Identification of Two Novel Mutations in the ATM Gene from Patients with Ataxia-Telangiectasia by Whole Exome Sequencing.

BACKGROUND: Ataxia telangiectasia (AT) is one of the most common autosomal recessive hereditary ataxia presenting in childhood. The responsible gene for AT designated ATM (AT, mutated) encodes a protein which is involved in cell cycle checkpoints and other responses to genotoxicity. We describe two novel disease-causing mutations in two unrelated Iranian families with Ataxia-telangiectasia. METHODS: The probands including a 6-year-old female and an 18-year-old boy were diagnosed with Ataxia-telangiectasia among two different Iranian families. In this study, Whole-Exome Sequencing (WES) was employed for the detection of genetic changes in probands. The analysis of the co-segregation of the variants with the disease in families was conducted using PCR direct sequencing. RESULTS: Two novel frameshift mutations, (c.4236_4236del p. Pro1412fs) and (c.8907T>G p. Tyr2969Ter) in the ataxia telangiectasia mutated ATM gene were detected using Whole-Exome Sequencing (WES) in the probands. These mutations were observed in two separate A-T families. CONCLUSION: Next-generation sequencing successfully identified the causative mutation in families with ataxia-telangiectasia. These novel mutations in the ATM gene reported in the present study could assist genetic counseling, Preimplantation Genetic Diagnosis (PGD) and prenatal diagnosis (PND) of AT.

Efficient route to pre-organized and linear polyaminopolycarboxylates: Cy-TTHA, Cy-DTPA and mono/di- reactive, tert-butyl protected TTHA/Cy-TTHA.

Pre-organized polyaminopolycarboxylate chelators Cy-TTHA and Cy-DTPA were synthesized via modular five-step syntheses from commercially available starting materials in ~ 62% and 47% overall yields, respectively. Furthermore, strategies are reported for the efficient preparation of mono- and di-reactive, tert-butyl-protected TTHA/Cy-TTHA to selectively functionalize central chelators' carboxylic acids.

Brightly Luminescent and Kinetically Inert Lanthanide Bioprobes Based on Linear and Preorganized Chelators.

The synthesis, photophysical properties, and kinetic stability of a series of water-soluble, highly emissive Tb(III) and Eu(III) complexes featuring triethylenetetraamine hexaacetic acid (TTHA) and cyclohexyl triethylenetetraamine hexaacetic acid (cyTTHA) chelator scaffolds and carbostyril sensitizers are reported. The unique and modular design of the chelators gives rise to striking quantum yields of emission in aqueous solutions (up to 54%) as well as the characteristic lanthanides' photophysical properties (long excited-state lifetimes, large effective Stokes shifts, and narrow emission peaks). Furthermore, the preorganized chelators (L3, L4, and L6) bind metal within minutes at ambient temperature yet exhibit substantial resistance to transchelation in the presence of a challenge solution (EDTA, 1 mM). Moreover, the Eu(III) complex of L4 remains stably luminescent in HeLa cells over hours, demonstrating the suitability of these compounds for live-cell imaging applications. Representative chelators suitable for derivatization and protein bioconjugation were also prepared that were functionalized with clickable azide and alkyne moieties, biotin, and trimethoprim (TMP). With exceptional long-wavelength brightness, enhanced kinetic inertness, and an adaptable synthetic route, the reported lanthanide complexes are promising probes and labels for time-gated bioanalysis, biosensing, and optical microscopy.

Photographic Evaluation of Burn Depth via Telemedicine: Insights from Iranian Surgeons.

Background: The accurate assessment of burn depth is crucial for determining appropriate treatment. Telemedicine has emerged as a promising tool for supporting burn diagnosis and decision-making, as it allows for remote consultation with burn specialists and access to high-quality imaging. The aim of this study was to evaluate the diagnostic capabilities of telemedicine in diagnosing burn depth. Methods: A total of 15 Iranian surgeons participated in this study; they were presented with 13 images of partial thickness burn ulcers located on the extremities and trunk of patients. The participating surgeons were required to provide their diagnoses of burn type and depth, as well as the necessity of surgical intervention, and their responses were recorded. Results: Data from 11 participants and 143 responses were analyzed. The average diagnostic accuracy for superficial burns was 79.3%, while for deep burns, it was 13.72%. The mean total diagnostic accuracy was 75.2%. Conclusion: The results of this study suggest that photographs can be a reliable diagnostic tool for evaluating superficial burns. However, photographs are neither valid nor reliable for assessing burn depth. These findings have important implications for the use of telemedicine in burn diagnosis and indicate that additional diagnostic tools may be necessary for accurate assessment of deep burns.

