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GLH/Vasa/DDX4 helicases are core germ-granule proteins that promote germline development and fertility. A yeast-two-hybrid screen using Caenorhabditis elegans GLH-1 as bait identified BYN-1, the homolog of human bystin/BYSL. In humans, bystin promotes cell adhesion and invasion in gliomas, and, with its binding partner trophinin, triggers embryonic implantation into the uterine wall. C. elegans embryos do not implant and lack a homolog of trophinin, but both trophinin and GLH-1 contain unique decapeptide phenylalanine-glycine (FG)-repeat domains. In germ cells, we find endogenous BYN-1 in the nucleolus, partitioned away from cytoplasmic germ granules. However, BYN-1 enters the cytoplasm during spermatogenesis to colocalize with GLH-1. Both proteins become deposited in residual bodies (RBs), which are then engulfed and cleared by the somatic gonad. We show that BYN-1 acts upstream of CED-1 to drive RB engulfment, and that removal of the FG-repeat domains from GLH-1 and GLH-2 can partially phenocopy byn-1 defects in RB clearance. These results point to an evolutionarily conserved pathway whereby cellular uptake is triggered by the cytoplasmic mobilization of bystin/BYN-1 to interact with proteins harboring FG-repeats. [ABSTRACT FROM AUTHOR]

Alphaviruses, such as chikungunya virus (CHIKV), are mosquito-borne viruses that represent a significant threat to human health due to the current context of global warming. Efficient alphavirus infection relies on the activity of the non-structural protein 3 (nsP3), a puzzling multifunctional molecule whose role in infection remains largely unknown. NsP3 is a component of the plasma membrane-bound viral RNA replication complex (vRC) essential for RNA amplification and is also found in large cytoplasmic aggregates of unknown function. Here, we report the cryo-electron microscopy (cryo-EM) structure of the CHIKV nsP3 at 2.35 Å resolution. We show that nsP3 assembles into tubular structures made by a helical arrangement of its alphavirus unique domain (AUD). The nsP3 helical scaffolds are consistent with crown structures found on tomographic reconstructions of the mature viral RCs. In addition, nsP3 helices assemble into cytoplasmic granules organized in a network of tubular structures that contain viral genomic RNA and capsid as well as host factors required for productive infection. Structure-guided mutagenesis identified residues that prevent or disturb nsP3 assemblies, resulting in impaired viral replication or transcription. Altogether, our results reveal an unexpected nsP3-dependent molecular organization essential for different phases of alphavirus infection. The role of alphavirus nsP3 protein is not entirely clear. Here, through structural analysis the authors show that CHIKV nsP3 polymerizes to form tubular scaffolds in both replication complexes and alpha-granules and that these scaffolds are important for virus RNA synthesis and infectivity. [ABSTRACT FROM AUTHOR]

Summary: Cytoplasmic ribonucleoprotein (RNP) granules are membraneless structures composed of various RNAs and proteins that play important roles in post‐transcriptional regulation. While RNP granules are known to regulate the meiotic entry in some organisms, little is known about their roles in plants.In this study, we observed the cytoplasmic granular structures of rice RNA‐binding protein MEIOSIS ARRESTED AT LEPTOTENE2 (MEL2), which contributes to the control of meiotic entry timing, in leaf protoplasts and spore mother cells. We performed colocalization analysis with known cytoplasmic RNP factors, and domain deletion analysis to assess their impact on granule formation and meiosis progression. Conservation of MEL2 domains across plant species was also explored.Our results indicated that MEL2 granules colocalized with processing body and stress granule factors. The maintenance of granule properties modulated by LOTUS domain and the intrinsically disordered region (IDR) is essential for proper MEL2 function in meiosis progression. MEL2‐like proteins widely found in plant kingdom conserved LOTUS domain followed by the IDR despite their diverse domain structures, suggesting the functional conservation of these domains among plant species.This study highlights the role of MEL2 granule dynamics and its impact on meiotic transition and progression. [ABSTRACT FROM AUTHOR]

Somatic mitochondrial DNA (mtDNA) mutation accumulation has been observed in individuals with retinal degenerative disorders. To study the effects of aging and mtDNA mutation accumulation in the retina, a polymerase gamma (POLG) exonuclease‐deficient model, the PolgD257A mutator mice (D257A), was used. POLG is an enzyme responsible for regulating mtDNA replication and repair. Retinas of young and older mice with this mutation were analyzed in vivo and ex vivo to provide new insights into the contribution of age‐related mitochondrial (mt) dysfunction due to mtDNA damage. Optical coherence tomography (OCT) image analysis revealed a decrease in retinal and photoreceptor thickness starting at 6 months of age in mice with the D257A mutation compared to wild‐type (WT) mice. Electroretinography (ERG) testing showed a significant decrease in all recorded responses at 6 months of age. Sections labeled with markers of different types of retinal cells, including cones, rods, and bipolar cells, exhibited decreased labeling starting at 6 months. However, electron microscopy analysis revealed differences in retinal pigment epithelium (RPE) mt morphology beginning at 3 months. Interestingly, there was no increase in oxidative stress and parkin‐mediated mitophagy in the ages analyzed in the retina or RPE of D257A mice. Additionally, D257A RPE exhibited an accelerated rate of autofluorescence cytoplasmic granule formation and accumulation. Mt markers displayed different abundance in protein lysates obtained from retina and RPE samples. These findings suggest that the accumulation of mtDNA mutations leads to impaired mt function and accelerated aging, resulting in retinal degeneration. [ABSTRACT FROM AUTHOR]

Background: The global outbreak of COVID-19 caused by the SARS-CoV-2 has led to millions of deaths. This unanticipated emergency has prompted virologists across the globe to delve deeper into the intricate dynamicity of the host-virus interface with an aim to identify antiviral targets and elucidate host and viral determinants of severe disease. Aim: The present study was undertaken to analyse the role of histone deacetylase 6 (HDAC6) in regulating SARS-CoV-2 infection. Results: Gradual increase in HDAC6 expression was observed in different SARS-CoV-2-permissive cell lines following SARS-CoV-2 infection. The SARS-CoV-2 nucleocapsid protein (N protein) was identified as the primary viral factor responsible for upregulating HDAC6 expression. Downregulation of HDAC6 using shRNA or a specific inhibitor tubacin resulted in reduced viral replication suggesting proviral role of its deacetylase activity. Further investigations uncovered the interaction of HDAC6 with stress granule protein G3BP1 and N protein during infection. HDAC6-mediated deacetylation of SARS-CoV-2 N protein was found to be crucial for its association with G3BP1. Conclusion: This study provides valuable insights into the molecular mechanisms underlying the disruption of cytoplasmic stress granules during SARS-CoV-2 infection and highlights the significance of HDAC6 in the process. [ABSTRACT FROM AUTHOR]

Neutrophil elastase (HNE), like other members of the so-called GASPIDs (Granule-Associated Serine Peptidases of Immune Defense), is activated during protein biosynthesis in myeloid precursors and stored enzymatically active in cytoplasmic granules of resting neutrophils until secreted at sites of host defense and inflammation. Inhibitors thus could bind to the fully formed active site of the protease intracellularly in immature progenitors, in circulating neutrophils, or to HNE secreted into the extracellular space. Here, we have compared the ability of a panel of diverse inhibitors to inhibit HNE in the U937 progenitor cell line, in human blood-derived neutrophils, and in solution. Most synthetic inhibitors and, surprisingly, even a small naturally occurring proteinaceous inhibitor inhibit HNE intracellularly, but the extent and dynamics differ markedly from classical enzyme kinetics describing extracellular inhibition. Intracellular inhibition of HNE potentially affects neutrophil functions and has side effects, but it avoids competition of inhibitors with extracellular substrates that limit its efficacy. As both intra- and extracellular inhibition have advantages and disadvantages, the quantification of intracellular inhibition, in addition to classical enzyme kinetics, will aid the design of novel, clinically applicable HNE inhibitors with targeted sites of action. [ABSTRACT FROM AUTHOR]

