The microRNA pathway has been implicated in the regulation of synaptic protein synthesis and ultimately in dendritic spine morphogenesis, a phenomenon associated with long-lasting forms of memory. ...However, the particular microRNAs (miRNAs) involved are largely unknown. Here we identify specific miRNAs that function at synapses to control dendritic spine structure by performing a functional screen. One of the identified miRNAs, miR-138, is highly enriched in the brain, localized within dendrites and negatively regulates the size of dendritic spines in rat hippocampal neurons. miR-138 controls the expression of acyl protein thioesterase 1 (APT1), an enzyme regulating the palmitoylation status of proteins that are known to function at the synapse, including the alpha(13) subunits of G proteins (Galpha(13)). RNA-interference-mediated knockdown of APT1 and the expression of membrane-localized Galpha(13) both suppress spine enlargement caused by inhibition of miR-138, suggesting that APT1-regulated depalmitoylation of Galpha(13) might be an important downstream event of miR-138 function. Our results uncover a previously unknown miRNA-dependent mechanism in neurons and demonstrate a previously unrecognized complexity of miRNA-dependent control of dendritic spine morphogenesis.
Accumulating evidence suggests that oxidation-specific epitopes (OSEs) constitute a novel class of damage-associated molecular patterns (DAMPs) generated during high oxidative stress but also in the ...physiological process of apoptosis. To deal with the potentially harmful consequences of such epitopes, the immune system has developed several mechanisms to protect from OSEs and to orchestrate their clearance, including IgM natural antibodies and both cellular- and membrane-bound receptors. Here, we focus on malondialdehyde (MDA) epitopes as prominent examples of OSEs that trigger both innate and adaptive immune responses. First, we review the mechanisms of MDA generation, the different types of adducts on various biomolecules and provide relevant examples for physiological carriers of MDA such as apoptotic cells, microvesicles, or oxidized low-density lipoproteins. Based on recent insights, we argue that MDA epitopes contribute to the maintenance of homeostatic functions by acting as markers of elevated oxidative stress and tissue damage. We discuss multiple lines of evidence that MDA epitopes are proinflammatory and thus important targets of innate and adaptive immune responses. Finally, we illustrate the relevance of MDA epitopes in human pathologies by describing their capacity to drive inflammatory processes in atherosclerosis and highlighting protective mechanisms of immunity that could be exploited for therapeutic purposes.
The gold standard for the diagnosis of cholera is stool culture, but this requires laboratory facilities and takes at least 24 hours. A rapid diagnostic test (RDT) that can be used by minimally ...trained staff at treatment centers could potentially improve the reporting and management of cholera outbreaks.
We evaluated the Crystal VC™ RDT under field conditions in Zanzibar in 2009. Patients presenting to treatment centers with watery diarrhea provided a stool sample for rapid diagnostic testing. Results were compared to stool culture performed in a reference laboratory. We assessed the overall performance of the RDT and evaluated whether previous intake of antibiotics, intravenous fluids, location of testing, and skill level of the technician affected the RDT results.
We included stool samples from 624 patients. Compared to culture, the overall sensitivity of the RDT was 93.1% (95%CI: 88.7 to 96.2%), specificity was 49.2% (95%CI: 44.3 to 54.1%), the positive predictive value was 47.0% (95%CI: 42.1 to 52.0%) and the negative predictive value was 93.6% (95%CI: 89.6 to 96.5%). The overall false positivity rate was 50.8% (213/419); fieldworkers frequently misread very faint test lines as positive.
The observed sensitivity of the Crystal VC RDT evaluated was similar compared to earlier versions, while specificity was poorer. The current version of the RDT could potentially be used as a screening tool in the field. Because of the high proportion of false positive results when field workers test stool specimens, positive results will need to be confirmed with stool culture.
