Temporary Conjunctival Flap Plus Medical Treatment Versus Medical Treatment Only for High-Risk Fungal Keratitis: A Randomized Controlled Trial.

PURPOSE: The purpose of this study was to evaluate the efficacy of temporary conjunctival flap with topical natamycin and oral voriconazole compared with medical treatment only in reducing the rate of perforation in high-risk fungal keratitis. METHODS: Sixty-two eyes of 62 patients with severe fungal keratitis were examined; only 54 patients were enrolled and divided randomly into 2 groups. The first group received medical treatment only in the form of topical natamycin 5% together with oral voriconazole 200 mg, while the second group received the same medical treatment plus a temporary conjunctival flap that was removed after 2 weeks. Five patients were lost during the follow-up, and only 49 patients were statistically analyzed. All patients were examined frequently until reepithelialization or the development of perforation. RESULTS: Fifteen perforations were reported, with a higher rate among the medical group (48%) compared with the conjunctival flap group (12.5%), with P value <0.05. A significant delay was noted in reepithelialization time in the medical group compared with the conjunctival flap group (mean time was 21.69 ± 5.41 and 15.36 ± 2.2 days, respectively), with P value = 0.001. Significant improvement in visual acuity was reported over time when comparing baseline versus 3-month visual acuity in the same group using paired t sample test (P value was 0.003 and <0.001 in the first and second group, respectively). CONCLUSIONS: Temporary conjunctival flap is associated with a lower perforation rate compared with medical treatment only in severe fungal keratitis, which can provide a cheap and available alternative to therapeutic penetrating keratoplasty.

Investigating Forkhead Box O Transcription Factor 1 Gene's Relation to Immunoglobulin E in House Dust Mite-Allergic Asthma Patients.

House dust mite (HDM)-allergic asthma is an abnormal immune response to extrinsic aeroallergens found in human vicinities. Studying the role of the associated immunity biomarkers and their interplay helps in discovering novel therapeutic strategies that can be used in adjunct with effective long-term immunotherapy. This study investigates the total serum IgE, FoxO1, and Sirtuin 1 (SIRT1) gene expressions in HDM-allergic asthma patients. We enrolled 40 patients for each of the following three groups: an HV group of healthy volunteers and HDM/AA and HDM/SCIT groups of HDM-allergic asthma patients who did not and who did receive immunotherapy before recruitment in this study, respectively. The results elucidated that total IgE was strikingly elevated in the HDM/AA group and showed little decline in the HDM/SCIT group. Both FoxO1 and SIRT1 gene expressions showed the highest levels in the HDM/SCIT group. There was a negative correlation between total IgE and both FoxO1 and SIRT1 in the HDM/AA group while there was a positive correlation with SIRT1 in the HDM/SCIT group. In conclusion, the interplay of the three immunity biomarkers related to HDM-allergic asthma after the course of immunotherapy treatment suggests further, broader studies on the feasibility of their role as immunity biomarkers in the control and remission of HDM-allergic asthma.

Incidence of Food allergy among patients with allergic conjunctivitis.

The prevalence of ocular allergy is increasing worldwide. Skin prick test is widely recognized as the most reliable method for diagnosing the incriminating allergen as regards type I hypersensitivity reactions. Food allergy results as immunological response to food protein which leads to occurrence of allergic conjunctivitis (AC), allergic rhinitis, asthma, atopic dermatitis, and eosinophilic esophagitis. There is a scarcity of research investigating the association between food allergy and AC. This retrospective cohort study aimed to determine the incidence of food allergy within AC patients and its linkage to disease intensity and to compare the response to sublingual immunotherapy after 4 months of therapy. The study included 240 individuals diagnosed with AC. Of these patients, only 214 (89.16%) cases exhibited positive skin prick test results and showed incidence of food allergy of 29.6 %. After 4 months of sublingual allergen immunotherapy, the total serum IgE level and the grades of severity decreased significantly (p 0.001 for each). On comparing patients with food allergy on sublingual immunotherapy and patients without food allergy and on sublingual immunotherapy, the change in total serum IgE concentration and the grade of severity did not differ among the two groups (p value was 0.63 and 1.00 respectively). In conclusion, food allergies can contribute to the development of AC. Sublingual allergen immunotherapy can be proposed as a promising therapeutic option for AC patients.

