Punctate Signals Research Articles

Immunoexpression Pattern of Autophagy-Related Proteins in Human Congenital Anomalies of the Kidney and Urinary Tract.

The purpose of this study was to evaluate the spatiotemporal immunoexpression pattern of microtubule-associated protein 1 light chain 3 beta (LC3B), glucose-regulated protein 78 (GRP78), heat shock protein 70 (HSP70), and lysosomal-associated membrane protein 2A (LAMP2A) in normal human fetal kidney development (CTRL) and kidneys affected with congenital anomalies of the kidney and urinary tract (CAKUT). Human fetal kidneys (control, horseshoe, dysplastic, duplex, and hypoplastic) from the 18th to the 38th developmental week underwent epifluorescence microscopy analysis after being stained with antibodies. Immunoreactivity was quantified in various kidney structures, and expression dynamics were examined using linear and nonlinear regression modeling. The punctate expression of LC3B was observed mainly in tubules and glomerular cells, with dysplastic kidneys displaying distinct staining patterns. In the control group's glomeruli, LAMP2A showed a sporadic, punctate signal; in contrast to other phenotypes, duplex kidneys showed significantly stronger expression in convoluted tubules. GRP78 had a weaker expression in CAKUT kidneys, especially hypoplastic ones, while normal kidneys exhibited punctate staining of convoluted tubules and glomeruli. HSP70 staining varied among phenotypes, with dysplastic and hypoplastic kidneys exhibiting stronger staining compared to controls. Expression dynamics varied among observed autophagy markers and phenotypes, indicating their potential roles in normal and dysfunctional kidney development.

PIP2 regulating calcium signal modulates actin cytoskeleton-dependent cytoadherence and cytolytic capacity in the protozoan parasite Trichomonas vaginalis.

Trichomonas vaginalis is a prevalent causative agent that causes trichomoniasis leading to uropathogenic inflammation in the host. The crucial role of the actin cytoskeleton in T. vaginalis cytoadherence has been established but the associated signaling has not been fully elucidated. The present study revealed that the T. vaginalis second messenger PIP2 is located in the recurrent flagellum of the less adherent isolate and is more abundant around the cell membrane of the adherent isolates. The T. vaginalis phosphatidylinositol-4-phosphate 5-kinase (TvPI4P5K) with conserved activity phosphorylating PI(4)P to PI(4, 5)P2 was highly expressed in the adherent isolate and partially colocalized with PIP2 on the plasma membrane but with discrete punctate signals in the cytoplasm. Plasma membrane PIP2 degradation by phospholipase C (PLC)-dependent pathway concomitant with increasing intracellular calcium during flagellate-amoeboid morphogenesis. This could be inhibited by Edelfosine or BAPTA simultaneously repressing parasite actin assembly, morphogenesis, and cytoadherence with inhibitory effects similar to the iron-depleted parasite, supporting the significance of PIP2 and iron in T. vaginalis colonization. Intriguingly, iron is required for the optimal expression and cell membrane trafficking of TvPI4P5K for in situ PIP2 production, which was diminished in the iron-depleted parasites. TvPI4P5K-mediated PIP2 signaling may coordinate with iron to modulate T. vaginalis contact-dependent cytolysis to influence host cell viability. These observations provide novel insights into T. vaginalis cytopathogenesis during the host-parasite interaction.

Independent and Complementary Functions of Caf1b and Hir1 for Chromatin Assembly in Tetrahymena thermophila.