Respiratory viruses such as influenza do not typically cause viremia; however, SARS-CoV-2 has been detected in the blood of COVID-19 patients with mild and severe symptoms. Detection of SARS-CoV-2 in ...blood raises questions about its role in pathogenesis as well as transfusion safety concerns. Blood donor reports of symptoms or a diagnosis of COVID-19 after donation (post-donation information, PDI) preceded or coincided with increased general population COVID-19 mortality. Plasma samples from 2,250 blood donors who reported possible COVID-19-related PDI were tested for the presence of SARSCoV-2 RNA. Detection of RNAemia peaked at 9%-15% of PDI donors in late 2020 to early 2021 and fell to approximately 4% after implementation of widespread vaccination in the population. RNAemic donors were 1.2- to 1.4-fold more likely to report cough or shortness of breath and 1.8-fold more likely to report change in taste or smell compared with infected donors without detectable RNAemia. No infectious virus was detected in plasma from RNAemic donors; inoculation of permissive cell lines produced less than 0.7-7 plaque-forming units (PFU)/mL and in susceptible mice less than 100 PFU/mL in RNA-positive plasma based on limits of detection in these models. These findings suggest that blood transfusions are highly unlikely to transmit SARS-CoV-2 infection.
Diet‐related health issues such as nonalcoholic fatty liver disease and cardiovascular disorders are known to have a major inflammatory component. However, the exact pathways linking diet‐induced ...changes (e.g., hyperlipidemia) and the ensuing inflammation have remained elusive so far. We identified biological processes related to innate immunity and oxidative stress as prime response pathways in livers of low‐density lipoprotein receptor‐deficient mice on a Western‐type diet using RNA sequencing and in silico functional analyses of transcriptome data. The observed changes were independent of the presence of microbiota and thus indicative of a role for sterile triggers. We further show that malondialdehyde (MDA) epitopes, products of lipid peroxidation and markers for enhanced oxidative stress, are detectable in hepatic inflammation predominantly on dying cells and stimulate cytokine secretion as well as leukocyte recruitment in vitro and in vivo. MDA‐induced cytokine secretion in vitro was dependent on the presence of the scavenger receptors CD36 and MSR1. Moreover, in vivo neutralization of endogenously generated MDA epitopes by intravenous injection of a specific MDA antibody results in decreased hepatic inflammation in low‐density lipoprotein receptor‐deficient mice on a Western‐type diet. Conclusion: Accumulation of MDA epitopes plays a major role during diet‐induced hepatic inflammation and can be ameliorated by administration of an anti‐MDA antibody. (Hepatology 2017;65:1181‐1195)
Otavi Group is a 1.5–3.5-km-thick epicontinental marine carbonate succession of Neoproterozoic age, exposed in an 800-km-long Ediacaran−Cambrian fold belt that rims the SW cape of Congo craton in ...northern Namibia. Along its southern margin, a contiguous distally tapered foreslope carbonate wedge of the same age is called Swakop Group. Swakop Group also occurs on the western cratonic margin, where a crustal-scale thrust cuts out the facies transition to the platformal Otavi Group.
Subsidence accommodating Otavi Group resulted from S−N crustal stretching (770–655 Ma), followed by post-rift thermal subsidence (655–600 Ma). Rifting under southern Swakop Group continued until 650–635 Ma, culminating with breakup and a S-facing continental margin. No hint of a western margin is evident in Otavi Group, suggesting a transform margin to the west, kinematically consistent with S−N plate divergence. Rift-related peralkaline igneous activity in southern Swakop Group occurred around 760 and 746 Ma, with several rift-related igneous centres undated. By comparison, western Swakop Group is impoverished in rift-related igneous rocks.
Despite low paleoelevation and paleolatitude, Otavi and Swakop groups are everywhere imprinted by early and late Cryogenian glaciations, enabling unequivocal stratigraphic division into five epochs (period divisions): (1) non-glacial late Tonian, 770–717 Ma; (2) glacial early Cryogenian/Sturtian, 717–661 Ma; (3) non-glacial middle Cryogenian, 661–646 ± 5 Ma; (4) glacial late Cryogenian/Marinoan, 646 ± 5–635 Ma; and (5) non-glacial early Ediacaran, 635–600 ± 5 Ma. Odd numbered epochs lack evident glacioeustatic fluctuation; even numbered ones were the Sturtian and Marinoan snowball Earths. This study aimed to deconstruct the carbonate succession for insights on the nature of Cryogenian glaciations. It focuses on the well-exposed southwestern apex of the arcuate fold belt, incorporating 585 measured sections (totaling >190 km of strata) and > 8764 pairs of δ13C/δ18Ocarb analyses (tabulated in Supplementary On-line Information).
