Canine multiple primary tumours: Mammary tubular carcinoma, uterine leiomyosarcoma, and facial sebaceous epithelioma.

Multiple primary malignant tumours (MPMTs) are multiple neoplasms with independent pathogenetic origins, placing great importance on the tumorigenesis and clinical treatment. However, due to the rare occurrence and diagnostic confusion, MPMTs have rarely been investigated in veterinary medicine. In this report, a 10-year-old intact female Maltese dog had MPMTs, consisting of two malignant tumours and one benign tumour each derived from a topographically different site: tubular carcinoma in the mammary glands, leiomyosarcoma in the uterus and sebaceous epithelioma in the cheek. The unique combination of MPMTs would be the first case in veterinary research to give insight into the diagnosis, disease characteristics, and surgical treatment.

A Noble Extract of Pseudomonas sp. M20A4R8 Efficiently Controlling the Influenza Virus-Induced Cell Death.

Epidemic diseases that arise from infectious RNA viruses, particularly influenza viruses, pose a constant threat to the global economy and public health. Viral evolution has undermined the efficacy of acquired immunity from vaccines and the antiviral effects of FDA-approved drugs. As such, there is an urgent need to develop new antiviral lead agents. Natural compounds, owing to their historical validation of application and safety, have become a promising solution. In this light, a novel marine bacterium, Pseudomonas sp. M20A4R8, has been found to exhibit significant antiviral activity [half maximal inhibitory concentration (IC50) = 1.3 µg/mL, selectivity index (SI) = 919.4] against influenza virus A/Puerto Rico/8/34, surpassing the activity of chloroquine. The antiviral response via M20A4R8 extract was induced during post-entry stages of the influenza virus, indicating suitability for post-application after the establishment of viral infection. Furthermore, post-treatment with M20A4R8 extract protected the host from virus-induced apoptosis, suggesting its potential use in acute respiratory disease complexes resulting from immune effectors' overstimulation and autophagy-mediated self-apoptosis. The extract demonstrated an outstanding therapeutic index against influenza virus A/Wisconsin/15/2009 (IC50 = 8.1 µg/mL, SI = 146.2) and B/Florida/78/2015 Victoria lineage (IC50 = 3.5 µg/mL, SI = 343.8), indicating a broad anti-influenza virus activity with guaranteed safety and effectiveness. This study provides a new perspective on mechanisms for preventing a broad spectrum of viral infections through antiviral agents from novel and natural origins. Future studies on a single or combined compound from the extract hold promise, encouraging its use in preclinical challenge tests with various influenza virus strains.

Concurrent primary splenic lymphoma and mammary gland tumour with polycystic ovaries in a dog.

Here, we report a rare case of concurrent primary splenic lymphoma and mammary gland tumour (MGT) with polycystic ovaries in a 10-year-old, intact female Jindo dog. The dog was presented with multiple masses in the fourth left mammary gland, the largest of which measured 6 cm in diameter, along with enlargement of the left inguinal lymph node on physical examination. Ultrasonography, radiography, and computed tomography scans revealed polycystic ovaries and a mass in the tail of the spleen, after total splenectomy and mastectomy with ovariohysterectomy, histopathological examination identified splenic diffuse large B cell lymphoma and malignant myoepithelioma of the mammary gland was found. To our knowledge, this is the first report of the concurrent occurrence of splenic lymphoma, MGT, and polycystic ovaries in a dog.

Identification and characterization of a marine bacterium extract from Mameliella sp. M20D2D8 with antiviral effects against influenza A and B viruses.

