Optimal sparsity allows reliable system-aware restoration of fluorescence microscopy images

Incluye: artículo, material suplementario, videos y software.Fluorescence microscopy is one of the most indispensable and informative driving forces for biological research, but the extent of observable biological phenomena is essentially determined by the content and quality of the acquired images. To address the different noise sources that can degrade these images, we introduce an algorithm for multiscale image restoration through optimally sparse representation (MIRO). MIRO is a deterministic framework that models the acquisition process and uses pixelwise noise correction to improve image quality. Our study demonstrates that this approach yields a remarkable restoration of the fluorescence signal for a wide range of microscopy systems, regardless of the detector used (e.g., electron-multiplying charge-coupled device, scientific complementary metal-oxide semiconductor, or photomultiplier tube). MIRO improves current imaging capabilities, enabling fast, low-light optical microscopy, accurate image analysis, and robust machine intelligence when integrated with deep neural networks. This expands the range of biological knowledge that can be obtained from fluorescence microscopy.We acknowledge the support of the National Institutes of Health grants R35GM124846 (to S.J.) and R01AA028527 (to C.X.), the National Science Foundation grants BIO2145235 and EFMA1830941 (to S.J.), and Marvin H. and Nita S. Floyd Research Fund (to S.J.). This research project was supported, in part, by the Emory University Integrated Cellular Imaging Microscopy Core and by PHS Grant UL1TR000454 from the Clinical and Translational Science Award Program, National Institutes of Health, and National Center for Advancing Translational Sciences.S

Comprehensive Characterization of COVID-19 Patients with Repeatedly Positive SARS-CoV-2 Tests Using a Large U.S. Electronic Health Record Database.

In the absence of genome sequencing, two positive molecular tests for severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) separated by negative tests, prolonged time, and symptom resolution remain the best surrogate measure of possible reinfection. Using a large electronic health record database, we characterized clinical and testing data for 23 patients with repeatedly positive SARS-CoV-2 PCR test results ≥60 days apart, separated by ≥2 consecutive negative test results. The prevalence of chronic medical conditions, symptoms, and severe outcomes related to coronavirus disease 19 (COVID-19) illness were ascertained. The median age of patients was 64.5 years, 40% were Black, and 39% were female. A total of 83% smoked within the prior year, 61% were overweight/obese, 83% had immunocompromising conditions, and 96% had ≥2 comorbidities. The median interval between the two positive tests was 77 days. Among the 19 patients with 60 to 89 days between positive tests, 17 (89%) exhibited symptoms or clinical manifestations consistent with COVID-19 at the time of the second positive test and 14 (74%) were hospitalized at the second positive test. Of the four patients with ≥90 days between two positive tests (patient 2 [PT2], PT8, PT14, and PT19), two had mild or no symptoms at the second positive test and one, an immunocompromised patient, had a brief hospitalization at the first diagnosis, followed by intensive care unit (ICU) admission at the second diagnosis 3 months later. Our study demonstrated a high prevalence of compromised immune systems, comorbidities, obesity, and smoking among patients with repeatedly positive SARS-CoV-2 tests. Despite limitations, including a lack of semiquantitative estimates of viral load, these data may help prioritize suspected cases of reinfection for investigation and continued surveillance

MicroRNA 506 regulates expression of PPAR alpha in hydroxycamptothecin-resistant human colon cancer cells

AbstractChemotherapeutic drug resistance remains a major obstacle to the successful treatment of colon cancer. Here, we show that 77 differentially expressed miRNAs were identified in SW1116/HCPT versus SW1116, and over-expressed miR-506 in SW1116/HCPT cells was validated. Then it was indicated that PPARα is a common target of miR-506 by using a luciferase reporter assay. Our results also demonstrated that cytotoxic ability of HCPT requires the concomitant presence of PPARα, and that loss of PPARα expression imparts resistance to HCPTs anti-tumor effects. All together, our studies indicate that miR-506 over-expression in established HCPT-resistant colon cancer cell line confers resistance to HCPT by inhibiting PPARα expression, then providing a rationale for the development of miRNA-based strategies for reversing resistance in HCPT-resistant colon cancer cells

Dynamic diffusion tensor imaging reveals structural changes in the bilateral pyramidal tracts after brain stem hemorrhage in rats

Background and Purpose: Few studies have concentrated on pyramidal tract (PY) changes after brain stem hemorrhage (BSH). In this study, we used a diffusion tensor imaging (DTI) technique and histologic identification to investigate longitudinal PY changes on both the contralateral and ipsilateral sides after experimental BSH. Methods: BSH was induced in 61 Sprague-Dawley rats by infusing 30 μl of autogenous tail blood into each rat’s right pons. DTI and motor function examinations were performed repeatedly on days 1, 3, 7, 14, and 28 after surgery. Fractional anisotropy, mean diffusivity, axial diffusivity, and radial diffusivity were measured in the bilateral PYs. The axon and myelin injury in the PY were evaluated by histologic study. Results: As compared with normal controls, the bilateral PYs in rats with induced BSH showed an early decrease and a late increase in fractional anisotropy and an early increase and a late decrease in mean diffusivity. A progressive decrease in axial diffusivity with dramatic axon loss from day 1 to day 28 after BSH was found bilaterally. The bilateral PYs showed an early increase and a late decrease in radial diffusivity. Early myelin injury and late repair were also detected pathologically in the bilateral PYs of rats with BSH. Thus, the early motor function deficits of rats with BSH began to improve on day 14 and had almost completely disappeared by day 28. Conclusions: DTI revealed dynamic changes in the bilateral PYs after BSH, which was confirmed by histologic findings and which correlated with motor function alteration. These findings support the idea that quantitative DTI can track structural changes in the bilateral PYs and that DTI may serve as a noninvasive tool to predict the prognoses of patients with BSH

