Interleukin 17F Gene Polymorphism as a Potential Protective Factor in the Immunopathology of Ocular Toxoplasmosis.

Ocular toxoplasmosis (OT) is characterised by intraocular inflammation due to Toxoplasma gondii infection. Studies have found that interleukin 17 (IL-17) plays a central role in the pathology of OT. However, nucleotide variability in IL17 and interleukin 17 receptor (IL17R) genes has not been characterised in OT. As cytokine gene polymorphisms may influence the expression of these molecules, the aim of this study was to verify whether IL17A (rs2275913), IL17F (rs763780), IL17RA (rs4819554) and IL17RC (rs708567) polymorphisms are associated with OT in a Brazilian population. This study enrolled 214 patients seropositive for T. gondii (110 with OT and 104 without) and 107 controls. Polymorphisms were identified by PCR-restriction fragment length polymorphism analysis, validated by DNA sequencing with chi-square and multivariate analyses being used to assess possible associations between polymorphisms and OT. Logistic regression under the dominant model revealed a protection factor against OT of the C mutant allele of the IL17F (rs763780) polymorphism. The T/C-C/C genotypes were significantly more common in patients without OT compared to those with OT (p value = 0.0066) and controls (p value = 0.014). Findings from this study suggest that the IL17F polymorphism may have an influence in the immunopathology of OT in Brazilian individuals.

HLA-G, LILRB1 and LILRB2 Variants in Zika Virus Transmission from Mother to Child in a Population from South and Southeast of Brazil.

During the 2015-2016 epidemic, Brazil was the country with the highest rate of Zika virus (ZIKV) infection in the Americas. Twenty-nine percent of pregnant women positive for ZIKV exhibited ultrasound scans with fetus anomalies. Human leukocyte antigen-G (HLA-G) exerts immunoregulatory effects by binding to inhibitory receptors, namely LILRB1 and LILRB2, thus preventing mother-fetus rejection and vertical pathogen transmission. The binding of HLA-G to one of its receptors modulates both innate and adaptive immunity. However, in a viral infection, these molecules may behave as pathogenic mediators shifting the pregnancy environment from an anti-inflammatory profile to a pro-inflammatory phenotype. Genetic mutations might be associated with the change in phenotype. This study aimed to explore the possible role of polymorphic sites in HLA-G, LILRB1 and LILRB2 in mother-fetus ZIKV transmission. Polymorphisms were detected by direct sequencing. Differences in allele and/or genotype frequencies for each SNP analyzed among ZIKV non-transmitting and transmitting mother-child pairs, among ZIKV-transmitting and non-transmitting mothers and between ZIKV-infected and non-infected children were compared by Mid-P exact test or Yates' correction. Significant susceptibility of ZIKV vertical transmission is suggested in ZIKV-transmitting and non-transmitting mothers and ZIKV-infected and non-infected children for LILRB1_rs1061684 T/T (p = 0.03, Pc = 0.06, OR = 12.4; p = 0.008, Pc = 0.016, OR = 16.4) and LILRB1_rs16985478 A/A (p = 0.01, Pc = 0.02, OR = 19.2; p = 0.008, Pc = 0.016, OR = 16.4). HLA-G_rs1710 (p = 0.04, Pc = 0.52, OR = 4.30) was also a susceptibility factor. LILRB2_rs386056 G/A (p = 0.02, Pc = 0.08, OR = 0.07), LILRB2_rs7247451 G/G (p = 0.01, Pc = 0.04, OR = 0.04) and HLAG_rs9380142 T/T (p = 0.04, Pc = 0.52, OR = 0.14) were suggested as protective factors against vertical transmission. The current study suggests that polymorphic sites in the LILRB1 and HLA-G genes might be associated with mother-to-child ZIKV transmission while LILRB2 might be associated with protection against ZIKV transmission in the womb in a population from the south and southeast of Brazil.

TNFα rs1799964 TT genotype may be a susceptibility factor for vertical transmission of Toxoplasma gondii and clinical signs in newborns from pregnant women with acute toxoplasmosis.