NLRP3 inflammasome (rs10754558) gene polymorphism in patients with atopic dermatitis.

The nucleotide-binding oligomerization domain-like receptor 3 (NLRP3) inflammasome is a high molecular weight protein complex that has been linked to a variety of allergic and inflammatory disorders in humans, including atopic dermatitis (AD). Polymorphisms in NLRP3 genes could lead to immune dysregulation. This case-control study aimed to assess the association between NLRP3 inflammasome (rs10754558) gene polymorphism in AD and the incidence and severity of the disease. We included 62 subjects in each of the AD and control groups. Serum total IgE levels and NLRP3 inflammasome (rs10754558) gene polymorphism were assessed and compared between the two study groups and among the AD group as arranged by disease severity. The AD group showed significantly higher levels of serum total IgE compared to controls (p˂0.001). Serum IgE levels were also significantly associated with AD severity. The (rs10754558) G allele was significantly predominant among AD participants (OR: 2.33; 95% CI: 1.1 -4.92) and 51.6% of the AD group was carriers of the GG genotype. Moreover, there was a substantial correlation between NLRP3 (rs10754558) G allele and AD score index for disease severity (OR: 7.17; 95% CI: 1.47 - 35.7). In conclusion, NLRP3 inflammasome (rs10754558) gene polymorphism G allele could be an important factor in the predisposition and exacerbation of AD.

LncRNA-GAS5 and ß-Catenin as Independent Predictors of Asymptomatic Organ Damage in Nondiabetic Hypertensive Patients.

Hypertension is a serious medical condition that can increase the risk of developing heart, brain, kidney, and other diseases. Many asymptomatic hypertension patients experience asymptomatic organ damage (AOD). The purpose of this study was to determine the roles of LncRNA-GAS5 and ß-catenin in predicting AOD in hypertensive nondiabetic patients. This study included 256 subjects, 128 hypertension patients (75 of whom had AOD, and 53 of whom did not) and 128 healthy controls. qRT-PCR was used to assess LncRNA-GAS5, and ELISA was used to assess ß-catenin. The LncRNA-GAS5 expression level was decreased in hypertensive patients compared to controls (p-value < 0.001). On the other hand, ß-catenin levels showed higher levels in the patients in comparison with controls (p-value < 0.001). A 0.38-fold change in LncRNA-GAS5 expression predicted AOD with 86.6% sensitivity and 88.7% specificity. ß-Catenin > 80.5 pg/mL predicted AOD with a sensitivity of 82.6% and specificity of 69.8%. LncRNA-GAS5 expression was a better diagnostic predictor of AOD than ß-catenin. According to multivariate logistic regression analysis, decreased LncRNA-GAS5 expression independently increased the risk of AOD (adjusted odds ratio = 0.03 (95% CI: 0.01-0.1) (p < 0.001). Furthermore, elevated ß-catenin levels may be an independent risk factor for AOD (adjusted odds ratio = 14.3 (95% confidence interval, 3.3-61.9) (p < 0.001). Collectively, in hypertensive patients, LncRNA GAS5 and ß-catenin can distinguish patients with AOD from those who do not have AOD. LncRNA GAS5 and ß-catenin can be used as independent predictors of AOD in hypertensive patients.

H19 and TUG1 lncRNAs as Novel Biomarkers for Irritable Bowel Syndrome in Diabetic Patients.

