Study of oral microbiota diversity among groups of families originally from different countries

The diversity of oral microbiota is affected by diets habits, gender, age, ethnic group, and environment. The acquisition of oral microbiota and the role of family on oral microbiota development is poorly understood. This study aims to characterize and compare the oral bacterial microbiota among families using 16S rRNA gene sequencing. This work was conducted in Jeddah city from 2020 to 2021, in which four families composed of 20 members of different ethnicity and lifestyle were recruited. After the collection of saliva samples, the DNA was extracted and processed for 16S rRNA gene metagenomics sequencing. Among 378 OUTs generated, 39 (10.3%) were unique in group A, 13 (3.4%) unique in group B, and 11 (2.9%) were unique in groups C and D. We observed a significant variation at the level of top abundance phylum (14), families (23), genera (24), and species (22) of bacteria among family members. Within family groups, different bacterial species were reported to be more dominant among certain family members than the other; Prevotella melaninogenica, Prevotella histicola and Haemophilus parainfluenzae, Veillonella atypica, Porphyromonas pasteri and Haemophilus pittmaniae were more dominant in parents of some families than the other family member. In summary, this study highlights the precise and perceptible association of oral microbial between family members. Our findings documented the clustering of certain bacterial species in family groups, supporting the role of community in the development of oral microbiota.     

Gene and protein interaction network analysis in the epithelial-mesenchymal transition of Hertwig's Epithelial Root Sheath reveals periodontal regenerative drug targets – An in silico study

Background and aimHertwig’s Εpithelial Root Sheath (HΕRS) has a major function in the developing tooth roots. Earlier research revealed that it undergoes epithelial–mesenchymal transition, a vital process for the morphogenesis and complete development of the tooth and its surrounding periodontium. Few studies have demonstrated the role of HERS in cementogenesis through ΕMΤ. The background of this in-silico system biology approach is to find a hub protein and gene involved in the EMT of HERS that may uncover novel insights in periodontal regenerative drug targets.Materials and methodsThe protein and gene list involved in epithelial–mesenchymal transition were obtained from literature sources. The protein interaction was constructed using STRING software and the protein interaction network was analyzed. Molecular docking simulation checks the binding energy and stability of protein-ligand complex.ResultsResults revealed the hub gene to be DYRK1A(Hepcidin), and the ligand was identified as isoetharine. SΤRIΝG results showed a confidence cutoff of 0.9 in sensitivity analysis with a condensed protein interaction network. Overall, 98 nodes from 163 nodes of expected edges were found with an average node degree of 11.9. Docking results show binding energy of −4.70, and simulation results show an RMSD value of 5.6 Å at 50 ns.ConclusionIsoetharine could be a potential drug for periodontal regeneration.     

Mechanically unfolding protein L using a laser-feedback-controlled cantilever

Force spectroscopy using the atomic force microscope (AFM) can yield important information on the strength and lifetimes of the folded states of single proteins and their complexes when they are loaded with force. For example, by mechanically unfolding concatenated proteins at different velocities, a dynamic force spectrum can be built up that allows reconstruction of the energy landscape that the protein traverses during unfolding. To characterize fully the unfolding landscape, however, it is necessary both to explore the entire force spectrum and to characterize each species populated during unfolding. In the conventional AFM apparatus, force is applied to the protein construct through a compliant cantilever. This limits the dynamic range of the force spectrum that can be probed, and the cantilever recoil after unfolding may mask the presence of metastable intermediates. Here, we describe to our knowledge a new technique—constant-deflection AFM—in which the compliance of the AFM cantilever is removed. Using this technique, we show that protein L exhibits a more complex unfolding energy landscape than previously detected using the conventional technique. This technique is also able to detect the presence of a refolding intermediate whose formation is otherwise prevented by cantilever recoil.     

Recessive mutations in ELOVL4 cause ichthyosis, intellectual disability, and spastic quadriplegia

Very-long-chain fatty acids (VLCFAs) play important roles in membrane structure and cellular signaling, and their contribution to human health is increasingly recognized. Fatty acid elongases catalyze the first and rate-limiting step in VLCFA synthesis. Heterozygous mutations in ELOVL4, the gene encoding one of the elongases, are known to cause macular degeneration in humans and retinal abnormalities in mice. However, biallelic ELOVL4 mutations have not been observed in humans, and murine models with homozygous mutations die within hours of birth as a result of a defective epidermal water barrier. Here, we report on two human individuals with recessive ELOVL4 mutations revealed by a combination of autozygome analysis and exome sequencing. These individuals exhibit clinical features of ichthyosis, seizures, mental retardation, and spasticity—a constellation that resembles Sjögren-Larsson syndrome (SLS) but presents a more severe neurologic phenotype. Our findings identify recessive mutations in ELOVL4 as the cause of a neuro-ichthyotic disease and emphasize the importance of VLCFA synthesis in brain and cutaneous development.     