Despite significant improvements in vaccines and chemotherapeutic drugs, pathogenic RNA viruses continue to have a profound impact on the global economy and pose a serious threat to animal and human health through emerging and re-emerging outbreaks of diseases. To overcome the challenge of viral adaptation and evolution, increased vigilance is required. Particularly, antiviral drugs derived from new, natural sources provide an attractive strategy for controlling problematic viral diseases. In this antiviral study, we discovered a previously unknown bacterium, Mameliella sp. M20D2D8, by conducting an antiviral screening of marine microorganisms. An extract from M20D2D8 exhibited antiviral activity with low cytotoxicity and was found to be effective in vitro against multiple influenza virus strains: A/PR8 (IC50 = 2.93 µg/mL, SI = 294.85), A/Phil82 (IC50 = 1.42 µg/mL, SI = 608.38), and B/Yamagata (IC50 = 1.59 µg/mL, SI = 543.33). The antiviral action was found to occur in the post-entry stages of viral replication and to suppress viral replication by inducing apoptosis in infected cells. Moreover, it efficiently suppressed viral genome replication, protein synthesis, and infectivity in MDCK and A549 cells. Our findings highlight the antiviral capabilities of a novel marine bacterium, which could potentially be useful in the development of drugs for controlling viral diseases.

CREB5 promotes the proliferation and self-renewal ability of glioma stem cells.

Glioblastoma multiforme (GBM) is the most fatal form of brain cancer in humans, with a dismal prognosis and a median overall survival rate of less than 15 months upon diagnosis. Glioma stem cells (GSCs), have recently been identified as key contributors in both tumor initiation and therapeutic resistance in GBM. Both public dataset analysis and direct differentiation experiments on GSCs have demonstrated that CREB5 is more highly expressed in undifferentiated GSCs than in differentiated GSCs. Additionally, gene silencing by short hairpin RNA (shRNA) of CREB5 has prevented the proliferation and self-renewal ability of GSCs in vitro and decreased their tumor forming ability in vivo. Meanwhile, RNA-sequencing, luciferase reporter assay, and ChIP assay have all demonstrated the closely association between CREB5 and OLIG2. These findings suggest that targeting CREB5 could be an effective approach to overcoming GSCs.

Genome-wide association study of mammary gland tumors in Maltese dogs.

Background: A genome-wide association study (GWAS) is a valuable tool for investigating genetic and phenotypic variation in many diseases. Objective: The objective of this study was to identify variations in the genomes of Maltese dogs that are associated with the mammary gland tumor (MGT) phenotype and to assess the association between each biological condition and MGT phenotype in Maltese dogs. Methods: DNA was extracted from 22 tumor samples and 11 whole blood samples from dogs with MGTs. Genome-wide single-nucleotide polymorphism (SNP) genotyping was performed, and the top 20 SNPs associated with various conditions and genetic variations were mapped to their corresponding gene locations. Results: The genotyping process successfully identified 173,662 loci, with an overall genotype completion rate of 99.92%. Through the quality control analysis, 46,912 of these SNPs were excluded. Allelic tests were conducted to generate Manhattan plots, which showed several significant SNPs associated with MGT phenotype in intergenic region. The most prominent SNP, located within a region associated with transcription and linked to the malignancy grade of MGT, was identified on chromosome 5 (p = 0.00001) though there may be lack of statistical significance. Other SNPs were also found in several genes associated with oncogenesis, including TNFSF18, WDR3, ASIC5, STAR, and IL1RAP. Conclusion: To our knowledge, this is the first GWAS to analyze the genetic predisposition to MGT in Maltese dogs. Despite the limited number of cases, these analyzed data could provide the basis for further research on the genetic predisposition to MGTs in Maltese dogs.

Targeting the multidrug and toxin extrusion 1 gene (SLC47A1) sensitizes glioma stem cells to temozolomide.