Each glaciation began and ended abruptly, and each was followed by anomalously thick ‘catch-up’ depositional sequences that filled accommodation space created by synglacial tectonic subsidence accompanied by very low average rates of sediment accumulation. Net subsidence was 38% larger on average for the younger glaciation, despite its 3.5–9.3-times shorter duration. Average accumulation rates were subequal, 4.0 vs 3.3–8.8 m Myr−1, despite syn-rift tectonics and topography during Sturtian glaciation, versus passive-margin subsidence during Marinoan. Sturtian deposits everywhere overlie an erosional disconformity or unconformity, with depocenters ≤1.6 km thick localized in subglacial rift basins, glacially carved bedrock troughs and moraine-like buildups. Sturtian deposits are dominated by massive diamictite, and the associated fine-grained laminated sediments appear to be local subglacial meltwater deposits, including a deep subglacial rift basin. No marine ice-grounding line is required in the 110 Sturtian measured sections in our survey.
In contrast, the newly-opened southern foreslope was occupied by a Marinoan marine ice grounding zone, which became the dominant repository for glacial debris eroded from the upper foreslope and broad shallow troughs on the Otavi Group platform, which was glaciated but left nearly devoid of glacial deposits. On the distal foreslope, a distinct glacioeustatic falling-stand carbonate wedge is truncated upslope by a glacial disconformity that underlies the main lowstand grounding-zone wedge, which includes a proximal 0.60-km-high grounding-line moraine. Marinoan deposits are recessional overall, since all but the most distal overlie a glacial disconformity. The Marinoan glacial record is that of an early ice maximum and subsequent slow recession and aggradation, due to tectonic subsidence. Terminal deglaciation is recorded by a ferruginous drape of stratified diamictite, choked with ice-rafted debris, abruptly followed by a syndeglacial-postglacial cap-carbonate depositional sequence. Unlike its Sturtian counterpart, the post-Marinoan sequence has a well-developed basal transgressive (i.e., deepening-upward) cap dolomite (16.9 m regional average thickness, n = 140) with idiosyncratic sedimentary features including sheet-crack marine cements, tubestone stromatolites and giant wave ripples. The overlying deeper-water calci-rhythmite includes crystal-fans of former aragonite benthic cement ≤90 m thick, localized in areas of steep sea-floor topography. Marinoan sequence stratigraphy is laid out over ≥0.6 km of paleobathymetric relief.
Late Tonian shallow-neritic δ13Ccarb records were obtained from the 0.4-km-thick Devede Fm (~770–760 Ma) in Otavi Group and the 0.7-km-thick Ugab Subgroup (~737–717 Ma) in Swakop Group. Devede Fm is isotopically heavy, +4–8‰ VPDB, and could be correlative with Backlundtoppen Fm (NE Svalbard). Ugab Subgroup post-dates 746 Ma volcanics and shows two negative excursions bridged by heavy δ13C values. The negative excursions could be correlative with Russøya and Garvellach CIEs (carbon isotope excursions) in NE Laurentia.
Middle Cryogenian neritic δ13C records from Otavi Group inner platform feature two heavy plateaus bracketed by three negative excursions, correlated with Twitya (NW Canada), Taishir (Mongolia) and Trezona (South Australia) CIEs. The same pattern is observed in carbonate turbidites in distal Swakop Group, with the sub-Marinoan falling-stand wedge hosting the Trezona CIE recovery. Proximal Swakop Group strata equivalent to Taishir CIE and its subsequent heavy plateau are shifted bidirectionally to uniform values of +3.0–3.5‰.
Early Ediacaran neritic δ13C records from Otavi Group inner platform display a deep negative excursion associated with the post-Marinoan depositional sequence and heavy values (≤ + 11‰) with extreme point-to-point variability (≤10‰) in the youngest Otavi Group formation. Distal Swakop Group mimics older parts of the early Ediacaran inner platform δ13C records, but after the post-Marinoan negative excursion, proximal Swakop Group values are shifted bidirectionally to +0.9 ± 1.5‰. Destruction of positive and negative CIEs in proximal Swakop Group is tentatively attributed to early seawater-buffered diagenesis (dolomitization), driven by geothermal porewater convection that sucks seawater into the proximal foreslope of the platform. This hypothesis provocatively implies that CIEs originating in epi-platform waters and shed far downslope as turbidites are decoupled from open-ocean DIC (dissolved inorganic carbon), which is recorded by the altered proximal Swakop Group values closer to DIC of modern seawater.
