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Novel Morbillivirus as Putative Cause of Fetal Death and Encephalitis among Swine

Emerging infectious diseases

2021 Jul 01

Arruda, B;Shen, H;Zheng, Y;Li, G;
PMID: 34152961 | DOI: 10.3201/eid2707.203971

Morbilliviruses are highly contagious pathogens. The Morbillivirus genus includes measles virus, canine distemper virus (CDV), phocine distemper virus (PDV), peste des petits ruminants virus, rinderpest virus, and feline morbillivirus. We detected a novel porcine morbillivirus (PoMV) as a putative cause of fetal death, encephalitis, and placentitis among swine by using histopathology, metagenomic sequencing, and in situ hybridization. Phylogenetic analyses showed PoMV is most closely related to CDV (62.9% nt identities) and PDV (62.8% nt identities). We observed intranuclear inclusions in neurons and glial cells of swine fetuses with encephalitis. Cellular tropism is similar to other morbilliviruses, and PoMV viral RNA was detected in neurons, respiratory epithelium, and lymphocytes. This study provides fundamental knowledge concerning the pathology, genome composition, transmission, and cellular tropism of a novel pathogen within the genus Morbillivirus and opens the door to a new, applicable disease model to drive research forward.

Transcriptome profiling of the Olig2-expressing astrocyte subtype reveals their unique molecular signature

iScience

2021 Jul 01

Ohayon, D;Aguirrebengoa, M;Escalas, N;Jungas, T;Soula, C;
| DOI: 10.1016/j.isci.2021.102806

Astrocytes are recognized to be a heterogeneous population of cells that differ morphologically, functionally and molecularly. Whether this heterogeneity results from generation of distinct astrocyte cell lineages, each functionally specialized to perform specific tasks, remains an open question. In this study, we used RNA-seq analysis to determine the global transcriptome profile of the Olig2-expressing astrocyte subtype (Olig2-AS), a specific spinal astrocyte subtype which segregates early during development from Olig2 progenitors and differs from other spinal astrocytes by the expression of Olig2. We identified 245 differentially expressed genes. Among them, 135 exhibit higher levels of expression when compared to other populations of spinal astrocytes, indicating that these genes can serve as a ‘unique’ functional signature of Olig2-AS. Among them, we identify two genes, inka2 and kcnip3, as specific molecular markers of the Olig2-AS in the P7 spinal cord. Our work thus reveals that Olig2 progenitors produce a unique spinal astrocyte subtype.

Translatomic analysis of regenerating and degenerating spinal motor neurons in injury and ALS

iScience

2021 Jul 01

Shadrach, J;Stansberry, W;Milen, A;Ives, R;Fogarty, E;Antonellis, A;Pierchala, B;
| DOI: 10.1016/j.isci.2021.102700

The neuromuscular junction is a synapse critical for muscle strength and coordinated motor function. Unlike CNS injuries, motor neurons mount robust regenerative responses after peripheral nerve injuries. Conversely, motor neurons selectively degenerate in diseases such as amyotrophic lateral sclerosis (ALS). To assess how these insults affect motor neurons in vivo, we performed ribosomal profiling of mouse motor neurons. Motor neuron-specific transcripts were isolated from spinal cords following sciatic nerve crush, a model of acute injury and regeneration, and in the SOD1G93A ALS model. Of the 267 transcripts upregulated after nerve crush, 38% were also upregulated in SOD1G93A motor neurons. However, most upregulated genes in injured and ALS motor neurons were context specific. Some of the most significantly upregulated transcripts in both paradigms were chemokines such as Ccl2 and Ccl7, suggesting an important role for neuroimmune modulation. Collectively these data will aid in defining pro-regenerative and pro-degenerative mechanisms in motor neurons.

