Identification of a Novel Variant in CC2D1A Gene Linked to Autosomal Recessive Intellectual Disability 3 in an Iranian Family and Investigating the Structure and Pleiotropic Effects of this Gene.

Objectives: Intellectual disability (ID) represents a significant health challenge due to its diverse and intricate nature. A multitude of genes play a role in brain development and function, with defects in these genes potentially leading to ID. Considering that many of these genes have yet to be identified, and those identified have only been found in a small number of patients, no complete description of the phenotype created by these genes is available. CC2D1A is one of the genes whose loss-of-function mutation leads to a rare form of non-syndromic ID-3(OMIM*610055), and four pathogenic variants have been reported in this gene so far. Materials & Methods: n the current study, two affected females were included with an initial diagnosis of ID who were from an Iranian family with consanguineous marriage. Whole-exome sequencing was used to identify the probable genetic defects. The Genotypic and phenotypic characteristics of the patients were compared with a mutation in the CC2D1A gene, and then the structure of the gene and its reported variants were investigated. Results: The patients carried a novel homozygous splicing variant (NM_017721, c.1641+1G>A) in intron 14, which is pathogenic according to the ACMG guideline. Loss-of-function mutations in CC2D1A have severe phenotypic consequences such as ID, autism spectrum disorder (ASD), and seizures. However, missense mutations lead to ASD with or without ID, and in some patients, they cause ciliopathy. Conclusion: This study reports the fifth novel, probably pathogenic variant in the CC2D1A gene. Comparing the clinical and molecular genetic features of the patients with loss-of-function mutation helped to describe the phenotype caused by this gene more precisely. Investigating the CC2D1A gene's mutations and structure revealed that it performs multiple functions. The DM14 domain appears more pivotal in triggering severe clinical symptoms, including ID, than the C2 domain.

Bond strength of conventional versus modified methods for Class IV restorations in primary incisors: An in vitro study.

Background. Aesthetic restorations of severely decayed anterior primary teeth are challenging due to the small size of the teeth and the large pulp chambers. Therefore, this study evaluated and compared the tensile and shear bond strengths of conventional versus modified methods (slot technique) of Cl IV restorations in primary incisors. Methods. A total of 120 extracted intact maxillary primary central and lateral incisors were divided into two groups. In group A, conventional Cl IV cavities were prepared. In group B, after conventional cavity preparation, four and three incisal slots were prepared on the incisal edges of the central and lateral incisors, respectively. All the teeth were restored using composite resin, and then the tensile and shear bond strengths were evaluated. Results. A statistically significant increase in the tensile bond strength of restorations was recorded in the modified technique compared to the conventional method (P=0.001). Although an increase in the shear bond strengths was found in the modified method, the value did not reach a statistically significant level (P=0.158). The most frequent fracture type was adhesive in the conventional group and cohesive in the modified group, considering the tensile and shear bond strength tests. In both groups, the highest average tensile bond strength was recorded in teeth with the cohesive fracture in dentin, and the lowest average was seen in the adhesive type. Conclusions. Incisal slots could increase the tensile bond strengths of Cl IV composite resin restorations in primary teeth.

Screening for differentially expressed microRNAs in BALF and blood samples of infected COVID-19 ARDS patients by small RNA deep sequencing.

BACKGROUND: The pandemic COVID-19 has caused a high mortality rate and poses a significant threat to the population of the entire world. Due to the novelty of this disease, the pathogenic mechanism of the disease and the host cell's response are not yet fully known, so lack of evidence prevents a definitive conclusion about treatment strategies. The current study employed a small RNA deep-sequencing approach for screening differentially expressed microRNA (miRNA) in blood and bronchoalveolar fluid (BALF) samples of acute respiratory distress syndrome (ARDS) patients. METHODS: In this study, BALF and blood samples were taken from patients with ARDS (n = 5). Control samples were those with suspected lung cancer candidates for lung biopsy (n = 3). Illumina high-throughput (HiSeq 2000) sequencing was performed to identify known and novel miRNAs differentially expressed in the blood and BALFs of ARDS patients compared with controls. RESULTS: Results showed 2234 and 8324 miRNAs were differentially expressed in blood and BALF samples, respectively. In BALF samples, miR-282, miR-15-5p, miR-4485-3p, miR-483-3p, miR-6891-5p, miR-200c, miR-4463, miR-483-5p, and miR-98-5p were upregulated and miR-15a-5p, miR-548c-5p, miR-548d-3p, miR-365a-3p, miR-3939, miR-514-b-5p, miR-513a-3p, miR-513a-5p, miR-664a-3p, and miR-766-3p were downregulated. On the contrary, in blood samples miR-15b-5p, miR-18a-3p, miR-486-3p, miR-486-5p, miR-146a-5p, miR-16-2-3p, miR-6501-5p, miR-365-3p, miR-618, and miR-623 were top upregulated miRNAs and miR-21-5p, miR-142a-3p, miR-181-a, miR-31-5p, miR-99-5p, miR-342-5p, miR-183-5p, miR-627-5p, and miR-144-3p were downregulated miRNAs. Network functional analysis for Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG), in ARDS patients' blood and BALF samples, showed that the target genes were more involved in activating inflammatory and apoptosis process. CONCLUSION: Based on our results, the transcriptome profile of ARDS patients would be a valuable source for understanding molecular mechanisms of host response and developing clinical guidance on anti-inflammatory medication.