Histones and DNA associate to form the nucleosomes of eukaryotic chromatin. Chromatin assembly factor 1 (CAF-1) complex and histone regulatory protein A (HIRA) complex mediate replication-couple (RC) and replication-independent (RI) nucleosome assembly, respectively. CHAF1B and HIRA share a similar domain but play different roles in nucleosome assembly by binding to the different interactors. At present, there is limited understanding for the similarities and differences in their respective functions. Tetrahymena thermophila contains transcriptionally active polyploid macronuclei (MAC) and transcriptionally silent diploid micronuclei (MIC). Here, the distribution patterns of Caf1b and Hir1 exhibited both similarities and distinctions. Both proteins localized to the MAC and MIC during growth, and to the MIC during conjugation. However, Hir1 exhibited additional signaling on parental MAC and new MAC during sexual reproduction and displayed a punctate signal on developing anlagen. Caf1b and Hir1 only co-localized in the MIC with Pcna1 during conjugation. Knockdown of CAF1B impeded cellular growth and arrested sexual reproductive development. Loss of HIR1 led to MIC chromosome defects and aborted sexual development. Co-interference of CAF1B and HIR1 led to a more severe phenotype. Moreover, CAF1B knockdown led to the up-regulation of HIR1 expression, while knockdown of HIR1 also led to an increase in CAF1B expression. Furthermore, Caf1b and Hir1 interacted with different interactors. These results showed that CAF-1 and Hir1 have independent and complementary functions for chromatin assembly in T. thermophila.

Assessment of cheese sign and its association with vascular risk factors: Data from PUMCH dementia cohort.

In the clinic, practitioners encounter many patients with an abnormal pattern of dense punctate magnetic resonance imaging (MRI) signal in the basal ganglia, a phenomenon known as "cheese sign". This sign is reported as common in cerebrovascular diseases, dementia, and old age. Recently, cheese sign has been speculated to consist of dense perivascular space (PVS). This study aimed to assess the lesion types of cheese sign and analyze the correlation between this sign and vascular disease risk factors. A total of 812 patients from Peking Union Medical College Hospital (PUMCH) dementia cohort were enrolled. We analyzed the relationship between cheese sign and vascular risk. For assessing cheese sign and defining its degree, the abnormal punctate signals were classified into basal ganglia hyperintensity (BGH), PVS, lacunae/infarctions and microbleeds, and counted separately. Each type of lesion was rated on a four-level scale, and then the sum was calculated; this total was defined as the cheese sign score. Fazekas and Age-Related White Matter Changes (ARWMC) scores were used to evaluate the paraventricular, deep, and subcortical gray/white matter hyperintensities. A total of 118 patients (14.5%) in this dementia cohort were found to have cheese sign. Age (odds ratio [OR]: 1.090, 95% confidence interval [CI]: 1.064-1.120, P <0.001), hypertension (OR: 1.828, 95% CI: 1.123-2.983, P = 0.014), and stroke (OR: 1.901, 95% CI: 1.092-3.259, P = 0.025) were risk factors for cheese sign. There was no significant relationship between diabetes, hyperlipidemia, and cheese sign. The main components of cheese sign were BGH, PVS, and lacunae/infarction. The proportion of PVS increased with cheese sign severity. The risk factors for cheese sign were hypertension, age, and stroke. Cheese sign consists of BGH, PVS, and lacunae/infarction.

Genome-Wide Analysis of Exocyst Complex Subunit Exo70 Gene Family in Cucumber.

Cucumber (Cucumis sativus L.) is an important vegetable worldwide, but its yield is affected by a wide range of pathogens and pests. As the major subunit of the exocyst complex, the roles of Exo70 members have been shown in Arabidopsis and rice, but their function are unknown in cucumber. Here, we identified 18 CsExo70 members in cucumber, which were divided into three groups (Exo70.1-Exo70.3) and nine subgroups (Exo70A-Exo70I) based on the phylogenetic tree. Subsequently, systematical analyses were performed, including collinearity, gene structure, cis-acting elements, conserved motifs, expression patterns, and subcellular localization. Our results showed that CsExo70 genes were generally expressed in all tissues, and CsExo70C1 and CsExo70C2 were highly expressed in the stamen. Moreover, the expression levels of most CsExo70 genes were induced by Pseudomonas syringae pv. lachrymans (Psl) and Fusarium oxysporum f. sp. cucumerinum Owen (Foc), especially CsExo70E2 and CsExo70H3. In addition, these CsExo70s displayed similar location patterns with discrete and punctate signals in the cytoplasm. Together, our results indicate that CsExo70 members may be involved in plant development and resistance, and provide a reference for future in-depth studies of Exo70 genes in cucumber.

Histopathologic and molecular characterization of Erethizon dorsatum papillomavirus 1 and Erethizon dorsatum papillomavirus 2 infection in North American porcupines (Erethizon dorsatum).