SUMMARY: The RNA regulatory network is a complex and dynamic regulation in plant cells involved in mRNA modification, translation, and degradation. Ras‐GAP SH3 domain‐binding protein (G3BP) is a scaffold protein for the assembly of stress granules (SGs) and is considered an antiviral component in mammals. However, the function of G3BP during virus infection in plants is still largely unknown. In this study, four members of the G3BP‐like proteins (NtG3BPLs) were identified in Nicotiana tabacum and the expression levels of NtG3BPL1 were upregulated during chilli veinal mottle virus (ChiVMV) infection. NtG3BPL1 was localized in the nucleus and cytoplasm, forming cytoplasmic granules under transient high‐temperature treatment, whereas the abundance of cytoplasmic granules was decreased under ChiVMV infection. Overexpression of NtG3BPL1 inhibited ChiVMV infection and delayed the onset of symptoms, whereas knockout of NtG3BPL1 promoted ChiVMV infection. In addition, NtG3BPL1 directly interacted with ChiVMV 6K2 protein, whereas 6K2 protein had no effect on NtG3BPL1‐derived cytoplasmic granules. Further studies revealed that the expression of NtG3BPL1 reduced the chloroplast localization of 6K2‐GFP and the NtG3BPL1‐6K2 interaction complex was localized in the cytoplasm. Furthermore, NtG3BPL1 promoted the degradation of 6K2 through autophagy pathway, and the accumulation of 6K2 and ChiVMV was affected by autophagy activation or inhibition in plants. Taken together, our results demonstrate that NtG3BPL1 plays a positive role in tobacco resistance against ChiVMV infection, revealing a novel mechanism of plant G3BP in antiviral strategy. Significance Statement: G3BP is a key component of cytoplasmic stress granules and exerts antiviral effects in mammals, but its function during plant virus infection remains unclear. Here, we revealed that NtG3BPL1 is induced upon ChiVMV infection and interacts with ChiVMV 6K2 to disturb the chloroplast localization of 6K2, and promote the degradation of 6K2, thereby limiting ChiVMV infection. This study enhances our understanding of the function of G3BP in plant virus infection. [ABSTRACT FROM AUTHOR]

Zika virus (ZIKV) has become a global health problem over the past decade due to the extension of the geographic distribution of the Asian/American genotype. Recent epidemics of Asian/American ZIKV have been associated with developmental disorders in humans. There is mounting evidence that African ZIKV may be associated with increased fetal pathogenicity necessitating to pay a greater attention towards currently circulating viral strains in sub-Saharan Africa. Here, we generated an infectious molecular clone GUINEA-18 of a recently transmitted human ZIKV isolate from West Africa, ZIKV-15555. The available infectious molecular clone MR766MC of historical African ZIKV strain MR766-NIID was used for a molecular clone-based comparative study. Viral clones GUINEA-18 and MR766MC were compared for their ability to replicate in VeroE6, A549 and HCM3 cell lines. There was a lower replication rate for GUINEA-18 associated with weaker cytotoxicity and reduced innate immune system activation compared with MR766MC. Analysis of chimeric viruses between viral clones stressed the importance of NS1 to NS4B proteins, with a particular focus of NS4B on GUINEA-18 replicative properties. ZIKV has developed strategies to prevent cytoplasmic stress granule formation which occurs in response to virus infection. GUINEA-18 was greatly efficient in inhibiting stress granule assembly in A549 cells subjected to a physiological stressor, with NS1 to NS4B proteins also being critical in this process. The impact of these GUINEA-18 proteins on viral replicative abilities and host-cell responses to viral infection raises the question of the role of nonstructural proteins in the pathogenicity of currently circulating ZIKV in sub-Saharan Africa. Author summary: Most studies with the objective to understand the biology of Zika virus (ZIKV) were carried out using the epidemic Asian genotype of this pathogen. It is currently being discussed whether ZIKV of African genotype may have epidemic potential associated a high risk of fetal pathogenicity. It is thus urgent to improve our knowledge on recently isolated ZIKV strains from West Africa. In this study, we used the sequence of a viral strain from an individual infected by ZIKV in Guinea in 2018 to generate an infectious molecular clone. Analysis of this viral clone highlighted the preponderant role of NS1 to NS4B proteins in virus replication and cell interactions with a particular focus on ZIKV-specific stress granule formation blocking activity. We believe that our data will improve our understanding of the biology of contemporary West Africa ZIKV, opening perspectives towards a better understanding of the pathogenicity of African viral strains. [ABSTRACT FROM AUTHOR]

Cytologic evaluation of aspirate slides from a small, <1-cm, interdigital mass on a 9-y-old, spayed female Yorkshire Terrier revealed a proliferation of discrete, round cells containing few-to-many, variably sized, round, eosinophilic, cytoplasmic inclusions. The top differentials based on the cytologic findings were either a plasma cell tumor or a B-cell lymphoma with Mott cell differentiation. The unencapsulated, well-demarcated, multilobulated round-cell neoplasm was completely excised. Immunohistochemical stains were performed to further characterize the neoplasm, which had immunolabeling for multiple myeloma oncogene 1 and vimentin, but did not react with CD3, CD20, melan A, or ionized calcium–binding adapter molecule 1, nor with a Giemsa special stain. Ultrastructurally, the cytoplasmic granules had Russell body–like morphology. A solitary, cutaneous plasmacytoma with Mott cell differentiation has not been described previously in veterinary medicine, to our knowledge. [ABSTRACT FROM AUTHOR]

Background: The escalation of global population aging has accentuated the prominence of senile diabetes mellitus (SDM) as a consequential public health concern. Oxidative stress and chronic inflammatory cascades prevalent in individuals with senile diabetes significantly amplify disease progression and complication rates. Traditional Chinese Medicine (TCM) emerges as a pivotal player in enhancing blood sugar homeostasis and retarding complication onset in the clinical management of senile diabetes. Nonetheless, an evident research gap persists regarding the integration of TCM's renal tonification pharmacological mechanisms with experimental validation within the realm of senile diabetes therapeutics. Aims: The objective of this study was to investigate the mechanisms of action of New Shenqi Pills (SQP) in the treatment of SDM and make an experimental assessment. Methods: Network analysis is used to evaluate target pathways related to SQP and SDM. Mitochondrial-related genes were obtained from the MitoCarta3.0 database and intersected with the common target genes of the disease and drugs, then constructing a protein-protein interaction (PPI) network making use of the GeneMANIA database. Representative compounds in the SQP were quantitatively measured using high performance liquid chromatographytandem mass spectrometry (HPLC-MS/MS) to ensure quality control and quantitative analysis of the compounds. A type 2 diabetes mice (C57BL/6) model was used to investigate the pharmacodynamics of SQP. The glucose lowering efficacy of SQP was assessed through various metrics including body weight and fasting blood glucose (FBG). To elucidate the modulatory effects of SQP on pancreatic beta cell function, we measured oral glucose tolerance test (OGTT), insulin histochemical staining and tunel apoptosis detection, then assessed the insulin-mediated phosphoinositide 3-kinase (PI3K)/protein kinase A (Akt)/glycogen synthase kinase-3ß (GSK-3ß) pathway in diabetic mice via Western blotting. Additionally, we observe the structural changes of the nucleus, cytoplasmic granules and mitochondria of pancreatic islet ß cells. Results: In this investigation, we identified a total of 1876 genes associated with senile diabetes, 278 targets of SQP, and 166 overlapping target genes, primarily enriched in pathways pertinent to oxidative stress response, peptide response, and oxygen level modulation. Moreover, an intersection analysis involving 1,136 human mitochondrial genes and comorbidity targets yielded 15 mitochondria-related therapeutic targets. Quality control assessments and quantitative analyses of SQP revealed the predominant presence of five compounds with elevated concentrations: Catalpol, Cinnamon Aldehyde, Rehmanthin D, Trigonelline, and Paeonol Phenol. Vivo experiments demonstrated notable findings. Relative to the control group, mice in the model group exhibited significant increases in body weight and fasting blood glucose levels, alongside decreased insulin secretion and heightened islet cell apoptosis. Moreover, ß-cells nuclear condensation and mitochondrial cristae disappearance were observed, accompanied by reduced expression levels of p-GSK-3ß protein in islet cells (p < 0.05 or p < 0.01). Conversely, treatment groups administered SQP and Rg displayed augmented expressions of the aforementioned protein markers (p < 0.05 or p < 0.01), alongside preserved mitochondrial cristae structure in islet ß cells. Conclusion: Our findings suggest that SQP can ameliorate diabetes by reducing islet cell apoptosis and resist oxidative stress. These insulin-mediated PI3K/AKT/GSK-3ß pathway plays an important regulatory role in this process. [ABSTRACT FROM AUTHOR]