Glioblastoma multiforme (GBM) is the most aggressive type of brain tumor, with an extremely poor prognosis due to resistance to standard-of-care treatments. Strong evidence suggests that the small population of glioma stem cells (GSCs) contributes to the aggressiveness of GBM. One of the mechanisms that promote GSC progression is the dysregulation of membrane transporters, which mediate the influx and efflux of substances to maintain cellular homeostasis. Here, we investigated the role of multidrug and toxin extrusion transporter gene SLC47A1 in GSCs. Results show that SLC47A1 is highly expressed in GSCs compared to non-stem cell glioma cells, and non-tumor cells. Additionally, in-silico analysis of public datasets showed that high SLC47A1 expression is linked to malignancy and a poor prognosis in glioma patients. Further, SLC47A1 expression is correlated with important biological processes and signaling pathways that support tumor growth. Meanwhile, silencing SLC47A1 by short-hairpin RNA (shRNA) influenced cell viability and self-renewal activity in GSCs. Interestingly, SLC47A1 shRNA knockdown or pharmacological inhibition potentiates the effect of temozolomide (TMZ) in GSC cells. The findings suggest that SLC47A1 could serve as a useful therapeutic target for gliomas.

Flow Diverter Treatment Using a Flow Re-Direction Endoluminal Device for Unruptured Intracranial Vertebral Artery Dissecting Aneurysm: Single-Center Case Series and Technical Considerations.

PURPOSE: This study aimed to evaluate the effectiveness, safety, and technical considerations of flow diverter (FD) treatment using a Flow Re-direction Endoluminal Device (FRED) for unruptured intracranial vertebral artery dissecting aneurysms (VADAs). MATERIALS AND METHODS: We conducted a retrospective study of 23 patients with unruptured intracranial VADAs who underwent FD treatment using a FRED between June 2017 and August 2021. Symptoms, imaging findings, treatment strategies, and angiographic and clinical outcomes were evaluated. Dissections were categorized according to the dominance of the VA in which they occurred: dominant VA, co-dominant VA, and non-dominant VA. RESULTS: All patients successfully underwent FD treatment with either a FRED (n=11) or FRED Jr. (n=12). Complete occlusion rates were 78.3% at 6-month follow-up magnetic resonance angiography and 91.3% at 12-month. There were no instances of complications, recurrence, or retreatment during a median follow-up of 20 months. Dissections occurred in the dominant VA in 3 cases (13.0%), the co-dominant VA in 13 cases (56.5%), and the non-dominant VA in 7 cases (30.4%). Intimal flap and true lumen stenosis were observed in 39.1% and 30.4% of cases, respectively. Four cases required a bilateral VA approach due to technical difficulties, all in the non-dominant VA. CONCLUSION: Flow diversion treatment using a FRED for unruptured intracranial VADAs proved feasible and safe, yielding satisfactory occlusion rates. Technical challenges were more likely in lesions involving non-dominant VAs in the acute or subacute stage, mainly due to associated intraluminal lesions compromising the arterial lumen.

Clinical feasibility of CS-VIBE accelerates MRI techniques in diagnosing intracranial metastasis.

Our objective was to evaluate and compare the diagnostic performance of post-contrast 3D compressed-sensing volume-interpolated breath-hold examination (CS-VIBE) and 3D T1 magnetization-prepared rapid-acquisition gradient-echo (MPRAGE) in detecting intracranial metastasis. Additionally, we analyzed and compared the image quality between the two. We enrolled 164 cancer patients who underwent contrast-enhanced brain MRI. Two neuroradiologists independently reviewed all the images. The signal-to-noise ratio (SNR), contrast-to noise ratio (CNR) were compared between two sequences. For patients with intracranial metastasis, we measured enhancement degree and CNRlesion/parenchyma of the lesion. The overall image quality, motion artifact, gray-white matter discrimination and enhancing lesion conspicuity were analyzed. Both MPRAGE and CS-VIBE showed similar performance in diagnosing intracranial metastasis. Overall image quality of CS-VIBE was better with less motion artifact; however conventional MPRAGE was superior in enhancing lesion conspicuity. Overall, the SNR and CNR of conventional MPRAGE were higher than those of CS-VIBE. For 30 enhancing intracranial metastatic lesions, MPRAGE showed a lower CNR (p = 0.02) and contrast ratio (p = 0.03). MPRAGE and CS-VIBE were preferred in 11.6 and 13.4% of cases, respectively. In comparison with conventional MPRAGE, CS-VIBE achieved comparable image quality and visualization, with the scan time being half of that of MPRAGE.