Erethizon dorsatum papillomavirus 1 (EdPV1) and Erethizon dorsatum papillomavirus 2 (EdPV2) are associated with cutaneous papillomas in North American porcupines (Erethizon dorsatum). This study defined gross, histopathologic, and molecular characteristics of viral papillomas in 10 North American porcupines submitted to the New York State Animal Health Diagnostic Center. Investigation for the presence of EdPV1 and EdPV2 DNA via polymerase chain reaction (PCR) was performed in 9 of the 10 (90.0%) porcupines, and all porcupines were investigated for the detection and localization of EdPV1 and EdPV2 E6 and E7 nucleic acid via chromogenic in situ hybridization (CISH). Next-generation sequencing (NGS) was performed in 2 porcupines. Papillomas were diagnosed on the muzzle (n = 4), caudal dorsum (n = 1), upper lip (n = 1), chin (n = 1), gingiva (n = 2), and nasal planum (n = 1). Histologically, the lesions consisted of hyperplastic epidermis or epithelium with orthokeratotic keratin, prominent keratohyalin granules, and intranuclear inclusion bodies. PCR identified EdPV1 in 6 of 9 samples and EdPV2 in the remaining 3 samples. NGS resulted in 100% genome coverage of EdPV1 and 76.20% genome coverage of EdPV2 compared with GenBank reference sequences, with 99.8% sequence identity to the complete EdPV2 L1 gene of a novel subtype recently identified in France. Hybridization patterns in 9 of the 10 (90.0%) porcupines were characterized by strong nuclear signals in the superficial epidermis, with strong nuclear and punctate cytoplasmic signals in the stratum spinosum and basale. In one animal, CISH suggested dual EdPV1 and EdPV2 infection.

Establishment of a rabbit liver metastasis model by percutaneous puncture of the spleen and implantation of the VX2 tumor strain under CT guidance

This study aimed to compare the feasibility, success rate, and safety of establishing a rabbit VX2 liver metastasis model by percutaneous splenic implantation under CT guidance and open splenic implantation of the VX2 tumor strain. Fifty-two New Zealand white rabbits were randomly divided into group A (the percutaneous puncture group) (n = 26) and group B (the laparotomy group) (n = 26). In group A, 26 New Zealand white rabbits were implanted with tumor strains by percutaneous splenic puncture under CT guidance. In group B, 26 New Zealand white rabbits were implanted with tumor strains in the spleen by laparotomy. After 2–3 weeks of implantation, both group A and group B underwent MRI to confirm tumor growth in the spleen and metastasis to the liver. Two experimental rabbits randomly selected from groups A and B were killed for pathological examination. The success rate, complication rate, and operation time in groups A and B were compared and analyzed. A total of 23 rabbits in group A were successfully induced, and the success rate was 88.5% (23/26). The average time of operation was 14.42 ± 3.26 min. A total of 22 rabbits in group B were successfully induced, and the success rate was 84.6% (22/26). The average time of operation in group B was 23.69 ± 5.27 min. There was no significant difference in the success rate of induction between the two groups (P > 0.05). The MRI manifestations of liver metastases were multiple nodular and punctate abnormal signal shadows in the liver. Hematoxylin–eosin (HE) staining showed a large number of tumor cells in the tumor area. CT-guided percutaneous splenic implantation of the VX2 tumor strain to establish a rabbit liver metastasis model is a minimally invasive and feasible inducing method. The success rate of this technique is not lower than that of open splenic implantation, with low incidence of complications, and short operation time.