Identification of Intestinal Fungal Microflora and Bacterial Pathogens in the Collected Adult Ixodes ricinus from the Northern Provinces of Iran.

Background: Ticks are vectors of many pathogens that involve various important diseases in humans and animals, they have several diverse hosts consequently can retain a diverse group of indigenous microbes, from bacteria to fungi. Little is known about the prevalence and diversity of tick microflora colonizing the midgut and their effects on ticks and their interaction. This information is important for development of vector control strategies. Methods: This study was carried out in northern Iran during autumn 2019. Ticks, Ixodes ricinus caught alive on the bodies of domestic animals in the fall. The tick homogenate was prepared. The identification of fungal isolates was carried out according to a combination of macro and microscopic morphology and molecular sequencing. Pathogenic bacteria of the family Borreliaceae, Francisella tularensis, Borrelia burgdorferi and Coxiella burnetii were tested by real-time PCR. Results: A total of 133 mature I. ricinus ticks were collected from domestic animals, including 71.5% cattle and 28.5% sheep. The tick frequency rates were 87.21% for Mazandaran, 8.28% for Golestan and 4.51% for Gilan Provinces. Total prevalence of fungal tick contamination was 53.4% (75/133) of which Trichoderma harzianum (57%) was the most prevalent species followed by Aspergillus spp. (42%), Mortierella alpine (19%) and Penicillium polonicum (14%). All tick samples were negative for three pathogenic bacteria including Francisella tularensis, Coxiella burnetii, and Borrelia burgdorferi by real-time PCR analysis. Conclusion: These results show a first picture of the microbial diversity of ticks and highlight the importance of microbiota and their role in host-pathogen interaction.

Study the adsorption process of 5-Fluorouracil drug on the pristine and doped graphdiyne nanosheet.

The electronic properties of pristine graphdiyne nanosheet (GDY) and boron-doped graphdiyne (BGDY) were scrutinized using the first-principles calculation. Furthermore, the adsorption energy, charge transfer, and electrical conductivity variation of the 5-fluorouracil (5FU) drugs on both the GDY and BGDY sheet surfaces were reported and were employed to investigate the binding among them. The tendency of pristine GDY to 5FU drug was already identified to be negligible. Moreover, the band gap energy was changed only around 3.31% after 5FU adsorption on the GDY sheet. The adsorption energy of 5FU on the BGDY was computed in both gas and water solvent media and was about - 32.72 and - 41.96 kcal/mol, respectively. The moderate amount of solvation energy indicates the good solubility of the implemented drugs in the aqueous medium. Moreover, significant transfer of charge from the 5FU to the BGDY sheet results in a substantially positive charge for B, which is a prerequisite of the adsorption of the 5FU molecule with the suitable binding energy. In addition, after 5FU adsorption, the electrical conductivity of BGDY was increased by about 25.5%, and based on this result, the BGDY is a suitable electronic sensor for 5FU detection unlike to pristine GDY.

A Review on Important Zoonotic Bacterial Tick-Borne Diseases in the Eastern Mediterranean Region.