Deep learning referral suggestion and tumour discrimination using explainable artificial intelligence applied to multiparametric MRI.

OBJECTIVES: An appropriate and fast clinical referral suggestion is important for intra-axial mass-like lesions (IMLLs) in the emergency setting. We aimed to apply an interpretable deep learning (DL) system to multiparametric MRI to obtain clinical referral suggestion for IMLLs, and to validate it in the setting of nontraumatic emergency neuroradiology. METHODS: A DL system was developed in 747 patients with IMLLs ranging 30 diseases who underwent pre- and post-contrast T1-weighted (T1CE), FLAIR, and diffusion-weighted imaging (DWI). A DL system that segments IMLLs, classifies tumourous conditions, and suggests clinical referral among surgery, systematic work-up, medical treatment, and conservative treatment, was developed. The system was validated in an independent cohort of 130 emergency patients, and performance in referral suggestion and tumour discrimination was compared with that of radiologists using receiver operating characteristics curve, precision-recall curve analysis, and confusion matrices. Multiparametric interpretable visualisation of high-relevance regions from layer-wise relevance propagation overlaid on contrast-enhanced T1WI and DWI was analysed. RESULTS: The DL system provided correct referral suggestions in 94 of 130 patients (72.3%) and performed comparably to radiologists (accuracy 72.6%, McNemar test; p = .942). For distinguishing tumours from non-tumourous conditions, the DL system (AUC, 0.90 and AUPRC, 0.94) performed similarly to human readers (AUC, 0.81~0.92, and AUPRC, 0.88~0.95). Solid portions of tumours showed a high overlap of relevance, but non-tumours did not (Dice coefficient 0.77 vs. 0.33, p < .001), demonstrating the DL's decision. CONCLUSIONS: Our DL system could appropriately triage patients using multiparametric MRI and provide interpretability through multiparametric heatmaps, and may thereby aid neuroradiologic diagnoses in emergency settings. CLINICAL RELEVANCE STATEMENT: Our AI triages patients with raw MRI images to clinical referral pathways in brain intra-axial mass-like lesions. We demonstrate that the decision is based on the relative relevance between contrast-enhanced T1-weighted and diffusion-weighted images, providing explainability across multiparametric MRI data. KEY POINTS: • A deep learning (DL) system using multiparametric MRI suggested clinical referral to patients with intra-axial mass-like lesions (IMLLs) similar to radiologists (accuracy 72.3% vs. 72.6%). • In the differentiation of tumourous and non-tumourous conditions, the DL system (AUC, 0.90) performed similar with radiologists (AUC, 0.81-0.92). • The DL's decision basis for differentiating tumours from non-tumours can be quantified using multiparametric heatmaps obtained via the layer-wise relevance propagation method.

Feasibility and intra-and interobserver reproducibility of quantitative susceptibility mapping with radiomic features for intracranial dissecting intramural hematomas and atherosclerotic calcifications.

Quantitative susceptibility mapping (QSM) for 61 patients with dissecting intramural hematomas (n = 36) or atherosclerotic calcifications (n = 25) in intracranial vertebral arteries were collected to assess intra- and interobserver reproducibility in a 3.0-T MR system between January 2015 and December 2017. Two independent observers each segmented regions of interest for lesions twice. The reproducibility was evaluated using intra-class correlation coefficients (ICC) and within-subject coefficients of variation (wCV) for means and concordance correlation coefficients (CCC) and ICC for radiomic features (CCC and ICC > 0.85) were used. Mean QSM values were 0.277 ± 0.092 ppm for dissecting intramural hematomas and - 0.208 ± 0.078 ppm for atherosclerotic calcifications. ICCs and wCVs were 0.885-0.969 and 6.5-13.7% in atherosclerotic calcifications and 0.712-0.865 and 12.4-18.7% in dissecting intramural hematomas, respectively. A total of 9 and 19 reproducible radiomic features were observed in dissecting intramural hematomas and atherosclerotic calcifications, respectively. QSM measurements in dissecting intramural hematomas and atherosclerotic calcifications were feasible and reproducible between intra- and interobserver comparisons, and some reproducible radiomic features were demonstrated.

