Microglial Neuroinflammation-Independent Reversal of Demyelination of Corpus Callosum by Arsenic in a Cuprizone-Induced Demyelinating Mouse Model.

Demyelination is the loss of myelin in CNS, resulting in damaged myelin sheath. Oxidative stress and neuroinflammation play a key role in inducing demyelinating diseases like MS; hence, controlling oxidative stress and neuroinflammation is important. Cuprizone (CPZ), a copper chelator, generates oxidative stress and neuroinflammation, thereby inducing demyelination. Therefore, the CPZ-induced demyelinating mouse model (CPZ model) is widely used in research. The present study was intended to unravel a mechanism of inhibition of demyelination by arsenic in a CPZ model, which is otherwise known for its toxicity. We investigated an alternative mechanism of inhibition of demyelination by arsenic through the reversal of SOD1 activity employing in silico analysis, analytical chemistry techniques, and in vitro and in vivo experiments. In vivo experiments showed protection of body weight, survivability, and myelination of the corpus callosum in CPZ and arsenic-co-exposed animals, where neuroinflammation was apparently not involved. In vitro experiments revealed that arsenic-mediated reversal of impaired SOD1 activity leads to reduced cellular ROS levels and better viability of primary oligodendrocytes. Reversal of SOD1 activity was also observed in the corpus callosum tissue isolated from experimental animals. In silico and analytical chemistry studies revealed that similar to copper, arsenic can potentially bind to CPZ and thereby make the copper freely available for SOD1 activity. Suitable neurobehavior tests further validated the protective effect of arsenic. Taken together, the present study revealed that arsenic protects oligodendrocytes and demyelination of corpus callosum by reversing CPZ-induced impaired SOD1 activity.

Sperm intrusion into the implantation-stage blastocyst and its potential biological significance.

The human embryo derives from fusion of oocyte and sperm, undergoes growth and differentiation, resulting in a blastocyst. To initiate implantation, the blastocyst hatches from the zona pellucida, allowing access from external inputs. Modelling of uterine sperm distribution indicates that 200-5000 sperm cells may reach the implantation-stage blastocyst following natural coitus. We show ultrastructural evidence of sperm cells intruding into trophectoderm cells of zona-free blastocysts obtained from the uterus of rhesus monkeys. Interaction between additional sperm and zona-free blastocyst could be an evolutionary feature yielding adaptive processes influencing the developmental fate of embryos. This process bears potential implications in pregnancy success, sperm competition and human health.

Heparin Biofunctionalized Selenium Nanoparticles as Potential Antiangiogenic-Chemotherapeutic Agents for Targeted Doxorubicin Delivery.

Combining antiangiogenic and chemotherapeutic agents has shown promising clinical benefits in cancer cures when the therapeutic intervention takes into account the tissue and molecular targets. Moreover, the risk of induced drug resistance is minimized when multiple pathways are involved in the treatment regimen, yielding a better therapeutic outcome. Nanodrug delivery systems have proven to be a prudent approach to treating complex disease pathologies. As such, combining antiangiogenic and chemotherapeutic drugs within multimodal nanocarriers synergistically augments the clinical efficiency of the drugs. This study reports the combinatorial efficacy of heparin (Hep), selenium NPs (SeNPs), and doxorubicin (Dox) to inhibit tumor growth and progression. Both Se@Hep-NPs and Se@Hep-Dox-NPs with excellent water dispersity having a size and charge in the range of 250 ± 5 and 253 ± 5 nm and -53 ± 0.4 and -48.4 ± 6.4 mV, respectively, showed strong anticancer potential assessed through in vitro assays like cell viability, specificity, colony formation, and wound scratch in MCF7 cells. Strong synergistic interactions among SeNPs, Hep, and Dox in Se@Hep-Dox-NPs render it to be an antiangiogenic and proapoptotic cancer cell death inducers. In vivo imaging highlights the dual-mode attributes of Se@Hep-NPs with desirable passive tumor targeting and biomedical imaging ability when tagged with Cy7.5, while Se@Hep-Dox-NPs significantly reduce the tumor burden and prolong the longevity of subcutaneous EAC-bearing mice. Histopathology studies reveal no signs of toxicity in major organs. Collectively, these results qualify Se@Hep-Dox-NPs as a plausible clinical therapeutic candidate.

Incentive conflict and supply contracts under carbon cap policy.