Background: Zoonotic diseases as health concerns worldwide account for more than half of the emerging infectious diseases. Arachnids are powerful vectors to transmit several diseases to humans. Additionally, these emerging zoonotic diseases have been a considerable health threat in the Eastern Mediterranean Region of the WHO (EMRO) due to the large population living close to farms and international trade with nearby countries. Methods: This review study is based on the reported three tick-borne diseases, Lyme disease, Tularemia, and Q fever, from Iran and other EMRO countries. To this end, we searched PubMed central, ISI web of Science, and Google with the related keywords in English at any time. The reported data are then sorted by countries for each disease. Results: According to the published data, 15 countries in the region have one/more emerging infectious diseases. Q fever has been the most frequent infection in EMRO countries, while Lyme was less recorded. Furthermore, Iran is among the countries with documented history of all three investigated diseases. Conclusion: Tick-borne disease is popular among EMRO countries, indicating that they have natural conditions for infections in animals and humans. It appears necessary to develop a disease management strategy and control programs against tick-borne diseases (TBDs). Moreover, the disease-resistant animal could be bred instead of susceptible livestock. Therefore, research studies to control TBDs should be regarded as a top priority plan.

Quantum chemical studies of mercaptan gas detection with calcium oxide nanocluster.

The electronic sensitivity and adsorption behavior for mercaptan natural gas of a Ca12O12 nanocluster were studied via ab initio computations. To be more specific, to fully grasp the influence of mercaptan molecules on the chemical and electronic features of Ca12O12 nanocluster, some parameters, namely, charge transfer of natural bond orbital, molecular electrostatic potential, binding energies, and frontier molecular orbitals, are computed. The interaction between CH4S molecule and calcium atoms of Ca12O12 nanocluster through the sulfur head is strong. This strong interaction leads to a considerable transfer of charge from CH4S to the nanocluster. After mercaptan adsorption, the existing energy gap between two levels, the highest occupied molecular orbital (HOMO) and the lowest unoccupied molecular orbital (LUMO) of the nanostructure, dropped by 2.21 eV, illustrating that the dissociation process has extensively increased the electrical conductance of nanostructure. This electrical signal can help to detect CH4S molecules. Moreover, it could be concluded that Ca12O12 nanocluster has a short recovery time. In addition, solvent considerably influences the geometry factors and electronic features of CH4S/Ca12O12 complexes, and the interactions between species are significantly weaker in the aqueous medium compared with those in the vacuum.

Medical and developmental impact of transition from subcutaneous insulin to oral glyburide in a 15-yr-old boy with neonatal diabetes mellitus and intermediate DEND syndrome: extending the age of KCNJ11 mutation testing in neonatal DM.

Mutations in the KCNJ11 gene, which encodes the Kir6.2 subunit of the ATP-sensitive potassium channel, often result in neonatal diabetes. Patients with this mutation have been successfully transitioned from insulin to sulfonylurea (SU) therapy without compromise in their glycemic control. Among patients with neonatal diabetes due to KCNJ11 mutations, approximately 25% have neurological findings including developmental delay, motor dysfunction, and epilepsy, known as DEND syndrome. There have been rare cases of juvenile patients with intermediate DEND syndrome (iDEND) reporting variable improvement in neurological function following transition from insulin to SU treatment. We describe the response to glyburide in a 15-yr-old boy with severe global developmental delays resulting from the KCNJ11 mutation V59M. The patient was discovered to have diabetes mellitus at 11.5 months of age, making this the oldest age at diagnosis of a KCNJ11 mutation-related case of neonatal diabetes. Because consensus has been to screen patients for this mutation only if younger than 6 months at the time of diagnosis, we suggest that all patients under the age of 12 months at diagnosis should receive genetic testing for monogenic causes of diabetes.

Single-Chain Lanthanide Luminescence Biosensors for Cell-Based Imaging and Screening of Protein-Protein Interactions.

Lanthanide-based, Förster resonance energy transfer (LRET) biosensors enabled sensitive, time-gated luminescence (TGL) imaging or multiwell plate analysis of protein-protein interactions (PPIs) in living cells. We prepared stable cell lines that expressed polypeptides composed of an alpha helical linker flanked by a Tb(III) complex-binding domain, GFP, and two interacting domains at each terminus. The PPIs examined included those between FKBP12 and the rapamycin-binding domain of m-Tor (FRB) and between p53 (1-92) and HDM2 (1-128). TGL microscopy revealed dramatic differences (>500%) in donor- or acceptor-denominated, Tb(III)-to-GFP LRET ratios between open (unbound) and closed (bound) states of the biosensors. We observed much larger signal changes (>2,500%) and Z'-factors of 0.5 or more when we grew cells in 96- or 384-well plates and analyzed PPI changes using a TGL plate reader. The modular design and exceptional dynamic range of lanthanide-based LRET biosensors will facilitate versatile imaging and cell-based screening of PPIs.