Background: Lipofuscin-like cytoplasmic inclusions have been reported in human blood neutrophils and monocytes but have not been described in dogs. In people, these “green granules of death” have been associated with moderate to severe hepatocellular injury and high mortality. Objectives: To describe clinicopathologic abnormalities, diagnoses, and outcomes of dogs with greenish inclusions in blood neutrophils or monocytes, and to determine if the inclusions have features of lipofuscin. Methods: Clinical cases were identified prospectively through routine evaluation of CBC samples. Leukocyte inclusions were characterized with routine staining and assessed for iron and autofluorescence. Additional cases were identified by examination of archived blood smears from dogs meeting search criteria for hepatocellular injury, and clinicopathologic findings were recorded. Results: All 7 prospectively identified dogs with inclusions had inflammation and moderate to marked increases in serum alanine aminotransferase (ALT) activity, as did the 4 dogs identified from the 97 meeting retrospective search criteria. The inclusions were Prussian blue–negative (5/5) with broad-spectrum autofluorescence (5/5) and the appearance of lipofuscin with and without Wright staining. Most clinical diagnoses involved hepatic disorders (5/7 prospective and 3/4 retrospective cases) or pancreatitis (3/7 prospective and 2/4 retrospective cases), and some involved both; 8 of 11 dogs died within 7 days of admission. Conclusions: Blue-green cytoplasmic inclusions uncommonly found in blood neutrophils ± monocytes of routine canine blood smears have stained and unstained properties of lipofuscin and suggest the presence of hepatocellular injury, often severe. Reporting these inclusions is recommended to guide clinical management. [ABSTRACT FROM AUTHOR]

The functions of mammalian Atg8 proteins (mATG8s) expand beyond canonical autophagy and include processes collectively referred to as Atg8ylation. Global modulation of protein synthesis under stress conditions is governed by MTOR and liquid-liquid phase separated condensates containing ribonucleoprotein particles known as stress granules (SGs). We report that lysosomal damage induces SGs acting as a hitherto unappreciated inhibitor of protein translation via EIF2A/eIF2α phosphorylation while favoring an ATF4-dependent integrated stress response. SGs are induced by lysosome-damaging agents, SARS-CoV-2 open reading frame 3a protein (ORF3a) expression, Mycobacterium tuberculosis infection, and exposure to proteopathic MAPT/tau. Proteomic studies revealed recruitment to damaged lysosomes of the core SG proteins NUFIP2 and G3BP1 along with the GABARAPs of the mATG8 family. The recruitment of these proteins is independent of SG condensates or canonical autophagy. GABARAPs interact directly with NUFIP2 and G3BP1 whereas Atg8ylation is needed for their recruitment to damaged lysosomes. At the lysosome, NUFIP2 contributes to MTOR inactivation together with LGALS8 (galectin 8) via the Ragulator-RRAGA-RRAGB complex. The separable functions of NUFIP2 and G3BP1 in SG formation vis-a-vis their role in MTOR inactivation are governed by GABARAP and Atg8ylation. Thus, cells employ membrane Atg8ylation to control and coordinate SG and MTOR responses to lysosomal damage.Abbreviations: Atg8: autophagy related 8; ATG: autophagy related; ATF4: activating transcription factor 4; EIF2A/eIF2α: eukaryotic translation initiation factor 2A; GABARAP: GABA type A receptor-associated protein; G3BP1: G3BP stress granule assembly factor 1; LLOMe: L-leucyl-L-leucine methyl ester; LysoIP: lysosome immunopurification; mRNA: messenger ribonucleic acid; MTOR: mechanistic target of rapamycin kinase; NUFIP2: nuclear FMR1 interacting protein 2; ORF3a: open reading frame 3a protein; SARS-CoV-2: severe acute respiratory syndrome coronavirus 2; SG: stress granule; TIA1: TIA1 cytotoxic granule associated RNA binding protein.

Double-stranded RNA produced during viral replication and transcription activates both protein kinase R (PKR) and ribonuclease L (RNase L), which limits viral gene expression and replication through host shutoff of translation. In this study, we find that APOBEC3B forms a complex with PABPC1 to stimulate PKR and counterbalances the PKR-suppressing activity of ADAR1 in response to infection by many types of viruses. This leads to translational blockage and the formation of stress granules. Furthermore, we show that APOBEC3B localizes to stress granules through the interaction with PABPC1. APOBEC3B facilitates the formation of protein-RNA condensates with stress granule assembly factor (G3BP1) by protecting mRNA associated with stress granules from RNAse L-induced RNA cleavage during viral infection. These results not only reveal that APOBEC3B is a key regulator of different steps of the innate immune response throughout viral infection but also highlight an alternative mechanism by which APOBEC3B can impact virus replication without editing viral genomes.

This article discusses a case of feline apocrine carcinoma, a rare type of skin tumor in cats. The case involves a 12-year-old male cat with multiple dermal masses that were surgically removed and diagnosed as apocrine carcinomas. The article provides detailed descriptions of the cytologic and histopathologic features of the tumors. It also mentions that feline apocrine carcinomas are typically solitary and locally invasive, but in this case, multiple masses developed within a short period of time. The article concludes that surgical excision with wide margins is the recommended treatment for these tumors. [Extracted from the article]

Intracellular aggregation of fused in sarcoma (FUS) is associated with the pathogenesis of familial amyotrophic lateral sclerosis (ALS). Under stress, FUS forms liquid droplets via liquid–liquid phase separation (LLPS). Two types of wild-type FUS LLPS exist in equilibrium: low-pressure LLPS (LP-LLPS) and high-pressure LLPS (HP-LLPS); the former dominates below 2 kbar and the latter over 2 kbar. Although several disease-type FUS variants have been identified, the molecular mechanism underlying accelerated cytoplasmic granule formation in ALS patients remains poorly understood. Herein, we report the reversible formation of the two LLPS states and the irreversible liquid–solid transition, namely droplet aging, of the ALS patient-type FUS variant R495X using fluorescence microscopy and ultraviolet–visible absorption spectroscopy combined with perturbations in pressure and temperature. Liquid-to-solid phase transition was accelerated in the HP-LLPS of R495X than in the wild-type variant; arginine slowed the aging of droplets at atmospheric conditions by inhibiting the formation of HP-LLPS more selectively compared to that of LP-LLPS. Our findings provide new insight into the mechanism by which R495X readily forms cytoplasmic aggregates. Targeting the aberrantly formed liquid droplets (the HP-LLPS state) of proteins with minimal impact on physiological functions could be a novel therapeutic strategy for LLPS-mediated protein diseases. [ABSTRACT FROM AUTHOR]