Environmental regulations, once promulgated, can cause incentive conflict between manufacturers and suppliers. A manufacturer facing the regulation may undertake choices that can affect his sourcing decisions with the supplier. To analyze this, we develop a game-theoretic model considering a manufacturer who faces a per-unit carbon emissions cap and sources from a supplier. The manufacturer operates in a carbon sensitive market. We analyze the responses of the manufacturer and supplier and show that since the burden of carbon emissions cap is borne by the manufacturer, the first-best outcomes are not reached. Therefore, the supplier may offer different contracts to incentivize the manufacturer. We study two mechanisms: the two-part tariff and the revenue-and-investment sharing contracts. We show how such contracts achieve coordination and deliver efficient supply chain outcomes. Interestingly, we find that the contract preferences of the manufacturer and the supplier may not be the same and vary under different market conditions. Summarily, we highlight important considerations for the supply chain players in designing suitable incentives.

Absorption Dominated Directional Electromagnetic Interference Shielding through Asymmetry in a Multilayered Construct with an Exceptionally High Green Index.

Fabricating green electromagnetic interference (EMI) shields is the need of the hour because strong secondary reflections in the vicinity of the shield adversely affect the environment and the reliability of the neighboring devices. To this end, the present work aims to maximize the absorption-based EMI shielding through a multilayered construct comprising a porous structure (pore size less than λ/5), a highly conducting entity, and a layer to match the impedance. The elements of this construct were positioned so that the incoming electromagnetic (EM) radiation interacts with the other layers of the construct before the conducting entity. This positioning of the layers in the construct offers a high green shielding index (gs) and low reflection coefficient (R ∼ 0.1) with an exceptionally high percent absorption (up to 99%). Polyurethane (PU) foams were fabricated using the salt-leaching technique and strategically positioned with carbon nanotube (CNT) papers and polycarbonate (PC)-based films to obtain symmetric and asymmetric constructs. These structures were then employed to gain mechanistic insight into the directional dependency of shielding performance, gs, and heat dissipation ability. Interestingly, maximum total shielding effectiveness (SET) of -52 dB (88% absorption @8.2 GHz) and specific shielding effectiveness/thickness (SSEt) of -373 dB/cm2g were achieved for a symmetric construct whereas, for the asymmetric construct, the SET and SSEt were -37 dB and -280 dB/cm2g, respectively, with an exceptionally high gs of 8.6, the highest reported so far. The asymmetricity in the construct led to directional dependence of the absorption component (% SEA, shielding effectiveness due to absorption) and heat dissipation, primarily governed by the electrical and thermal conductivity gradient, respectively. This study opens new avenues in this field and reports constructs with an exceptionally high green index.

Understanding systemic disruption from the Covid-19-induced semiconductor shortage for the auto industry.

Covid-19 has allowed us to study systemic disruptions that impact entire industries. This paper explores how disruptions start, propagate, and continue over time by examining the semiconductor chip shortage faced by the auto industry during the years following Covid-19 in 2020. First, we carried out a thematic analysis of 209 pertinent newspaper articles. The analysis resulted in a thematic model of such disruptions with the interplay of various factors leading to the prolonged disruption to the auto sector. Second, we present the results from a stylized supply chain planning model run at different times to show how disruptions propagate to the auto and other sectors, causing systemic shortages. Overall, we contribute to the supply chain risk literature by focusing on system disruptions impacting entire industries versus normal disruptions affecting a particular company's supply chain.

Perinatal arsenic exposure-induced sustained microglial activation leads to impaired cognitive response in BALB/c mice.

Arsenic is infamous for its adverse health effects worldwide. It is known to induce cognitive impairment in experimental model animals and children in the arsenic-affected area. Although the effect of arsenic on neuronal health is well studied, but the involvement of the brain immune component, microglia, has not been well explored. The present study is focused on examining the role of microglia in arsenic-induced cognitive impairment. We have used balb/c mice for the study. Pregnant dams were gavaged with sodium arsenite (0.38 mg/kg body weight) from gestational day 5 (GD5) till postnatal day 22 (PND22). Mice were sacrificed on PND 7, 14, 22 and isolated brains were used for various assays. The study reveals that perinatal arsenic exposure keeps the microglia activated and skews them towards the M1 phenotype. Increased microglial proliferation, ROS, NO, higher levels of proinflammatory cytokines and chemokines were observed in the arsenic exposed group. Enhanced phagocytosis and phagocytic receptor TREM2, along with decreased expression of SNAP25 and PSD95, were correlated for enhanced neuronal pruning leading to impaired learning and memory response. Taken together, the study reveals an association between arsenic exposure and altered cognitive response where enhanced neuronal pruning by arsenic-activated microglia plays an important role in developing mice.