Association of a novel homozygous mutation in the HMGCS2 gene with an HMGCSD in an Iranian patient.

BACKGROUND: 3-Hydroxy-3-methylglutaryl-CoA (HMG-CoA) synthase 2 gene (HMGCS2) encodes a mitochondrial enzyme catalyzing the first reaction of ketogenesis metabolic pathway which provides lipid-derived energy for various organs during times of carbohydrate deprivation, such as fasting. Mutations in this gene are responsible for HMG-CoA synthase deficiency (HMGCSD). The aim of present study was to investigate the association of mutation in the HMGCS2 gene with HMGCSD in a patient with atypical symptoms. METHODS: The clinical and genetic features of an 8-months-old girl with HMGCSD were evaluated. Molecular genetic testing was conducted using whole-exome sequencing (WES) in order to identify potential disease-causing mutation. The WES finding was confirmed by the polymerase chain reaction (PCR) amplification of the target sequence carried out for the patient and her parents. The PCR products were subjected to direct sequencing using forward and reverse specific primers corresponding to the HMGCS2 gene. RESULTS: A novel homozygous missense mutation (c.266G>A p.Gly89Asp) was detected in the HMGCS2 gene. Sanger sequencing along with co-segregation analysis of all family members confirmed this novel pathogenic germline mutation. The mutant gene was found to be pathogenic by bioinformatics analysis. CONCLUSION: To our best knowledge, this is the first report of HMGCSD in Iran which would expand our knowledge about the mutational spectrum of the HMGCS2 gene and the phenotype variations of the disease.

Chemical composition and antibacterial properties of Peganum harmala L.

OBJECTIVE: The present study was conducted to investigate antibacterial properties of fruit and flower of Peganum harmala. MATERIAL AND METHODS: Column chromatography, followed by preparative thin layer chromatography (TLC) was used for final purification. The structure of pure alkaloids was determined using spectroscopic methods (1H-NMR, 13C-NMR, UV and MS). Smoke and extract of total alkaloids were investigated for antimicrobial activity against five different microorganisms (standards and hospital isolates). The antibacterial activity was evaluated using disc diffusion assay and minimum inhibitory concentration (MIC) was determined by serial dilution methods. RESULTS: Chemical investigation of the chloroform extract of ripe fruit and flower of P. harmala led to identification of three alkaloids in ripe fruit and two alkaloids in the flower and leaves of this plant. Alkaloids identified in ripe fruit were harmine, peganine (vasicine) and harmaline. Two alkaloids, harmine and peganine, were detected in the flower of P. harmala, while harmaline was only found in the ripe fruit. The total alkaloids of flower were compared with total alkaloids of ripe fruit by TLC method. Fruits and flowers had 3.12 and 3.27% alkaloid contents, respectively. CONCLUSION: Our results showed that the alkaloids and smoke were specifically more effective on Candida albicans and Gram- positive bacteria (Micrococcus luteus and Staphylococcus aureus), while Gram- negative bacteria, especially Pseudomonas aeruginosa, were less sensitive.

Relationship of Serum Klotho Level With ACE Gene Polymorphism in Stable Kidney Allograft Recipients.

INTRODUCTION: The kidney is the main source of serum Klotho production. Immunosuppressive agents could affect the kidney in this regard. The effect of the ACE gene polymorphism on Klotho production is a less studied area. This study aimed to assess serum Klotho and ACE gene in a group of stable kidney transplant recipients. MATERIALS AND METHODS: In a cross-sectional study, 30 kidney transplant recipients with stable allograft function and 27 healthy young individuals were assessed for their serum Klotho levels. The ACE gene polymorphisms were studied in both groups. RESULTS: Klotho level was higher in kidney transplant recipients than the controls, but the difference was not significant (2.76 ± 2.41 ng/mL versus 2.01 ± 1.41 ng/mL, respectively). In both groups, serum Klotho level was higher in those with the I>I polymorphism, the men, those with higher glomerular filtration rate, and younger individuals, but the differences did not reach a significant level. Higher body mass index was significantly associated with lower serum Klotho level in both groups. CONCLUSIONS: Klotho level after kidney transplantation meets the range in healthy individuals, and it is not affected by the ACE gene polymorphism.