Insulin treatment enhances the myocardial angiogenic response in diabetes

ObjectiveGrowth factor and cell-based angiogenesis are attractive therapeutic options for diabetic patients with end-stage coronary disease. Reduced collateral vessel formation observed in diabetes is associated with increased expression of anti-angiogenic proteins, angiostatin and endostatin. The aim of this study was to determine the effects of insulin treatment on the diabetic angiogenic response to chronic myocardial ischemia.MethodsYucatan miniswine were treated with alloxan (pancreatic β-cell specific toxin, 150 mg/kg) and divided into two groups. In the diabetic group (DM, n = 8), blood glucose levels were kept greater than 250 mg/dL, and in the insulin-treated group (IDM, n = 6), intramuscular insulin was administered daily to keep blood glucose less than 150 mg/dL. A third group of age-matched swine served as nondiabetic controls (ND; n = 8). Eight weeks later, all animals underwent circumflex artery ameroid constrictor placement to induce chronic ischemia. Myocardial perfusion was assessed at 3 and 7 weeks after ameroid placement using microspheres. Microvascular function, capillary density, and myocardial expression of anti-angiogenic mediators were evaluated.ResultsDiabetic animals exhibited significant impairments in endothelium-dependent microvessel relaxation to adenosine diphosphate and substance P, which were reversed in insulin-treated animals. Collateral-dependent perfusion in the ischemic circumflex territory, which was profoundly reduced in diabetic animals (−0.18 ± 0.02 vs +0.23 ± 0.07 mL · min−1 · g−1; P < .001), improved significantly with insulin treatment (0.12 ± 0.05 mL · min−1 · g−1; P < .01). Myocardial expression of anti-angiogenic proteins, angiostatin and endostatin, showing a 4.3- and 3.6-fold increase in diabetic animals respectively (both P < .01 vs ND), was markedly reduced in insulin-treated animals (2.3- and 1.8-fold vs ND; both P < .01).ConclusionsInsulin treatment successfully reversed diabetic coronary endothelial dysfunction and significantly improved the endogenous angiogenic response. These pro-angiogenic effects may be mediated through downregulation of anti-angiogenic mediators. Insulin therapy appears to be a promising modality to enhance the angiogenic response in diabetic patients

Pholidota chinensis alleviates azoxymethane/dextran sulfate sodium-induced colorectal carcinogenesis through inhibition of TLR4 and COX-2

Background: Inflammatory bowel disease (IBD) always progresses to colorectal cancer (CRC) which is the second most frequent cause of death by cancer. It is about 2% of population in the lifetime worldwide who at the risk for development of CRC. Oxaliplatin is an effective anticancer drug used for the treatment of advanced CRC; however, it always causes a robust painful neuropathy. Pholidota chinensis is a Chinese folk herbal medicine which was used for treatment of inflammation such as gastroenteritis, duodenal ulcer and bronchitis.Materials and Methods: The azoxymethane (AOM) and dextran sulfate sodium (DSS) were used to induce the colon tumor of mice. The effect of Pholidota chinensis on colon tumorigenesis was evaluated. Immunohistochemistry and semi-quantitative RT-PCR were used to detect the expression of toll-like receptor 4 (TLR4) and cyclooxygenase-2 (COX-2) in colon.Results: Pholidota chinensis can alleviate the colon tumorigenesis. The prevention effects of Pholidota chinensis are similar to oxaliplatin. Specifically, administration of Pholidota chinensis solution suppresses the expression of the proliferating cell nuclear antigen (PCNA), toll-like receptor 4 (TLR4) and cyclooxygenase-2 (COX-2).Conclusion: Our findings suggested that Pholidota chinensis participate in the regulation of colon cancer development through inhibiting the expression of TLR4 and COX-2.Keywords: Pholidota chinensis; colorectal cancer; Toll-like receptor 4; Cyclooxygenase-

Intermolecular Failure of L-type Ca(2+) Channel and Ryanodine Receptor Signaling in Hypertrophy