BACKGROUND: One of the main impacts of Toxoplasma gondii infection occurs during pregnancy and is related to the vertical transmission of the parasite (congenital toxoplasmosis), which can cause severe clinical outcomes and fetal death. During acute infection, in order to control the rapid replication of tachyzoites, different host immune response genes are activated, and these include cytokine-encoding genes. Considering that polymorphisms in cytokine genes may increase susceptibility to vertical transmission of T. gondii by determining the immune status of the pregnant woman, this study evaluated the influence of polymorphisms of tumor necrosis factor alpha (TNFα) rs1799964 (- 1031) and interleukin 1 beta (IL1ß) rs16944 (- 511) genes on gestational toxoplasmosis and on the vertical transmission of the parasite and verified the allele and genotype frequency of these polymorphisms in pregnant patients whose respective newborn did or did not present clinical abnormalities suggestive of congenital toxoplasmosis. METHODS AND RESULTS: A total of 204 pregnant patients with (n = 114) or without (n = 90) infection by T. gondii were enrolled. No associations were found involving the polymorphisms rs1799964 (- 1031) of the TNFα gene and rs16944 (- 511) of the IL1ß gene with the increased chance of T. gondii infection during pregnancy. However, it was observed that the maternal TT genotype referring to the polymorphism of the TNFα gene seems to influence the vertical transmission of the parasite (P = 0.01; χ2 = 6.05) and the presence of clinical manifestation in newborns from pregnancies with acute toxoplasmosis (P = 0.007; χ2 = 9.68). CONCLUSION: The TNFα rs1799964 TT genotype may act as a susceptibility factor for the vertical transmission of parasite and for the presence of clinical signs in newborns from pregnant women with acute toxoplasmosis.

Lack of association of the KIR and HLA class I ligands with ZIKV infection in south and southeast of Brazil.

BACKGROUND: Zika virus (ZIKV) is an emerging arbovirus associated with foetal malformations and neurological complications. The infection is usually associated with mild symptoms. The comparison between the allelic frequency of polymorphic genes in symptomatic infected individuals in the population can clarify the pathogenic mechanisms of ZIKV. During ZIKV infection, cytokines are produced and natural killer (NK) cells are recruited, whose activation depends on signaling pathways activated by specific receptors, such as killer cell immunoglobulin-like receptors (KIR). These molecules interact with human leukocyte antigen (HLA) class I ligands and are encoded by polymorphic genes. OBJECTIVES: This study aimed to evaluate the frequency of allelic variants of the genes encoding the KIR receptors and their HLA class I ligands in 139 symptomatic ZIKV-patients and 170 controls negative for the virus, and to evaluate the role of these variants for ZIKV susceptibility. METHODS: KIR and HLA class I genes were genotyped using the polymerase chain reaction-sequence specific oligonucleotide (PCR-SSO) technique. FINDINGS: No significant differences in the frequency distribution of KIRs and KIR-HLA in patients compared to controls were observed. MAIN CONCLUSIONS: KIR and its HLA ligands might play a minor role in ZIKV infection in the south and southeast Brazilian individuals.

Influence of interleukin 17 A and 17 F polymorphisms in keratoconus.

BACKGROUND: Until a few years ago, keratoconus was defined as a noninflammatory degenerative disease. However, recent studies have shown that the altered balance between inflammatory cytokines, proteases, and protease inhibitors, as well as free radicals and oxidants, have a crucial role in the pathogenesis of this disease. The aim of this study is to investigate whether interleukin 17 A G197A (rs2275913) and interleukin 17 F T7488C (rs763780) polymorphisms are associated with keratoconus in patients from a population of the northwestern region of the State of São Paulo, Brazil. METHODS AND RESULTS: 35 patients and 61 controls were enrolled. Genotyping of interleukin 17 A G197A and interleukin 17 F T7488C polymorphisms was carried out using the polymerase chain reaction-restriction fragment length polymorphism technique. Statistical analyses were conducted using the chi-square test, and an odds ratio with a 95% confidence interval was also calculated to evaluate the association between polymorphisms and disease. Evaluating interleukin 17 F T7488C, we found that the TT genotype is associated as a risk factor for keratoconus (P = 0.04; OR = 3.01; CI 1.11-8.14). As for evaluating interleukin 17 A G197A, the allele and genotype frequencies between patients and controls were compared and no statistically significant differences were found. CONCLUSIONS: Our data showed that the interleukin 17 F T7488C polymorphisms may exert an influence in keratoconus.