Effect of heat-reinforcing needling on expression of P2X7R, NLRP3 and Caspase-1 in synovial tissues of knee joint in rabbits with cold syndrome rheumatoid arthritis

To observe the effects of heat-reinforcing needling on the expression of purinergic ligand-gated ion channel 7 receptor(P2X7R)，Nodlike receptor protein 3(NLRP3) and Caspase-1 in synovium tissues of knee joint of rabbits with cold syndrome rheumatoid arthritis(RA)，so as to explore the anti-inflammatory mechanism of heat reinforcing needling in the treatment of RA. Thirty male New Zealand white rabbits were randomly divided into normal group, model group, reinforcing-reducing needling group(RRG), heat-reinforcing needling group(HRG), and antagonist group(AG), with 6 rabbits in each group.The model of cold syndrome RA was established by ovalbumin combined with Freund's adjuvant and cryogenic freezing. Rabbits of RRG and HRG were treated with corresponding acupuncture techniques on both sides of "Zusanli"(ST36) for 30 min, once a day for 7 days; Rabbits of the AG was intraperitoneally injected with A438079(2.5 mg/kg), once a day for 7 days. After intervention, color Doppler ultrasound was used to observe the joint cavity effusion, synovial thickness and internal blood flow signal.The histomorphological changes of synovial tissues were observed by HE staining. Quantitative real-time PCR method was used to detect the mRNA expression levels of P2X7R, NLRP3, and Caspase-1 in synovial tissues. Western blot was used to detect the protein expression of interleukin(IL)-1β, IL-6 and tumor necrosis factor(TNF)-α in synovial tissues. Compared with the normal group, the synovial tissues in the model group was thickened, linear and punctate blood flow signals were increased, joint effusion was obvious, synovial coating cells were enlarged, the synovial matrix was severely hyperplasia, inflammatory cells infiltration was obvious.The mRNAs expression levels of P2X7R, NLRP3, Caspase-1 and IL-1β, IL-6, TNF-α proteins in synovial tissues were significantly increased(P<0.01). Compared with model group, abnormal blood flow signals, synovial thickness, joint effusion, proliferation of synovial matrix, inflammatory cell infiltration and synovial coating cells in the RRG, HRG and AG were improved. The mRNAs expression levels of P2X7R, NLRP3, Caspase-1 and IL-1β, IL-6, TNF-α proteins in synovial tissues were decreased in the RRG, HRG and AG (P<0.05, P<0.01). Compared with the RRG, the above indexes were lower in the HRG and AG (P<0.05, P<0.01).There was no significant difference in all indexes above between the HRG and AG (P<0.01, P<0.05). Heat reinforcing needling can improve synovial inflammation of RA, which may be related to regulating the expressions of P2X7R/NLRP3/Caspase-1 signaling pathway.

Logbook of a Retinal Autograft.

This study aimed to evaluate the prognosis of a retinal autograft that was used to treat a macular hole that occurred after repeated vitreoretinal surgeries. A patient underwent repeated vitreoretinal surgery due to retinal detachment in the right eye, and the internal limiting membrane was also removed during the surgeries. After silicone oil removal, the patient developed recurrent retinal detachment and macular hole, and for this reason a retinal autograft was applied to the macular hole and silicone tamponade was administered. The silicone oil was removed at postoperative 7 months, and the macular hole was observed to be closed on fundus examination at 18 months. The final visual acuity was 5/100. On optical coherence tomography (OCT), the hole in the detached retina was measured as 600 µm in diameter preoperatively, 1020 µm on the first postoperative day, gradually narrowed to 765 µm, and graft integration occurred. During follow-up, the accumulation of hyperreflective spots persisted on the inner surface of the graft tissue and in all vertically extending sections. In en face sections, it appeared as a hyperreflective arc between the graft and host retina with a shadowing artefact. In OCT angiography evaluation, a punctate multiple blood flow signal in the vertical axis of the graft was detected in the early phase at 3 months. This finding persisted at 1 month after silicone removal, and the flow signal disappeared with resorption of the cystic edema. These flow signals were in the same location as the areas of hyperreflective spots on structural OCT. In conclusion, structural OCT and OCT angiography are effective methods for the follow-up of retinal autograft integration into host tissue.

Visualizing Influenza A Virus vRNA Replication.