Introduction: Irritable bowel syndrome (IBS) is a gastrointestinal disorder due to enteric nervous system impairment that produces different patterns of digestion. IBS is a common finding in diabetic patients. The functions of lncRNAs in IBS are still not clear and need to be further investigated. The aim of this study was to assess the diagnostic roles of lncRNA H19 and TUG1 for IBS associated with diabetes and to evaluate their association with clinical and laboratory findings. Subjects and Methods: Samples from 42 diabetic patients, 42 diabetic patients with IBS, and 42 healthy controls were obtained. The LncRNA H19 and TUG1 expressions were measured by quantitative real-time PCR. Results: The patients with IBS had significantly lower levels of lncRNA H19 and TUG1 expression than the healthy controls and diabetic-only patients (p < 0.001). LncRNA H19 and TUG1 can discriminate between diabetic-only patients and those with IBS (areas under the ROC curves of 0.95 and 0.722, respectively). The TUG1 expression levels were significantly different among types of IBS (IBS-D lower than IBS-M and IBS-C lower than IBS-M; p = 0.0165 and p = 0.043, respectively). H19 and TUG1 were downregulated in patients with poor glycemic control. lncRNA H19 and TUG1 expression in diabetic patients with IBS significantly negatively correlated with the IBS severity scoring system. Both lncRNAs' expression significantly predicted the disease severity. LncRNA H19 expression can be an independent predictor for disease severity (adjusted odds ratio = 0.00001, 95% CI = 0−0.5, p = 0.045). Conclusions: Diabetic patients with IBS had significantly lower levels of lncRNA H19 and TUG1 expression than healthy controls and diabetic-only patients. LncRNA H19 had better diagnostic performance criteria for IBS. LncRNA H19 expression can be an independent predictor for IBS severity.

Toll-Like Receptor-4 Gene (Asp299Gly) polymorphism in allergic conjunctivitis.

Allergic conjunctivitis (AC) is an allergic reaction that causes inflammation of the conjunctiva. Toll-like receptors (TLRs) are essential innate immune receptors that contribute to developing various allergic diseases. This case-control study aims to determine the correlation between TLR-4 gene (Asp299Gly) polymorphism and AC incidence and severity. The study included 70 AC patients and 70 non-allergic controls. All included subjects were subjected to a skin prick test, total immunoglobulin E (IgE) measurement, and TLR-4 gene (Asp299Gly) polymorphism detection by PCR restriction fragment length polymorphism (PCR-RFLP) technique. AC patients had significantly higher total IgE levels than controls (P ≤ 0.001). The frequency of the wild-type AA and heterozygous AG genotype were significantly lower in AC patients compared to controls (60 % vs. 80 % and 8.6% vs. 12.9 %, respectively). In contrast, the homozygous mutant GG genotype was significantly more prevalent among AC patients than controls (31.4 % vs. 7.1 %). Furthermore, the wild AA genotype was strongly associated with mild disease (68.2%); nonetheless, the homozygous mutant GG genotype was linked to severe disease (53.8%). The heterozygous AG genotype was only found in moderate AC patients (17.1%). AC patients with the mutant G allele may be more likely to have a severe course of AC.

Toll-Like Receptor 4 Polymorphisms (896A/G and 1196C/T) as an Indicator of COVID-19 Severity in a Convenience Sample of Egyptian Patients.

BACKGROUND: The clinical spectrum of COVID-19 is extremely variable. Thus, it is likely that the heterogeneity in the genetic make-up of the host may contribute to disease severity. Toll-like receptor (TLR)-4 plays a vital role in the innate immune response to SARS-CoV-2 infection. The susceptibility of humans to severe COVID-19 concerning TLR-4 single nucleotide polymorphisms (SNPs) has not been well examined. OBJECTIVE: The goal of this research was to investigate the association between TLR-4 (Asp299Gly and Thr399Ile) SNPs and COVID-19 severity and progression as well as the cytokine storm in Egyptian patients. METHODS: We genotyped 300 adult COVID-19 Egyptian patients for TLR-4 (Asp299Gly and Thr399Ile) SNPs using PCR-restriction fragment length polymorphism (PCR-RFLP). We also measured interleukin (IL)-6 levels by enzyme-linked immunosorbent assay (ELISA) as an indicator of the cytokine storm. RESULTS: The minor 299Gly (G) and 399Ile (T) alleles were associated with a significant (P < 0.001) positive risk of severe COVID-19 (OR = 3.14; 95% CI = 2.02-4.88 and OR = 2.75; 95% CI = 1.66-4.57), their frequency in the severe group were 71.8% (84/150) and 70.7% (58/150), respectively. We detected significant differences between TLR-4 (Asp299Gly, Thr399Ile) genotypes with regard to serum levels of IL-6. Levels of IL-6 increased significantly with the presence of the mutant 299Gly (G) and 399Ile (T) alleles to reach the highest levels in the Gly299Gly (GG) and the Ile399Ile (TT) genotypes (170 pg/mL (145-208.25) and 112 pg/mL (24-284.75), respectively). CONCLUSION: The TLR-4 (Asp299Gly and Thr399Ile) minor alleles 299Gly (G) and 399Ile (T) are associated with COVID-19 severity, mortality, and the cytokine storm.