Evaluation of plasma circ_0006282 as a novel diagnostic biomarker in colorectal cancer.

BACKGROUND: Nowadays, non-invasive and rapid detection of cancers through molecular biomarkers has received much attention. Therefore, this study investigated the non-invasive and rapid diagnosis of colorectal cancer through one of the newest biomarkers (circular RNA). METHODS: For this purpose, we collected tumoral, adjacent normal tissue, and plasma samples from 100 colorectal cancer (CRC) patients, 25 postoperative CRC patients, 28 colitis patients, and 108 healthy donors. First Illumina high-throughput (Hi Seq 2000) sequencing was performed to identify known and novel differentially expressed circRNAs in the cancerous and adjacent normal tissues (n = 3). We used quantitative real-time fluorescent polymerase chain reaction (qRT-PCR) to detect the expression level of hsa_circ_0006282 among the different samples. Moreover, inter- and intra-assays were performed to evaluate the potential of hsa_circ_0006282 as being a biomarker. The receiver operating characteristic curve (ROC) was drawn to appraise its diagnostic efficacy, and the sensitivity of this circ RNA was evaluated. RESULTS: Based on RNA-sequencing results circ_0006282, cirs7, circ-0001313, circ_0055625, circ_000984, circ_0055625, circ_0001178, circ_0071589, circ-001569 were upregulated, and circ-ITGA7, circ-CDYL, circITCH, circ_0026344, circ_0000038, circ_0002220, circ_0067480, circIGHV3-20-1, circ_104916, circ_0009361 were downregulated circRNA. The hsa_circ_0006282 was the highest upregulated differentially expressed circRNA. Expression evaluation of this circRNA on different samples showed upregulation in CRC tissues (p < 0.0001) and plasma samples of CRC patients in comparison to healthy controls (p < 0.0001), while the area under the curve (AUC) was 0.831 (95% CI: 0.779-0.883). Expression of hsa_circ_0006282 in CRC patients decreased to normal after surgery (p < 0.0001). Our results showed high specificity and sensitivity of CRC detection when hsa_circ_0006282, carcinoembryonic antigen (CEA), and carbohydrate antigen199 (CA199) are combined. CONCLUSION: Plasma hsa_circ_0006282 can be used as a novel diagnostic and dynamic monitoring biomarker in CRC patients.

Effect of Layering Technique on Push-Out Bond Strength of Composite Resin to Intracanal Dentin of Primary Anterior Teeth.

OBJECTIVES: This in-vitro study aimed to compare the push-out bond strength of composite resin posts packed into the root canal of primary anterior teeth using two different layering techniques. MATERIALS AND METHODS: Thirty-two primary anterior teeth were randomly divided into two groups. In group 1, after the preparation of post spaces, a posterior composite resin (Filtek P60) was packed in three horizontal layers by a composite condenser instrument with a cylindrical tip using the horizontal layering technique (HLT). In group 2, this was done using a condenser with a conical tip in three funnel-shaped layers according to the funnel-shaped layering technique (FSLT). Next, the specimens were subjected to push-out bond strength testing. Data were analyzed using t-test and the Kaplan-Meier curves. RESULTS: The mean±standard deviation (SD) bond strengths of composite resin posts were 8.46±3.45 MPa and 7.7±2.24 MPa for the HLT and FSLT, respectively; the difference was not statistically significant (P=0.46). CONCLUSIONS: The layering technique by which composite resin was packed into the root canal of primary anterior teeth (HLT versus FSLT) had no significant effect on the push-out bond strength of composite resin posts.

Fracture resistance of three different posts in restoration of severely damaged primary anterior teeth: An in vitro study.

BACKGROUND: Restoration of anterior primary teeth with severe caries lesion is a big challenge. The aim of this study was to compare the fracture resistance of three types of post, including composite resin, customized quartz fiber and prefabricated glass fiber in restoration of severely damaged primary anterior teeth. MATERIALS AND METHODS: Sixty extracted human primary maxillary incisors were randomly divided into three groups: Group 1: Customized quartz fiber post, Group 2: Composite post and Group 3: Prefabricated glass fiber post. Due to the effect of bonded area on the fracture resistance, the bonded surface of each sample was measured 1 mm above cementoenamel junction. An increasing force was subjected with a crosshead speed of 0.5 mm/min by a universal testing machine until fracture occurred, and the failure mode was assessed afterwards. Data were analyzed using One-way analysis of variance and Kruskal-Wallis tests. The level of significance was considered at P < 0.05. RESULTS: The mean fracture resistance values of three groups were 343.28 N, 278.70 N and 284.76 N, respectively. Although customized quartz fiber post showed the greatest fracture resistance, statistical analysis revealed no significant difference between groups (P = 0.21). The mean fracture strength values of three groups were 12.82 N/mm(-2), 11.93 N/mm(-2) and 11.31 N/mm(-2), respectively; however, the differences were not statistically significant (P = 0.72). Favorable failure mode was more frequent in all groups (P = 0.12). CONCLUSION: Within the limitations of this study, it can be concluded that all three types of studied posts can be successfully used to restore badly destructed primary anterior teeth.