Epidemiological features of hepatitis B and C infection in a high risk population: results of screening programs.

AIM: The aim of this study was to report the epidemiological features of HBV & HCV infection in an Iranian high risk population. BACKGROUND: Hepatitis B and hepatitis C infections are worldwide serious public health problems. Iran has an intermediate prevalence of infection and a screening program was started in 2010 among high risk individuals. PATIENTS AND METHODS: This cross-sectional study was conducted on 4455 new patients during two past years. Demographic information, age, gender, occupational status, medical history, history of vaccination against HBV, high risk exposure and laboratory findings were collected for each patient. Then distribution of demographic and risk factors was evaluated in each type of hepatitis. RESULTS: The mean age of patients was 45.6±17.3 years. More than two-thirds of the diagnosed cases were infected with HBV. 74% of patients were carriers of hepatitis virus. 60% of patients had no symptoms at diagnosis. Illicit intravenous drug use was most common high risk exposure in patients under study (n=366, 8.2%). High risk behaviors including illicit intravenous drug use and unprotected sex were relatively higher in patients infected with hepatitis C compared to patients with hepatitis B infection. CONCLUSION: Findings of this study suggest that illicit intravenous drug use, contact with an infected household member and unprotected sex are the most common high risk exposure in Iranian patients infected with viral hepatitis. Therefore, preventive strategies such as health education, vaccination and screening programs should be directed to these groups. The results also show that a majority of patients have no symptoms at the time of diagnosis, therefore periodic screening tests in high risk groups is required.

Comparison of intradermal injection of autologous epidermal cell suspension vs. spraying of these cells on dermabraded surface of skin of patients with post-burn hypopigmentation.

INTRODUCTION: One of the most important complications after burning is hypo/depigmentation. This study was designed to compare two methods of cell spray and intradermal injection of epidermal cell suspension for treatment of burn induced hypopigmentation. MATERIAL AND METHODS: In this study, 28 patients with post burn hypo/depigmentation were selected and divided in 2 groups. A small skin biopsy was taken from normal skin of patients in operation room and epidermal cell suspension was prepared using NaBr 4N and trypsin. In the first group, the epidermal cell suspension was sprayed on the wound surface and then the area was dressed with amniotic membrane and gauze. In the second group, the cell suspension was injected in intradermal manner in the hypopigmented area. The patients were followed up and to evaluate the effect of the cells, photos were taken from the area before operation and also at follow-up. Clinical evaluation was done by the surgeon and a clinical score between "0" to "4" was used to demonstrate the clinical status from poor to excellent pigmentation. Skin biopsies were taken from depigmented area before and after interventions. Melanocytes were stained using anti S100 antibody and were counted in ×400 magnification fields. RESULTS: Eighteen patients were in cell spray and 10 were in cell injection groups. Mean change of pigmentation in two group showed that there was no statistical significant differences in pigmentation between two groups, (P value = 0.52) although a limited improvement in pigmentation status was observed in both groups. Regarding melanocyte numbers per field, there was not a significant difference between two groups and also before and after interventions, but melanocyte number increased after treatment in both groups. CONCLUSION: We did not find noticeable differences between cell spray and intradermal injection methods. Although both methods showed a limited effect on pigmentation of depigmented skin, the clinical results were not satisfactorily for both patients and clinicians.

Effect of an integrated case-based nutrition curriculum on medical education at Qazvin University of Medical Sciences, Iran.

