Immature Morphology Research Articles

Systematics of Dynastor Doubleday, [1849] (Lepidoptera: Nymphalidae, Satyrinae, Brassolini)

Dynastor Doubleday, [1849] (Lepidoptera: Nymphalidae: Satyrinae: Brassolini) is an exclusively Neotropical genus, occurring from Mexico to Argentina. Adults have crepuscular habits, while the larvae are unique in Brassolini in feeding on bromeliads (Bromeliaceae). Until now, Dynastor contained three species, nine subspecies, and five synonyms (two of them generic synonyms). The present study analyzed information on the immature stages and adult morphology, with molecular and distribution data, to provide a revised taxonomic hypothesis for Dynastor species and subspecies. Analyses of approximately 380 specimens, including all species and subspecies known for the genus, supported the reinstatement of the statuses of three species: Dynastor stygianus Butler, 1872 stat. rest., Dynastor hannibal Oberthür, 1881 stat. rest., and Dynastor strix (Bates, 1864) stat. rest. One subspecies is here treated as a species: Dynastor pharnaces Stichel, 1908 stat. nov. Three new synonyms are proposed: Papilio anaxarete Cramer, 1776 syn. nov., Dynastor darius ictericus Stichel, 1904 syn. nov. and Dynastor darius faenius Fruhstorfer, 1912 syn. nov. of Dynastor darius (Fabricius, 1775). To ensure unambiguous identification of names, two neotypes were designated for: Dynastor napoleon f. maculatus Niepelt, 1922 and Dynastor populus Röber, 1927, and nine lectotypes for: Dynastor napoleon Doubleday, [1849], Papilio darius Fabricius, 1775, Papilio anaxarete, Dynastor mardonius Fruhstorfer, 1911, Dynastor darius faenius, Brassolis macrosiris Westwood, 1851, Dynastor hannibal, Brassolis strix, and Dynastor pharnaces stat. nov. The proposed taxonomic checklist for Dynastor includes seven species, more than double the previous proposals, and eight subjective synonyms (two of them generic synonyms).

Sella turcica and facial bones: Morphological integration in the human fetal cranium.

The cranial base plays a significant role in facial growth, and closer analyses of the morphological relationship between these two regions are needed to understand the morphogenesis of the face. Here, we aimed to study morphological integration between the sella turcica (ST) and facial bones during the fetal period using geometric morphometrics. Magnetic resonance images of 47 human fetuses in the Kyoto Collection, with crown-rump lengths of 29.8-225 mm, were included in this study. Anatomical homologous landmarks and semilandmarks were registered on the facial bones and the midsagittal contour of the ST, respectively. The shape variations in the craniofacial skeleton and the ST were statistically investigated by reducing dimensionality using principal component analysis (PCA). Subsequently, the morphological integration between the facial bones and ST was investigated using two-block partial least squares (2B-PLS) analysis. PCA showed that small specimens represented the concave facial profile, including the mandibular protrusion and maxillary retrusion. The 2B-PLS showed a strong integration (RV coefficient = 0.523, r = .79, p < .01) between the facial bones and ST. The curvature of the anterior wall of the ST was highly associated with immature facial morphology characterized by a concave profile. The strong integration between the two regions suggested that the anterior ST may be associated with facial morphology. This result quantitatively confirms previous studies reporting ST deformities in facial anomalies and induces further research using postnatal subjects.

Geomorphology of craters located at Mercury’s north pole

Geomorphology of craters located at Mercury’s north pole

Structural, Functional, and Genetic Changes Surrounding Electrodes Implanted in the Brain.

