Aqueous angiopoietin-like levels correlate with optical coherence tomography angiography metrics in diabetic macular edema

AIM: To quantitatively detect aqueous levels of angiopoietin-like (ANGPTL)3, ANGPTL4, and ANGPTL6 and investigate their correlation with optical coherence tomography angiography (OCTA) findings in patients with diabetic macular edema (DME). METHODS: This cross-sectional study included 23 patients (27 eyes) with type 2 diabetes and 16 control subjects (20 eyes). All patients underwent OCTA imaging and ultra-wide field fundus photography. Diabetic patients were categorized into two groups according to the presence or absence of diabetic retinopathy (DME group, 14 patients, 16 eyes); and non-diabetic retinopathy (NDR) group, 9 patients, 11 eyes, respectively. Aqueous levels of ANGPTL3, ANGPTL4, and ANGPTL6 were assessed using suspension array technology, and foveal-centered 3×3 mm2 OCTA scans were automatically graded to determine the central, inner, and full vessel density (CVD, IVD, FVD); central, inner, and full perfusion density (CPD, IPD, FPD), foveal avascular zone (FAZ) area, FAZ perimeter, and FAZ circularity index (FAZ-CI) on superficial capillary plexuses. Additionally, central subfield thickness (CST), cube volume (CV), and cube average thickness (CAT) were measured in a model of macular cube 512×128. RESULTS: Aqueous ANGPTL3 levels were not significantly different among the three groups (P>0.05). ANGPTL4 levels were significantly higher in the DME group than the control and NDR groups (P<0.0001 and P<0.001), while ANGPTL6 levels were significantly higher in the DME group than the control group (P<0.05). In the whole cohort, the aqueous ANGPTL3 levels correlated negatively with the IVD, FVD, IPD, and FPD, and positively with the CV and CAT. The aqueous ANGPTL4 levels correlated negatively with the CVD, IVD, FVD, CPD, IPD, and FPD, and positively with the FAZ perimeter, CST, CV, and CAT. The aqueous ANGPTL6 levels correlated negatively with the IVD, FVD, IPD, FPD, FAZ-CI and positively with CST, CV, CAT. CONCLUSION: ANGPTL4 and ANGPTL6 may be associated with vascular leakage in DME and may represent good targets for DME therapy. In addition, OCTA metrics may be useful for evaluating macular ischemia in DME.

New insights into colorectal cancer liver metastasis carcinogenesis and its effect with moxibustion

Background: Chemotherapeutic drugs creates severe adverse reactions for colorectal cancer. Moxibustion confers clinical benefits for postoperative patients undergoing chemotherapy, it will fill the blank period of western medicine treatment and provide useful help for tumor patients to prevent recurrence and metastasis, but the physiological mechanisms behind the antitumor effects are unclear. This study was aimed to determine the effect and characterize the differential cytokines and gene expression profiles in intrasplenic transplanted GFP-HCT-116 cells-induced tumors model by Pre-Mox, Post-Mox and Pre-Post-Mox intervention. Methods: Human CRC cells with GFP fluorescence were implanted via intrasplenic injection into Balb/c nude mice spleens. Moxibustion stimulation was applied to the BL18 and ST36 acupoints. The model control (MC) group were given no treatment. Pre-Mox mice were received moxibustion for 2 weeks before HCT-116 cell injection. Post-Mox mice received moxibustion for 3 weeks after CRC cell injection. Pre-Post-Mox mice received moxibustion for 5 weeks (2 weeks before and 3 weeks after CRC cell injection). Peripheral bloods were collected, pooled and assayed using a RayBio mouse inflammation antibody array. Multi-Analyte Suspension Array was opted for verification. RNA isolated from liver paracancerous tissues from the control group and the experimental groups was subjected to RNA-seq, and then screened out significant differences for in-depth verification. RESULTS: The results showed that moxibustion stimulation increased the survival rate and decreased CRC liver metastasis. With the help of Multi-Analyte Suspension Array and RNA-seq, we screened significant differential expression of cytokines and RNA, then further verified them. The metastasis rate decreased significantly from 100% (10/10, MC group) to 50% (6/12, Pre-Mox group), to 46% (6/13, Post-Mox group), and further to 25% (3/12, Pre-Post-Mox group). Cytokine chips were used significant differences were found in MIP-3α, MDC, IL-6, and IL-1a. Transcriptomic analysis suggested that the low-dose combination of Pre-Mox and Post-Mox modulated larger gene sets than the single treatment. We identified a small subset of genes, like APOA4, IGFBP5, IGFBP6, TIMP1, and MGP, as potential molecular targets involved in the preventive action of the combination of Pre-Mox and Post-Mox. Conclusions: Taken together, the current results provide the first evidence in support of the chemopreventive effect of a combination of Pre-Mox and Post-Mox in CRC. Moreover, the cytokines and transcriptional profile obtained in our study may provide a framework for identifying the mechanisms underlying the carcinogenesis process from colonic cancer to liver metastasis as well as the cancer inhibitory effects and potential molecular targets of Pre-Post-Mox.