Ocular toxoplasmosis associated with up-regulation of miR-155-5p/miR-29c-3p and down-regulation of miR-21-5p/miR-125b-5p.

Ocular toxoplasmosis (OT) is one of the most common manifestations of Toxoplasma gondii infection and can be related with congenital or acquired infections. OT cause posterior uveitis that cause serious sequelae as complete loss of vision. microRNAs (miRNAs) are small non-coding RNAs, which have regulatory roles in cells by silencing messenger RNA. This study evaluated gene expression of miR-155-5p, miR-146a-5p, miR-21-5p, miR-29c-3p and miR-125b-5p in plasma of 51 patients with ocular toxoplasmosis (OT Group), 26 individuals with asymptomatic toxoplasmosis (AT Group), and 25 healthy individuals seronegative for toxoplasmosis (NC Group). Peripherical blood samples were collected in tube with EDTA for plasma isolation, laboratorial diagnosis for toxoplasmosis and RNA extraction. miRNA expression of each sample was performed by qPCR and values were expressed in Relative Quantification (RQ). Results showed that miR-155-5p and miR-29c-3p were up-expressed in OT patients than AT individuals. On the other hand, miR-21-5p and miR-125b-5p were down-expressed in OT patients. Differences were statistically significant. miR-146a-5p expression was similar in OT patients and AT individuals, without significant difference. In addition, comparative analysis for miRNA levels between AT and OT groups confirms these results. So far, this is the first study to evaluate circulating miRNA levels in ocular toxoplasmosis. These findings may contribute to further studies evaluating the exact role of these miRNAs in the course of infection, which may help in understanding the complex parasite-host interaction and future use in diagnosis, prognosis and therapeutic control in ocular toxoplasmosis.

Anti-A and anti-A,B monoclonal antisera with high titers favor the detection of A weak phenotypes.

OBJECTIVES: This study aimed to evaluate the reactivity and the titers of commercial anti-A and anti-A,B antisera in the detection of A weak antigen expression in human red blood cells. BACKGROUND: Commercial monoclonal antisera for ABO phenotyping are useful reagents allowing the identification of the four main ABO phenotypes (A, B, AB, and O). However, the reactivity of these commercial reagents can not be evident when the A or B antigens are weakly expressed, and these antisera have low titers. METHODS/MATERIALS: Six samples from blood donors and five samples from patients with ABO forward and reverse discrepant phenotyping were evaluated. The ABO phenotyping was carried out with different commercial monoclonal anti-A and anti-A,B antisera under different temperatures, using test tubes and gel column agglutination. RESULTS: Monoclonal anti-A antisera with titers less than 256 and anti-A,B with titers less than 128 failed to detect the weak expression of A antigen in 73% and 67% of the A weak phenotypes, respectively. Titres equal to or higher than 2048 (anti-A) and 1024 (anti-A,B) showed better reactivity, independent of the cell clone. CONCLUSION: Our data indicate that anti-A and anti-A,B antisera with high titers give better reactivity with red blood cells carrying A weak antigen expression.

Human platelet antigen polymorphisms and the risk of chronic Chagas disease cardiomyopathy.