Implantable neurotechnology enables monitoring and stimulating of the brain signals responsible for performing cognitive, motor, and sensory tasks. Electrode arrays implanted in the brain are increasingly used in the clinic to treat a variety of sources of neurological diseases and injuries. However, the implantation of a foreign body typically initiates a tissue response characterized by physical disruption of vasculature and the neuropil as well as the initiation of inflammation and the induction of reactive glial states. Likewise, electrical stimulation can induce damage to the surrounding tissue depending on the intensity and waveform parameters of the applied stimulus. These phenomena, in turn, are likely influenced by the surface chemistry and characteristics of the materials employed, but further information is needed to effectively link the biological responses observed to specific aspects of device design. In order to inform improved design of implantable neurotechnology, we are investigating the basic science principles governing device-tissue integration. We are employing multiple techniques to characterize the structural, functional, and genetic changes that occur in the cells surrounding implanted electrodes. First, we have developed a new "device-in-slice" technique to capture chronically implanted electrodes within thick slices of live rat brain tissue for interrogation with single-cell electrophysiology and two-photon imaging techniques. Our data revealed several new observations of tissue remodeling surrounding devices: (a) there was significant disruption of dendritic arbors in neurons near implants, where losses were driven asymmetrically on the implant-facing side. (b) There was a significant loss of dendritic spine densities in neurons near implants, with a shift toward more immature (nonfunctional) morphologies. (c) There was a reduction in excitatory neurotransmission surrounding implants, as evidenced by a reduction in the frequency of excitatory postsynaptic currents (EPSCs). Lastly, (d) there were changes in the electrophysiological underpinnings of neuronal spiking regularity. In parallel, we initiated new studies to explore changes in gene expression surrounding devices through spatial transcriptomics, which we applied to both recording and stimulating arrays. We found that (a) device implantation is associated with the induction of hundreds of genes associated with neuroinflammation, glial reactivity, oligodendrocyte function, and cellular metabolism and (b) electrical stimulation induces gene expression associated with damage or plasticity in a manner dependent upon the intensity of the applied stimulus. We are currently developing computational analysis tools to distill biomarkers of device-tissue interactions from large transcriptomics data sets. These results improve the current understanding of the biological response to electrodes implanted in the brain while producing new biomarkers for benchmarking the effects of novel electrode designs on responses. As the next generation of neurotechnology is developed, it will be increasingly important to understand the influence of novel materials, surface chemistries, and implant architectures on device performance as well as the relationship with the induction of specific cellular signaling pathways.

Canonical Wnt activator Chir99021 prevents epileptogenesis in the intrahippocampal kainate mouse model of temporal lobe epilepsy

The Wnt signaling pathway mediates the development of dentate granule cell neurons in the hippocampus. These neurons are central to the development of temporal lobe epilepsy and undergo structural and physiological remodeling during epileptogenesis, which results in the formation of epileptic circuits. The pathways responsible for granule cell remodeling during epileptogenesis have yet to be well defined, and represent therapeutic targets for the prevention of epilepsy. The current study explores Wnt signaling during epileptogenesis and for the first time describes the effect of Wnt activation using Wnt activator Chir99021 as a novel anti-epileptogenic therapeutic approach.Focal mesial temporal lobe epilepsy was induced by intrahippocampal kainate (IHK) injection in wild-type and POMC-eGFP transgenic mice. Wnt activator Chir99021 was administered daily, beginning 3 h after seizure induction, and continued up to 21-days. Immature granule cell morphology was quantified in the ipsilateral epileptogenic zone and the contralateral peri-ictal zone 14 days after IHK, targeting the end of the latent period. Bilateral hippocampal electrocorticographic recordings were performed for 28-days, 7-days beyond treatment cessation. Hippocampal behavioral tests were performed after completion of Chir99021 treatment.Consistent with previous studies, IHK resulted in the development of epilepsy after a 14 day latent period in this well-described mouse model. Activation of the canonical Wnt pathway with Chir99021 significantly reduced bilateral hippocampal seizure number and duration. Critically, this effect was retained after treatment cessation, suggesting a durable antiepileptogenic change in epileptic circuitry. Morphological analyses demonstrated that Wnt activation prevented pathological remodeling of the primary dendrite in both the epileptogenic zone and peri-ictal zone, changes in which may serve as a biomarker of epileptogenesis and anti-epileptogenic treatment response in pre-clinical studies. These findings were associated with improved object location memory with Chir99021 treatment after IHK.This study provides novel evidence that canonical Wnt activation prevents epileptogenesis in the IHK mouse model of mesial temporal lobe epilepsy, preventing pathological remodeling of dentate granule cells. Wnt signaling may therefore play a key role in mesial temporal lobe epileptogenesis, and Wnt modulation may represent a novel therapeutic strategy in the prevention of epilepsy.