Development of a Molecular Serotyping Scheme for Morganella morganii

Morganella morganii, which is often regarded as a human commensal organism, can be an opportunistic pathogen, causing a variety of clinical infections with serious morbidity and mortality. An efficient and convenient method for subtyping and identifying M. morganii strains in epidemiological surveillance and control is urgently needed. Serotyping based on bacterial surface polysaccharide antigens (O-antigen or K-antigens) is a standard subtyping method for many gram-negative bacteria. Here, through whole genome sequencing and comparative genomics analysis of 27 strains, we developed a molecular serotyping scheme based on the genetic variation of O-antigen gene clusters (O-AGC) in M. morganii, and 11 distinct O-AGC types were identified. A conventional serotyping scheme was also developed by the production of antisera and agglutination experiments, which was shown to be perfectly consistent with the molecular serotyping scheme, confirming that the variation in M. morganii O-AGC correlated with phenotypic O-antigen diversification. Furthermore, a microsphere-based suspension array (MSA) with high specificity was developed based on the specific genes within each O-AGC type. The sensitivity of MSA was determined to be 0.1 ng of genomic DNA and 103 CFU of pure culture. We further analyzed 104 M. morganii genomes available in GenBank, and an additional six novel O-AGC types were identified, indicating that the extension of this molecular serotyping scheme is convenient. Our work provides an important tool for the detection and epidemiological surveillance of M. morganii, and this method has the potential to be widely utilized, especially for bacterial genera/species without an efficient typing approach.

Suspension array platform based on aptamer for high-throughput detection of five environmental hormones

Background As a common small molecule substance, environmental hormones widely exist in nature, especially water sources, which have a profound effects in humans. Highly efficient and sensitive method for estrogens in the environment are essential. Results In this paper, a novel high-throughput platform was established based on five small hormones molecules specificity aptamer and magnetic beads (MBs). The results showed that the sensitivity of the proposed method are greatly improved. The limit of detection(LOD) of this method for atrazine(Atz), profenofos, bisphenolA(BPA), estradiol(E2), and polychlorinated biphenyls(PCBs) were 9.46, 20.75, 23.81, 8.97, 6.27 pg/mL, respectively. The Recovery rate of the diluted environmental hormones spiked in the samples of Haihe river were in the range of 87.5-111.02% with relative standard deviations (RSDs) lower than 28.44%. Conclusion This platform based on new complementary strand fragments can simultaneously rapid detection five environmental hormones. The whole procedure completed within 1.5h including sample treatment, incubation and detection, greatly improving the detection efficiency.

Aqueous Flare, Functional-Morphological Parameters, and Cytokines in Age-Related Macular Degeneration after Anti-VEGF Treatment