Human platelet antigen (HPA) polymorphisms are considered to be a risk factor for cardiac and vascular diseases, but the role of HPA in chronic Chagas disease cardiomyopathy (CCC) is not available. Therefore, the aim of this study was to investigate the association of HPA polymorphisms, HPA-1, HPA-2, HPA-3, HPA-5 and HPA-15, in the severity of left ventricular systolic dysfunction (LVSD) in CCC patients. For this, 229 CCC patients were separated into three groups: without LVSD, mild/moderate LVSD and severe LVSD. PCR-SSP was performed for HPA genotyping and the risk was assessed using SNPStats software. HPA-1 allele and genotype frequencies were lower in mild/moderate LVSD patients compared to other groups, without statistical significance. After stratified analyzes, the HPA-3a/3b genotype frequency was lower in women with severe LVSD compared to those without LVSD (OR:0.29; 95% CI: 0.10-0.84). In conclusion, HPA-3 variant could be a protection factor for CCC in the female patients.

Effect of ABO blood group on asymptomatic, uncomplicated and placental Plasmodium falciparum infection: systematic review and meta-analysis.

BACKGROUND: Malaria clinical outcomes vary by erythrocyte characteristics, including ABO blood group, but the effect of ABO blood group on asymptomatic, uncomplicated and placental Plasmodium falciparum (P. falciparum) infection remains unclear. We explored effects of ABO blood group on asymptomatic, uncomplicated and placental falciparum infection in the published literature. METHODS: A systematic review and meta-analysis was performed using the preferred reporting items for systematic reviews and meta-analyses guidelines. Articles in Pubmed, Embase, Web of Science, CINAHL and Cochrane Library published before February 04, 2017 were searched without restriction. Studies were included if they reported P. falciparum infection incidence or prevalence, stratified by ABO blood group. RESULTS: Of 1923 articles obtained from the five databases (Embase = 728, PubMed = 620, Web of Science = 549, CINAHL = 14, Cochrane Library = 12), 42 met criteria for systematic review and 37 for meta-analysis. Most studies (n = 30) were cross-sectional, seven were prospective cohort, and five were case-control studies. Meta-analysis showed similar odds of uncomplicated P. falciparum infection among individuals with blood group A (summary odds ratio [OR] 0.96, 15 studies), B (OR 0.89, 15 studies), AB (OR 0.85, 10 studies) and non-O (OR 0.95, 17 studies) as compared to those with blood group O. Meta-analysis of four cohort studies also showed similar risk of uncomplicated P. falciparum infection among individuals with blood group non-O and those with blood group O (summary relative risk [RR] 1.03). Meta-analysis of six studies showed similar odds of asymptomatic P. falciparum infection among individuals with blood group A (OR 1.05), B (OR 1.03), AB (OR 1.23), and non-O (OR 1.07) when compared to those with blood group O. However, odds of active placental P. falciparum infection was significantly lower in primiparous women with non-O blood groups (OR 0.46, 95% confidence interval [CI] 0.23 - 0.69, I2 0.0%, three studies), particularly in those with blood group A (OR 0.41, 95% CI 0.003 - 0.82, I2 1.4%, four studies) than those with blood group O. CONCLUSIONS: This study suggests that ABO blood group may not affect susceptibility to asymptomatic and/or uncomplicated P. falciparum infection. However, blood group O primiparous women appear to be more susceptible to active placental P. falciparum infection.

Hematopoietic Chimera in a Male Blood Donor and His Dizygotic Twin Sister.

Twin hematopoietic chimera in humans is a phenomenon that was discovered accidentally and the prevalence of which remains unclear. The resolution of chimera cases requires studying family medical records, data analysis, and investigations of hematopoietic cells and cells from other tissues. The interactions among ABO, Lewis, and secretor histo-blood group systems are explored to resolve cases of hematopoietic chimera. Here we report a rare case of hematopoietic chimera where twins present a mixed field reaction in the ABO, Rh, and Kidd red blood cell phenotyping. Using red blood cells separated from the mixed field as well as molecular approaches and investigations of family members, we identify inconsistent genotypes with the Mendelian inheritance pattern when comparing the peripheral blood with the buccal epithelium of the male twin and his twin sister. Analysis of the ABO, Lewis, and secretor phenotypes, and genomic DNA from buccal epithelium showed the genotypes ABO*A1.01/ABO*B.01 and FUT2*01N.02/ FUT2*01N.02 in the male twin and the genotypes ABO*O.01.01/ABO*O.01.02 and FUT2*01/FUT2*01 in the female twin. The results of the HLA-DRB1 genotyping showed inconsistency between the male and his twin sister. We conclude that the serological analyses combined with molecular approaches used in this study are good tools to resolve cases of hematopoietic chimera.