Effects of Vigna angularis extract and its active compound hemiphloin against atopic dermatitis-like skin inflammation.

Vigna angularis is an edible crop and herbal medicine that is known to have antipyretic, anti-inflammatory, and anti-edema effects. Many studies have been conducted on the 95% ethanol extract of V. angularis, but there is little research on the 70% ethanol extract and hemiphloin, which is a new indicator component of the 70% ethanol extract of V. angularis. To investigate the in vitro anti-atopic effect and verify the mechanism action of 70% ethanol extract of V. angularis (VAE), TNF-α/IFN-γ-induced HaCaT keratinocytes were used. The VAE treatment alleviated TNF-α/IFN-γ-induced IL-1ß, IL-6, IL-8, CCL17/TARC, and CCL22/MDC gene expressions and productions. VAE also inhibited the phosphorylation of MAPKs, including p38, ERK, JNK, STAT1, and NF-κB in TNF-α/IFN-γ-induced HaCaT cells. 2,4-dinitochlorobenzene (DNCB)-induced skin inflammation mice model, and HaCaT keratinocytes were used. In the DNCB-induced mouse model, VAE treatment alleviated ear thicknesses and IgE levels. Furthermore, VAE decreased IL-1ß, IL-6, IL-8, CCL17/TARC, and CCL22/MDC gene expressions of DNCB-applied ear tissue. Additionally, we investigated the anti-atopic and anti-inflammatory effects of hemiphloin using TNF-α/IFN-γ-induced HaCaT keratinocytes and LPS-induced J774 macrophages. Treatment hemiphloin decreased gene expressions and productions of IL-1ß, IL-6, IL-8, CCL17/TARC, and CCL22/MDC in TNF-α/IFN-γ-induced HaCaT cells. The phosphorylations of p38, ERK, STAT1, and NF-κB were inhibited by hemiphloin in TNF-α/IFN-γ-induced HaCaT cells. Finally, hemiphloin showed anti-inflammatory activities in LPS-induced J774 cells. It decreased LPS-induced NO productions and iNOS and COX-2 expressions. Treatment of hemiphloin also inhibited LPS-induced TNF-α, IL-1ß, and IL-6 gene expressions. These results suggest that VAE is an anti-inflammatory agent for inflammatory skin diseases and that hemiphloin could be a therapeutic candidate for inflammatory skin diseases.

Addendum: Outpatient Day-Care Management of Unruptured Intracranial Aneurysm: A Retrospective Cohort Study.

This corrects the article on p. 828 in vol. 23, PMID: 35762182.

Downregulation of ADAMTS3 Suppresses Stemness and Tumorigenicity in Glioma Stem Cell.

AIMS: Glioblastoma multiforme (GBM) is the most aggressive type of human brain tumor, with a poor prognosis and a median overall survival of fewer than 15 months. Glioma stem cells (GSCs) have recently been identified as a key player in tumor initiation and therapeutic resistance in GBM. ADAMTS family of metalloproteinases is known to cleave a wide range of extracellular matrix substrates and has been linked to tissue remodeling events in tumor development. Here, we investigate that ADAMTS3 regulates GSC proliferation and self-renewal activities, and tumorigenesis in orthotopic xenograft models. METHODS: ADAMTS3 mRNA expression levels in normal human astrocyte (NHA), glioma, and GSCs cell lines were compared. After knockdown of ADAMTS3, alamarBlue assay, in vitro limiting dilution, and orthotopic xenograft assays were performed. To investigate the tumor-associated roles of ADAMTS3, several statistical assays were conducted using publicly available datasets. RESULTS: ADAMTS3 level was remarkably higher in GSCs than in NHA, glioma cell lines, and their matched differentiated tumor cells. Interestingly, knockdown of ADAMTS3 disrupted GSC's proliferation, self-renewal activity, and tumor formation in vivo. Furthermore, ADAMTS3 could be used as an independent predictor of malignancy progression in GBM. CONCLUSION: We identified ADAMTS3 as a potential therapeutic target for GBM.

