Optical Monitoring of In Situ Iron Loading into Single, Native Ferritin Proteins.

Ferritin is a protein that stores and releases iron to prevent diseases associated with iron dysregulation in plants, animals, and bacteria. The conversion between iron-loaded holo-ferritin and empty apo-ferritin is an important process for iron regulation. To date, studies of ferritin have used either ensemble measurements to quantify the characteristics of a large number of proteins or single-molecule approaches to interrogate labeled or modified proteins. Here we demonstrate the first real-time study of the dynamics of iron ion loading and biomineralization within a single, unlabeled ferritin protein. Using optical nanotweezers, we trapped single apo- and holo-ferritins indefinitely, distinguished one from the other, and monitored their structural dynamics in real time. The study presented here deepens the understanding of the iron uptake mechanism of ferritin proteins, which may lead to new therapeutics for iron-related diseases.

Structural Flexibility and Disassembly Kinetics of Single Ferritin Molecules Using Optical Nanotweezers.

Ferritin, a spherical protein shell assembled from 24 subunits, functions as an efficient iron storage and release system through its channels. Understanding how various chemicals affect the structural behavior of ferritin is crucial for unravelling the origins of iron-related diseases in living organisms including humans. In particular, the influence of chemicals on ferritin's dynamics and iron release is barely explored at the single-protein level. Here, by employing optical nanotweezers using double-nanohole (DNH) structures, we examined the effect of ascorbic acid (reducing reagent) and pH on individual ferritin's conformational dynamics. The dynamics of ferritin increased as the concentration of ascorbic acid approached saturation. At pH 2.0, ferritin exhibited significant structural fluctuations and eventually underwent a stepwise disassembly into fragments. This work demonstrated the disassembly pathway and kinetics of a single ferritin molecule in solution. We identified four critical fragments during its disassembly pathway, which are 22-mer, 12-mer, tetramer, and dimer subunits. Moreover, we present single-molecule evidence of the cooperative disassembly of ferritin. Interrogating ferritin's structural change in response to different chemicals holds importance for understanding their roles in iron metabolism, hence facilitating further development of medical treatments for its associated diseases.

Prevalence of bifid variations of the mandibular canal in an Iranian population using cone-beam computed tomography.

OBJECTIVES: Bifid mandibular canal (MC) is an anatomical variation of the MC. This study aimed to assess the prevalence and shape of bifid MC in an Iranian population. MATERIALS AND METHODS: A total of 681 patients who had undergone cone-beam computed tomography (CBCT) for different purposes between 2018 and 2020 were evaluated. After detection, bifid MCs were classified into four types forward, buccolingual, dental, and retromolar. CBCT images were assessed by two oral and maxillofacial radiologists. Data were analyzed by SPSS using an independent t-test and Chi-square test. RESULTS: Bifid MC was found in 23 (3.4%) out of 681 patients, with a mean age of 32.21 years. Ten patients (1.5%) had a bifid MC on the right side, 6 (0.9%) on the left side, and 7 (1%) bilaterally. However, no significant correlation was found between laterality and the prevalence of bifid MC (P > 0.05). Bifid MC was found in 8 males (34.8%) and 15 females (65.2%). Gender had no significant correlation with the prevalence of bifid MC (P > 0.05). Forward type was the most common (n = 8, 1.2%) followed by buccolingual (n = 5, 0.73%), dental (n = 2, 0.3%), and retromolar (n = 1, 0.14%) types. CONCLUSION: According to the present results, bifid MC was not uncommon in the Iranian population of the present study, and forward type was the most common, followed by buccal and then dental bifid MCs. There was no significant correlation between sex and age with bifid MC but bifid MC was detected more frequently in females than males, and it was seen unilaterally in a higher percentage of the cases.

Efficient targeted cancer cell detection, isolation and enumeration using immuno-nano/hybrid magnetic microgels.