The prevalence of diseases characterised by eosinophilia is on the rise, emphasising the importance of understanding the role of eosinophils in these conditions. Eosinophils are a subset of granulocytes that contribute to the body's defence against bacterial, viral, and parasitic infections, but they are also implicated in haemostatic processes, including immunoregulation and allergic reactions. They contain cytoplasmic granules which can be selectively mobilised and secrete specific proteins, including chemokines, cytokines, enzymes, extracellular matrix, and growth factors. There are multiple biological and emerging functions of these specialised immune cells, including cancer surveillance, tissue remodelling and development. Several oral diseases, including oral cancer, are associated with either tissue or blood eosinophilia; however, their exact mechanism of action in the pathogenesis of these diseases remains unclear. This review presents a comprehensive synopsis of the most recent literature for both clinicians and scientists in relation to eosinophils and oral diseases and reveals a significant knowledge gap in this area of research. [ABSTRACT FROM AUTHOR]

Objective(s): Age-related macular degeneration (AMD) is one of the eye diseases that can affect a person's central vision. Retinal pigment epithelium (RPE) cells are damaged in this medical condition and some pigments are presented in these cells. Here, we aimed to investigate melanin and lipofuscin granules of RPE cells as a precursor of AMD. Materials and Methods: Hooded rats (n=18) were divided into two groups and received 100 µl of sodium iodate (SI) into the retro-orbital sinus of their eyes at 40 and 60 mg/kg doses. The total number of melanin and lipofuscin granules, different types of granules, cytoplasmic dispersion of granules as well as morphological changes in the shape and number of nuclei of RPE cells were evaluated over the course of 1-30 days. Results: The total number of melanin pigments increases over time at a dose of 40 mg/kg and decreases at a dose of 60 mg/kg. Also, the total number of lipofuscin granules in 40 mg/kg increases over time and decreases in 60 mg/kg. Autofluorescent intensity (AF) is also increased at 40 mg/kg, but at 60 mg/kg, the highest intensity is on day 7. Also, the highest number of multinucleated giant cells was on day 7 at 60 mg/kg and the most changes in cell appearance due to sodium iodate injection were seen on the first day after injection. Conclusion: We demonstrated that granules and autofluorescent intensity appear to decrease at high doses of sodium iodate, which is similar to the advanced stage of the AMD disease, where the number of granules and AF intensity increase in the middle and even early stages of the disease. [ABSTRACT FROM AUTHOR]

Three types of flash calcined powders with different specific surfaces and phases were selected to make alumina granules. The granulation was performed in a pan granulator as the spherical form. The hydrothermal and calcination conditions were applied to the granules. Physical, chemical and microstructural studies were applied to the hydrated and calcined granules. This study focussed on the thermal historical effects of the initial powders derived from bauxite on the granules. The least physical changes and the highest specific surface area were obtained for the granules prepared with flash calcined powder at 650°C. The hydrated granules can have boehmite, gamma and bayerite phases with rod, porous and plate morphologies, respectively. The presence of rod boehmite phase increases the strength of hydrated granules and plays an important role in the final strength. In the calcination stage, the phases transformed into the transition alumina and gamma phases with the growth of nanometer-sized crystals. [ABSTRACT FROM AUTHOR]

We report that lysosomal damage is a hitherto unknown inducer of stress granule (SG) formation and that the process termed membrane atg8ylation coordinates SG formation with mTOR inactivation during lysosomal stress. SGs were induced by lysosome-damaging agents including SARS-CoV-2ORF3a, Mycobacterium tuberculosis, and proteopathic tau. During damage, mammalian ATG8s directly interacted with the core SG proteins NUFIP2 and G3BP1. Atg8ylation was needed for their recruitment to damaged lysosomes independently of SG condensates whereupon NUFIP2 contributed to mTOR inactivation via the Ragulator-RagA/B complex. Thus, cells employ membrane atg8ylation to control and coordinate SG and mTOR responses to lysosomal damage.

This letter reports a case of a 66-year-old woman who presented with a fever and was diagnosed with Philadelphia chromosome-positive B-cell acute lymphoblastic leukemia (B-ALL) with pseudo Chediak-Higashi anomaly and complex karyotype. The patient had abnormal granules and inclusion bodies in the blasts, and the cytogenetic test showed a translocation of the BCR::ABL P190 fusion gene. Treatment with dasatinib and chemotherapy was initiated, but the patient's family refused further treatment and she left the hospital. The letter also provides information on the characteristics of pseudo Chediak-Higashi granules and discusses the rarity of this anomaly in B-ALL. The prognosis for this patient was poor due to her age, the high-risk subtype of ALL, and the presence of complex karyotypes. Further studies are needed to understand the relationship between cellular morphology and prognosis in ALL with pseudo Chediak-Higashi anomaly. [Extracted from the article]

Granular cell tumors are rare soft tissue neoplasms derived from Schwann cells and are characterized by their infiltrative, non-encapsulated nests and sheets of polygonal cells with fine eosinophilic cytoplasmic granules on histology. Herin, we report a case of a 10-year-old Saudi female who presented to the hospital with multiple asymptomatic skin lesions, the largest located on the right shoulder and left foot. Preoperative workup revealed the absence of liver metastasis, and the patient underwent complete surgical excision successfully. Histopathology revealed ill-defined proliferation of large bland cells with prominent eosinophilic granular cytoplasm and mild epithelial hyperplasia consistent with granular cell tumors. Granular cell tumors are a rare entity that represent only 0.5% of all soft tissue tumors. They have characteristic histological features and can present with both malignant and being features. Due to the rarity of this disease, further research is needed to enhance our understanding and improve recognition in future practice. [ABSTRACT FROM AUTHOR]

This article, published in Veterinary Record Case Reports, describes a case study of a 5-year-old female Cairn terrier with marked mastocythemia and an abdominal mass. The dog presented with vomiting, poor appetite, polydipsia, lethargy, discomfort, and reluctance to jump off furniture. Diagnostic tests revealed neutrophilia, basophilia, and mastocythemia. Abdominal radiographs and ultrasound identified two masses in the mid-abdomen suspected to be enlarged mesenteric lymph nodes. Aspiration of one of the masses revealed a diagnosis of round cell neoplasia, either large cell lymphoma or mast cell neoplasia. The study highlights the limitations of popular dip stains, such as Diff-Quik, in staining mast cells accurately and emphasizes the importance of including unstained slides for evaluation by a clinical pathologist. The authors conclude that clinicians should be aware of these limitations to avoid misdiagnosis. [Extracted from the article]

Summary: Oocyte cytoplasmic evaluation is based on homogeneity and granular appearance. Our study investigated if a granular cytoplasm, highly heterogeneous, would affect oocyte competence in bovine. In two experiments, bovine cumulus–oocyte complexes (COCs) with homogeneous cytoplasm (control, CC) and granulated cytoplasm (granular, GC) were selected from a regular pool of COCs. Experiment 1 was performed with slaughterhouse ovaries, and Experiment 2 was carried out in Girolando COCs obtained from ovum pick-up. Granular oocytes had higher caspase 3 levels (66.17 ± 11.61 vs 172.08 ± 16.95, P < 0.01) and similar GAP junction activity (5.64 ± 0.45 vs 6.29 ± 0.29). ZAR1 relative mRNA amount was lower in granular oocytes (178.27 ± 151.63 vs 0.89 ± 0.89, P = 0.01) and no effect was detected for MATER , PPP2R1A , ENY2 , IGF2R , and BMP15 genes. Despite molecular differences, no detrimental effect was detected on oocyte competence in GC oocytes. Cleavage (Experiment 1: 59.52 ± 7.21% vs 59.79 ± 6.10% and Experiment 2: 68.88 ± 4.82 vs 74.41 ± 5.89%) and blastocyst (Experiment 1: 29.28 ± 4.14% vs 23.15 ± 2.96% and Experiment 2: 21.11 ± 3.28% vs 21.02 ± 6.08%) rates were similar between CC and GC (Experiments 1 and 2, respectively). Post-transfer embryo development revealed that pregnancy (CC: 24.27 ± 9.70% vs GC: 26.31 ± 7.23%) and calving (23.68% vs 33.33%) rates and fetal growth were not affected by the presence of cytoplasmic granules. Our results demonstrated that oocytes with granular cytoplasm present equivalent efficiency for IVF and calf production compared with homogenous cytoplasm oocytes. This could be observed through similar cleavage, blastocyst rates, and fetal growth development. In addition to differences in oocyte gene expression related to oocyte quality, it seems not to affect oocyte developmental competence. [ABSTRACT FROM AUTHOR]