Exposure of human fetal kidneys to mild analgesics interferes with early nephrogenesis

FASEB journal : official publication of the Federation of American Societies for Experimental Biology

2021 Jul 01

Leverrier-Penna, S;Michel, A;Lecante, LL;Costet, N;Suglia, A;Desdoits-Lethimonier, C;Boulay, H;Viel, R;Chemouny, JM;Becker, E;Lavoué, V;Rolland, AD;Dejucq-Rainsford, N;Vigneau, C;Mazaud-Guittot, S;
PMID: 34105801 | DOI: 10.1096/fj.202100050R

Acetaminophen, aspirin, and ibuprofen are mild analgesics commonly used by pregnant women, the sole current recommendation being to avoid ibuprofen from the fifth month of gestation. The nephrotoxicity of these three analgesics is well documented in adults, as is their interference with prostaglandins biosynthesis. Here we investigated the effect of these analgesics on human first trimester kidneys ex vivo. We first evaluated prostaglandins biosynthesis functionality by performing a wide screening of prostaglandin expression patterns in first trimester human kidneys. We demonstrated that prostaglandins biosynthesis machinery is functional during early nephrogenesis. Human fetal kidney explants aged 7-12 developmental weeks were exposed ex vivo to ibuprofen, aspirin or acetaminophen for 7 days, and analyzed by histology, immunohistochemistry, and flow cytometry. This study has revealed that these analgesics induced a spectrum of abnormalities within early developing structures, ranging from cell death to a decline in differentiating glomeruli density. These results warrant caution for the use of these medicines during the first trimester of pregnancy.

Visualization of HIV-1 reservoir: an imaging perspective

Current opinion in HIV and AIDS

2021 Jul 01

Chapon, C;Moysi, E;Naninck, T;Mayet, C;Petrovas, C;
PMID: 34039844 | DOI: 10.1097/COH.0000000000000691

The persistence of HIV-1-infected cells, despite the introduction of the combinatorial antiretroviral therapy, is a major obstacle to HIV-1 eradication. Understanding the nature of HIV reservoir will lead to novel therapeutic approaches for the functional cure or eradication of the virus. In this review, we will update the recent development in imaging applications toward HIV-1/simian immunodeficiency virus (SIV) viral reservoirs research and highlight some of their limitations.CD4 T cells are the primary target of HIV-1/SIV and the predominant site for productive and latent reservoirs. This viral reservoir preferentially resides in lymphoid compartments that are difficult to access, which renders sampling and measurements problematical and a hurdle for understanding HIV-1 pathogenicity. Novel noninvasive technologies are needed to circumvent this and urgently help to find a cure for HIV-1. Recent technological advancements have had a significant impact on the development of imaging methodologies allowing the visualization of relevant biomarkers with high resolution and analytical capacity. Such methodologies have provided insights into our understanding of cellular and molecular interactions in health and disease.Imaging of the HIV-1 reservoir can provide significant insights for the nature (cell types), spatial distribution, and the role of the tissue microenvironment for its in vivo dynamics and potentially lead to novel targets for the virus elimination.

Assessing proviral competence: current approaches to evaluate HIV-1 persistence

Current opinion in HIV and AIDS

2021 Jul 01

Cicilionytė, A;Berkhout, B;Pasternak, AO;
PMID: 33993171 | DOI: 10.1097/COH.0000000000000687

Despite decades of suppressive antiretroviral therapy (ART), HIV-1 reservoirs persist and fuel viral rebound if therapy is interrupted. The persistence of viral reservoirs in infected individuals is the main obstacle to achieving HIV-1 eradication or a long-term remission. Accurate assessment of the viral reservoir size is necessary for monitoring the effectiveness of the curative interventions. Here, we review the recent progress in the development of assays to measure HIV-1 persistence, highlighting their key advantages and limitations.To estimate the viral reservoir size, a number of assays have been developed that assess different aspects of HIV-1 persistence in ART-treated individuals. These include viral outgrowth assays to measure proviral replication competence, sequencing-based assays to measure genetic intactness of HIV-1 proviruses, and diverse techniques that measure the ability of proviruses to produce viral RNA and/or proteins (transcription and translation competence), with or without ex vivo stimulation. Recent years have seen the development of next-generation reservoir assays that, in addition to measuring viral persistence markers, assess the proviral integration sites and characterize the HIV-1 reservoir cells on the single-cell level.Although no assay yet can measure the HIV-1 reservoir with 100% accuracy, recent technical advances allow reliable estimation of its size and composition.