Magnetic nanomaterials have drawn ample attention in the field of biotechnology due to their excellent magnetic properties and biocompatibility. These materials have been widely used for exosome isolation, DNA separation, magnetic resonance imaging, and drug delivery. However, their application in cell isolation has been limited due to the lack of efficient antibody conjugation and instability in aqueous solutions. In this study, we produced hybrid maghemite nanorod/immuno-microgels with high capturing capacity for cell isolation and enumeration. Lepidocrocite (γ-FeOOH) and maghemite (γ-Fe2O3) nanorods with controlled morphology are synthesized using hydrolysis method. The effects of the different synthesis conditions on morphology, phase composition, and magnetic properties of lepidocrocite are studied to determine the best synthesis conditions. We coat the nanorods with chitosan and attach them to the poly(N-isopropylacrylamide-co-acrylic acid) (PNIPAM-AA) microgel through chemical bonding to form a nano/hybrid microstructure. Our results suggest that the hybrid magnetic microgels have more antibody binding capacity and higher cancer cell capturing rate compared to pristine maghemite nanorods. In addition, new cell magnetometery method was applied for cancer cell quantification after capturing step in which different magnetized labelled cells were correlated to the saturation magnetization. In this method, higher concentrations of the primary cell suspension resulted in more binding of the magnetic immuno-microgels to the cells which was shown as saturation magnetization drop in the microgel-cell complex.

Developmental Screening of Children with Congenital Hypothyroidism Using Ages and Stages Questionnaires Test.

OBJECTIVES: Congenital hypothyroidism (CH) is one of the most common causes of mental retardation in children. We investigated the developmental status of children with CH screened by Ages & Stages Questionnaires (ASQ) measurement scores. MATERIALS & METHODS: In this retrospective study, neurodevelopmental status of 78 children diagnosed with CH followed up at the Outpatient Pediatric Endocrinology Clinic of Besat Hospital, Hamadan, Iran from May 2006 to Mar 2013, was evaluated by ASQ method. Data on age, sex, birth weight, birth length, head circumference, residency location, parental education level, primary venous TSH and T4 levels, age at diagnosis, treatment start age and initial levothyroxine dosage were extracted from medical records. Data were analyzed using statistical software SPSS. P-value less than 0.05 was considered statistically significant. RESULTS: Of the 78 patients, 34 (43.6%) were female, 32 (41%) had developmental disorder, and 56 (71.8%) were living in urban areas. Types of developmental impairments included: global motor delay in 13 (40.6%) patients, problem-solving in 11 (34.3%), impaired communication skills in 5 (15.6%), impaired fine motor skills in 2 (6.2%), and impairment of personal social skills in 1 (3.1%). The average ages for diagnosis and treatment were 25.65 days in patients with developmental impairment and 17.99 days in those without developmental delay. ASQ results showed significant statistical correlation with initial dose of levothyroxine (P=0.017), age of hypothyroidism diagnosis (P=0.002) and age of treatment initiation (P=0.018). CONCLUSION: Early diagnosis and treatment along with initial levothyroxine dose were most important factors of ASQ scores of children with CH. Higher dose of the levothyroxine is required at onset.

Evaluation of the Effect of Light and Scanning Time Delay on The Image Quality of Intra Oral Photostimulable Phosphor Plates.

BACKGROUND: Nowadays, digital radiography is widely used in dental practice. One of the most common types is Photo Stimulated Phosphor Plate (PSP). OBJECTIVE: The aims of this experimental study were to evaluate the impacts of different combinations of storage conditions and varying delays in reading of digital images captured using PSPs. METHODS: Standardized images of a step wedges were obtained using PSPs from the Digora digital systems. Plates were exposed and immediately scanned to produce the baseline gold standard. The plates were re-exposed and stored in four different storage conditions: white light, yellow light, natural light environment and dark room, then scanned after 10 and 30 minutes and 4 and 8 hours. Objective analysis was conducted by density measurements and the data were analyzed statistically using GEE test. Subjective analysis was performed by two oral and maxillofacial radiologists and the results were analyzed using McNemar's test. RESULTS: The results from GEE analysis show that in the natural light environment, the densities in 10 minutes did not differ from the baseline. The mean densities decreased significantly during the time in all environments. The mean densities in step 2 for the dark room environment decreased with a slighter slope in comparison to yellow environment significantly. CONCLUSION: PSP images showed significant decrease in the density in plates scanned for 10 minutes or longer after exposure which may not be detected clinically. The yellow light environment had a different impact on the quality of PSP images. The spatial resolution did not change significantly with time.