This document presents the results of a three-year field experiment evaluating the use of Sardar Amin Granules (SAG) and Bentonite Sulfur (BS) in different maize-based cropping systems. The study found that the application of SAG and BS significantly increased the yield of maize, wheat, onion, and summer moong. However, the effects varied depending on the specific crop and fertilizer treatment. The study also compared different cropping systems and found that the maize-potato-summer moong system was the most productive, while the maize-wheat system was the least productive. The document also includes two scientific articles discussing soil organic matter determination and advances in fertilizer use efficiency. [Extracted from the article]

Mast cell tumour is one of the most common cutaneous neoplasms in dogs, ranging from well differentiated to aggressive tumors with metastases. Currently, prognosis and therapeutic definitions of canine cutaneous MCTs are based on the histological grade of malignancy. Clinical examination and treatment were performed in Dnipro veterinary clinics and pathological analysis in the Department of Animal Anatomy, Histology and Pathomorphology, Dnipro State Agrarian and Economic University (Dnipro, Ukraine). Thirty-six dogs with a history of skin neoplasia were examined. Skin MCTs were diagnosed in 23 males and 13 females, aged between 5 and 15 years. No breed predisposition was found out in tested cohort. MCTs were located on the trunk (44.4%), limbs (33.4%) and less frequently on the head and neck, axillary, inguinal and perineal areas (22.2%). MCT was detected in 8 animals where the malignancy was systemic and develops metastatic spread to internal organs. Pathological diagnosis included cytopathological examination of tumor material, pathological analysis of samples obtained by incisional biopsy using haematoxylin/eosin and toluidine blue staining. According to the diagnostic criteria (smear cellularity, presence of cells with basophilic cytoplasmic granularity, size and number of cytoplasmic granules, presence or absence of mitotic figures, nuclear pleomorphism, presence or absence of binucleations or multinucleations, anisokaryosis), benign mast cell tumour (high grade) was diagnosed in 23 animals and malignant mast cell tumour (low grade) in 13 animals. These results were confirmed by histopathological methods using haematoxylin and eosin staining. Additional staining of histological sections with toluidine blue allowed differentiation of benign mast cell tumours into high-grade (38.9%) and intermediate-grade (5%). [ABSTRACT FROM AUTHOR]

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The present study investigated the behavioural response of bone-like cells towards dense and porous dicalcium phosphate dihydrate (DCPD)-coated β-tricalcium phosphate (β-TCP) granules. The surfaces of the dense and porous β-TCP granules were coated with a layer of DCPD through the dissolution-precipitation process by exposing them to an acidic calcium phosphate solution for 30 min at 25 °C. Subsequently, the specimens were characterised with X-ray diffraction (XRD), scanning electron microscope (SEM), dissolution and pH tests, and evaluation of initial cell attachment. The results demonstrated that the porous β-TCP granule surfaces coated with a DCPD layer contributed to the high concentration of calcium ions released and decreased pH values. Furthermore, the DCPD-coated porous β-TCP granules produced good initial cell attachment and enhanced cell viability for up to three days. The results would provide new insights to bioceramic researchers regarding the impacts of pore presence in accelerating DCPD-coated β-TCP granule cell responses. [ABSTRACT FROM AUTHOR]

Granular cell tumor was described in the testis of two rabbits. Testis from each rabbit was surgically removed and submitted for histopathological diagnosis. Both testes were about 2.0 cm in diameter, firm, and tan. Microscopically, testicular mass consisted of compact sheets of round to polygonal and occasional spindle-shaped cells. The neoplastic cells contain a large amount of eosinophilic granular material in the cytoplasm. The cytoplasmic eosinophilic granules were positive for periodic acid Schiff stain. Immunohistochemically, the neoplastic cells were immunoreactive to Melan-A and vimentin. Based on these results, the testicular mass was diagnosed as a granular cell tumor. [ABSTRACT FROM AUTHOR]

Persistent cytoplasmic aggregates containing RNA binding proteins (RBPs) are central to the pathogenesis of late-onset neurodegenerative disorders such as amyotrophic lateral sclerosis (ALS). These aggregates share components, molecular mechanisms, and cellular protein quality control pathways with stress-induced RNA granules (SGs). Here, we assess the impact of stress on the global mRNA localization landscape of human pluripotent stem cell-derived motor neurons (PSC-MNs) using subcellular fractionation with RNA sequencing and proteomics. Transient stress disrupts subcellular RNA and protein distributions, alters the RNA binding profile of SG- and ALS-relevant RBPs and recapitulates disease-associated molecular changes such as aberrant splicing of STMN2. Although neurotypical PSC-MNs re-establish a normal subcellular localization landscape upon recovery from stress, cells harboring ALS-linked mutations are intransigent and display a delayed-onset increase in neuronal cell death. Our results highlight subcellular molecular distributions as predictive features and underscore the utility of cellular stress as a paradigm to study ALS-relevant mechanisms.

Background: Smoking is currently the most preventable cause of diseases and death worldwide and is one of the causative risk factors for developing cancer in different organs. Hence, it is necessary to detect potentially malignant lesions at their incipient stage. Oral cancers commonly affects floor of the mouth, soft palate, lateral border of the tongue and other areas of the mouth. Aim and Objective: The present study was the Cytomorphometric Changes In The Buccal Mucosal Cell In Smokers In Central Indian Populations. Material and Methods:A comparative study conducted among smokers of Central India population, was performed on 200 males under the age group of 20-60 years. The subjects were chosen randomly from IPD and OPD patients of Medicine Department of Index Medical College and Hospital, Indore. Results: There was significant difference observed between non smoking and smoking for cells with binucleation pyknosis, perinuclear halo, cytoplasmic granules, karyolsis, karyorrhexis, and micronuclei, in buccal mucosal cells but non significance difference was found for cytoplasmic vacuoles between nonsmoking and smokers. similar study of non-smoking populations. Conclusion: The present study indicates that almost all cytomorphological findings were high in smokers than non-smokers. Early detection of oral cancers becomes complex as they are mostly innocuous and asymptomatic during their initial stages, Cytomorphometric analysis can be used regularly to detect these cell alterations. Currently, use of exfoliative cytology has increased as an adjunct to screening of precancerous lesions and malignancies of the oral cavity. [ABSTRACT FROM AUTHOR]

Neutrophils are a specialized subset of white blood cells, which have the ability to store pre-formed mediators in their cytoplasmic granules. Neutrophils are wellknown effector cells involved in host protection against pathogens through diverse mechanisms such as phagocytosis, degranulation, extracellular traps, and oxidative burst. In this study, we provide evidence highlighting the significance of the SNARE proteins syntaxin-4 and synaptosomal-associated protein (SNAP) 23 in the release of azurophilic granules, specific granules, and the production of reactive oxygen species in human neutrophils. In contrast, the specific blockade of either syntaxin-4 or SNAP23 did not prevent the release of mitochondrial dsDNA in the process of neutrophil extracellular trap (NET) formation. These findings imply that degranulation and the release of mitochondrial dsDNA involve at least partially distinct molecular pathways in neutrophils. [ABSTRACT FROM AUTHOR]