Morphology of the immature stages of Agrotis canities and A. robusta and an identification key of Agrotis species associated to economic crops in South America

Agrotis Ochsenheimer (Lepidoptera: Noctuidae) is a widespread, cosmopolitan genus, with 20 species occurring in South America. Two of those species are considered major pests of different crops, Agrotis ipsilon (Hufnagel) and Agrotis robusta (Blanchard). The latter is often confused with Agrotis malefida Guenée in the literature and, despite its economic importance, its immature stages have been only superficially described. Agrotis canities (Grote), although never regarded as a pest, is known to use some crops as hosts and is commonly captured in light traps. However, its immature stages are unknown, making the evaluation of its true economic importance difficult. In this study, we examine in detail the external morphology and ultrastructure of the egg, larva and pupa of A. robusta and A. canities using light and scanning electron microscopy. Furthermore, we provide an identification key for those four species including all immature stages and up-to-date maps of their distribution in South America. Finally, we discussed the application of immature stages morphology of Agrotis species in systematic and phylogenetic studies. Particularly, we examine the morphology of the trichobothria, or “tonosensillum”, through larval development and the progressive and differential development of abdominal legs in each species.

Switching of hypertrophic signalling towards enhanced cardiomyocyte identity and maturity by a GATA4-targeted compound.

The prevalence of heart failure is constantly increasing, and the prognosis of patients remains poor. New treatment strategies to preserve cardiac function and limit cardiac hypertrophy are therefore urgently needed. Human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs) are increasingly used as an experimental platform for cardiac in vitro studies. However, in contrast to adult cardiomyocytes, hiPSC-CMs display immature morphology, contractility, gene expression and metabolism and hence express a naive phenotype that resembles more of a foetal cardiomyocyte. A library of 14 novel compounds was synthesized in-house and screened for GATA4-NKX2-5 reporter activity and cellular toxicity. The most potent compound, 3i-1262, along with previously reported GATA4-acting compounds, were selected to investigate their effects on hypertrophy induced by endothelin-1 or mechanical stretch. Morphological changes and protein expression were characterized using immunofluorescence staining and high-content analysis. Changes in gene expression were studied using qPCR and RNA sequencing. The prototype compound 3i-1262 inhibited GATA4-NKX2-5 synergy in a luciferase reporter assay. Additionally, the isoxazole compound 3i-1262 inhibited the hypertrophy biomarker B-type natriuretic peptide (BNP) by reducing BNP promoter activity and proBNP expression in neonatal rat ventricular myocytes and hiPSC-CMs, respectively. Treatment with 3i-1262 increased metabolic activity and cardiac troponin T expression in hiPSC-CMs without affecting GATA4 protein levels. RNA sequencing analysis revealed that 3i-1262 induces gene expression related to metabolic activity and cell cycle exit, indicating a change in the identity and maturity status of hiPSC-CMs. The biological processes that were enriched in upregulated genes in response to 3i-1262 were downregulated in response to mechanical stretch, and conversely, the downregulated processes in response to 3i-1262 were upregulated in response to mechanical stretch. There is currently a lack of systematic understanding of the molecular modulation and control of hiPSC-CM maturation. In this study, we demonstrated that the GATA4-interfering compound 3i-1262 reorganizes the cardiac transcription factor network and converts hypertrophic signalling towards enhanced cardiomyocyte identity and maturity. This conceptually unique approach provides a novel structural scaffold for further development as a modality to promote cardiomyocyte specification and maturity.

MYELOID-DERIVED SUPPRESSOR CELLS TWO YEARS AFTER HEPATITIS C VIRUS ERADICATION USING DIRECTLY ACTING ANTIVIRALS.