Pressure overload–induced hypertrophy is a key step leading to heart failure. The Ca(2+)-induced Ca(2+) release (CICR) process that governs cardiac contractility is defective in hypertrophy/heart failure, but the molecular mechanisms remain elusive. To examine the intermolecular aspects of CICR during hypertrophy, we utilized loose-patch confocal imaging to visualize the signaling between a single L-type Ca(2+) channel (LCC) and ryanodine receptors (RyRs) in aortic stenosis rat models of compensated (CHT) and decompensated (DHT) hypertrophy. We found that the LCC-RyR intermolecular coupling showed a 49% prolongation in coupling latency, a 47% decrease in chance of hit, and a 72% increase in chance of miss in DHT, demonstrating a state of “intermolecular failure.” Unexpectedly, these modifications also occurred robustly in CHT due at least partially to decreased expression of junctophilin, indicating that intermolecular failure occurs prior to cellular manifestations. As a result, cell-wide Ca(2+) release, visualized as “Ca(2+) spikes,” became desynchronized, which contrasted sharply with unaltered spike integrals and whole-cell Ca(2+) transients in CHT. These data suggested that, within a certain limit, termed the “stability margin,” mild intermolecular failure does not damage the cellular integrity of excitation-contraction coupling. Only when the modification steps beyond the stability margin does global failure occur. The discovery of “hidden” intermolecular failure in CHT has important clinical implications

Dietary inclusion of fermented ginger straw effect on the growth performance, gastrointestinal tract development and caecal fermentation of fattening rabbits

[EN] This experiment was conducted to evaluate the effects of dietary inclusion of fermented ginger straw on the growth performance, gastrointestinal tract development and caecal fermentation of fattening rabbits. A total of 160 45-d-old Laiwu black rabbits were randomly divided into 4 groups and fed 0% (Control), 5, 10 or 15% fermented ginger straw in their diet as a replacement for peanut straw powder. The trial lasted for 7 d of adaptation and 43 d for testing. Growth performance was recorded from 52 to 95 d of age (n=5 per treatment with 30 rabbits, 3 males and 3 females per replicate), TTAD of nutrients from 91 to 95 d of age, and gastrointestinal tract development, caecum fermentation and carcass traits were determined at 95 d of age (n=5 per treatment with 10 rabbits, 1 males and 1 females in per replicate).The results showed that the average daily gain and final body weight in the experimental groups (5, 10 and 15% fermented ginger straw) were higher than in the control group (P&lt;0.05). However, the average daily feed intake in the 15% group was higher than in the other groups, while the total tract apparent digestibility of crude protein, ether extract, neutral detergent fibre and acid detergent fibre were lower than in the control group (P&lt;0.05), and the relative weights of the stomach, small intestine and caecum content in the 15% substitution group were higher than those in the control group (P&lt;0.05). In addition, the thickness of the muscle layer in the 15% substitution group was higher than that in the other groups (P&lt;0.05). Moreover, pH and total volatile fatty acids concentration in the caecal content were similar among the 4 groups (P&gt;0.05). The current work shows that fermented ginger straw could be used as roughage material in fattening rabbit production up to a dietary dose of 10%.This study was supported by Earmarked Fund for Modern Agro-industry Technology Research System (CARS-43-G-7); Shandong Province Modern Agricultural Industry Technology System (SDAIT-21); Scientific and Technological Problems Project Unveiled by Shandong Academy of Agricultural Sciences (SHJB2021-43).Sun, HT.; Wang, Y.; Bai, LY.; Liu, C.; Xu, YH.; Gao, SX.; Jiang, WX.... (2022). Dietary inclusion of fermented ginger straw effect on the growth performance, gastrointestinal tract development and caecal fermentation of fattening rabbits. World Rabbit Science. 30(4):267-276. https://doi.org/10.4995/wrs.2022.1609326727630

Alterations of Serum Levels of BDNF-Related miRNAs in Patients with Depression

Depression is a serious and potentially life-threatening mental disorder with unknown etiology. Emerging evidence shows that brain-derived neurotrophic factor (BDNF) and microRNAs (miRNAs) play critical roles in the etiology of depression. Here this study was aimed to identify and characterize the roles of BDNF and its putative regulatory miRNAs in depression. First, we identified that miR-182 may be a putative miRNA that regulates BDNF levels by bioinformatic studies, and characterized the effects of miR-182 on the BDNF levels using cell-based studies, side by side with miR-132 (a known miRNA that regulates BDNF expression). We showed that treatment of miR-132 and miR-182 respectively decreased the BDNF protein levels in a human neuronal cell model, supporting the regulatory roles of miR-132 and miR-182 on the BDNF expression. Furthermore, we explored the roles of miR-132 and miR-182 on the BDNF levels in depression using human subjects by assessing their serum levels. Compared with the healthy controls, patients with depression showed lower serum BDNF levels (via the enzyme-linked immunosorbent assays) and higher serum miR-132 and miR-182 levels (via the real-time PCR). Finally, the Pearson’s (or Spearman’s) correlation coefficient was calculated to study whether there was a relationship among the Self-Rating Depression Scale score, the serum BDNF levels, and serum BDNF-related miRNA levels. Our results revealed that there was a significant negative correlation between the SDS scores and the serum BDNF levels, and a positive correlation between the SDS scores and miR-132 levels. In addition, we found a reverse relationship between the serum BDNF levels and the miR-132/miR-182 levels in depression. Collectively, we provided evidence supporting that miR-182 is a putative BDNF-regulatory miRNA, and suggested that the serum BDNF and its related miRNAs may be utilized as important biomarkers in the diagnosis or as therapeutic targets of depression