Therapeutic strategy targeting host lipolysis limits infection by SARS-CoV-2 and influenza A virus.

The biosynthesis of host lipids and/or lipid droplets (LDs) has been studied extensively as a putative therapeutic target in diverse viral infections. However, directly targeting the LD lipolytic catabolism in virus-infected cells has not been widely investigated. Here, we show the linkage of the LD-associated lipase activation to the breakdown of LDs for the generation of free fatty acids (FFAs) at the late stage of diverse RNA viral infections, which represents a broad-spectrum antiviral target. Dysfunction of membrane transporter systems due to virus-induced cell injury results in intracellular malnutrition at the late stage of infection, thereby making the virus more dependent on the FFAs generated from LD storage for viral morphogenesis and as a source of energy. The replication of SARS-CoV-2 and influenza A virus (IAV), which is suppressed by the treatment with LD-associated lipases inhibitors, is rescued by supplementation with FFAs. The administration of lipase inhibitors, either individually or in a combination with virus-targeting drugs, protects mice from lethal IAV infection and mitigates severe lung lesions in SARS-CoV-2-infected hamsters. Moreover, the lipase inhibitors significantly reduce proinflammatory cytokine levels in the lungs of SARS-CoV-2- and IAV-challenged animals, a cause of a cytokine storm important for the critical infection or mortality of COVID-19 and IAV patients. In conclusion, the results reveal that lipase-mediated intracellular LD lipolysis is commonly exploited to facilitate RNA virus replication and furthermore suggest that pharmacological inhibitors of LD-associated lipases could be used to curb current COVID-19- and future pandemic outbreaks of potentially troublesome RNA virus infection in humans.

Clinical Manifestation and Radiologic Patterns of Spontaneous Cervicocephalic Dissection According to the Anatomic Location: A Single-Center Analysis in Korean Patients.

PURPOSE: Spontaneous cervicocephalic dissection (SCAD) is an important cause of stroke and shows various lesion locations and clinical features. The purpose of this study was to analyze the location of SCAD and its clinical and radiologic patterns in Korean patients. MATERIALS AND METHODS: Patients with SCAD who were evaluated between 2013 and 2018 at a tertiary center in Korea were reviewed. We classified and compared the morphological (aneurysm or steno-occlusion) and presenting (hemorrhage or infarction) patterns according to the lesion locations (anterior circulation [AC] vs. posterior circulation [PC]; intradural [ID] vs. extradural [ED]). RESULTS: A total of 166 patients were included in this study. The SCAD most commonly occurred in the PC-ID location (65.1%), followed by AC-ID (13.3%), AC-ED (13.3%), and PC-ED (8.4%). Aneurysm and steno-occlusion patterns were observed in 66.9% and 57.8% of the cases, respectively. The aneurysm pattern was significantly more common in the PC-ID location (78.7%) than in other locations. As for the presenting pattern, cerebral infarction was the most common pattern (39.8%), and intracranial hemorrhage was observed only in the ID location (7.2%). CONCLUSION: In Korean patients, PC-ID, especially ID vertebral artery, was the most common location of SCAD, and most cases were accompanied by an aneurysm. It also suggested that these location trends differ by population or ethnicity.

Outpatient Day-Care Management of Unruptured Intracranial Aneurysm: A Retrospective Cohort Study.