Plasma concentrations of CCL3 and CCL4 in the cardiac and digestive clinical forms of chronic Chagas disease.

The aim of this study was to investigate the plasma levels of the CCL3 and CCL4 chemokines in patients with the cardiac and digestive clinical forms of chronic Chagas disease and in cardiac patients with and without left ventricular systolic dysfunction (LVSD). Plasma samples from 75 patients were evaluated by enzyme-linked immunosorbent assay (ELISA) to confirm infection by T. cruzi. Plasma levels of the CCL3 and CCL4 chemokines were measured using Milliplex® MAP assay (Millipore). There were no significant differences in the levels of CCL3 and CCL4 between patients with the digestive and cardiac clinical forms of Chagas disease. Moreover, no significant differences were found between patients without LVSD and those with LVSD. Higher CCL3 and CCL4 plasma levels were found in patients with LVSD compared to those with the digestive form of the disease. The CCL3 and CCL4 chemokines might not be involved in differential susceptibility to the digestive and cardiac clinical forms of chronic Chagas disease, and it seems they do not influence the development of LVSD.

Fetal Infection by Zika Virus in the Third Trimester: Report of 2 Cases.

Zika virus (ZIKV) infection acquired during pregnancy is associated with congenital microcephaly. We describe 2 cases of ZIKV infection in women in their 36th week of pregnancy whose fetuses had preserved head circumference at birth and findings of subependymal cysts and lenticulostriate vasculopathy in postnatal imaging. These represent the first signs of congenital brain injury acquired due to ZIKV in the third trimester.

Association of the Functional MICA-129 Polymorphism With the Severity of Chronic Chagas Heart Disease.

MICA-129 polymorphism affects the binding affinity of MICA molecules with the NKG2D receptor and influences effector cell function. The genotype met/met was associated with the severity of left ventricular systolic dysfunction (LVSD) in patients with chronic Chagas heart disease, while the val/val genotype was associated with the absence of LVSD.

Seroepidemiology of Toxoplasma gondii infection in blood donors in a population from the northwestern region of São Paulo state, Brazil.

BACKGROUND: Toxoplasmosis is one of the most common parasitic infections worldwide with varying prevalence between human populations. These variations are mainly associated with human exposure to risk factors. In this article, the seroprevalence of Toxoplasma gondii infection and the risk factors associated with infection in 1729 blood donors from São José do Rio Preto, São Paulo, Brazil were analysed. METHODS: The serological tests for detecting immunoglobulin M (IgM) and immunoglobulin G (IgG) anti-T. gondii were used. The risk factors associated with the infection were identified through the application of an epidemiological questionnaire. RESULTS: The prevalence of T. gondii infection was 48.0%. The following factors were identified in the final model after multiple logistic regression analysis: drinking raw milk (p=0.003; odds ratio [OR] 1.364 [confidence interval {CI} 1.1 to 1.7]), residing in a rural area (p<0.0001; OR 2.764 [CI 1.7 to 4.6]) and receiving a blood transfusion (p=0.015; OR 1.856 [CI 1.1 to 3.0]). CONCLUSIONS: The data obtained in this study showed that the blood donor population is exposed to risk factors related to infection by T. gondii. These data allow the establishment of control programs to contribute to public health in northwestern São Paulo state.

Portable color retinography findings in COVID-19 patients admitted to the ward.