Biopolymeric composite hydrogel loaded with silver NPs and epigallocatechin gallate (EGCG) effectively manages ROS for rapid wound healing in type II diabetic wounds.

Delayed wound healing in patients having type-II diabetes mellitus (T2DM) often results in a high rate of amputation. We report an innovative Guar Gum-based macroporous hydrogel (HG) infused with an antibacterial agent (Ag NPs), and antioxidant, epigallocatechin gallate (EGCG) to address rapid wound healing and interestingly could inhibit the associated pathophysical bone infection in a high-fat-diet-induced T2DM C57BL/6 mice model. The HG-Ag-EGCG elicits scar-free wound healing in subcutaneous wounds and histopathological evidence confirmed HG-Ag-EGCG hydrogel patch elicits better wound healing through enhanced cell proliferation, mature connecting tissue fiber formation, minimum void spaces formation, and better re-epithelialization when compared with a market available hydrogel patch material (Luofucon®). Supportive of the in vivo outcomes, in vitro experiments delineated better-wound closure due to improved management of ROS by the HG-Ag-EGCG. Additionally, a favorable non-toxicity outcome assessed through both in vitro and in vivo conditions confirmed its potential applicability in clinical wound care management.

Minocycline reverses developmental arsenic exposure-induced microglia activation and functional alteration in BALB/c mice.

Arsenic activates microglia and exerts bystander effects on neuron. The present study is focused to test whether minocycline, a second generation antibiotic, can reverse the effect of developmental arsenic exposure on microglial activation and function. Pregnant Balb/c dams were gavaged with sodium arsenite (0.38 mg/kg bd wt) from gestational day 5 (GD5) till post natal day 21 (PND21) and then one group of pups continued till PND59 with arsenic gavage. Minocycline (33 mg/kg bd wt) was administered intraperitoneally two weeks till sacrifice, every alternate day. Mice were sacrificed on PND22 and PND60 and used for various assays. Primary microglial were isolated (ex vivo microglia) from experimental animals and used to measure reactive oxygen species (ROS), nitric oxide (NO), cytokine production and phagocytosis. The whole brain lysate was used for western blot analysis of microglial marker CD68 and synaptic marker, post synaptic density protein 95 (PSD95). For real-time PCR analysis of triggering receptor expressed on myeloid cells 2 (TREM2) and PSD95, RNA isolated from whole brain was used. The study reveals that minocycline administration reversed arsenic-induced increased expression of CD68, ROS, NO, cytokine production, phagocytosis and TREM2 expression. Arsenic-induced reduced expression of PSD95 protein was reversed by minocycline, although the mRNA of PSD95 was unaltered among different groups. Finally, we have checked the learning and memory response of the experimental animals using Y-maze test to correlate the arsenic-induced altered level of synaptic protein. Taken together, the present study finds minocycline to reduce arsenic-induced microglial activation and function which in turn reverses the arsenic-induced impaired learning and memory response.

MicroRNA-129-5p-regulated microglial expression of the surface receptor CD200R1 controls neuroinflammation.

CD200R1 is an inhibitory surface receptor expressed in microglia and blood macrophages. Microglial CD200R1 is known to control neuroinflammation by keeping the microglia in resting state, and therefore, tight regulation of its expression is important. CCAAT/enhancer-binding protein ß (CEBPß) is the known regulator of CD200R1 transcription. In the present study, our specific intention was to find a possible posttranscriptional regulatory mechanism of CD200R1 expression. Here we investigated a novel regulatory mechanism of CD200R1 expression following exposure to an environmental stressor, arsenic, combining in silico analysis, in vitro, and in vivo experiments, as well as validation in human samples. The in silico analysis and in vitro studies with primary neonatal microglia and BV2 microglia revealed that arsenic demethylates the promoter of a microRNA, miR-129-5p, thereby increasing its expression, which subsequently represses CD200R1 by binding to its 3'-untranslated region and shuttling the CD200R1 mRNA to the cytoplasmic-processing body in mouse microglia. The role of miR-129-5p was further validated in BALB/c mouse by stereotaxically injecting anti-miR-129. We found that anti-miR-129 reversed the expression of CD200R1, as well as levels of inflammatory molecules IL-6 and TNF-α. Experiments with a CD200R1 siRNA-induced loss-of-function mouse model confirmed an miR-129-5p→CD200R1→IL-6/TNF-α signaling axis. These main findings were replicated in a human cell line and validated in human samples. Taken together, our study revealed miR-129-5p as a novel posttranscriptional regulator of CD200R1 expression with potential implications in neuroinflammation and related complications.