The active human immunodeficiency virus reservoir during antiretroviral therapy: emerging players in viral persistence

Current opinion in HIV and AIDS

2021 Jul 01

Astorga-Gamaza, A;Buzon, MJ;
PMID: 33973900 | DOI: 10.1097/COH.0000000000000685

To discuss the role of CD4+ T cells with active Human immunodeficiency virus (HIV), meaning infected cells with transcriptional and/or translational viral activity during antiretroviral therapy (ART), focusing on new technologies for its detection, potential cell markers for its characterization, and evidences on the contribution of the active HIV reservoir to long-term viral persistence.HIV-infected cells expressing viral ribonucleic acid are systematically detected in subjects on long-term ART. In recent years, powerful new tools have provided significant insights into the nature, quantification, and identification of cells with active HIV, including the identification of new cell markers, and the presence of viral activity in specific cell populations located in different cellular and anatomical compartments. Moreover, studies on viral sequence integrity have identified cell clones with intact viral genomes and active viral transcription that could potentially persist for years. Together, new investigations support the notion that the active reservoir could represent a relevant fraction of long-term infected cells, and therefore, the study of its cell sources and mechanisms of maintenance could represent a significant advance in our understanding of viral persistence and the development of new curative strategies.The presence of HIV-infected cells with viral expression during ART has been traditionally overlooked for years. Based on recent investigations, this active viral reservoir could play an important role in HIV persistence.

Monocyte and macrophage derived myofibroblasts: Is it fate? A review of the current evidence

Wound repair and regeneration : official publication of the Wound Healing Society [and] the European Tissue Repair Society

2021 Jul 01

Vierhout, M;Ayoub, A;Naiel, S;Yazdanshenas, P;Revill, SD;Reihani, A;Dvorkin-Gheva, A;Shi, W;Ask, K;
PMID: 34107123 | DOI: 10.1111/wrr.12946

Since the discovery of the myofibroblast over 50 years ago, much has been learned about its role in wound healing and fibrosis. Its origin, however, remains controversial, with a number of progenitor cells being proposed. Macrophage-myofibroblast transition (MMT) is a recent term coined in 2014 that describes the mechanism through which macrophages, derived from circulating monocytes originating in the bone marrow, transformed into myofibroblasts and contributed to kidney fibrosis. Over the past years, several studies have confirmed the existence of MMT in various systems, suggesting that MMT could potentially occur in all fibrotic conditions and constitute a reasonable therapeutic target to prevent progressive fibrotic disease. In this perspective, we examined recent evidence supporting the notion of MMT in both human disease and experimental models across organ systems. Mechanistic insight from these studies and information from in vitro studies is provided. The findings substantiating plausible MMT showcased the co-expression of macrophage and myofibroblast markers, including CD68 or F4/80 (macrophage) and α-SMA (myofibroblast), in fibroblast-like cells. Furthermore, fate-mapping experiments in murine models exhibiting myeloid-derived myofibroblasts in the tissue further provide direct evidence for MMT. Additionally, we provide some evidence from single cell RNA sequencing experiments confirmed by fluorescent in situ hybridisation studies, showing monocyte/macrophage and myofibroblast markers co-expressed in lung tissue from patients with fibrotic lung disease. In conclusion, MMT is likely a significant contributor to myofibroblast formation in wound healing and fibrotic disease across organ systems. Circulating precursors including monocytes and the molecular mechanisms governing MMT could constitute valid targets and provide insight for the development of novel antifibrotic therapies; however, further understanding of these processes is warranted.

PD-L1 AND FOXP3 EXPRESSION IN ORAL DYSPLASTIC TISSUES AND ORAL SQUAMOUS CELL CARCINOMA

Oral Surgery, Oral Medicine, Oral Pathology and Oral Radiology

2021 Jul 01

Arora, S;Wan, Z;Dong, F;Kalmadin, N;De Silva, H;Seo, B;Hussaini, H;Rich, A;
| DOI: 10.1016/j.oooo.2021.03.043

Background Oral squamous cell carcinoma (OSCC) is an aggressive, highly immunosuppressive cancer with a high mortality rate. Interactions between programmed cell death protein 1 (PD-1; on T cells) and programmed death ligand 1 (PD-L1; on tumor cells) within the tumor microenvironment facilitates T-lymphocyte exhaustion. Regulatory T cells (Treg) are a distinct lymphocyte population, expressing the transcription factor forkhead homeobox protein-3 (FoxP3), which downregulates immune responses in OSCC. PD-L1+ tumor cells and FoxP3+ Treg expression in OSCC has been associated with poor prognosis. This research investigates the expression of PD-L1+ cells and Tregs in control, dysplastic, and OSCC tissues. Objective To investigate and compare the expression of PD-L1+ tumor cells and FoxP3+ Tregs in nondysplastic tisssue, dysplastic tissue, and OSCC using immunohistochemistry. Methods Immunohistochemistry was performed on formalin-fixed, paraffin-embedded, archival tissues. Qualitative and quantitative analyses of positively stained cells were undertaken and the dysplastic (n = 20) and OSCC groups (n = 20) were compared against the non-dysplastic control group (n = 20), using image analysis Results A higher proportion score and immunoreactive score for PD-L1+ and FoxP3+ Tregs was found in OSCC and dysplastic groups when compared to the nondysplastic control group (P < .05). There was no significant difference between the OSCC and dysplastic tissues. Conclusions Significantly more PD-L1+ cells and Tregs were detected in dysplastic and OSCC tissues. An increase in PD-L1 and FoxP3 expression may serve as an indicator of progression from normal to a potentially malignant lesion.