Epidemiological studies show an association between inflammatory bowel disease (IBD) and increased risk of thrombosis. However, how IBD influences thrombosis remains unknown. The current study shows that formation of neutrophil extracellular traps (NETs) significantly increased in the dextran sulfate sodium (DSS)‐induced IBD mice, which in turn, contributes to thrombus formation in a NETs‐dependent fashion. Furthermore, the exosomes isolated from the plasma of the IBD mice induce arterial and venous thrombosis in vivo. Importantly, proinflammatory factors‐exposed intestinal epithelial cells (inflamed IECs) promote neutrophils to release NETs through their secreted exosomes. RNA sequencing revealed that LINC00668 is highly enriched in the inflamed IECs‐derived exosomes. Mechanistically, LINC00668 facilitates the translocation of neutrophil elastase (NE) from the cytoplasmic granules to the nucleus via its interaction with NE in a sequence‐specific manner, thereby inducing NETs release and thrombus formation. Importantly, berberine (BBR) suppresses the nuclear translocation of NE and subsequent NETs formation by inhibiting the interaction of LINC00668 with NE, thus exerting its antithrombotic effects. This study provides a novel pathobiological mechanism linking IBD and thrombosis by exosome‐mediated NETs formation. Targeting LINC00668 can serve as a novel molecular treatment strategy to treat IBD‐related thrombosis. [ABSTRACT FROM AUTHOR]

The immune activity of polysaccharides increases after structural modification, whereas the structure of dietary fiber is similar to that of polysaccharides, and research on the chemical modification of dietary fiber is lacking. Therefore, in this study, we used soluble dietary fiber from millet to chelate metal selenium ions, named the selenium-enriched additive (SDF-Se), and investigated its effects on macrophages and natural killer (NK) cells in vitro. The results showed that SDF-Se could activate RAW 264.7 cells through NF-κB and mitogen-activated protein kinase (MAPK) signaling and promote the accumulation of the reactive oxygen species, thereby increasing the levels of TNF-α, IL-6, and IL-10 and releasing numerous NO molecules. Furthermore, SDF-Se treatment induced NK cells to secrete cytokines (IFN-γ and TNF-α), cytoplasmic granules (granzyme-B), surface-activating receptors (NKp44), and increased CD56 and CD107a expressions, which promote NK cell cytotoxic responses. These results suggest that SDF-Se can promote macrophage and NK cell activation and be used as a potent immunostimulant. [ABSTRACT FROM AUTHOR]

Background: Ultrastructural information regarding the peripheral blood cells of local (Zovawk) pigs from Mizoram, India, is not available in the scientific literature. Objectives: The present study was designed to reveal the fine structural details of the blood cells from these local pigs using a transmission electron microscope (TEM). Methods: Blood samples were collected from 12 healthy Zovawk pigs of either sex and processed according to a standard protocol. Processed blood samples were then sent to the All India Institute of Medical Sciences, New Delhi, for further processing and imaging under TEM. Different types of blood cells were viewed under TEM, and different characteristics of these cells were assessed. Results: In the present study, erythrocytes are elongated, biconcave, and nucleated without cytoplasmic organelles. Neutrophils are round with 2‐5 lobed nuclei surrounded by cytoplasm with an indistinct bilayered nuclear membrane. The cytoplasm is packed with membrane bound round, oval, and elongated cytoplasmic granules. Eosinophils are round to oval with 2‐3 lobed nuclei with distinct nuclear membranes. Basophils are spherical and contained small, medium, and large electron‐dense granules. Lymphocytes are small, medium, and large and contained all cellular components. Monocytes are irregularly spherical with slight nuclear indentations. The platelets are elongated, oval, or rounded, with a few pseudopods at the cell surface. Conclusions: From the present study, we can conclude that the ultrastructural morphology of blood cells from Zovawk pigs resembles those of other domestic animals. However, a few differences have been observed. [ABSTRACT FROM AUTHOR]

Chronic cellular stress associated with neurodegenerative disease can result in the persistence of stress granule (SG) structures, membraneless organelles that form in response to cellular stress. In Huntington's disease (HD), chronic expression of mutant huntingtin generates various forms of cellular stress, including activation of the unfolded protein response and oxidative stress. However, it has yet to be determined whether SGs are a feature of HD neuropathology. We examined the miRNA composition of extracellular vesicles (EVs) present in the cerebrospinal fluid (CSF) of patients with HD and show that a subset of their target mRNAs were differentially expressed in the prefrontal cortex. Of these targets, SG components were enriched, including the SG-nucleating Ras GTPase-activating protein-binding protein 1 (G3BP1). We investigated localization and levels of G3BP1 and found a significant increase in the density of G3BP1-positive granules in the cortex and hippocampus of R6/2 transgenic mice and in the superior frontal cortex of the brains of patients with HD. Intriguingly, we also observed that the SG-associated TAR DNA-binding protein 43 (TDP43), a nuclear RNA/DNA binding protein, was mislocalized to the cytoplasm of G3BP1 granule-positive HD cortical neurons. These findings suggest that G3BP1 SG dynamics may play a role in the pathophysiology of HD.

Cells organize biochemical processes into biological condensates. P-bodies are cytoplasmic condensates that are enriched in enzymes important for mRNA degradation and have been identified as sites of both storage and decay. How these opposing outcomes can be achieved in condensates remains unresolved. mRNA decapping immediately precedes degradation, and the Dcp1/Dcp2 decapping complex is enriched in P-bodies. Here, we show that Dcp1/Dcp2 activity is modulated in condensates and depends on the interactions promoting phase separation. We find that Dcp1/Dcp2 phase separation stabilizes an inactive conformation in Dcp2 to inhibit decapping. The activator Edc3 causes a conformational change in Dcp2 and rewires the protein-protein interactions to stimulate decapping in condensates. Disruption of the inactive conformation dysregulates decapping in condensates. Our results indicate that the regulation of enzymatic activity in condensates relies on a coupling across length scales ranging from microns to ångstroms. We propose that this regulatory mechanism may control the functional state of P-bodies and related phase-separated compartments.

Purpose: To examine the specific changes that occur in the expression levels of extracellular vesicle-derived microRNAs (miRNAs) in intracranial cerebrospinal fluid (CSF) in moyamoya disease. Methods: Patients with arteriosclerotic cerebral ischemia were used as controls to eliminate the effects of cerebral ischemia. Intracranial CSF was collected from moyamoya disease and control patients during bypass surgery. Extracellular vesicles (EVs) were extracted from the CSF. Comprehensive expression analysis of miRNAs extracted from EVs by next-generation sequencing (NGS) and validation by quantitative reverse transcription-polymerase chain reaction (qRT-PCR) was performed. Results: Experiments were conducted on eight cases of moyamoya disease and four control cases. In the comprehensive miRNA expression analysis, 153 miRNAs were upregulated, and 98 miRNAs were downregulated in moyamoya disease compared to the control cases (q-value < 0.05 and |log2 fold change|> 1). qRT-PCR performed on the four most variable miRNAs (hsa-miR-421, hsa-miR-361-5p, hsa-miR-320a, and hsa-miR-29b-3p) associated with vascular lesions among the differentially expressed miRNAs gave the same results as miRNA sequencing. On gene ontology (GO) analysis for the target genes, cytoplasmic stress granule was the most significant GO term. Conclusions: This study is the first comprehensive expression analysis of EV-derived miRNAs in the CSF of moyamoya disease patients using NGS. The miRNAs identified here may be related to the etiology and pathophysiology of moyamoya disease. [ABSTRACT FROM AUTHOR]