Serum zinc level and its association with multidrug-resistant tuberculosis.

Background: Zinc deficiency is evident in chronic diseases, but little is known about its association with multi-drug resistant tuberculosis (MDRTB). We aimed to measure serum zinc level in MDRTB patients and explore its association with MDRTB compared to drug-sensitive tuberculosis (DSTB). Methods: We recruited 107 MDRTB and 87 DSTB patients from a tuberculosis referral hospital in Bangladesh. After overnight fasting, 5 ml venous blood was collected from each patient to measure serum zinc level through graphite furnace atomic absorption spectrophotometry method. Multivariate logistic regression was done to measure its association with MDRTB. Results: The mean age of all patients was 36 years, where 70% were male. About 27% MDRTB patients and 2.3% DSTB patients had low serum zinc level (P < 0.0001). An inverse correlation was observed between serum zinc level and duration of anti-TB therapy (r-value: -0.252, P < 0.01). Reduced serum zinc level (odds ratio, 0.957; 95% confidence interval 0.923-0.992) was found as a significant associating factor for MDRTB after adjusted with age, sex, occupation, residence, tobacco consumption, Bacillus Calmette-Guérin-vaccination, and duration of anti-TB therapy. Conclusion: A lower serum zinc level is significantly associated with MDRTB. The tuberculosis control program should address this in the MDRTB control strategy.

Control Release of Adenosine Potentiate Osteogenic Differentiation within a Bone Integrative EGCG-g-NOCC/Collagen Composite Scaffold toward Guided Bone Regeneration in a Critical-Sized Calvarial Defect.

The regeneration of critical-sized bone defects with biomimetic scaffolds remains clinically challenging due to avascular necrosis, chronic inflammation, and altered osteogenic activity. Two confounding mechanisms, efficacy manipulation, and temporal regulation dictate the scaffold's bone regenerative ability. Equally critical is the priming of the mesenchymal stromal cells (MSCs) toward lineage-specific differentiation into bone-forming osteoblast, which particularly depends on varied mechanochemical and biological cues during bone tissue regeneration. This study sought to design and develop an optimized osteogenic scaffold, adenosine/epigallocatechin gallate-N,O-carboxymethyl chitosan/collagen type I (AD/EGCG-g-NOCC@clgn I), having osteoinductive components toward swift bone regeneration in a calvarial defect BALB/c mice model. The ex vivo findings distinctly establish the pro-osteogenic potential of adenosine and EGCG, stimulating MSCs toward osteoblast differentiation with significantly increased expression of alkaline phosphatase, calcium deposits, and enhanced osteocalcin expression. Moreover, the 3D matrix recapitulates extracellular matrix (ECM) properties, provides a favorable microenvironment, structural support against mechanical stress, and acts as a reservoir for sustained release of osteoinductive molecules for cell differentiation, proliferation, and migration during matrix osteointegration observed. Evidence from in vivo experiments, micro-CT analyses, histology, and histomorphometry signify accelerated osteogenesis both qualitatively and quantitatively: effectual bone union with enhanced bone formation and new ossified tissue in 4 mm sized defects. Our results suggest that the optimized scaffold serves as an adjuvant to guide bone tissue regeneration in critical-sized calvarial defects with promising therapeutic efficacy.

IUPS Physiology Education Workshop series in India: organizational mechanics, outcomes, and lessons.