Isolation and Molecular Characterization of Heavy Metal-Resistant Alcaligenes faecalis from Sewage Wastewater and Synthesis of Silver Nanoparticles

Environmental pollution with toxic heavy metals is increasing throughout the world alongside industrial development. Microorganisms and microbial products can be highly efficient bioaccumulators of soluble and particulate forms of metals, especially dilute external solutions. Microbe related technologies (Biotechnology) may provide an alternative or additive conventional method for metal removal or metal recovery. This study dealt with isolation, identification and characterization of heavy metal-resistant (Pb²⁺, Cd²⁺, Al³⁺, Cu²⁺, Ag²⁺ and Sn²⁺) bacteria from sewage wastewater at Taif Province, Saudi Arabia. Nine bacterial isolates were selected by using an enrichment isolation procedure based on high level of heavy metal resistance. All the isolates showed high resistance to heavy metals with Minimum Inhibitor Concentration (MIC) ranging from 800 μg/ml to 1400 μg/ml. All nine resistant isolates showed multiple tolerances to heavy metals. On the basis of morphological, biochemical and 16S rRNA characterization, the most potent isolates (Cd1-1, Ag1-1, Ag1-3 and Sn1-1) were identified as Alcaligenes faecalis. Scanning electron microscope analysis showed that the morphology of Alcaligenes faecalis Ag1-1 was unchanged after growth in medium without and with addition of Ag²⁺ indicative Ag²⁺ is not toxic to the isolate under the conditions tested. The ability of Alcaligenes faecalis Ag1-1 to synthesize sliver nanoparticles was examined. The heavy metal-resistant bacteria obtained could be useful for the bioremediation of heavy metal-contaminated environment.     

The association between two genetic polymorphisms in ITGB3 and increase risk of venous thromboembolism in cancer patients in Eastern Province of Saudi Arabia

Venous thromboembolism (VTE) is one of the major complications in most cancer patients leading to poor prognosis and short survival. Several common clinical risk factors coexist in cancer patients are used as risk predictive biomarkers to help in the management and prevention of VTE. These include cancer site and stage, chemotherapy regimen and elevated biological markers. However, Genetic polymorphisms in genes controlling coagulation and fibrinolysis are significantly associated with VTE if detected, then they might be more sensitive individual predictive biomarkers for VTE risk assessment. This study was conducted to evaluate the association between ITGB3 rs3809865 and rs5918 with VTE risk as well as monitor the effect of VTE on overall survival of these cancer patients. In this retrospective case-control study, 195 cancer patients’ formalin-fixed paraffin embedded tissue (FFPE) samples were collected (controls n = 157, case n = 38) using the stored data through Jan 2010 to Sep 2018 from King Fahad Specialist Hospital in Dammam. Samples were genotyped using TaqMan genotyping assay, then logistic regression analysis and Chi-square were used to predict the association between risk factors and VTE. Survival Comparison was tested by the log-rank test. Genetic polymorphisms in ITGB3 (rs3809865 and rs5918) found not to be associated with VTE increasing risk in cancer patients (p>0.05). While the advanced stage was potentially increasing the risk of VTE events (OR 5.1 CI 2.01–12.9p = 0.001). Patients with VTE showed a poor overall survival reflected by the median survival rate of only three years compared to seven years for cancer patients without VTE. This study highlighted the potential influence of VTE on prognosis and survival of cancer patients and raised the importance of exploring risk predictive biomarkers in our population. This will improve the risk prediction biomarkers leading to implementing safe and effective thrombosis prophylaxis strategies.     

Postharvest disease management of Alternaria spots on tomato fruit by Annona muricata fruit extracts

The aim of this study was to assess Annona muricata L. fruit extracts as an alternative to synthetic fungicide against Alternaria alternata (Fries) Keissler, the causative agent of black spots of tomato fruit. Antifungal activities of A. muricata pulp and seed extracts were tested both in vitro and in vivo. The seed extracts were more potent at inhibiting A. alternata than the pulp extracts. The in vitro assay showed maximum inhibition of radial mycelial growth of A. alternata (90%) by methanol seed extracts, at the highest concentration of 6%. Similarly, the in vivo assay showed marked reduction in lesion diameter (2.1 mm) and consequent disease inhibition (84%) on the tomato fruit treated with methanol seed extracts. Scanning electron microscopy showed that A. muricata extracts significantly damaged the morphology of hyphae and conidial structures. The FT-IR spectrum obtained from methanol extracts showed bands representing important bioactive compounds that possess antifungal activity. Based on our findings, Annona muricata fruit extracts can be further explored as a potential, excellent alternative approach to control the postharvest Alternaria spots of tomato fruit.