Severe forms of dilated cardiomyopathy (DCM) are associated with point mutations in the alternative splicing regulator RBM20 that are frequently located in the arginine/serine-rich domain (RS-domain). Such mutations can cause defective splicing and cytoplasmic mislocalization, which leads to the formation of detrimental cytoplasmic granules. Successful development of personalized therapies requires identifying the direct mechanisms of pathogenic RBM20 variants. Here, we decipher the molecular mechanism of RBM20 mislocalization and its specific role in DCM pathogenesis. We demonstrate that mislocalized RBM20 RS-domain variants retain their splice regulatory activity, which reveals that aberrant cellular localization is the main driver of their pathological phenotype. A genome-wide CRISPR knockout screen combined with image-enabled cell sorting identified Transportin-3 (TNPO3) as the main nuclear importer of RBM20. We show that the direct RBM20-TNPO3 interaction involves the RS-domain, and is disrupted by pathogenic variants. Relocalization of pathogenic RBM20 variants to the nucleus restores alternative splicing and dissolves cytoplasmic granules in cell culture and animal models. These findings provide proof-of-principle for developing therapeutic strategies to restore RBM20's nuclear localization in RBM20-DCM patients. The authors show that loss-of-interaction with the nuclear importer, TNPO3, causes cytoplasmic mislocalization of RBM20 variants linked to severe cases of dilated cardiomyopathy. Restoring their nuclear localization alleviates the disease phenotype. [ABSTRACT FROM AUTHOR]

The genome sequencing of the tardigrade Ramazzottius varieornatus revealed a unique nucleosome-binding protein named damage suppressor (Dsup), which was discovered to be crucial for the extraordinary abilities of tardigrades in surviving extreme stresses, such as UV. Evidence in Dsup-transfected human cells suggests that Dsup mediates an overall response in DNA damage signaling, DNA repair, and cell cycle regulation, resulting in an acquired resistance to stress. Given these promising outcomes, our study attempts to provide a wider comprehension of the molecular mechanisms modulated by Dsup in human cells and to explore the Dsup-activated molecular pathways under stress. We performed a differential proteomic analysis of Dsup-transfected and control human cells under basal conditions and at 24 h recovery after exposure to UV-C. We demonstrate via enrichment and network analyses, for the first time, that even in the absence of external stimuli, and more significantly, after stress, Dsup activates mechanisms involved with the unfolded protein response, the mRNA processing and stability, cytoplasmic stress granules, the DNA damage response, and the telomere maintenance. In conclusion, our results shed new light on Dsup-mediated protective mechanisms and increases our knowledge of the molecular machineries of extraordinary protection against UV-C stress. [ABSTRACT FROM AUTHOR]

Mammalian male germ cell differentiation relies on complex RNA biogenesis events, many of which occur in non-membrane bound organelles termed RNA germ cell granules that are rich in RNA binding proteins (RBPs). Though known to be required for male germ cell differentiation, we understand little of the relationships between the numerous granule subtypes. ADAD2, a testis specific RBP, is required for normal male fertility and forms a poorly characterized granule in meiotic germ cells. This work aimed to understand the role of ADAD2 granules in male germ cell differentiation by clearly defining their molecular composition and relationship to other granules. Biochemical analyses identified RNF17, a testis specific RBP that forms meiotic male germ cell granules, as an ADAD2-interacting protein. Phenotypic analysis of Adad2 and Rnf17 mutants identified a rare post-meiotic chromatin defect, suggesting shared biological roles. ADAD2 and RNF17 were found to be dependent on one another for granularization and together form a previously unstudied set of germ cell granules. Based on co-localization studies with well-characterized granule RBPs and organelle-specific markers, a subset of the ADAD2-RNF17 granules are found to be associated with the intermitochondrial cement and piRNA biogenesis. In contrast, a second, morphologically distinct population of ADAD2-RNF17 granules co-localized with the translation regulators NANOS1 and PUM1, along with the molecular chaperone PDI. These large granules form a unique funnel-shaped structure that displays distinct protein subdomains and is tightly associated with the endoplasmic reticulum. Developmental studies suggest the different granule populations represent different phases of a granule maturation process. Lastly, a double Adad2-Rnf17 mutant model suggests the interaction between ADAD2 and RNF17, as opposed to loss of either, is the likely driver of the Adad2 and Rnf17 mutant phenotypes. These findings shed light on the relationship between germ cell granule pools and define new genetic approaches to their study. Author summary: To differentiate successfully, male germ cells tightly regulate their RNA pools. To do so, they rely on RNA binding proteins, which often localize to cytoplasmic granules. The majority of studies have focused on a single granule type, the intermitochondrial cement, which regulates piRNA biogenesis. As a result, there is limited knowledge about the other granules. Here, we identify an interaction between two RNA binding proteins, ADAD2 and RNF17, and demonstrate mutants share a rare germ cell phenotype. Further, ADAD2 and RNF17 colocalize to the same germ cell granules, which are observed as two morphologically unique types. The first subset of ADAD2-RNF17 granules associate with known granules and likely play a role in the piRNA pathway. The second granule type forms a unique shape with distinct protein subdomains and may play a role in regulating translation. This second population is in close proximity to the endoplasmic reticulum and appears to form by relocalization and molecular remodeling of the first. Genetic models further suggest the interaction between ADAD2 and RNF17 likely drive the mutant phenotypes. These findings identify a novel population of germ cell granules that undergo maturation across germ cell differentiation and appear to regulate distinct aspects of RNA biology. [ABSTRACT FROM AUTHOR]

Granule cell-selective toxicity of methylmercury in the cerebellum is one of the main unresolved issues in the pathogenesis of Minamata disease. Rats were orally administered methylmercury chloride (10 mg/kg/day) for 5 consecutive days, and their brains were harvested on days 1, 7, 14, 21, or 28 after the last administration for histological examination of the cerebellum. It was found that methylmercury caused a marked degenerative change to the granule cell layers but not to the Purkinje cell layers. The generative change of the granule cell layer was due to cell death, including apoptosis, which occurred at day 21 and beyond after the methylmercury administration. Meanwhile, cytotoxic T-lymphocytes and macrophages had infiltrated the granule cell layer. Additionally, granule cells are shown to be a cell type susceptible to TNF-α. Taken together, these results suggest that methylmercury causes small-scale damage to granule cells, triggering the infiltration of cytotoxic T-lymphocytes and macrophages into the granule cell layer, which secrete tumor necrosis factor-a (TNF-α) to induce apoptosis in granule cells. This chain is established based on the susceptibility of granule cells to methylmercury, the ability of cytotoxic T lymphocytes and macrophages to synthesize and secrete TNF-α, and the sensitivity of granule cells to TNF-α and methylmercury. We propose to call the pathology of methylmercury-induced cerebellar damage the "inflammation hypothesis". [ABSTRACT FROM AUTHOR]

Lipopolysaccharide (LPS) has dose-dependent biphasic functions (cell protective versus cell toxic). To clarify the different effects of LPS on liver homeostasis or liver diseases, comparisons were made between low and high doses of LPS, in terms of the mutual relation of hepatic macrophages, autophagy, and damage-associated molecular patterns (DAMPs) in male F344/DuCrlCrlj rats. Rats injected with low dose (0.1 mg/kg) or high dose (2.0 mg/kg) of LPS were examined at 6, 10, and 24 hours following single injections. Histologically, focal hepatocellular necrosis was occasionally present in high-dose animals, whereas there were no significant changes in low-dose animals. In low-dose animals, Kupffer cells reacting to CD163 and CD204 were hypertrophic and regarded as M2 macrophages, which promote resolution of inflammation and tissue repair, whereas in high-dose animals, infiltration of M1 macrophages expressing CD68 and major histocompatibility complex class II, which enhance cell injury, was seen. Hepatocytes with high-mobility-group box-1 (HMGB1) (one of DAMPs)-positive cytoplasmic granules appeared more frequently in high-dose animals than in low-dose animals, indicating the translocation of nuclear HMGB1 into the cytoplasm. However, although light-chain 3 beta–positive autophagosomes in hepatocytes increased in both doses, abnormally vacuolated autophagosomes were only seen in injured hepatocytes in the high-dose group, indicating possible extracellular release of HMGB1, which might result in cell injury and inflammation. These findings suggested that low-dose LPS induced a favorable mutual relationship among hepatic macrophages, autophagy, and DAMPs leading to cytoprotection of hepatocytes, whereas failures of the relationship in high-dose LPS caused hepatocyte injury. [ABSTRACT FROM AUTHOR]