Active learning promotes the capacity of problem solving and decision making among learners. Teachers who apply instructional processes toward active participation of learners help their students develop higher order thinking skills. Due to the recent paradigm shift toward adopting competency-based curricula in the education of healthcare professionals in India, there is an emergent need for physiology instructors to be trained in active-learning methodologies and to acquire abilities to promote these curriculum changes. To address these issues, a series of International Union of Physiological Sciences (IUPS) workshops on physiology education techniques in four apex centers in India was organized in November 2018 and November 2019. The "hands-on" workshops presented the methodologies of case-based learning, problem-based learning, and flipped classroom; the participants were teachers of basic sciences and human and veterinary medicine. The workshop series facilitated capacity building and creation of a national network of physiology instructors interested in promoting active-learning techniques. The workshops were followed by a brainstorming meeting held to assess the outcomes. The aim of this report is to provide a model for implementing a coordinated series of workshops to support national curriculum change and to identify the organizational elements essential for conducting an effective Physiology Education workshop. The essential elements include a highly motivated core organizing team, constant dialogue between core organizing and local organizing committees, a sufficient time frame for planning and execution of the event, and opportunities to engage students at host institutions in workshop activities.

Low-dose naltrexone rescues inflammation and insulin resistance associated with hyperinsulinemia.

The incidence of diabetes, obesity, and metabolic diseases has reached an epidemic status worldwide. Insulin resistance is a common link in the development of these conditions, and hyperinsulinemia is a central hallmark of peripheral insulin resistance. However, how hyperinsulinemia leads to systemic insulin resistance is less clear. We now provide evidence that hyperinsulinemia promotes the release of soluble pro-inflammatory mediators from macrophages that lead to systemic insulin resistance. Our observations suggest that hyperinsulinemia induces sirtuin1 (SIRT1) repression and stimulates NF-κB p65 nuclear translocation and transactivation of NF-κB to promote the extracellular release of pro-inflammatory mediators. We further showed that low-dose naltrexone (LDN) abrogates hyperinsulinemia-mediated SIRT1 repression and prevents NF-κB p65 nuclear translocation. This, in turn, attenuates the hyperinsulinemia-induced release of pro-inflammatory cytokines and reinstates insulin sensitivity both in in vitro and in vivo diet-induced hyperinsulinemic mouse model. Notably, our data indicate that Sirt1 knockdown or inhibition blunts the anti-inflammatory properties of LDN in vitro Using numerous complementary in silico and in vitro experimental approaches, we demonstrated that LDN can bind to SIRT1 and increase its deacetylase activity. Together, these data support a critical role of SIRT1 in inflammation and insulin resistance in hyperinsulinemia. LDN improves hyperinsulinemia-induced insulin resistance by reorienting macrophages toward anti-inflammation. Thus, LDN treatment may provide a novel therapeutic approach against hyperinsulinemia-associated insulin resistance.

Endometrial stromal cell inflammatory phenotype during severe ovarian endometriosis as a cause of endometriosis-associated infertility.

RESEARCH QUESTION: Do endometrial stromal cells from primary infertile patients with severe ovarian endometriosis display differential secretory profiles of inflammation-associated cytokines during the implantation window that may cause infertility? DESIGN: Forty-eight cytokines were measured in conditioned medium of isolated endometrial stromal cells obtained from primary infertile patients without endometriosis (control group, n = 12) or with stage IV ovarian endometriosis (ovarian endometriosis group, n = 14) using multiplex assays. Key cytokines showing differential secretory profiles were validated using Western immunoblotting. Cellular phenotypic validation was carried out in vitro by comparing proliferation and migration capacity between control (n = 6) and ovarian endometriosis (n = 7) groups. RESULTS: CCL3, CCL4, CCL5, CXCL10, FGF2, IFNG, IL1RN, IL5, TNFA, and VEGF could be detected only in the conditioned media of stromal cells obtained from the ovarian endometriosis group. Among other cytokines detected in the conditioned media of both groups, CCL2 (P = 0.0018), CSF3 (P = 0.0017), IL1B (P = 0.0066), IL4 (P = 0.036), IL6 (P = 0.0039) and IL13 (P = 0.036) were found to be higher, whereas the concentration of IL18 was lower (P = 0.023) in the ovarian endometriosis group. Concentrations of CCL2, IL1B, IL4 and IL13 in conditioned medium reflected significant diagnostic performance for predicting ovarian endometriosis. Cellular phenotypic validation in vitro revealed an enhanced proliferative phenotype (P = 0.046) with no change in cell migratory capacity of endometrial stromal cells from the ovarian endometriosis group. CONCLUSIONS: Endometrial stromal cells derived from severe ovarian endometriosis samples displayed a hyperinflammatory and hyperproliferative bias in the endometrial stroma during the 'window of implantation' putatively causing loss of fecundability.