Carbonate sedimentation ended when the cratonic margins collided with and were overridden by the Atlantic coast-normal Northern Damara and coast-parallel Kaoko orogens at 0.60–0.58 Ga. A forebulge disconformity separates Otavi/Swakop Group from overlying foredeep clastics. In the cratonic cusp, where the orogens meet at a right angle, the forebulge disconformity has an astounding ≥1.85 km of megakarstic relief, and km-thick mass slides were displaced gravitationally toward both trenches, prior to orogenic shortening responsible for the craton-rimming fold belt.
Abstract
STUDY QUESTION
What is the impact of variants in the genes INSL3 (Insulin Like 3) and RXFP2 (Relaxin Family Peptide Receptor 2), respectively, on cryptorchidism and male infertility?
SUMMARY ...ANSWER
Bi-allelic loss-of-function (LoF) variants in INSL3 and RXFP2 result in bilateral cryptorchidism and male infertility, whereas heterozygous variant carriers are phenotypically unaffected.
WHAT IS KNOWN ALREADY
The small heterodimeric peptide INSL3 and its G protein-coupled receptor RXFP2 play a major role in the first step of the biphasic descent of the testes, and variants in the INSL3 and RXFP2 genes have long been implicated in inherited cryptorchidism. However, only one single homozygous missense variant in RXFP2 has clearly been linked to familial bilateral cryptorchidism, so the effects of bi-allelic variants in INSL3 and heterozygous variants in both genes on cryptorchidism and male infertility remain unclear.
STUDY DESIGN, SIZE, DURATION
Exome data of 2412 men from the MERGE (Male Reproductive Genomics) study cohort including 1902 infertile men with crypto-/azoospermia, of whom 450 men had a history of cryptorchidism, were screened for high-impact variants in INSL3 and RXFP2.
PARTICIPANTS/MATERIALS, SETTING, METHODS
For patients with rare, high-impact variants in INSL3 and RXFP2, detailed clinical data were collected and the testicular phenotype was determined. Genotyping of family members was performed to analyse the co-segregation of candidate variants with the condition. Immunohistochemical staining for INSL3 in patient testicular tissue and measuring serum INSL3 concentration was performed to analyse the functional impact of a homozygous loss-of-function variant in INSL3. For a homozygous missense variant in RXFP2, its impact on the protein’s cell surface expression and ability to respond to INSL3 in CRE reporter gene assay was determined.
MAIN RESULTS AND THE ROLE OF CHANCE
This study presents homozygous high-impact variants in INSL3 and RXFP2 and clearly correlates these to bilateral cryptorchidism. Functional impact of the identified INSL3 variant was demonstrated by absence of INSL3-specific staining in patients’ testicular Leydig cells as well as undetectable blood serum levels. The identified missense variant in RXFP2 was demonstrated to lead to reduced RXFP2 surface expression and INSL3 mediated receptor activation.
LIMITATIONS, REASONS FOR CAUTION
Further investigations are needed to explore a potential direct impact of bi-allelic INSL3 and RXFP2 variants on spermatogenesis. With our data, we cannot determine whether the infertility observed in our patients is a direct consequence of the disruption of a possible function of these genes on spermatogenesis or whether it occurs secondarily due to cryptorchidism.
WIDER IMPLICATIONS OF THE FINDINGS
In contrast to previous assumptions, this study supports an autosomal recessive inheritance of INSL3- and RXFP2-related bilateral cryptorchidism while heterozygous LoF variants in either gene can at most be regarded as a risk factor for developing cryptorchidism. Our findings have diagnostic value for patients with familial/bilateral cryptorchidism and additionally shed light on the importance of INSL3 and RXFP2 in testicular descent and fertility.
STUDY FUNDING/COMPETING INTEREST(S)
This study was carried out within the frame of the German Research Foundation (DFG) funded by Clinical Research Unit ‘Male Germ Cells: from Genes to Function’ (DFG, CRU326). Research at the Florey was supported by an NHMRC grant (2001027) and the Victorian Government Operational Infrastructure Support Program. A.S.B. is funded by the DFG (‘Emmy Noether Programme’ project number 464240267). The authors declare no conflict of interest.