The functions of annexin A1 (ANXA1), which is expressed on membranes and in cytoplasmic granules, have been fully described. Nonetheless, the role of this protein in protecting against DNA damage in the nucleus is still emerging and requires further investigation. Here, we investigated the involvement of ANXA1 in the DNA damage response in placental cells. Placenta was collected from ANXA1 knockout mice (AnxA1−/−) and pregnant women with gestational diabetes mellitus (GDM). The placental morphology and ANXA1 expression, which are related to the modulation of cellular response markers in the presence of DNA damage, were analyzed. The total area of AnxA1−/− placenta was smaller due to a reduced labyrinth zone, enhanced DNA damage, and impaired base excision repair (BER) enzymes, which resulted in the induction of apoptosis in the labyrinthine and junctional layers. The placentas of pregnant women with GDM showed reduced expression of AnxA1 in the villous compartment, increased DNA damage, apoptosis, and a reduction of enzymes involved in the BER pathway. Our translational data provide valuable insights into the possible involvement of ANXA1 in the response of placental cells to oxidative DNA damage and represent an advancement in investigations into the mechanisms involved in placental biology. [ABSTRACT FROM AUTHOR]

Background: This is an observational study which aims to research morphological changes of white blood cells in patients with Systemic Inflammatory Response Syndrome (SIRS) with and without sepsis and evaluate morphological changes in white blood cells as predictors of sepsis. Methods: Patients aged 18 years or more with SIRS with sepsis and SIRS without sepsis were included and those with haematological disorders or pregnant patients were excluded. A total of 52 patients with SIRS with sepsis and 32 patients of SIRS without sepsis were included. Peripheral blood smear was prepared from the venous blood sample drawn. The presence of toxic granules, cytoplasmic vacuoles, and Dohle bodies in both cases of SIRS with sepsis without sepsis were assessed and it was compared with culture-positive sepsis and shock. Results: The difference in the presence of toxic granules (55.8% vs. 12.5%; p <0.001), cytoplasmic vacuoles (30.8% vs. 6.3%; p -0.012), and Dohle bodies (17.3% vs. 0%; p = 0.012) was significantly higher in the SIRS with sepsis group, compared to the SIRS without sepsis group. In the subgroup analysis of patients in the sepsis group, it was observed that patients with positive blood culture (9%) had a significantly higher proportion of toxic granules (100% vs. 51.1%; p=0.059), cytoplasmic vacuoles (40% vs. 29.8%; p=0.637) and Dohle bodies (40% vs. 14.9%; p=0.202). However, these differences were not statistically significant. Conclusion: Toxic granules and cytoplasmic vacuoles in the neutrophils of patients with SIRS with sepsis were found more frequently, compared to patients of SIRS without sepsis. Dohle bodies were found only in patients with sepsis and not in those with SIRS without sepsis. [ABSTRACT FROM AUTHOR]

Introduction: The thyroid gland is primarily made up of endoderm-derived follicular cells which produces thyroid hormone in all vertebrates. Thyroid hormone is best known for its role in controlling metabolism in adults, although it is also required for several processes throughout development. Aim of the Work: This study was conducted to compare the histology and ultrastructure of the thyroid gland in Rousettus aegyptiacus with age. Material and Methods: A total of 20 thyroid gland samples, including 10 for each stage (postnatal (seven days after birth) and adult (10 months age) were used, the entire thyroid gland was separated from the neck region and prepared for histological, histochemical and transmission electron microscopic examinations. Results: In the adult stage, thyroid gland made of vast number of follicles of various shapes and sizes which lined with follicular epithelial cells that were flattened squamous to cuboidal having rough endoplasmic reticulum with dilated cisternae. In postnatal bats, the thyroid gland consisted of smaller follicles with reduced colloid and lined with columnar follicular epithelial cells containing hyperactive rough endoplasmic reticulum with highly dilated cisternae and Golgi bodies that imparted a vacuolated appearance to the cytoplasm. Short and sparse microvilli were detected on the follicular cells, and their quantity increased in postnatal bats. Among the apical vesicles, secretory vesicles, colloid droplets, and lysosome-like structures were found with different sizes and electron densities, and their appearance changed in postnatal bats. Due to the presence of particular basal cells in adult bats, parafollicular cells were found away from the follicular lumen; however, in postnatal bats, it was found between follicular cells in the basal position with numerous dense cytoplasmic granules. Conclusion: The general histological and ultrastructural features of the thyroid gland of postnatal and adult R. aegyptiacus have similarities to that of other mammals. [ABSTRACT FROM AUTHOR]

Stress granules (SGs) are evolutionarily conserved condensates of ribonucleoproteins that assemble in response to metabolic stresses. Because aberrant SG formation is associated with amyotrophic lateral sclerosis (ALS), understanding the connection between metabolic activity and SG composition can provide therapeutic insights into neurodegeneration. Here, we identify 17 metabolic enzymes recruited to yeast SGs in response to physiological growth stress. Furthermore, the product of one of these enzymes, AdoMet, is a regulator of SG assembly and composition. Decreases in AdoMet levels increase SG formation, while chronic elevation of AdoMet produces SG remnants lacking proteins associated with the 5' end of transcripts. Interestingly, acute elevation of AdoMet blocks SG formation in yeast and motor neurons. Treatment of ALS-derived motor neurons with AdoMet also suppresses the formation of TDP-43-positive SGs, a hallmark of ALS. Together, these results argue that AdoMet is an evolutionarily conserved regulator of SG composition and assembly with therapeutic potential in neurodegeneration.

Pbp1 (poly(A)-binding protein—binding protein 1) is a cytoplasmic stress granule marker that is capable of forming condensates that function in the negative regulation of TORC1 signaling under respiratory conditions. Polyglutamine expansions in its mammalian ortholog ataxin-2 lead to spinocerebellar dysfunction due to toxic protein aggregation. Here, we show that loss of Pbp1 in S. cerevisiae leads to decreased amounts of mRNAs and mitochondrial proteins which are targets of Puf3, a member of the PUF (Pumilio and FBF) family of RNA-binding proteins. We found that Pbp1 supports the translation of Puf3-target mRNAs in respiratory conditions, such as those involved in the assembly of cytochrome c oxidase and subunits of mitochondrial ribosomes. We further show that Pbp1 and Puf3 interact through their respective low complexity domains, which is required for Puf3-target mRNA translation. Our findings reveal a key role for Pbp1-containing assemblies in enabling the translation of mRNAs critical for mitochondrial biogenesis and respiration. They may further explain prior associations of Pbp1/ataxin-2 with RNA, stress granule biology, mitochondrial function, and neuronal health. Author summary: The yeast protein Pbp1 and its mammalian ortholog ataxin-2 are both markers of stress granules. The function of such granules and what they do to mRNA transcripts has been unclear. Here we use budding yeast to show that Pbp1 regulates transcripts involved in mitochondrial biogenesis, specifically those that are targets of Puf3, a member of the PUF family of sequence-specific RNA-binding proteins. Loss of Pbp1 results in reduced amounts of these mRNAs and the proteins they encode. Intriguingly, many of these are components of cytochrome c oxidase. We further show Pbp1 associates with Puf3 under respiratory conditions to assist the translation of these mRNAs. These findings suggest Pbp1-containing granules regulate the translation of a specific class of mRNAs according to the metabolic demands of the cell. [ABSTRACT FROM AUTHOR]