Myeloid-derived suppressor cells (MDSCs) have immature morphology, relatively weak phagocytic activity, as well as some immunosuppressive functions. The capacity of MDSCs to inhibit T-cell-mediated immunological responses is their most notable functional characteristic. Down-regulating antitumor immune surveillance is one way that the expansion and activation of MDSCs contribute significantly to the occurrence and progression of tumors. Increased levels of MDSCs in patients with chronic hepatitis C virus (HCV) infection could suppress T-cell responses, promoting viral escape and hepatitis progression. This may make HCV-infected individuals more vulnerable to severe infections, hepatic and extra-hepatic tumors, and a diminished capacity to react to immunization. It is still unknown if effective HCV eradication with directly acting antivirals (DAAs) can restore immune functions and immune surveillance capacity. The purpose of this study was to observe the frequency of M-MDSCs (CD33+, CD11b+, and HLA-DR) in patients with a previous history of HCV, 2-3 years after virus eradication using DAA therapy. This study was conducted on 110 subjects: fifty-five subjects without liver cirrhosis who were treated with HCV using DAAs and attained SVR for a period of 2-3 years and 55 age- and gender-matched healthy controls. The study was conducted during the period from January to July 2022. Patients were recruited from the National Viral Hepatitis Treatment Unit, Alexandria University Hepatology outpatient clinic, and the Alexandria University Tropical Medicine outpatient clinic. The frequencies of MDSCs (CD33+CD11b + HLA-DR-) by flow cytometry were assessed. Even after the virus had been eradicated for longer than two years, MDSC levels in HCV-treated individuals were found to be considerably higher. In the HCV-treated group, the median number of MDSCs was 5, with an interquartile range (IQR) of 3.79-7.69. In contrast, the median for the control group was 3.1, with an IQR of 1.4-3.2 (P˂0.001). Successful DAA therapy leads to slow and partial immunological reconstitution, as demonstrated by the failure to attain normal levels of MDSC's 2 years after successful HCV eradication despite the normalization of laboratory parameters as well as the absence of liver fibrosis. The clinical implications of these findings should be thoroughly studied.

The oribatid mite superfamily Eutegaeoidea (Acari, Oribatida), with descriptions of new taxa from Australia and New Caledonia and a re-assessment of genera and families.

Twenty-four new species of Eutegaeoidea from Australia and New Caledonia are described, and two new genera are proposed. These are Eutegaeus woiwurrung sp. nov., E. nothofagi sp. nov., E. bidhawal sp. nov., E. ptilosus sp. nov., Humerotegaeus carinatus gen. et sp. nov., H. concentricus gen. et sp. nov., Atalotegaeus crobylus sp. nov., Neoeutegaeus torsteini sp. nov., N. melipsilon sp. nov. N. malcolmi sp. nov., N. corniculatus sp. nov. (Eutegaeidae), Compactozetes goongerah sp. nov., C. crenellatus sp. nov. (Compactozetidae) and Pterozetes lawrencei sp. nov. (Pterozetidae) from temperate rainforests in Victoria and Tasmania; Compactozetes bundjalung sp. nov., C. calderi sp. nov., C. duonodulus sp. nov., Sadocepheus remus sp. nov. (Compactozetidae) and Porrhotegaeus githabul sp. nov. (Porrhotegaeidae fam. nov.) from temperate and sub-tropical rainforests of the Great Dividing Range in central and northern New South Wales and southern Queensland, Porrhotegaeus catherinae sp. nov. from scalybark closed forest on Lord Howe Island, Eutegaeus odontatus sp. nov. and Compactozetes pirumorpha sp. nov. from moist upland forest on Norfolk Island and Neseutegaeus wardi sp. nov. and Atalotegaeus deficiens sp. nov. from tropical rainforest and moss forest in New Caledonia. Based on the predominantly Southern Hemisphere distribution of Eutegaeoidea, indicating strong Gondwanan affinities, and the morphology of adults and immatures, this taxon is treated as distinct from the Cepheoidea which has a distribution almost entirely within the Northern Hemisphere. Eutegaeoid species previously described from Australia (Eutegaeus soror P. Balogh, 1985, Atalotegaeus mensarosi J. & P. Balogh, 1983, Neseutegaeus monteithi J. & P. Balogh, 1983, Neoeutegaeus phyllophorus J. & P. Balogh, 1983 and Porrhotegaeus ornatus J. Balogh & Mahunka, 1966) are redescribed based on type material and new distribution records provided. Species have distribution patterns predominantly indicative of short-range endemics associated with remnant Gondwanan rainforest. Neseutegaeus monteithi is recombined with Atalotegaeus Luxton, 1988a. The definitions of genera and families of Eutegaeoidea are revised, and their relationships are reconsidered. Birotegaeus Luxton, 1988a and Pareutegaeus Woolley, 1965 are designated junior synonyms of Eutegaeus Berlese, 1916. Immatures are described for the genera Atalotegaeus, Eutegaeus, Neoeutegaeus Aoki, 1964 and Porrhotegaeus J. Balogh & Mahunka, 1966. Neoeutegaeidae fam. nov. is established for Neoeutegaeus Aoki, 1964 and Humerotegaeus gen. nov., Porrhotegaeidae fam. nov. for Porrhotegaeus and Bornebuschiidae fam. nov. for Bornebuschia Hammer, 1966 and Dicrotegaeus Luxton, 1988 which had previously been placed in Cerocepheidae or Compactozetidae. Eutegaeus aysenensis Ermilov, 2021 and E. queulatensis Ermilov, 2021 from Chile, are recombined with Atalotegaeus. A key is provided to the genera of the eight families of Eutegaeoidea, as recognised herein: Eutegaeidae, Neoeutegaeidae fam. nov., Cerocepheidae, Compactozetidae, Bornebuschiidae fam. nov., Pterozetidae and Porrhotegaeidae fam. nov.