OBJECTIVE: This study aimed to assess the outcomes of outpatient day-care management of unruptured intracranial aneurysm (UIA), and to present the risks associated with different management strategies by comparing the outcomes and adverse events between outpatient day-care management and management with longer admission periods. MATERIALS AND METHODS: This retrospective cohort study used prospectively registered data and was approved by a local institutional review board. We enrolled 956 UIAs from 811 consecutive patients (mean age ± standard deviation, 57 ± 10.7 years; male:female = 247:564) from 2017 to 2020. We compared the outcomes after embolization among the different admission-length groups (1, 2, and ≥ 3 days). The outcomes included pre- and post-modified Rankin Scale (mRS) scores and rates of adverse events, cure, recurrence, and reprocedure. Events were defined as any cerebrovascular problems, including minor and major stroke, death, or hemorrhage. RESULTS: The mean admission period was 2 days, and 175 patients (191 aneurysms), 551 patients (664 aneurysms), and 85 patients (101 aneurysms) were discharged on the day of the procedure, day 2, and day 3 or later, respectively. During the mean 17-month follow-up period (range 6-53 months; 2757 patient years), no change in post-mRS was observed compared to pre-mRS in 99.6% of patients. Cure was achieved in 95.6% patients; minimal recurrence that did not require re-procedure occurred in 3.5% patients, and re-procedure was required in 2.3% (22 of 956) patients due to progressive enlargement of the recurrent sac during follow up (mean 17 months, range, 6-53 months). There were eight adverse events (0.8%), including five cerebrovascular (two major stroke, two minor strokes and one transient ischemic stroke), and three non-cerebrovascular events. Statistical comparison between groups with different admission lengths (1, 2, and ≥ 3 days) revealed no difference in the outcomes. CONCLUSION: This study revealed no difference in outcomes and adverse events according to the admission period, and suggested that UIA could be managed by outpatient day-care embolization.

Rotavirus exploits SREBP pathway for hyper lipid biogenesis during replication.

Species A rotavirus (RVA) is one of the pathogens causing severe acute gastroenteritis in young children and animals worldwide. RVA replicates and assembles its immature particle within electron dense compartments known as viroplasm. Despite the importance of lipid droplet (LD) formation in the RVA viroplasm, the upstream molecules modulating LD formation have remained elusive. Here, we demonstrate that RVA infection reprogrammes sterol regulatory element binding proteins (SREBPs)-dependent lipogenic pathways in virus-infected cells. Interestingly, silencing of SREBPs significantly reduced RVA protein synthesis, genome replication and progeny virus production. Moreover, knockout of SREBP-1c gene conferred resistance to RVA-induced diarrhoea, reduction of RVA replication, and mitigation of small intestinal pathology in mice. This study identifies SREBPs-mediated lipogenic reprogramming in RVA-infected host cells for facilitating virus replication and SREBPs as a potential target for developing therapeutics against RVA infection.

Ethanol ablation for the treatment of thyroglossal duct cysts: follow-up results for longer than 2 years.

OBJECTIVES: To evaluate the long-term efficacy and safety of ethanol ablation (EA) for the treatment of thyroglossal duct cysts (TGDCs). METHODS: This retrospective study included 81 consecutive patients diagnosed with and treated for symptomatic TGDCs at two institutions between Jan 2008 and Oct 2018. Preprocedural evaluation included US assessment with calculation of the TGDC volume. EA was performed under US guidance using 99% ethanol. Post-treatment follow-up was scheduled within 3 months, 6 months, and then annually. Immediate success was defined as a volume reduction ratio (VRR; ratio of the volume difference after EA to the initial TGDC volume) > 50% within 3 months. Long-term success was defined as VRR > 50% or resolution or improvement of cosmetic problems and symptoms without recurrence at last follow-up. RESULTS: Seventy-seven patients underwent EA, and outcomes were assessed in 68 patients with available follow-up data. The immediate success rate of the first EA was 81% (55/68), with a mean VRR within 3 months of 73% ± 31%. One patient (1.5%, 1/68) developed wound inflammation after the first EA. Forty-two patients were followed up for longer than 2 years. For the median follow-up of 69 months (range, 24-131 months), the long-term success rate was 83% (35/42), with a mean VRR at last follow-up of 81% ± 35%. No patients developed malignancy from the ablated TGDCs. CONCLUSIONS: EA for treatment of TGDCs achieved acceptable rates of immediate and long-term efficacy with a low complication rate, and can be considered as a first-line treatment for the management of TGDCs. KEY POINTS: • The immediate success rate of EA for the treatment of TGDCs was 81% (55/68), with a mean VRR within 3 months of 73% ± 31%. • For the median follow-up of 69 months (range, 24-131 months), the long-term success rate was 83% (35/42), with a mean VRR at last follow-up of 81% ± 35%. • No patients developed malignancy from the ablated TGDCs but one patient (1.5%, 1/68) developed wound inflammation after the first EA.