Retinal lesions, including cotton-wool exudates, microbleeds, vascular occlusions and vasculitis, occur in a minority of Coronavirus Disease-19 (COVID-19) patients. Retinal assessments using retinography can help document these lesions. The objective of this work was to identify retinal changes in patients admitted to the ward with a positive Real Time Quantitative Polymerase Chain Reaction (RT-qPCR) exam for COVID-19. A cross-sectional, observational study was carried out of patients with mild and moderate symptoms admitted to the Hospital de Base in São José do Rio Preto. The Eyer® portable retinal camera (Phelcom® Technologies) was used to evaluate 30 male and 21 female patients. The ages ranged from 21 to 83 years (mean: 47 years). Systemic arterial hypertension was identified in 21 (41.2 %) and diabetes mellitus in 12 (23.5 %) patients. Six (11.7 %) reported worsening visual acuity, however, none of these patients had ocular findings to justify this complaint. Ten patients (19.6 %) had intraretinal hemorrhages; one (1.9 %) had cotton-wool exudates and seven (13.7 %) had dilations of veins. Thirteen patients (25.4 %) had vascular tortuosity and six (11.7 %) had pathological arteriovenous crossings. Portable retinography is useful to evaluate patients admitted to isolation wards due to COVID-19. It is important to remember that some of the patients investigated had comorbidities like diabetic maculopathy and systemic arterial hypertension. Hence, some care should be taken in attributing these observations uniquely to COVID-19 infection.

Clinical and epidemiological profile of patients with mental disorders in a specialized outpatient clinic and its role in the psychosocial care network.

Introduction: Mental health disorders (MHDs) are responsible for much impairment of quality of life in Brazil and worldwide. Early diagnosis and effective treatment strategies are required due to the heterogeneous symptoms and multifactorial etiology. Methods: A descriptive retrospective observational study was performed aiming to characterize the clinical and psychiatric profiles of patients with MHD attending a Brazilian public tertiary psychiatric outpatient clinic, which is a reference health service for more than 2 million inhabitants. Predominant clinical and sociodemographic aspects of patients were evaluated between March 2019 and March 2021. Results: A total of 8,384 appointments were analyzed. The majority of patients were female, and the mean age was 45 years old. Generalized anxiety disorder (GAD) was the most common MHD. The prevailing symptoms were sadness, anxiety, and irritability, with the most prescribed medications being selective serotonin reuptake inhibitors. Conclusion: The epidemiological characterization of mental disorders in specialized mental health outpatient clinics provides evidence for the establishment of more specific protocols and advocates a dimensional transdiagnostic approach as an aid to public mental health services.

miRNA 511_5p is a potential biomarker for ocular toxoplasmosis.

BACKGROUND: Ocular toxoplasmosis (OT) is a frequent clinical manifestation due to infection by Toxoplasma gondii. It is characterized by an inflammatory process involving macrophages activated by pro-inflammatory cytokines. The expression of microRNAs takes place during the inflammatory process and, among them, miRNA 511 regulates the activation of macrophages. This study evaluated the expression of miRNA 511_5p in patients with OT and healthy controls. METHODS: A total of 361 patients from the Hospital de Base of Fundação Faculdade de Medicina de São José do Rio Preto were enrolled and divided into four groups: G1-patients with active ocular lesions and reagent serology for T. gondii; G2-patients with scars and reagent serology for T. gondii; G3-patients without ocular lesions or scars and reagent serology for T. gondii; G4-patients without ocular lesions or scars and non-reagent serology for T. gondii. All patients underwent clinical and laboratory evaluation to confirm the diagnosis of OT. Serology tests, RNA extraction and cDNA synthesis were performed. RESULTS: The miRNA 511_5p levels were compared among the groups. The G1 group showed a high blood plasma concentration of miRNA 511_5p (mean 22.34) compared with the G2 (4.65), G3 (8.91) and G4 (3.52) groups (p<0.0001). CONCLUSION: These data suggest that miRNA 511_5p has significant potential as a biomarker for OT.

Role of interleukin 1ß and interleukin 10 variants on ocular toxoplasmosis in Brazilian individuals.