Engineered neurogenesis in naïve adult rat cortex by Ngn2-mediated neuronal reprogramming of resident oligodendrocyte progenitor cells.

Adult tissue stem cells contribute to tissue homeostasis and repair but the long-lived neurons in the human adult cerebral cortex are not replaced, despite evidence for a limited regenerative response. However, the adult cortex contains a population of proliferating oligodendrocyte progenitor cells (OPCs). We examined the capacity of rat cortical OPCs to be re-specified to a neuronal lineage both in vitro and in vivo. Expressing the developmental transcription factor Neurogenin2 (Ngn2) in OPCs isolated from adult rat cortex resulted in their expression of early neuronal lineage markers and genes while downregulating expression of OPC markers and genes. Ngn2 induced progression through a neuronal lineage to express mature neuronal markers and functional activity as glutamatergic neurons. In vivo retroviral gene delivery of Ngn2 to naive adult rat cortex ensured restricted targeting to proliferating OPCs. Ngn2 expression in OPCs resulted in their lineage re-specification and transition through an immature neuronal morphology into mature pyramidal cortical neurons with spiny dendrites, axons, synaptic contacts, and subtype specification matching local cytoarchitecture. Lineage re-specification of rat cortical OPCs occurred without prior injury, demonstrating these glial progenitor cells need not be put into a reactive state to achieve lineage reprogramming. These results show it may be feasible to precisely engineer additional neurons directly in adult cerebral cortex for experimental study or potentially for therapeutic use to modify dysfunctional or damaged circuitry.

Evaluation of BCAS1-positive immature oligodendrocytes after cerebral ischemic stroke and SVD

Ischemic cerebrovascular disease is an important cause of physical disability and dementia. Oligodendrocytes (OLGs), which differentiate from oligodendrocyte precursor cells (OPCs), are crucial for remyelination of the damaged brain and functional recovery. Breast carcinoma amplified sequence 1 (BCAS1) has recently been shown to be highly expressed in newly formed pre-myelinating oligodendrocytes (pre-mOLGs), while its expression level is reduced in mature OLGs. In this study, we analyzed BCAS1 expression by immunohistochemical analysis of human post-mortem brain tissue from six stroke patients (death within 2 months after stroke onset) and eight small vessel disease (SVD) patients. Control post-mortem brain tissue was from eight age-matched patients without any obvious central nervous system (CNS) pathology. The Olig2 expression in the area corresponding to the same section of the BCAS1-stained slice was analyzed to determine the total oligodendrocyte lineage. The percentage of differentiating OPCs in the oligodendrocyte lineage was calculated as the ratio of BCAS1+ to Olig2+ cells (BCAS1+/Olig2+). The stroke and SVD cases showed demyelination with decreased expression of myelin basic protein (MBP, a mature OLG marker). The stroke cases showed significantly increased numbers of early-stage BCAS1+ cells with an immature morphology and Olig2+ cells (pan-oligodendrocyte lineages) in the peri-infarct areas in both the cortex and white matter, but showed no increase in the number of late-stage BCAS1+ cells with a mature morphology. In contrast, the SVD cases showed no significant increase in Olig2+ and BCAS1+ cells. These results indicated that remyelination dysfunction could be attributed to insufficient maturation of OPCs in stroke and impaired recruitment of OPCs in SVD.

PlexinD1 signaling controls domain-specific dendritic development in newborn neurons in the postnatal olfactory bulb.