Opposite Effects of Apoptotic and Necroptotic Cellular Pathways on Rotavirus Replication.

Group A rotavirus (RVA), one of the leading pathogens causing severe acute gastroenteritis in children and a wide variety of young animals worldwide, induces apoptosis upon infecting cells. Though RVA-induced apoptosis mediated via the dual modulation of its NSP4 and NSP1 proteins is relatively well studied, the nature and signaling pathway(s) involved in RVA-induced necroptosis are yet to be fully elucidated. Here, we demonstrate the nature of RVA-induced necroptosis, the signaling cascade involved, and correlation with RVA-induced apoptosis. Infection with the bovine NCDV and human DS-1 RVA strains was shown to activate receptor-interacting protein kinase 1 (RIPK1), RIPK3, and mixed-lineage kinase domain-like protein (MLKL), the key necroptosis molecules in virus-infected cells. Using an immunoprecipitation assay, RIPK1 was found to bind phosphorylated RIPK3 (pRIPK3) and pMLKL. pMLKL, the major executioner molecule in the necroptotic pathway, was translocated to the plasma membrane of RVA-infected cells to puncture the cell membrane. Interestingly, transfection of RVA NSP4 also induced necroptosis through the RIPK1/RIPK3/MLKL necroptosis pathway. Blockage of each key necroptosis molecule in the RVA-infected or NSP4-transfected cells resulted in decreased necroptosis but increased cell viability and apoptosis, thereby resulting in decreased viral yields in the RVA-infected cells. In contrast, suppression of RVA-induced apoptosis increased necroptosis and virus yields. Our findings suggest that RVA NSP4 also induces necroptosis via the RIPK1/RIPK3/MLKL necroptosis pathway. Moreover, necroptosis and apoptosis-which have proviral and antiviral effects, respectively-exhibited cross talk in RVA-infected cells. These findings significantly increase our understanding of the nature of RVA-induced necroptosis and the cross talk between RVA-induced necroptosis and apoptosis. IMPORTANCE Viral infection usually culminates in cell death through apoptosis, necroptosis, and, rarely, pyroptosis. Necroptosis is a form of programmed necrosis that is mediated by signaling complexes of the receptor-interacting protein kinase 1 (RIPK1), RIPK3, and mixed-lineage kinase domain-like protein (MLKL). Although apoptosis induction by rotavirus and its NSP4 protein is well known, rotavirus-induced necroptosis is not fully understood. Here, we demonstrate that rotavirus and also its NSP4 protein can induce necroptosis in cultured cells through activation of the RIPK1/RIPK3/MLKL necroptosis pathway. Moreover, rotavirus-induced necroptosis and apoptosis have opposite effects on viral yield, i.e., they function as proviral and antiviral processes, respectively, and counterbalance each other in rotavirus-infected cells. Our findings provide important insights for understanding the nature of rotavirus-induced necroptosis and the development of novel therapeutic strategies against infection with rotavirus and other RNA viruses.