Influenza A virus (IAV) has caused recurrent epidemics and severe pandemics. In this study, we adapted an MS2-MCP live-cell imaging system to visualize IAV replication. A reporter plasmid, pHH-PB2-vMSL, was constructed by replacing a part of the PB2-coding sequence in pHH-PB2 with a sequence encoding 24 copies of a stem-loop structure from bacteriophage MS2 (MSL). Binding of MS2 coat protein (MCP) fused to green fluorescent protein (GFP) to MSL enabled the detection of vRNA as fluorescent punctate signals in live-cell imaging. The introduction of pHH-PB2-vMSL into A549 cells transduced to express an MCP-GFP fusion protein lacking the nuclear localization signal (MCP-GFPdN), subsequently allowed tracking of the distribution and replication of PB2-vMSL vRNA after IAV PR8 infection. Spatial and temporal measurements revealed exponential increases in vRNA punctate signal intensity, which was only observed after membrane blebbing in apoptotic cells. Similar signal intensity increases in apoptotic cells were also observed after MDCK cells, transduced to express MCP-GFPdN, were infected with IAV carrying PB2-vMSL vRNA. Notably, PB2-vMSL vRNA replication was observed to occur only in apoptotic cells, at a consistent time after apoptosis initiation. There was a lack of observable PB2-vMSL vRNA replication in non-apoptotic cells, and vRNA replication was suppressed in the presence of apoptosis inhibitors. These findings point to an important role for apoptosis in IAV vRNA replication. The utility of the MS2-imaging system for visualizing time-sensitive processes such as viral replication in live host cells is also demonstrated in this study.

Postmortem brain 7T MRI with minimally invasive pathological correlation in deceased COVID-19 subjects

BackgroundBrain abnormalities are a concern in COVID-19, so we used minimally invasive autopsy (MIA) to investigate it, consisting of brain 7T MR and CT images and tissue sampling via transethmoidal route with at least three fragments: the first one for reverse transcription polymerase chain reaction (RT-PCR) analysis and the remaining fixed and stained with hematoxylin and eosin. Two mouse monoclonal anti-coronavirus (SARS-CoV-2) antibodies were employed in immunohistochemical (IHC) reactions.ResultsSeven deceased COVID-19 patients underwent MIA with brain MR and CT images, six of them with tissue sampling. Imaging findings included infarcts, punctate brain hemorrhagic foci, subarachnoid hemorrhage and signal abnormalities in the splenium, basal ganglia, white matter, hippocampi and posterior cortico-subcortical. Punctate brain hemorrhage was the most common finding (three out of seven cases). Brain histological analysis revealed reactive gliosis, congestion, cortical neuron eosinophilic degeneration and axonal disruption in all six cases. Other findings included edema (5 cases), discrete perivascular hemorrhages (5), cerebral small vessel disease (3), perivascular hemosiderin deposits (3), Alzheimer type II glia (3), abundant corpora amylacea (3), ischemic foci (1), periventricular encephalitis foci (1), periventricular vascular ectasia (1) and fibrin thrombi (1). SARS-CoV-2 RNA was detected with RT-PCR in 5 out of 5 and IHC in 6 out 6 patients (100%).ConclusionsDespite limited sampling, MIA was an effective tool to evaluate underlying pathological brain changes in deceased COVID-19 patients. Imaging findings were varied, and pathological features corroborated signs of hypoxia, alterations related to systemic critically ill and SARS-CoV-2 brain invasion.

MRI Imaging Omics and Risk Factors Analysis of PWMD in Premature Infants Based on Fuzzy Clustering Algorithm.

The magnetic resonance imaging (MRI) characteristics of periventricular white matter damage (PWMD) in premature infants using the fuzzy c-means clustering algorithm (FCM) is explored, and the influencing factors are further clarified. A total of 100 premature infants admitted to the neonatal department of our hospital from February 2020 to February 2022 are selected for in-depth investigation. According to the occurrence of PWMD, they are divided into the PWMD group and the simple premature delivery group, with 50 cases in each group. All preterm infants are examined by MRI and the changes in image characteristics and apparent diffusion coefficient (ADC) values are analyzed. Clinical information of the subjects is collected and the influencing factors of PWMD in preterm infants are analyzed by multivariate regression analysis. In the first magnetic resonance imaging (MRI) examination, the cases of punctured, clustered, and linear lesions are 28 cases, 12 cases, and 10 cases, respectively. The experimental results showed that PWMD of preterm infants presented punctate, clustered, and high linear T1 signal MRI manifestations, which caused a downward trend of ADC value, and caused respiratory distress, low birth weight, premature rupture of membranes, respiratory tract infection, and other risk symptoms.