Introduction: Ocular toxoplasmosis (OT) is an intraocular inflammation caused by Toxoplasma gondii infection that affects the retina and choroid, giving rise to posterior uveitis. Genetic polymorphisms in cytokine genes may exert influence in the expression of these molecules and play a significant role in inflammatory responses and susceptibility to OT. The aim of this study was to evaluate the role of polymorphisms rs16944 (-511 C > T) of the interleukin (IL) 1ß gene and rs1800896 (-1082 G > A) of the IL10 gene on OT in Brazilian individuals with a serologic diagnosis of T. gondii and after conducting fundoscopic exams. Methods: Participants with a positive serology were classified into two distinct groups according to the presence (G1; n = 110) or absence (G2; n = 104) of OT. The control group (G3) consisted of individuals without the infection (n = 108). Results: It was observed that the C/C genotype of the IL1ß gene polymorphism was a protective factor for OT (p = 0.02, OR = 0.28, 95% CI 0.08-0.78 for G1 vs. G2; p = 0.03; OR = 0.29, 95% CI 0.09-0.82 for G1 vs. G3), according to the recessive inheritance model. Conclusions: The -511C.T polymorphisms of the IL1ß gene seems to play an important role in the pathogenesis of OT in Brazilian individuals.

Identification and validation of reference genes of circulating microRNAs for use as control in gestational toxoplasmosis.

Toxoplasmosis causes serious harm to the fetus, as tachyzoite dissemination, during pregnancy in women developing the primo-infection. The microRNAs (miRNAs) are small non-coding RNAs, which have regulatory roles in cells by silencing messenger RNA. Circulating miRNA are promising biomarkers for diagnosis and prognosis of numerous diseases. The miRNAs levels are estimated by quantitative real-time PCR (qPCR), however, the relative quantification of each miRNA expression requires proper normalization methods using endogenous miRNAs as control. This study analyzed the expression of three endogenous miRNAs (miR-484, miR -423-3p and miR-26b-5p) for use as normalizers in future studies of target miRNAs for gestational toxoplasmosis (GT). A total of 32 plasma samples were used in all assays divided in 21 from women with GT and 11 from healthy women. The stability of each endogenous miRNA was evaluated by the algorithm methods RefFinder that included GeNorm, Normfinder, BestKeeper and comparative delta-CT programs. The miR-484 was the most stably gene, and equivalently expressed in GT and NC groups. These results contribute to future studies of target miRNAs in clinical samples of women with gestational toxoplasmosis.

New artificial intelligence index based on Scheimpflug corneal tomography to distinguish subclinical keratoconus from healthy corneas.

PURPOSE: To assess the efficiency of an index derived from multiple logistic regression analysis (MLRA) to measure differences in corneal tomography findings between subclinical keratoconus (KC) in 1 eye, corneal ectasia, and healthy corneas. SETTING: 2 private Brazilian ophthalmological centers. DESIGN: Multicenter case-control study. METHODS: This study included 187 eyes with very asymmetric ectasia and with normal corneal topography and tomography (VAE-NTT) in the VAE-NTT group, 2296 eyes with healthy corneas in the control group (CG), and 410 eyes with ectasia in the ectasia group. An index, termed as Boosted Ectasia Susceptibility Tomography Index (BESTi), was derived using MLRA to identify a cutoff point to distinguish patients in the 3 groups. The groups were divided into 2 subgroups with an equal number of patients: validation set and external validation (EV) set. RESULTS: 2893 patients with 2893 eyes were included. BESTi had an area under the curve (AUC) of 0.91 with 86.02% sensitivity (Se) and 83.97% specificity (Sp) between CG and the VAE-NTT group in the EV set, which was significantly greater than those of the Belin-Ambrósio Deviation Index (BAD-D) (AUC: 0.81; Se: 66.67%; Sp: 82.67%; P < .0001) and Pentacam random forest index (PRFI) (AUC: 0.87; Se: 78.49%; Sp: 79.88%; P = .021). CONCLUSIONS: BESTi facilitated early detection of ectasia in subclinical KC and demonstrated higher Se and Sp than PRFI and BAD-D for detecting subclinical KC.