Multiscale and morphological analysis of microvascular patterns depicted in contrast-enhanced ultrasound images.

Purpose: Impaired insulin-induced microvascular recruitment in skeletal muscle contributes to insulin resistance in type 2 diabetic disease. Previously, quantification of microvascular recruitment at the capillary level has been performed with either the full image or manually selected region-of-interests. These subjective approaches are imprecise, time-consuming, and unsuitable for automated processes. Here, an automated multiscale image processing approach was performed by defining a vessel diameter threshold for an objective and reproducible analysis at the microvascular level. Approach: A population of C57BL/6J male mice fed standard chow and studied at age 13 to 16 weeks comprised the lean group and 24- to 31-week-old mice who received a high-fat diet were designated the obese group. A clinical ultrasound scanner (Acuson Sequoia 512) equipped with an 15L8-S linear array transducer was used in a nonlinear imaging mode for sensitive detection of an intravascular microbubble contrast agent. Results: By eliminating large vessels from the dynamic contrast-enhanced ultrasound (DCE-US) images (above 300 µ m in diameter), obesity-related changes in perfusion and morphology parameters were readily detected in the smaller vessels, which are known to have a greater impact on skeletal muscle glucose disposal. The results from the DCE-US images including all of the vessels were compared for three different-sized vessel groups, namely, vessels smaller than 300, 200, and 150 µ m in diameter. Conclusions: Our automated image processing provides objective and reproducible results by focusing on a particular size of vessel, thereby allowing for a selective evaluation of longitudinal changes in microvascular recruitment for a specific-sized vessel group between diseased and healthy microvascular networks.

Reference Value for Serum Zinc Level of Adult Population in Bangladesh.

BACKGROUND: Zinc is an essential trace element that has an enormous role in regulation of physiological processes whose deviant value leads to malfunction in the body. So, establishing a country specific reference value is needed to serve as a standard for the interpretation of laboratory results during clinical decision making. OBJECTIVE: The objective of this study was to determine the reference value of serum zinc level of adult population in Bangladesh. MATERIALS AND METHODS: The overnight fasting blood was collected from 154 apparently healthy individuals aged 18 to 65 years, from a rural community after considering several criteria. Graphite furnace atomic absorption spectrophotometry (GF-AAS) method was used for serum zinc analysis. The 2.5th and 97.5th percentiles of zinc level were calculated for the reference value according to the recommendations of the International Federation of Clinical Chemistry and Laboratory Medicine (IFCC). RESULTS: The estimated reference range of serum zinc level in sample population was 60-120 µg/dL, where the range was 59-125 µg/dL for male and 50-103 µg/dL for female. Significant differences of serum zinc level between male and female (p<0.001) was observed. However, there was no significant correlation between age of the respondents and serum zinc level (r=0.110, p>0.05). CONCLUSION: The estimated reference range for serum zinc level in adult population of Bangladesh can serve as a useful indicator for clinical decision making.

Deep Learning of Spatiotemporal Filtering for Fast Super-Resolution Ultrasound Imaging.

Super-resolution ultrasound (SR-US) imaging is a new technique that breaks the diffraction limit and allows visualization of microvascular structures down to tens of micrometers. The image processing methods for the spatiotemporal filtering needed in SR-US, such as singular value decomposition (SVD), are computationally burdensome and performed offline. Deep learning has been applied to many biomedical imaging problems, and trained neural networks have been shown to process an image in milliseconds. The goal of this study was to evaluate the effectiveness of deep learning to realize a spatiotemporal filter in the context of SR-US processing. A 3-D convolutional neural network (3DCNN) was trained on in vitro and in vivo data sets using SVD as ground truth in tissue clutter reduction. In vitro data were obtained from a tissue-mimicking flow phantom, and in vivo data were collected from murine tumors of breast cancer. Three training techniques were studied: training with in vitro data sets, training with in vivo data sets, and transfer learning with initial training on in vitro data sets followed by fine-tuning with in vivo data sets. The neural network trained with in vitro data sets followed by fine-tuning with in vivo data sets had the highest accuracy at 88.0%. The SR-US images produced with deep learning allowed visualization of vessels as small as [Formula: see text] in diameter, which is below the diffraction limit (wavelength of [Formula: see text] at 14 MHz). The performance of the 3DCNN was encouraging for real-time SR-US imaging with an average processing frame rate for in vivo data of 51 Hz with GPU acceleration.