Localization of AD biomarker phospho-tau proteins in the brains of an App knock-in mouse model of Aβ amyloidosis.

Amyloid-β (Aβ) plaques, neurofibrillary tangles composed of hyper-phosphorylated tau proteins and neurodegeneration are neuropathological hallmarks of Alzheimer's disease (AD). Recently, phospho-tau proteins in blood as well as cerebrospinal fluid have been reported as promising biomarkers in the preclinical stage of AD. However, physiological and pathological roles of these phosphorylated tau proteins in AD pathogenesis remain elusive. In this study, we investigated the localization of AD biomarker phospho-tau proteins in the brains of App knock-in (App NLGF ) mouse, which is thought to mimic important aspects in the early phase of AD. To investigate the localization of AD biomarker phospho-tau proteins in the brains of aged (24- month-old) App NLGF and wild-type mice, we prepared frozen brain sections and co-immunostained with phospho-tau specific antibodies and reactive gliosis or synaptic marker antibodies. We found that one AD biomarker phospho-tau signals were detected in the axonal structures in the brains of wild-type mice, and these structures were disrupted in the brains of App NLGF mice. In contrast, the other AD biomarker phospho-tau epitopes were only detected in the brains of App NLGF mice as punctate signals around Aβ plaques. These phospho-tau signals were co-localized with post-synaptic marker PSD95, but not with reactive gliosis or pre-synaptic markers. Our results suggest that AD biomarker phospho-tau epitopes may represent axonal degeneration or Aβ plaque-related post-synaptic pathology in the early phase of AD brains.

Detection of HPV and Human Chromosome Sites by Dual-Color Fluorescence In Situ Hybridization Reveals Recurrent HPV Integration Sites and Heterogeneity in Cervical Cancer.

Human papillomavirus (HPV) integration in the human genome is suggested to be an important cause of cervical cancer. With the development of sequencing technologies, an increasing number of integration “hotspots” have been identified. However, this HPV integration information was derived from analysis of whole cervical cancer tissue, and we know very little about the integration in different cancer cell subgroups or individual cancer cells. This study optimized the preparation of probes and provided a dual-color fluorescence in situ hybridization (FISH) method to detect HPV integration sites in paraffin-embedded cervical cancer samples. We used both HPV probes and site-specific probes: 3p14 (FHIT), 8q24 (MYC), 13q22 (KLF5/KLF12), 3q28 (TP63), and 5p15 (TERT). We detected HPV signals in 75 of the 96 cases of cervical cancer; 62 cases showed punctate signals, and 13 cases showed diffuse punctate signals. We identified 3p14 as a high-frequency HPV integration site in 4 cervical cancer cases. HPV integration at 8p14 occurred in 2 cases of cervical cancer. In the same cervical cancer tissue of sample No.1321, two distinct subgroups of cells were observed based on the HPV probe but showed no difference in cell and nucleus morphology. Our study provides a new method to investigate the frequent HPV integration sites in cervical cancer and reports the heterogeneity within cervical cancer from the perspective of HPV integration.

Involvement of Felis catus papillomavirus type 2 in the tumorigenesis of feline Merkel cell carcinoma

Merkel cell carcinoma (MCC) is a cutaneous neuroendocrine tumor. We recently demonstrated that cats with MCC often have other proliferative cutaneous lesions, such as squamous cell carcinoma (SCC) and basal cell carcinoma (BCC). Based on this finding, we hypothesize that Felis catus papillomavirus (FcaPV) is involved in the development of MCC in cats, similar to SCC and BCC. To investigate this hypothesis, the presence of FcaPV nucleic acid and immunoreactivity for tumor suppressor proteins were examined in 21 feline MCC cases. Polymerase chain reaction using FcaPV type-specific primers detected FcaPV2 DNA in 20/21 samples of MCC. The complete FcaPV2 sequence was characterized in one case. In situ hybridization for FcaPV2 E7 revealed punctate nuclear signals within tumor cells in 19/21 MCC. Increased immunoreactivity for p16CDKN2A protein and decreased immunoreactivity for retinoblastoma (pRb) and p53 proteins were observed in 20/21 MCC. These results suggest that feline MCC cases are infected with FcaPV2 and the subsequent inhibition of pRb and p53 induced by integrated viral oncogenes is associated with feline MCC tumorigenesis, similar to other PV-induced proliferative cutaneous lesions. On the other hand, the single case of FcaPV2-negative MCC showed strong p53 immunoreactivity, suggesting mutations in p53 caused by cancer inducers other than FcaPV2 infection in this case. The present study suggests FcaPV2 as a cause of feline MCC.