TRIAL REGISTRATION NUMBER
N/A.
A coronavirus antigen microarray (COVAM) was constructed containing 11 SARS-CoV-2, 5 SARS-1, 5 MERS, and 12 seasonal coronavirus recombinant proteins. The array is designed to measure immunoglobulin ...isotype and subtype levels in serum or plasma samples against each of the individual antigens printed on the array. We probed the COVAM with COVID-19 convalescent plasma (CCP) collected from 99 donors who recovered from a PCR+ confirmed SARS-CoV-2 infection. The results were analyzed using two computational approaches, a generalized linear model (glm) and random forest (RF) prediction model, to classify individual specimens as either Reactive or non-reactive against the SARS-CoV-2 antigens. A training set of 88 pre-COVID-19 specimens (PreCoV) collected in August 2019 and102 positive specimens from SARS-CoV-2 PCR+ confirmed COVID-19 cases was used for these analyses. Results compared with an FDA emergency use authorized (EUA) SARS-CoV2 S1-based total Ig chemiluminescence immunoassay (Ortho Clinical Diagnostics VITROS Anti-SARS-CoV-2 Total, CoV2T) and with a SARS-CoV-2 S1-S2 spike-based pseudovirus micro neutralization assay (SARS-CoV-2 reporter viral particle neutralization titration (RVPNT) showed high concordance between the three assays. Three CCP specimens that were negative by the VITROS CoV2T immunoassay were also negative by both COVAM and the RVPNT assay. Concordance between VITROS CoV2T and COVAM was 96%, VITROS CoV2T and RVPNT 93%, and RVPNT and COVAM 91%. The discordances were all weakly reactive samples near the cutoff threshold of the VITROS CoV2T immunoassay. The multiplex COVAM allows CCP to be grouped according to antibody reactivity patterns against 11 SARS-CoV-2 antigens. Unsupervised K-means analysis, via the gap statistics, as well as hierarchical clustering analysis revealed three main clusters with distinct reactivity intensities and patterns. These patterns were not recapitulated by adjusting the VITROS CoV2T or RVPNT assay thresholds. Plasma classified by COVAM reactivity patterns offers potential to improve CCP therapeutic efficacy CoV2T alone. The use of a SARS-CoV-2 antigen array can qualify CCP for administration as a treatment for acute COVID-19, and interrogate vaccine immunogenicity and performance in preclinical, clinical studies, and routine vaccination to identify antibody responses predictive of protection from infection and disease.
Early in the SARS-CoV-2 pandemic, there was a high level of optimism based on observational studies and small controlled trials that treating hospitalized patients with convalescent plasma from ...COVID-19 survivors (CCP) would be an important immunotherapy. However, as more data from controlled trials became available, the results became disappointing, with at best moderate evidence of efficacy when CCP with high titers of neutralizing antibodies was used early in infection. To better understand the potential therapeutic efficacy of CCP, and to further validate SARS-CoV-2 infection of macaques as a reliable animal model for testing such strategies, we inoculated 12 adult rhesus macaques with SARS-CoV-2 by intratracheal and intranasal routes. One day later, 8 animals were infused with pooled human CCP with a high titer of neutralizing antibodies (RVPN NT50 value of 3,003), while 4 control animals received normal human plasma. Animals were monitored for 7 days. Animals treated with CCP had detectable but low levels of antiviral antibodies after infusion. In comparison to the control animals, CCP-treated animals had similar levels of viral RNA in upper and lower respiratory tract secretions, similar detection of viral RNA in lung tissues by in situ hybridization, but lower amounts of infectious virus in the lungs. CCP-treated animals had a moderate, but statistically significant reduction in interstitial pneumonia, as measured by comprehensive lung histology. Thus overall, therapeutic benefits of CCP were marginal and inferior to results obtained earlier with monoclonal antibodies in this animal model. By highlighting strengths and weaknesses, data of this study can help to further optimize nonhuman primate models to provide proof-of-concept of intervention strategies, and guide the future use of convalescent plasma against SARS-CoV-2 and potentially other newly emerging respiratory viruses.
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