Exploration of the association between HIF3α mRNA and lncRNA MALAT1 in laryngeal squamous cell carcinoma by correlation analysis.

Laryngeal squamous cell carcinoma (LSCC) is a significant global health burden, for which there has been limited evidence of improved survival rates. Although the roles of hypoxia-inducible factor (HIF)1α and HIF2α have been well documented in hypoxia, the involvement of HIF3α, particularly in LSCC, has been inadequately explored. The present study aimed to investigate the correlation between HIFα subunits and the hypoxia-related long noncoding RNAs (lncRNAs) MALAT1 and HOTAIR in 63 patients diagnosed with LSCC. Total RNA was extracted from fresh-frozen laryngeal tumor and adjacent normal tissues, and was subjected to reverse transcription-quantitative PCR for target detection. Statistical analyses were conducted using SPSS software, with significance set at P<0.05. The present study is the first, to the best of our knowledge, to report a positive moderate monotonic correlation (rs=0.347) and moderately strong positive linear correlation (r=0.630) between HIF3α mRNA and lncRNA MALAT1 in LSCC. Regression analysis revealed a direct association between 39.6% of both variables. Additionally, a positive correlation was observed between lncRNAs MALAT1 and HOTAIR (rs=0.353); HIF2α mRNA and lncRNA MALAT1 (rs=0.431); HIF3α mRNA and lncRNA HOTAIR (rs=0.279); and HIF3α mRNA and HIF2α mRNA (rs=0.285). Notably, a significant negative correlation (rs=-0.341) was detected between HIF3α mRNA and HIF1α mRNA, potentially indicative of the HIF switch or negative regulation. In addition, the present study investigated the association between HIFα subunits and the clinicopathological characteristics of patients. The results revealed a notable association between HIF1α transcript levels and the location of LSCC; specifically, subglottic tumors exhibited elevated HIF1α levels compared with glottic and supraglottic LSCC. Furthermore, a significant association was identified between HIF3α transcript levels and patient age (P=0.032) and positive family history (P=0.047). In conclusion, the present findings suggested a pivotal role for HIF3α in LSCC development, potentially involving direct regulation of lncRNA MALAT1. However, further research is warranted to elucidate its precise mechanisms.

Analysis of Bacterial Biofilm Formation and MUC5AC and MUC5B Expression in Chronic Rhinosinusitis Patients.

Chronic rhinosinusitis (CRS) is a condition affecting as much as 16% of the adult population in developed countries with many factors attributed to its development, including the more recently proposed role of bacterial biofilm infections. Plenty of research has been conducted on biofilms in CRS and the causes behind the development of such an infection in the nasal cavity and sinuses. One such probable cause is the production of mucin glycoproteins by the mucosa of the nasal cavity. To investigate the possible link between biofilm formation and mucin expression levels and their relationship with CRS etiology, we examined samples from 85 patients by means of spinning disk confocal microscopy (SDCM) to establish their biofilm status and quantitative reverse transcription polymerase chain reaction (qRT-PCR) to determine MUC5AC and MUC5B expression levels. We observed a significantly higher prevalence of bacterial biofilms in the CRS patient group compared to the control group. In addition, we detected higher expression levels of MUC5B but not MUC5AC in the CRS group, which suggested a possible role for MUC5B in CRS development. Finally, we found no direct relationship between biofilm presence and mucin expression levels, thereby showing a multifaceted connection between these two major factors implicated in CRS etiology.

Cases of acute hemiparesis in childhood.

INTRODUCTION: Acute hemiparesis is an emergency of various etiologies and possible fatal outcome.

Global microRNA expression profile in laryngeal carcinoma unveils new prognostic biomarkers and novel insights into field cancerization.

Laryngeal carcinoma is still a worldwide burden that has shown no significant improvement during the last few decades regarding definitive treatment strategies. The lack of suitable biomarkers for personalized treatment protocols and delineating field cancerization prevents further progress in clinical outcomes. In the light of this perspective, MicroRNAs could be promising biomarkers both in terms of diagnostic and prognostic value. The aim of this prospective study is to find strong prognostic microRNA biomarkers for advanced laryngeal carcinoma and molecular signatures of field cancerization. Sixty patients were enrolled and four samples were collected from each patient: tumor surface and depth, peritumor normal mucosa, and control distant laryngeal mucosa. Initially, a global microRNA profile was conducted in twelve patients from the whole cohort and subsequently, we validated a selected group of 12 microRNAs with RT-qPCR. The follow-up period was 24 months (SD ± 13 months). Microarray expression profile revealed 59 dysregulated microRNAs. The validated expression levels of miR-93-5p (χ2(2) = 4.68, log-rank p = 0.03), miR-144-3p (χ2(2) = 4.53, log-rank p = 0.03) and miR-210-3p (χ2(2) = 4.53, log-rank p = 0.03) in tumor samples exhibited strong association with recurrence-free survival as higher expression levels of these genes predict worse outcome. Tumor suppressor genes miR-144-3p (mean rank 1.58 vs 2.14 vs 2.29, p = 0.000) and miR-145-5p (mean rank 1.57 vs 2.15 vs 2.28, p = 0.000) were significantly dysregulated in peritumor mucosa with a pattern of expression consistent with paired tumor samples thus revealing a signature of field cancerization in laryngeal carcinoma. Additionally, miR-1260b, miR-21-3p, miR-31-3p and miR-31-5p were strongly associated with tumor grade. Our study reports the first global microRNA profile specifically in advanced laryngeal carcinoma that includes survival analysis and investigates the molecular signature of field cancerization. We report two strong biomarkers of field cancerization and three predictors for recurrence in advance stage laryngeal cancer.