INTRODUCTION: Nutrition education is identified as an important part of medical education by organizations. Qazvin University of Medical Sciences, school of medicine (QUMS SOM), has a required basic nutrition course of 36 hr in second year of medical school, but education experts reports show that the course does not provide required therapeutic skills for graduate student. METHOD: We decided to organize an 8-hr diet therapy work shop in order to develop a patient experience clinical based case study teaching to enhance clinical skills at QUMS SOM. RESULTS: Students' perception score about adequacy of nutrition instruction increased from 20% (at baseline) to 70% (after intervention). The mean nutrition knowledge score of total students in clinical nutrition were different between before and one month after integration (8.3±2.5, 13.4±3.2, P < 0.001). And two groups of participants including staggers and interns had similar nutritional knowledge score at pre-test (7.9±2.6 and 8.9±2.3 respectively). CONCLUSION: Implemented nutrition curriculum at QUMS was appropriate method to enhance student's perception about nutrition integration and to increase and translate the knowledge to clinical practice.

Type 2 diabetes mellitus in children and adolescents.

The rising prevalence of obesity in the United States has resulted in an increased incidence of T2DM in children and adolescents. T2DM now accounts for up to 45% of new cases of diabetes in the pediatric population. Many new medications have been developed for the treatment of T2DM, although few have been investigated in children. Although we have learned much in the past decade, more knowledge is needed regarding the unique aspects of T2DM in pediatric patients to optimize the prevention and treatment of this disease in children and adolescents.

Partial rescue of growth failure in growth hormone (GH)-deficient mice by a single injection of a double-stranded adeno-associated viral vector expressing the GH gene driven by a muscle-specific regulatory cassette.

Growth hormone (GH) deficiency (GHD) causes somatic growth impairment. GH has a short half-life and therefore it must be administered by daily subcutaneous injections. Adeno-associated viral (AAV) vectors have been used to deliver genes to animals, and double-stranded AAV (dsAAV) vectors provide widespread and stable transgene expression. In the present study we tested whether an intramuscular injection of dsAAV vector expressing GH under the control of a muscle creatine kinase regulatory cassette would ensure sufficient systemic GH delivery in conjunction with muscle-specific expression. Virus-injected GHD mice showed a significant (p < 0.05) increase in body length and body weight, without reaching full normalization, and significant (p < 0.05) reduction in absolute and relative visceral fat. Quantitative RT-PCR showed preferential GH expression in skeletal muscles that was confirmed by qualitative fluorescence analysis in mice injected with a similar virus expressing green fluorescent protein. The present study shows that systemic GH delivery to GHD animals is possible via a single intramuscular injection of dsAAV carrying a muscle-specific GH-expressing regulatory cassette.

Mutation analysis of the muscarinic cholinergic receptor genes in isolated growth hormone deficiency type IB.

BACKGROUND: Isolated GH deficiency (IGHD) is familial in 5-30% of patients. The most frequent form (IGHD-IB) has autosomal recessive inheritance, and it is known that it can be caused by mutations in the GHRH receptor (GHRHR) gene or in the GH gene. However, most forms of IGHD-IB have an unknown genetic cause. In normal subjects, muscarinic cholinergic stimulation causes an increase in pituitary GH release, whereas its blockade has the opposite effect, suggesting that a muscarinic acetylcholine receptor (mAchR) is involved in stimulating GH secretion. Five types of mAchR (M(1)-M(5)) exist. A transgenic mouse in which the function of the M(3) receptor was selectively ablated in the central nervous system has isolated GH deficiency similar to animals with defective GHRH or GHRHR gene. OBJECTIVE: We hypothesized that mAchR mutations may cause a subset of familial IGHD. PATIENTS/METHODS: After confirming the expression of M(1)-M(5) receptor mRNA in human hypothalamus, we analyzed the index cases of 39 families with IGHD-IB for mutations in the genes encoding for the five receptors. Coding sequences for each of the five mAchRs were subjected to direct sequencing. RESULTS: In one family, an affected member was homozygous for a M(3) change in codon 65 that replaces valine with isoleucine (V65I). The V65I receptor was expressed in CHO cells where it had normal ability to transmit methacholine signaling. CONCLUSION: mAchR mutations are absent or rare (less than 2.6%) in familial IGHD type IB.