Purpose: The role of inflammation and cytokines in AMD and anti-Vascular Endothelial Growth Factor (anti-VEGF) treatment remains unclear. Therefore, this study aimed to examine whether anti-VEGF treatment for exudative Age-related Macular Degeneration (AMD) affects aqueous flare value (an indicator of inflammation), functional-morphologic parameters, and aqueous humor levels of cytokines or inflammatory mediators. Methods: We compared aqueous humor levels of 8 cytokines, growth factors (including VEGF), and inflammatory mediators in 43 patients who received anti-VEGF treatment with aflibercept for AMD and 24 healthy controls by the suspension array method. In addition, we measured aqueous flare values with a laser flare meter and Central Macular Thickness (CMT) and Macular Volume (MV) by optical coherence tomography. Results: The patient group had a significantly higher aqueous flare value than the control group. At baseline, CMT showed significant correlations with aqueous humor levels of soluble intercellular adhesion molecule-1 (sICAM-1), monocyte chemoattractant protein 1 (MCP-1), interleukin (IL)-6, and IL-8 and MV, with aqueous humor levels of VEGF, sICAM-1, MCP-1, IL-6, and IL-8. Moreover, we found significant correlations between aqueous flare value and aqueous humor levels of MCP-1, IL-6, IL-8, and interferon-gamma–inducible protein 10. One month after anti-VEGF treatment, the patient group showed a significant correlation between the change in MV and improvement in best-corrected visual acuity (BCVA); CMT showed no such correlation. Conclusion: Inflammation appears to be involved in AMD. Change in MV may be an index of improvement in BCVA in patients receiving anti-VEGF treatment for AMD.

Cytokine Profiles Associated With Worse Prognosis in a Hospitalized Peruvian COVID-19 Cohort

Cytokines, chemokines and growth factors present different expression profiles related to the prognosis of COVID-19. We analyzed clinical parameters and assessed the expression of these biomarkers in patients with different disease severity in a hospitalized Peruvian cohort to determine those associated with worse prognosis. We measured anti-spike IgG antibodies by ELISA and 30 cytokines by quantitative suspension array technology in 123 sera samples. We analyzed differences between patients with moderate, severe and fatal COVID-19 by logistic regression at baseline and in longitudinal samples. Significant differences were found among the clinical parameters: hemoglobin, neutrophils, lymphocytes and C-reactive protein (CRP), creatinine and D-dimer levels. Higher anti-spike IgG antibody concentrations were associated to fatal patient outcomes. At hospitalization, IL-10, IL-6, MIP-1α, GM-CSF, MCP-1, IL-15, IL-5, IL1RA, TNFα and IL-8 levels were already increased in fatal patients´ group. Meanwhile, multivariable analysis revealed that increased GM-CSF, MCP-1, IL-15, and IL-8 values were associated with fatal outcomes. Moreover, longitudinal analysis identified IL-6 and MCP-1 as the main risk factors related to mortality in hospitalized COVID-19 patients. In this Peruvian cohort we identified and validated biomarkers related to COVID-19 outcomes. Further studies are needed to identify novel criteria for stratification of SARS-CoV-2 infected patients at hospital entry. BackgroundIn the most severe forms of SARS-CoV-2 infection, large numbers of innate and adaptive immune cells become activated and begin to produce pro-inflammatory cytokines, establishing an exacerbated feedback loop of inflammation.MethodsA total of 55 patients with laboratory-confirmed COVID-19 admitted to the Hospital Nacional Guillermo Almenara Irigoyen in Lima, Peru were enrolled during August-October 2020. Of these, 21 had moderate disease, 24 severe diseases and 10 died. We measured 30 cytokines and chemokines by quantitative suspension array technology and anti-spike IgG antibodies using a commercial ELISA. We evaluated these parameters in peripheral blood every 2-5 days until patient discharge or death. Patient information and clinical parameters related were obtained from the respective clinical histories.ResultsThe frequency of obesity differed among the 3 groups, being most frequent in patients who died. There were also significant differences in clinical parameters: hemoglobin, segmented neutrophils, lymphocytes,C-reactive protein, creatinine and D-dimer levels. Greater anti-spike IgG antibody concentrations were associated to fatal outcomes. In univariate analyses, higher baseline concentrations of IL-6, MIP-1α, GM-CSF, MCP-1, IL-15, IL-5, IL1RA, TNFα, IL-8 and IL-12p70 correlated with severity, while multivariable analysis showed that increased concentrations in 4 biomarkers (GM-CSF, MCP-1, IL-15, IL-8) were associated with fatal outcomes. Longitudinal analysis showed IL-6 (hazard ratio [HR] 6.81, 95% confidence interval [CI] 1.6-28.7) and MCP-1 (HR 4.61, 95%CI 1.1-19.1) to be related to mortality in hospitalized COVID-19 patients.ConclusionsCytokine, chemokine and growth factor profiles were identified and validated related to severity and outcomes of COVID-19. Our findings may be useful to identify novel criteria for COVID-19 patient stratification at hospital entry.