Molecular characterization and cellular localization of a transmembrane C-type lectin receptor in hemocytes from the scallop Argopecten purpuratus

Recognition of potential pathogens is a crucial function of the immune system of every organism. In this context, great efforts have been made in the last years to characterize the diversity of pattern recognition receptors (PRRs) present in cultured aquatic species, such as bivalve mollusks. Among the great diversity of PRRs described so far, C-type lectin receptors (CLRs) comprise a large family of innate immune receptors that can recognize different carbohydrates present on the cell surface of microorganisms. CLRs can be found extracellularly in soluble form, or transmembrane, located on immune cell surfaces. To date, almost only soluble CLRs have been described in bivalves and so far, no transmembrane CLRs are identified in pectinids. In the present study, a sequence showing structural homologies to transmembrane CLRs (tmCLR) was characterized from scallop Argopecten purpuratus hemocytes, named ApCLR. The cDNA sequence displays 948 nucleotides, coding for a deduced protein of 316 amino acids with a molecular weight of 35.24 kDa. The protein sequence showed characteristic domains of a tmCLR, such as a signal peptide, an extracellular N-terminal carbohydrate recognition domain (CRD), one transmembrane domain and a cytoplasmic tail. Phylogenetic analysis revealed that ApCLR groups with soluble CLRs from scallops but also with transmembrane CLRs from vertebrates, such as chondrolectins and layilins. Gene expression analysis showed that ApCLR was constitutively present in gill, mantle, digestive gland, gonad and hemocyte tissues from non-immune challenged scallops and it was most strongly expressed in the gill and mantle tissues. Furthermore, ApCLR was significantly upregulated in scallop hemocytes at 6 h after scallop injection with heat-killed Vibrio splendidus. At the protein level, ApCLR is located at the plasma membrane of hemocytes, observed as punctate signals by immunofluorescence and confocal analysis. Results obtained in this study reveal the first transmembrane CLR in scallops. Also, results showed that ApCLR is present at the surface of hemocytes and increase its transcript availability during the immune challenge, suggesting a potential role in the recognition of microorganisms.

The efficiency of susceptibility-weighted MRI in the differentiation of endometriomas from haemorrhagic ovarian cysts.

The aim of the study was to investigate the efficiency of susceptibility-weighted magnetic resonance (MR) imaging (SWIs) in differentiating endometriomas from haemorrhagic ovarian cysts. Between July 2017 and January 2019, 89 ovarian cystic lesions (57 endometriomas and 32 haemorrhagic cysts) that were identified as complicated cystic lesions on ultrasonography (US) and underwent lower abdominal MRI with susceptibility weighting were retrospectively evaluated. Final diagnoses were obtained with surgical pathological correlation and radiological-clinical follow-up. Two radiologists blinded to the final diagnoses retrospectively reviewed the images in consensus. The signal intensity on T1- and T2-weighted images and curved linear or punctate signal void areas on SWI were noted for the presence of lesions. Forty of the 57 endometriomas demonstrated the defined MRI criteria, including a cystic hyperintensity on T1-weighted images and hypointensity on T2-weighted images. The remaining 17 lesions did not demonstrate these criteria on conventional MR images. SWI showed punctate or curved linear signal void areas in 53 of 57 endometriomas (92.9%) and none of the haemorrhagic cysts. The sensitivity, specificity and accuracy of SWI in differentiating endometrioma from haemorrhagic cyst were 92.9%, 100.0%, and 95.5%, respectively. The addition of the SWI sequence to conventional MRI can help distinguish endometriomas from haemorrhagic ovarian cysts.