DETERMINATION OF SINGLE NUCLEOTIDE POLYMORPHISM (RS566926) OF WNT5A IN NONSYNDROMIC CLEFT LIP AND PALATE IN A PAKISTANI POPULATION

Oral Surgery, Oral Medicine, Oral Pathology and Oral Radiology

2021 Jul 01

Anjum, R;Mehmood, S;Nagi, A;Shahzad, M;Chuadhry, S;
| DOI: 10.1016/j.oooo.2021.03.042

Background Orofacial clefts are the most common birth defects affecting 1 in 750 live births worldwide. Various genetic loci to be involved in nonsyndromic cleft lip and palate has been identified with a variation among populations. Wnt5a is expressed in the frontonasal prominence and maxillary process, which fuse to form the primary palate. Therefore, its dysregulation can lead to certain birth defects along with other diseases. Single nucleotide polymorphism (rs566926) in Wnt5A shows a significant association with nonsyndromic cleft lip and palate in Brazilian and European American populations. Objective The aim of the present study was to describe single nucleotide polymorphism (SNP; rs566926) in patients with nonsyndromic cleft lip and palate in a Pakistani population. Methods This study was conducted on 120 patients with nonsyndromic cleft lip and palate. Demographics and phenotypes were noted. Blood samples were collected in ethylenediaminetetraacetic acid vials. DNA was extracted followed by conventional polymerase chain reaction. SNP (566926) was determined by Sanger sequencing. Data were analyzed using NCBI Blast and SPSS (24.0). Results The mean age of n = 30 patients was 51.33 ± 61.33 months. Sixty percent were male and 40% were female. Regarding cleft types, 70% were both cleft lip and palate, 26% cleft lip only, and 3.3% cleft palate only. Heterozygous polymorphism (T/G) was seen in 33.3% of patients with both cleft lip and palate with bilateral involvement and heterozygous polymorphism (T) was seen in 16.6%. Conclusions SNP in the WNT5A gene is associated with cleft lip and palate, supporting its involvement in pathogenesis of cleft lip and palate. Further studies are recommended to determine the role of Wnt5a genes during craniofacial development.

ORAL SECONDARY SYPHILIS IN PEOPLE LIVING WITH HIV: A 16-YEAR EXPERIENCE IN MEXICO CITY

Oral Surgery, Oral Medicine, Oral Pathology and Oral Radiology

2021 Jul 01

Anaya-Saavedra, G;Castillejos-García, I;Maldonado-Mendoza, J;Ramírez-Amador, V;
| DOI: 10.1016/j.oooo.2021.03.041

Background The increase in syphilis rates worldwide, particularly in people living with HIV (PLWH), as well as the challenging diagnosis that secondary syphilis represents, make essential the accurate recognition of its manifestations, particularly in easy-access sites like the oral mucosa. Objective To describe the clinicopathologic spectrum of oral secondary syphilis (OSS) in PLWH. Methods A cross-sectional and descriptive study that included PLWH with OSS from 3 HIV referral centers in Mexico City (2004-2020). Demographic and clinical data were obtained. A comprehensive oral examination was done. OSS was diagnosed following established criteria. Histopathologic/cytological procedures were performed to rule out specific oral lesions. In all patients, Venereal Disease Research Laboratory tests were assessed and, if possible, a confirmatory fluorescent treponemal antibody test or biopsy was performed. Statistical analysis was performed using SPSS v25. Results Forty-seven PLWH with OSS (97.8% male, median age: 32 years, 63.8% with acquired immunodeficiency syndrome) were included. Thirty-five were receiving combination antiretroviral therapy (74.5%; median of 1146 [Q1-Q3: 337.5-1971] days) with a median CD4+ count of 385 (Q1-Q3: 223-664) cells/mm3 and a Log10 HIV viral load of 4.1 (Q1-Q3: 3.7-5.3) copies/mL. Forty had a complete clinical-serological diagnosis (85.1%; 17 had histopathologic confirmation) and 7 had a clinical-histopathologic diagnosis. Twenty-nine individuals presented 1 lesion (61.7%), and mucous patch was the most common type mainly on oropharyngeal mucosa, followed by ulcers and macular lesions. Ten patients presented maculopapular dermatosis (21.3%). Conclusions In PLWH, oral lesions, particularly mucous patch and/or ulcers on the oral and oropharyngeal mucosa, must alert specialists to consider a diagnosis of syphilis and perform a comprehensive panel of confirmatory tests.