Association of Variants in FCGR2A, PTPN2 and GM-CSF with Cerebral Cavernous Malformation: Potential Biomarkers for Symptomatic Disease

Backgrounds: Cerebral cavernous malformations (CCM) predispose patients to a lifetime risk of seizures and symptomatic hemorrhage. Only a small percentage of affected people will develop clinical symptoms, and molecular mechanisms underlying lesional activity remain unclear. We have analyzed a panel of Single Nucleotide Polymorphisms (SNPs) in CCM patients and looked for plasmatic inflammatory cytokines checking for a pattern of plasmatic expression heterogeneity and any correlation with the genetic variation identified with different CCM clinical phenotypes. Methods: This is a case-control study from a long-term follow-up cohort, including 23 CCM patients, 16 symptomatic, and 7 asymptomatic patients. A 200 SNP´s panel was performed through next-generation sequencing and eighteen different plasma molecules were assessed through a suspension array system. Results: Fcγreceptor IIa rs1801274 (FCGR2A) and Protein tyrosine phosphatase non-receptor type 2 rs72872125 PTPN2 were found statically different between groups. Patients who had the combination of the presence of FCGR2A and the absence of PTPN2 also had symptoms earlier in their lifetime. The combination of the genetic polymorphisms and serum level of GM-CSF had resulted in the best diagnostic biomarker to distinguish symptomatic patients as formulated: [0.296*(FCGR2A)] + [-0.788*(PTPN2)] + [-0.107*(GM-CSF)]. Conclusion: We have shown that SNPs in inflammation genes might be related to the symptomatic phenotype in CCM. We also demonstrated that a formula based on two of these polymorphisms (FCGR2A+ and PTPN2+) was possibly capable of predicting a symptomatic phenotype during a patient’s lifetime.

A Comprehensive View on the Human Antibody Repertoire Against Staphylococcus aureus Antigens in the General Population

Our goal was to provide a comprehensive overview of the antibody response to Staphylococcus aureus antigens in the general population as a basis for defining disease-specific profiles and diagnostic signatures. We tested the specific IgG and IgA responses to 79 staphylococcal antigens in 996 individuals from the population-based Study of Health in Pomerania. Using a dilution-based multiplex suspension array, we extended the dynamic range of specific antibody detection to seven orders of magnitude, allowing the precise quantification of high and low abundant antibody specificities in the same sample. The observed IgG and IgA antibody responses were highly heterogeneous with differences between individuals as well as between bacterial antigens that spanned several orders of magnitude. Some antigens elicited significantly more IgG than IgA and vice versa. We confirmed a strong influence of colonization on the antibody response and quantified the influence of sex, smoking, age, body mass index, and serum glucose on anti-staphylococcal IgG and IgA. However, all host parameters tested explain only a small part of the extensive variability in individual response to the different antigens of S. aureus.

Reduced Placental Transfer of Antibodies Against a Wide Range of Microbial and Vaccine Antigens in HIV-Infected Women in Mozambique

Transplacental transfer of antibodies is essential for conferring protection in newborns against infectious diseases. We assessed the impact of different factors, including gestational age and maternal infections such as HIV and malaria, on the efficiency of cord blood levels and placental transfer of IgG subclasses. We measured total IgG and IgG subclasses by quantitative suspension array technology against 14 pathogens and vaccine antigens, including targets of maternal immunization, in 341 delivering HIV-uninfected and HIV-infected mother-infant pairs from southern Mozambique. We analyzed the association of maternal HIV infection, Plasmodium falciparum exposure, maternal variables and pregnancy outcomes on cord antibody levels and transplacental transfer. Our results show that maternal antibody levels were the main determinant of cord antibody levels. Univariable and multivariable analysis showed that HIV reduced the placental transfer and cord levels of IgG and IgG1 principally, but also IgG2 to half of the antigens tested. P. falciparum exposure and prematurity were negatively associated with cord antibody levels and placental transfer, but this was antigen-subclass dependent. Our findings suggest that lower maternally transferred antibodies may underlie increased susceptibility to infections of HIV-exposed infants. This could affect efficacy of maternal vaccination, especially in sub-Saharan Africa, where there is a high prevalence of HIV, malaria and unfavorable environmental factors.