Mannose-binding lectin serum levels and (Gly54asp) gene polymorphism in recurrent aphthous stomatitis: A case-control study.

Objectives: Dysregulation of the immune response appears to play a significant role in recurrent aphthous stomatitis (RAS) development. The main objective of this case-control study is to investigate the blood levels of mannose-binding lectin (MBL) and the frequency of the MBL2 gene (gly54asp) polymorphism in RAS patients, including 40 RAS patients and 40 healthy controls. Methods: Serum MBL levels were determined by ELISA, while the PCR-restriction fragment length polymorphism was used in MBL2 genotyping. Results: The median serum MBL level was significantly lower in the RAS group than in the control group (975 ng/mL (545-1320) vs. 1760 ng/mL (1254-2134); p≤ 0.001). The MBL levels were significantly lower in the BB genotype, whereas they were significantly higher in the wild type AA with a median of 525 and 1340 ng/mL, respectively (p =0.005). The B allele was expressed in significantly higher percentages of RAS patients than in controls. There was no significant association between MBL serum levels (p=0.685) or MBL2 codon 54 genotypes (p=0.382) with the type of ulcers. Conclusion: There was an association between low MBL serum levels and the variant allele B of the MBL2 (gly54asp) gene, and the susceptibility to RAS. As a result, potential novel therapeutic options for RAS patients with MBL deficiency should be investigated.

Association between serum zonulin level and severity of house dust mite allergic asthma.

BACKGROUND: Increased intestinal permeability, either due to the exposure to antigens in asthmatic patients or due to a barrier defect, plays a critical role in susceptibility to environmental allergens. House dust mite allergy occurs more commonly than any other type of allergy among Egyptian asthmatic patients. AIM: To assess the relation between serum zonulin level as a marker of increased intestinal permeability and the severity of house dust mite allergic asthma. METHODS: A case-control study which included 48 patients with house dust mite allergic asthma and 48 healthy control subjects attending the Allergy and Immunology Unit, Microbiology and Immunology Department, Faculty of Medicine, Zagazig University. RESULTS: A statistically significant difference was detected between the two studied groups with respect to serum IgE and serum zonulin levels (p Ë‚ 0.001 and ˂ 0.001, respectively). The mean serum zonulin was equal to 258.3 ± 153.01 ng/ml in the asthmatic group and 80 ± 13 ng/ml in the control group. Serum zonulin level significantly increased with the increase of asthma severity (p ˂ 0.001). The cut off value of serum zonulin was ≥ 198 ng/ml, and the area under the curve was 0.76. It displayed sensitivity equal to 80% and specificity equal to 71.4%. Its negative predictive value was equal to 83.3%. CONCLUSION: Intestinal barrier dysfunction contributes to the pathogenesis of allergic asthma. Serum zonulin level reflects an increase in intestinal permeability. Zonulin acts as prognostic factor of severity in asthma. Correction of the gut barrier defect may have a potential positive prognostic effect in asthma.

Role of circulating microRNA-132 in allergic bronchopulmonary aspergillosis: A case-control study.