IN SITU DETECTION OF CRTC-MAML2 TRANSLOCATION EXPRESSION IN MUCOEPIDERMOID CARCINOMA

Oral Surgery, Oral Medicine, Oral Pathology and Oral Radiology

2021 Jul 01

Amoura, E;Hunter, K;Bingle, C;Bingle, L;
| DOI: 10.1016/j.oooo.2021.03.040

Background The heterogeneity of salivary gland neoplasms, within and between histologic types, presents a major diagnostic challenge. Mucoepidermoid carcinoma (MEC), the most common salivary gland cancer in adults, children, and adolescents, is associated with the presence of a novel CRTC1-MAML2 fusion gene. The translocation can be detected by fluorescence in situ hybridization or reverse transcription polymerase chain reaction but without information regarding transcript level, identification of the cell type(s) harboring the translocation and histologic architecture is not preserved. This study describes, for the first time, a novel in situ chromogenic assay, BaseScope, to detect CRTC1-MAML2 translocation expression. Objective Design a novel BaseScope probe targeting the novel exon-exon junction in the CRTC1-MAML2 fusion transcript, determine expression levels of the transcript, and identify specific cell types harboring the translocation. Methods Formalin-fixed paraffin-embedded tissues and known fusion-positive and -negative human MEC cells were subjected to the assay. Results The CRTC1-MAML2 RNA transcript was detected in known fusion-positive cells but not in fusion-negative cells. In MEC tissues distinct fusion events, in the form of punctate red dots, were detected in all tumor grades and all cell types. Interestingly, the translocation was specifically identified in tumor cells that had direct contact with tumor stroma or nerve invasion. No positive staining was seen in normal tissue or surrounding stroma and no unique morphological features were noted in negative cases. Conclusions The BaseScope assay accurately detects the CRTC1-MAML2 fusion translocation and thus provides an alternative chromogenic technique, for easy use in routine clinical labs, to aid accurate diagnosis of MEC.

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	Description
sense Example: Hs-LAG3-sense	Standard probes for RNA detection are in antisense. Sense probe is reverse complent to the corresponding antisense probe.
Intron# Example: Mm-Htt-intron2	Probe targets the indicated intron in the target gene, commonly used for pre-mRNA detection
Pool/Pan Example: Hs-CD3-pool (Hs-CD3D, Hs-CD3E, Hs-CD3G)	A mixture of multiple probe sets targeting multiple genes or transcripts
No-XSp Example: Hs-PDGFB-No-XMm	Does not cross detect with the species (Sp)
XSp Example: Rn-Pde9a-XMm	designed to cross detect with the species (Sp)
O# Example: Mm-Islr-O1	Alternative design targeting different regions of the same transcript or isoforms
CDS Example: Hs-SLC31A-CDS	Probe targets the protein-coding sequence only
EnEm	Probe targets exons n and m
En-Em	Probe targets region from exon n to exon m
Retired Nomenclature
tvn Example: Hs-LEPR-tv1	Designed to target transcript variant n
ORF Example: Hs-ACVRL1-ORF	Probe targets open reading frame
UTR Example: Hs-HTT-UTR-C3	Probe targets the untranslated region (non-protein-coding region) only
5UTR Example: Hs-GNRHR-5UTR	Probe targets the 5' untranslated region only
3UTR Example: Rn-Npy1r-3UTR	Probe targets the 3' untranslated region only
Pan Example: Pool	A mixture of multiple probe sets targeting multiple genes or transcripts

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