Newborn neurons show immature bipolar morphology and continue to migrate toward their destinations. After the termination of migration, newborn neurons undergo spatially controlled dendrite formation and change into a complex morphology. The mechanisms of dendritic development of newborn neurons have not been fully understood. Here, we show that in the postnatal olfactory bulb (OB), the Sema3E-PlexinD1 signaling, which maintains bipolar morphology of newborn neurons, also regulates their dendritic development after the termination of migration in a dendritic domain-specific manner. Genetic ablation of Sema3E or PlexinD1 enhanced dendritic branching in the proximal domain of the apical dendrites of OB newborn granule cells, whereas PlexinD1 overexpression suppressed it in a Rho binding domain (RBD)-dependent manner. Furthermore, RhoJ, a small GTPase that directly binds to PlexinD1RBD in vascular endothelial cells, is expressed in migrating and differentiating newborn granule cells in the OB and is also involved in the suppression of proximal branching of their apical dendrites. These results suggest that the Sema3E-PlexinD1-RhoJ axis regulates domain-specific dendrite formation of newborn neurons in the postnatal OB.

Anurocampa (Lepidoptera: Notodontidae): two new species, systematics and immature stages.

Notodontidae (Lepidoptera, Noctuoidea) comprise over 4,000 described species distributed worldwide, among which nearly half are restricted to the Neotropics. Morphology of adults and immatures of Notodontidae have been broadly investigated and many larval, pupal, and adult characters were found to be synapomorphies of subfamilies and tribes. Despite this, the current classification of Notodontidae remains unsettled as most recent classification systems are contradictory due to reliance on incomplete global sampling and, many taxa, especially in the Neotropics, are still informally classified as incertae sedis. Anurocampa Herrich-Shäffer was recently treated as an incertae sedis genus, and immature and adult characters may provide further evidence for its systematic position among the Notodontidae. With this goal in mind, the present study describes the immature stages of Anurocampa mingens Herrich-Shäffer from Brazil and describes two new species in the genus from Costa Rica based on morphology and mitochondrial DNA: Anurocampa markhastingsi Chacón and St Laurent sp. nov. and Anurocampa abelardochaconi Chacón and St Laurent sp. nov. and discusses the systematic position of Anurocampa.

One generalist or several specialists? Comparative analysis of the polyphagous butterfly Adelpha serpa celerio and its serpa‐group relatives

AbstractPatterns of larval food plant use and immature stage morphology help clarify Neotropical butterfly species diversity, and, in many cases, lineages identified with molecular data are corroborated by morphology and ecology of immature stages. Here, we investigate cryptic species in Adelpha butterflies (Nymphalidae), a group known for challenging adult wing similarity coupled with mimicry, and for its remarkably wide larval food plant breadth. We focus on the Adelpha serpa‐group and in particular Adelpha serpa celerio, whose immatures have been documented feeding on 11 plant families. Our analyses of A. serpa, and among serpa‐group species across the Neotropics, revealed evidence of cryptic species, but the pattern within A. serpa celerio showed relatively weak mtDNA and morphological differences, and no differentiation in food plant use. We conclude that A. serpa should be revised and recognize A. serpa to include the nominate subspecies and recognize A. celerio (revised status) as a geographically widespread species and a larval food plant generalist that contains A. c. celerio (revised status), A. c. duiliae (revised status) and A. c. diadochus (revised status). Four additional species within the serpa‐group show strong evidence of harbouring allopatric cryptic species, and further research should be done to clarify their species limits.

Cytokine-Induced Myeloid-Derived Suppressor Cells Demonstrate Their Immunoregulatory Functions to Prolong the Survival of Diabetic Mice.

Type 1 diabetes is an inflammatory state. Myeloid-derived suppressive cells (MDSCs) originate from immature myeloid cells and quickly expand to control host immunity during infection, inflammation, trauma, and cancer. This study presents an ex vivo procedure to develop MDSCs from bone marrow cells propagated from granulocyte-macrophage-colony-stimulating factor (GM-CSF), interleukin (IL)-6, and IL-1β cytokines expressing immature morphology and high immunosuppression of T-cell proliferation. The adoptive transfer of cytokine-induced MDSCs (cMDSCs) improved the hyperglycemic state and prolonged the diabetes-free survival of nonobese diabetic (NOD) mice with severe combined immune deficiency (SCID) induced by reactive splenic T cells harvested from NOD mice. In addition, the application of cMDSCs reduced fibronectin production in the renal glomeruli and improved renal function and proteinuria in diabetic mice. Moreover, cMDSCs use mitigated pancreatic insulitis to restore insulin production and reduce the levels of HbA1c. In conclusion, administering cMDSCs propagated from GM-CSF, IL-6, and IL-1β cytokines provides an alternative immunotherapy protocol for treating diabetic pancreatic insulitis and renal nephropathy.