Diagnostic value of susceptibility-weighted imaging for endometrioma: preliminary results from a retrospective analysis.

Endometrioma is a common manifestation of endometriosis that can be difficult to diagnose with conventional magnetic resonance imaging (MRI). Susceptibility-weighted imaging (SWI) may be more sensitive than conventional MRI in the detection of chronic, local hemorrhagic disease. To investigate whether signal voids in SWI sequences could be used in the preoperative diagnosis of endometrioma. This retrospective study included consecutive female patients with clinically suspected endometrioma. All patients underwent pelvic 3-T MRI (T1- and T2-weighted) and SWI within two weeks before laparoscopy. Two experienced radiologists blinded to the histopathologic/clinical diagnoses interpreted the images together, and any disagreements were resolved by consensus. The final analysis included 73 patients: 46 patients (mean age=37 years; age range=22-68 years) with 85 endometrioma lesions and 27 patients (mean age=34 years; age range=15-68 years) with 34 non-endometrioid cystic lesions (18 hemorrhagic corpus luteal cysts, three simple cysts, three mucinous cystadenomas, two mature teratomas, and one endometrioid cyst with corpus luteum rupture/hemorrhage). The presenting symptoms for patients with endometrioma were chronic pelvic pain (44.6%), dysmenorrhea (31.9%), infertility (12.8%), dyspareunia (6.4%), and menstrual irregularity (4.3%). MRI identified all 119 lesions observed laparoscopically. SWI visualized punctate or curvilinear signal voids along the cyst wall or within the lesion in 67 of 85 endometriomas (78.8%) and only 3 of 31 non-endometrioid cysts (8.8%). The use of SWI to look for signal voids in the cyst wall or within the lesion could facilitate the preoperative diagnosis of endometrioma.

QSM is an imaging biomarker for chronic glial activation in multiple sclerosis lesions.

BackgroundInflammation in chronic active lesions occurs behind a closed blood–brain barrier and cannot be detected with MRI. Activated microglia are highly enriched for iron and can be visualized with quantitative susceptibility mapping (QSM), an MRI technique used to delineate iron.ObjectiveTo characterize the histopathological correlates of different QSM hyperintensity patterns in MS lesions.MethodsMS brain slabs were imaged with MRI and QSM, and processed for histology. Immunolabeled cells were quantified in the lesion rim, center, and adjacent normal‐appearing white matter (NAWM). Iron+ myeloid cell densities at the rims were correlated with susceptibilities. Human‐induced pluripotent stem cell (iPSC)‐derived microglia were used to determine the effect of iron on the production of reactive oxygen species (ROS) and pro‐inflammatory cytokines.ResultsQSM hyperintensity at the lesion perimeter correlated with activated iron+ myeloid cells in the rim and NAWM. Lesions with high punctate or homogenous QSM signal contained no or minimally activated iron− myeloid cells. In vitro, iron accumulation was highest in M1‐polarized human iPSC‐derived microglia, but it did not enhance ROS or cytokine production.ConclusionA high QSM signal outlining the lesion rim but not punctate signal in the center is a biomarker for chronic inflammation in white matter lesions.

Combined kinesin-1 and kinesin-3 activity drives axonal trafficking of TrkB receptors in Rab6 carriers.

SummaryNeurons depend on proper localization of neurotrophic receptors in their distal processes for their function. The Trk family of neurotrophin receptors controls neuronal survival, differentiation, and remodeling and are well known to function as retrograde signal carriers transported from the distal axon toward the cell body. However, the mechanism driving anterograde trafficking of Trk receptors into the axon is not well established. We used microfluidic compartmental devices and inducible secretion assay to systematically investigate the retrograde and anterograde trafficking routes of TrkB receptor along the axon in rat hippocampal neurons. We show that newly synthesized TrkB receptors traffic through the secretory pathway and are directly delivered into axon. We found that these TrkB carriers associate and are regulated by Rab6. Furthermore, the combined activity of kinesin-1 and kinesin-3 is needed for the formation of axon-bound TrkB secretory carriers and their effective entry and processive anterograde transport beyond the proximal axon.