Novel insights into laryngeal squamous cell carcinoma from association study of aberrantly expressed miRNAs, lncRNAs and clinical features in Bulgarian patients.

PURPOSE: Laryngeal cancer is one of most common and aggressive head and neck cancers with poor prognosis and great necessity for improvement of treatment modalities. MicroRNAs (miRs) are among the most investigated molecules recently due to their potential as diagnostic and prognostic biomarkers in cancer. The purpose of our study was to explore the association of certain clinicopathological features with the expression levels of some known cancer associated non-coding (nc) RNAs: miR-21 and miR-31 in both of their isoforms, miR-145-5p, miR-55-5p, miR-196a-5p, miR-210-3p, miR-221-3p, miR-222-3p, miR-424-5p, lncRNA MALAT1 and lncRNA HOTAIR. METHODS: Expression levels of the chosen markers were investigated in laryngeal squamous cell carcinoma (LSCC) and normal samples in 82 Bulgarian patients via RT-qPCR, and the results were analyzed with SPSS v23.0 statistical software. RESULTS: All of the explored ncRNAs were significantly deregulated in LSCC samples, suggesting their involvement in laryngeal carcinogenesis. New significant association were found between the expression levels of miR-21-5p, miR-222-3p, HOTAIR and family history. Moreover, miR-424-5p showed potential as marker for subglottic LSCC location, and "passenger" miR-31-3p was significantly upregulated in well and moderately differentiated LSCC. CONCLUSION: Our results enrich the knowledge about ncRNA involvement in LSCC tumorigenesis. Further studies are needed to evaluate the clinical utility of the differently expressed ncRNAs as potential diagnostic and prognostic biomarkers in LSCC.

Magnetic Resonance Imaging of Hemophilic Joints: Correlations with the Bleeding Phenotype and Physical Examination.

INTRODUCTION: Blood-induced joint damage as a hallmark of haemophilia continues to occur despite the widespread prophylaxis. Pre-cise assessment and follow-up of joint status are crucial for tailoring their treatment. AIM: To study the correlation between the bleeding phenotype, the functional joint status, and the magnetic resonance imaging score in pediatric patients with haemophilia. MATERIALS AND METHODS: Eighty-six joints (ankles, knees, and elbows) in patients aged 10.7±0.5 (range 4 - 20) years with severe/moderate haemophilia A, severe haemophilia B and haemophilia A with inhibitors were included in the study. The joints were assessed by Haemophilia Joint Health Score 2.1 (HJHS2.1) one month after the last hemarthrosis in a non-bleeding state. The magnetic reso-nance imaging was performed on 40 (46.5%) of the examined hemophilic joints (16 ankles, 11 knees and 13 elbows). RESULTS: Joint bleeds were present in 37 (38.9%) of the joints with ankles being the most commonly affected. Sixty joints (69.8%) had normal HJHS2.1 score. Only the loss of flexion score differed significantly between the joints and the ankles had highest score. The cumulative number of hemarthrosis in the joint correlated moderately with hemosiderin deposition and strongly with the formation of subchondral cysts on magnetic resonance imaging. The magnetic resonance imaging scores for soft tissue and osteochondral domains correlated moderately with the cumulative number of hemarthrosis in the joint and only with the presence of pain and crepitus of mo-tion from the physical examination. CONCLUSIONS: Magnetic resonance imaging is more sensitive than the bleeding phenotype and physical examination in detecting early signs of haemophilic arthropathy.

BPIFA1 Gene Expression in the Human Middle Ear Mucosa.

OBJECTIVE: The bactericidal/permeability-increasing, fold-containing family member A1 (BPIFA1) gene codes a secretory protein (BPIFA1), which is present in the respiratory tract mucosa, and is part of the innate immune system. This study aimed to prove that BPIFA1 gene expression exists in the human middle ear mucosa. MATERIALS AND METHODS: In total, 32 patients participated in the study between March 2016 and September 2016. Seventeen patients had chronic otitis media with cholesteatoma (COMC) and 15 had bilateral sensorineural hearing loss (BSHL). The patients with COMC underwent radical mastoidectomy with cholesteatoma removal and those with BSHL underwent cochlear implantation. Part of the processus mastoideus mucosa was examined for BPIFA1 gene expression and the two groups were compared. RESULTS: For the first time, BPIFA1 gene expression was examined in the mucosa of the human middle ear, and it was verified in 100% (n=32) of the participants. We confirmed that there is a difference in the BPIFA1 expression in 83.33% of the patients with COMC compared to the patients with BSHL but this difference was not statistically significant (p=0.947; probably due to the low number of participants in this group). CONCLUSION: It is highly likely that the BPIFA1 protein participates in the non-specific immune defense of the middle ear and is relevant to the pathogenesis of the inflammatory diseases of the middle ear.

