma, multiple sclerosis, and metastatic cancer. Images were evaluated in terms of contrast-to-noise ratio and lesion-to-brain ratio, and qualitative scores. In the preclinical study, virtual double-dose images showed high degrees of similarity to experimental double-dose images for both peak signal-to-noise ratio and structural similarity index (29.49 dB and 0.914 dB at 7 T, respectively, and 31.32 dB and 0.942 dB at 3 T) and significant improvement over standard contrast dose (ie, 0.1 mmol Gd/kg) images at both field strengths. In the clinical study, contrast-to-noise ratio and lesion-to-brain ratio increased by an average 155% and 34% in virtual contrast images compared with standard-dose images. Blind scoring of AI-enhanced images by 2 neuroradiologists showed significantly better sensitivity to small brain lesions compared with standard-dose images (4.46/5 vs 3.51/5). Synthetic data generated by a physical model of contrast enhancement provided effective training for a deep learning model for contrast amplification. Contrast above that attainable at standard doses of gadolinium-based CA can be generated through this approach, with significant advantages in the detection of small low-enhancing brain lesions.

SLC4A10 mutation causes a neurological disorder associated with impaired GABAergic transmission.

SLC4A10 is a plasma-membrane bound transporter that utilizes the Na+ gradient to drive cellular HCO3- uptake, thus mediating acid extrusion. In the mammalian brain, SLC4A10 is expressed in principal neurons and interneurons, as well as in epithelial cells of the choroid plexus, the organ regulating the production of CSF. Using next generation sequencing on samples from five unrelated families encompassing nine affected individuals, we show that biallelic SLC4A10 loss-of-function variants cause a clinically recognizable neurodevelopmental disorder in humans. The cardinal clinical features of the condition include hypotonia in infancy, delayed psychomotor development across all domains and intellectual impairment. Affected individuals commonly display traits associated with autistic spectrum disorder including anxiety, hyperactivity and stereotyped movements. In two cases isolated episodes of seizures were reported in the first few years of life, and a further affected child displayed bitemporal epileptogenic discharges on EEG without overt clinical seizures. While occipitofrontal circumference was reported to be normal at birth, progressive postnatal microcephaly evolved in 7 out of 10 affected individuals. Neuroradiological features included a relative preservation of brain volume compared to occipitofrontal circumference, characteristic narrow sometimes 'slit-like' lateral ventricles and corpus callosum abnormalities. Slc4a10 -/- mice, deficient for SLC4A10, also display small lateral brain ventricles and mild behavioural abnormalities including delayed habituation and alterations in the two-object novel object recognition task. Collapsed brain ventricles in both Slc4a10-/- mice and affected individuals suggest an important role of SLC4A10 in the production of the CSF. However, it is notable that despite diverse roles of the CSF in the developing and adult brain, the cortex of Slc4a10-/- mice appears grossly intact. Co-staining with synaptic markers revealed that in neurons, SLC4A10 localizes to inhibitory, but not excitatory, presynapses. These findings are supported by our functional studies, which show the release of the inhibitory neurotransmitter GABA is compromised in Slc4a10-/- mice, while the release of the excitatory neurotransmitter glutamate is preserved. Manipulation of intracellular pH partially rescues GABA release. Together our studies define a novel neurodevelopmental disorder associated with biallelic pathogenic variants in SLC4A10 and highlight the importance of further analyses of the consequences of SLC4A10 loss-of-function for brain development, synaptic transmission and network properties.

Effect of magnet strength on fetal brain biometry - a single-center retrospective MRI-based cohort study.

Abnormal fetal brain measurements might affect clinical management and parental counseling. The effect of between-field-strength differences was not evaluated in quantitative fetal brain imaging until now. Our study aimed to compare fetal brain biometry measurements in 3.0T with 1.5T scanners. A retrospective cohort of 1150 low-risk fetuses scanned between 2012 and 2021, with apparently normal brain anatomy, were retrospectively evaluated for biometric measurements. The cohort included 1.5T (442 fetuses) and 3.0T scans (708 fetuses) of populations with comparable characteristics in the same tertiary medical center. Manually measured biometry included bi-parietal, fronto-occipital and trans-cerebellar diameters, length of the corpus-callosum, vermis height, and width. Measurements were then converted to centiles based on previously reported biometric reference charts. The 1.5T centiles were compared with the 3.0T centiles. No significant differences between centiles of bi-parietal diameter, trans-cerebellar diameter, or length of the corpus callosum between 1.5T and 3.0T scanners were found. Small absolute differences were found in the vermis height, with higher centiles in the 3.0T, compared to the 1.5T scanner (54.6th-centile, vs. 39.0th-centile, p < 0.001); less significant differences were found in vermis width centiles (46.9th-centile vs. 37.5th-centile, p = 0.03). Fronto-occipital diameter was higher in 1.5T than in the 3.0T scanner (66.0th-centile vs. 61.8th-centile, p = 0.02). The increasing use of 3.0T MRI for fetal imaging poses a potential bias when using 1.5T-based charts. We elucidate those biometric measurements are comparable, with relatively small between-field-strength differences, when using manual biometric measurements. Small inter-magnet differences can be related to higher spatial resolution with 3T scanners and may be substantial when evaluating small brain structures, such as the vermis.

An Isolated CNN Architecture for Classification of Finger-Tapping Tasks Using Initial Dip Images: A Functional Near-Infrared Spectroscopy Study.

This work investigates the classification of finger-tapping task images constructed for the initial dip duration of hemodynamics (HR) associated with the small brain area of the left motor cortex using functional near-infrared spectroscopy (fNIRS). Different layers (i.e., 16-layers, 19-layers, 22-layers, and 25-layers) of isolated convolutional neural network (CNN) designed from scratch are tested to classify the right-hand thumb and little finger-tapping tasks. Functional t-maps of finger-tapping tasks (thumb, little) were constructed for various durations (0.5 to 4 s with a uniform interval of 0.5 s) for the initial dip duration using a three gamma functions-based designed HR function. The results show that the 22-layered isolated CNN model yielded the highest classification accuracy of 89.2% with less complexity in classifying the functional t-maps of thumb and little fingers associated with the same small brain area using the initial dip. The results further demonstrated that the active brain area of the two tapping tasks from the same small brain area are highly different and well classified using functional t-maps of the initial dip (0.5 to 4 s) compared to functional t-maps generated for delayed HR (14 s). This study shows that the images constructed for initial dip duration can be helpful in the future for fNIRS-based diagnosis or cortical analysis of abnormal cerebral oxygen exchange in patients.

Affibody Molecules Intended for Receptor-Mediated Transcytosis via the Transferrin Receptor.

The development of biologics for diseases affecting the central nervous system has been less successful compared to other disease areas, in part due to the challenge of delivering drugs to the brain. The most well-investigated and successful strategy for increasing brain uptake of biological drugs is using receptor-mediated transcytosis over the blood-brain barrier and, in particular, targeting the transferrin receptor-1 (TfR). Here, affibody molecules are selected for TfR using phage display technology. The two most interesting candidates demonstrated binding to human TfR, cross-reactivity to the murine orthologue, non-competitive binding with human transferrin, and binding to TfR-expressing brain endothelial cell lines. Single amino acid mutagenesis of the affibody molecules revealed the binding contribution of individual residues and was used to develop second-generation variants with improved properties. The second-generation variants were further analyzed and showed an ability for transcytosis in an in vitro transwell assay. The new TfR-specific affibody molecules have the potential for the development of small brain shuttles for increasing the uptake of various compounds to the central nervous system and thus warrant further investigations.