HIV-2 Immature Particle Morphology Provides Insights into Gag Lattice Stability and Virus Maturation

Retrovirus immature particle morphology consists of a membrane enclosed, pleomorphic, spherical and incomplete lattice of Gag hexamers. Previously, we demonstrated that human immunodeficiency virus type 2 (HIV-2) immature particles possess a distinct and extensive Gag lattice morphology. To better understand the nature of the continuously curved hexagonal Gag lattice, we have used the single particle cryo-electron microscopy method to determine the HIV-2 Gag lattice structure for immature virions. The reconstruction map at 5.5 Å resolution revealed a stable, wineglass-shaped Gag hexamer structure with structural features consistent with other lentiviral immature Gag lattice structures. Cryo-electron tomography provided evidence for nearly complete ordered Gag lattice structures in HIV-2 immature particles. We also solved a 1.98 Å resolution crystal structure of the carboxyl-terminal domain (CTD) of the HIV-2 capsid (CA) protein that identified a structured helix 12 supported via an interaction of helix 10 in the absence of the SP1 region of Gag. Residues at the helix 10–12 interface proved critical in maintaining HIV-2 particle release and infectivity. Taken together, our findings provide the first 3D organization of HIV-2 immature Gag lattice and important insights into both HIV Gag lattice stabilization and virus maturation.

LAG3+ erythroid progenitor cells inhibit HBsAg seroclearance during finite pegylated interferon treatment through LAG3 and TGF-β

HBsAg seroclearance, the ideal aim of anti-hepatitis B virus (HBV) treatment, cannot be achieved easily. Anemia is another common issue for chronic hepatitis B (CHB) patients, which leads to elevation of erythroid progenitor cells (EPCs) and immune suppression in cancer. This study investigated the role of EPCs in HBsAg seroclearance following pegylated interferon-α (PEG-IFN) treatment. CD45+EPC accumulation in CHB patients and an AAV/HBV mice model was found in the circulation and liver by flow cytometry and immunofluorescence tests. Wright-Giemsa staining showed that these pathological CD45+EPCs presented elevated erythroid cells with relative immature morphologies and atypical cells compared with the control cells. CD45+EPCs were associated with immune tolerance and decreased HBsAg seroclearance during finite PEG-IFN treatment. CD45+EPCs suppressed antigen non-specific T cell activation and HBV-specific CD8+T cells, partially through transforming growth factor β (TGF-β). RNA-seq revealed that CD45+EPCs in patients with CHB presented a distinct gene expression profile compared with CD45−EPCs and CD45+EPCs from cord blood. Notably, CD45+EPCs from patients with CHB expressed high level of Lymphocyte-activation gene 3 (LAG3), an immune checkpoint molecule, and were then defined as LAG3+EPCs. LAG3+EPCs diminished the function of antigen presenting cells through LAG3, which was another mechanism by which LAG3+EPCs’ suppressed HBV-specific CD8+T cells. Anti-LAG3 and anti-TGF-β combination treatment decreased serum HBeAg, HBV DNA levels and HBsAg level, as well as HBsAg-expression in hepatocytes during PEG-IFN treatment in the AAV/HBV mice model. Conclusions: LAG3+EPCs inhibited the efficacy of PEG-IFN treatment on HBsAg seroclearance induced by LAG3 and TGF-β. Anti-LAG3, anti-TGF-β and PEG-IFN combination treatment might facilitate HBV clearance.