Usage and Attitudes Towards Natural Remedies and Homeopathy in General Pediatrics: A Cross-Country Overview.

In order to better understand the global approach and country differences in physicians' usage, knowledge, and attitudes towards natural remedies and homeopathy in pediatric practice, an online survey involving 582 general pediatricians and general practitioners treating pediatric diseases was conducted in 6 countries. Overall, 17% of the pediatric prescriptions refer to phytotherapy and 15% refer to homeopathic preparations. Natural remedies and homeopathic preparations are more frequently used in upper respiratory tract infections, infant colic, sleep disturbances, and recurrent infections. In the majority of cases, they are used together with chemical drugs. Both treatment options are typically used if parents are concerned about side effects of conventional drugs or prefer natural remedies for themselves. Physicians express high interest in natural remedies and homeopathy; however, their knowledge is variable. Lack of proven efficacy, knowledge on mechanism of action, and information on indications are main factors that limit their usage.

Auditory outcome after cochlear implantation in patients with congenital nonsyndromic hearing loss: influence of the GJB2 status.

OBJECTIVE: To compare the audiologic outcome after cochlear implantation between 2 groups of patients with congenital nonsyndromic sensorineural hearing loss. STUDY DESIGN: Retrospective cohort study. SETTING: Department of Otorhinolaryngology, University hospital (tertiary referral center). PATIENTS: From a bigger pool of implanted patients, 2 groups, each numbering 30 were enrolled. The patients from the first group were diagnosed with a Connexin 26 mutation (GJB2), whereas all of the patients from the second cohort were with a wild type genotype. Both groups were age matched, 1 to 7 years old at the age of implantation, with diagnosed congenital nonsyndromic sensorineural hearing loss. MAIN OUTCOME MEASURES: Both groups were evaluated with the help evaluation of auditory responses to speech/EARS/test battery - LiP test (Listening Progress Profile), MTP tests 3,6,12 (Monosyllabic-Trochee-Polysyllabic Test), GASP test (Glendonald Auditory Screening Procedure), and others. Follow-up period was at least 36 months. RESULTS: Mean test scores were compared at the 1st, 6th, 12th, 24th, and 36th month. LiP outcome was significantly better (p < 0.05) for the GJB2-related cohort for the whole follow-up period except at the first month. MTP3, 6, and 12 tests displayed the same statistically significant outcome in favor of the first group. In the open-set test GASP, the difference was apparent: 1.22, 2.40, 5.59, and 7.40 mean scores at the 6th, 12th, 24, and 36th months for the first cohort versus 0.00, 0.07, 0.81, and 1.74 for the GJB2-unrelated patients. CONCLUSION: The results from our study suggest that children with GJB2-related deafness show better auditory performance after cochlear implantation than age-matched children with GJB2-nonrelated sensorineural hearing loss.

Prevalence of GBJ2 mutations in patients with severe to profound congenital nonsyndromic sensorineural hearing loss in Bulgarian population.

Objective of the study is to assess the prevalence of Connexin 26 (GJB2) mutation in patients with congenital nonsyndromic sensorineural hearing loss in Bulgarian population. Study design is done prospectively. Patient inclusion criteria for this study were diagnosis of congenital nonsyndromic hearing loss, and absence of potential sibling relationships between patients included in the study (anamnestic pedigree for at least three generations). Patients were excluded from the study group if one of the following conditions were present: secondary hearing loss (cytomegalovirus, rubella, meningo-encephalitis, mastoiditis, other infections, posterior fossa tumors, etc.), exposure to drugs or other prenatal or perinatal etiology of deafness, and congenital syndromic hearing loss. Genomic DNA samples from whole blood were tested with sequence analysis for mutations in the coding region of the GJB2. Results state that 51 patients were analyzed for GJB2 mutations. Twenty of the patients (39%) with mutant alleles were homozygous for the c.35delG mutation (c.35delG/c.35delG) and four patients (8%) presented as heterozygotes (c.35delG/WT). In one patient, who carried a heterozygous mutation c.35delG, a second mutation was found-312del114. Additionally, in two other patients were discovered the mutations Trp24X (W24X) and, respectively, Arg127His(R127H), both in heterozygous states. From the whole study group there was only one patient with compound heterozygous genotype-p.Leu90Pro(L90P)/p.Ile121Asn. The latter one has never been reported in the literature so far. In conclusion, this study determines the importance of connexin 26 mutations in Bulgarian children with severe to profound congenital nonsyndromic sensorineural hearing loss, the prevalence of the different mutation variants and their relationship with the ethnical background of the patients. In addition, we report for the first time a novel mutation in the GJB2 gene.