Allergic bronchopulmonary aspergillosis (ABPA) is an allergic respiratory disease. In the current study, we aimed to evaluate the roles of miRNA-21 and miRNA132 as biomarkers in the diagnosis of ABPA. A total of 30 controls, 30 allergic asthmatic patients, 30 severe asthma with fungal sensitization (SAFS) patients, and 30 ABPA patients were included. Real-time polymerase chain reaction was used to quantify the level of miRNAs expression. The expression level of miRNA-21 was significantly higher in allergic asthmatic, SAFS, and ABPA patients in comparison with controls (p < 0.001). However, no significant difference was detected in the expression level of miRNA-21 among the different patient groups (p > 0.05). The ABPA patients had significantly higher levels of miRNA-132 expression compared to controls, allergic asthmatic patients, and SAFS patients (p < 0.001), but there was a non-significant difference between controls and allergic asthmatic patients (p = 0.09). At a cut-off of 1.52, the sensitivity of miRNA-132 expression was 93.3% and the specificity was 100% different ABPA from healthy controls. At a cut-off of 6.5, miRNA-132 expression was found to reliably differentiate between ABPA and SAFS, with a sensitivity of 86.7% and a specificity of 80%. In ABPA patients, miRNA-132 expression positively correlation with the levels of serum IL-5 (r = 0.91, p < 0.001). miRNA-132 has a role in ABPA detection and distinguishing ABPA from allergic asthma and SAFS. These preliminary data from case-control study need further studies to confirm its finding.

Diagnostic and prognostic roles of peripheral blood Toll-like receptor-4 and stanniocalcin-1 genes expression in acute lung injury.

Acute lung injury (ALI) is an acute inflammatory disorder. Toll-like receptor-4 (TLR-4) and Stanniocalcin -1 (STC-1) had roles in lung endothelial protection. This study aims to assess TLR-4 and SCT-1 genes expressions in peripheral blood of ALI patients. Total RNA was extracted from peripheral blood of 48 subjects (20 healthy controls, 28 ALI patients) and expressions of genes were assessed by real-Time qRT-PCR. The expression levels of TLR-4 and SCT-1 genes were significantly lower in ALI patients compared to controls (P < 0.0001). After 10 days, the expression levels of TLR-4 and SCT-1 were increased compared to their baseline levels (p = 0.012 and 0.024, respectively). SCT-1 has 92.9% sensitivity and 100% specificity in ALI detection. SCT-1 gene expression was negatively correlated with severity score (r= -0.54, p = 0.003). The mortality pattern was higher in ALI patients with lower TLR-4 gene expression (p = 0.014). In conclusion, the peripheral blood expressions of TLR-4 and STC-1 genes were decreased in ALI patients. Both genes expressions were increased with patients' recovery. SCT-1 had higher sensitivity and specificity in ALI diagnosis. The peripheral blood expressions of SCT-1 and TLR-4 genes seem to be diagnostic and prognostic markers in ALI.

Assessment of circulating FCεRIa in Chronic Spontaneous Urticaria patients and its correlation with clinical and immunological variables.

Chronic spontaneous urticaria (CSU) is a chronic type characterized by episodes of wheals with or without angioedema. Autoantibody against the alpha subunit of Fc epsilon receptor (FcεRIa) was detected in CSU patients' sera. The study aims to evaluate the clinical utility of skin tests in CSU patients. In addition, it assesses the presence of circulating FcεRIa in CSU patients and their correlation with other clinical and immunological variables. The study includes 40 healthy controls and 40 CSU patients who had urticaria symptoms for at least 8 weeks. All subjects underwent the following tests: autologous serum skin test (ASST), autologous plasma skin test (APST), immunoglobulin E (IgE), antinuclear antibodies (ANA), antithyroid antibodies (ATA). An in-house enzyme-linked immunosorbent assay was used for FcεRIa detection. The prevalence of ANA and ATA in CSU was 7.5% and 20% respectively. Total IgE was significantly higher in CSU than in controls (p < 0.0001). The study detected circulating antibody to FcεRIα in 2.5% of controls and 52.5% of CSU patients (p < 0.0001). The prevalence of antibody to FcεRIa was 27.3% and 83.3% of ASST negative and positive patients respectively (p = 0.0004). But the prevalence was 17.6% and 78.3% of APST negative and positive patients respectively (p = 0.0002). In conclusion, Circulating antibody to FcεRIa has a role in the pathogenic mechanisms of CSU.