Altered striatal actin dynamics drives behavioral inflexibility in a mouse model of fragile X syndrome

The proteome of glutamatergic synapses is diverse across the mammalian brain and involved in neurodevelopmental disorders (NDDs). Among those is fragile X syndrome (FXS), an NDD caused by the absence of the functional RNA-binding protein FMRP. Here, we demonstrate how the brain region-specific composition of postsynaptic density (PSD) contributes to FXS. In the striatum, the FXS mouse model shows an altered association of the PSD with the actin cytoskeleton, reflecting immature dendritic spine morphology and reduced synaptic actin dynamics. Enhancing actin turnover with constitutively active RAC1 ameliorates these deficits. At the behavioral level, the FXS model displays striatal-driven inflexibility, a typical feature of FXS individuals, which is rescued by exogenous RAC1. Striatal ablation of Fmr1 is sufficient to recapitulate behavioral impairments observed in the FXS model. These results indicate that dysregulation of synaptic actin dynamics in the striatum, a region largely unexplored in FXS, contributes to the manifestation of FXS behavioral phenotypes.

The effect of single-cell knockout of Fragile X Messenger Ribonucleoprotein on synaptic structural plasticity.

Fragile X Syndrome (FXS) is the best-known form of inherited intellectual disability caused by the loss-of-function mutation in a single gene. The FMR1 gene mutation abolishes the expression of Fragile X Messenger Ribonucleoprotein (FMRP), which regulates the expression of many synaptic proteins. Cortical pyramidal neurons in postmortem FXS patient brains show abnormally high density and immature morphology of dendritic spines; this phenotype is replicated in the Fmr1 knockout (KO) mouse. While FMRP is well-positioned in the dendrite to regulate synaptic plasticity, intriguing in vitro and in vivo data show that wild type neurons embedded in a network of Fmr1 KO neurons or glia exhibit spine abnormalities just as neurons in Fmr1 global KO mice. This raises the question: does FMRP regulate synaptic morphology and dynamics in a cell-autonomous manner, or do the synaptic phenotypes arise from abnormal pre-synaptic inputs? To address this question, we combined viral and mouse genetic approaches to delete FMRP from a very sparse subset of cortical layer 5 pyramidal neurons (L5 PyrNs) either during early postnatal development or in adulthood. We then followed the structural dynamics of dendritic spines on these Fmr1 KO neurons by in vivo two-photon microscopy. We found that, while L5 PyrNs in adult Fmr1 global KO mice have abnormally high density of thin spines, single-cell Fmr1 KO in adulthood does not affect spine density, morphology, or dynamics. On the contrary, neurons with neonatal FMRP deletion have normal spine density but elevated spine formation at 1 month of age, replicating the phenotype in Fmr1 global KO mice. Interestingly, these neurons exhibit elevated thin spine density, but normal total spine density, by adulthood. Together, our data reveal cell-autonomous FMRP regulation of cortical synaptic dynamics during adolescence, but spine defects in adulthood also implicate non-cell-autonomous factors.

Insights into the systematics of Alluaudinella and allied Aethiopomyia and Ochromusca (Muscidae, Diptera)

AbstractThe muscid genera Alluaudinella Giglio‐Tos, 1895, Aethiopomyia Malloch, 1921 and Ochromusca Malloch, 1927 form a monophyletic group supported by immature and adult morphology and a highly specialised snail‐feeding strategy of immature stages. In contrast to the undoubted monophyly of the Alluaudinella‐Aethiopomyia‐Ochromusca clade, previous studies have provided contradictory hypotheses of the subfamiliar position within the Muscidae, and these three genera have been placed in the subfamily Muscinae, Dichaetomyiinae, Phaoniinae and Reinwardtiinae. The systematic position of Alluaudinella, as a representative of Alluaudinella, Aethiopomyia and Ochromusca group, is revised by means of larval morphology, biology and molecular data. Light microscopy (LM), confocal laser scanning microscopy (CLSM) and scanning electron microscopy (SEM) are used to study the egg and all larval instars of Alluaudinella flavicornis (Macquart, 1855) and a multilocus Sanger sequencing (mS‐seq) approach to examine position within Muscidae. Results are inconsistent with the traditional, morphology‐based concept of the Alluaudinella‐Aethiopomyia‐Ochromusca clade as closely related to Dichaetomyia Malloch, 1921, and the phylogenetic analysis revealed no support for inclusion within subfamily Phaoniinae. Larval morphology in Alluaudinella differs significantly from that of Dichaetomyia (and other Phaoniinae), but resembles that of genera nested in Reinwardtiinae. Based on molecular data and larval morphology a transfer of Alluaudinella, Aethiopomyia and Ochromusca to the subfamily Reinwardtiinae is proposed.