Synthesis of imidazo[1,2-a]benzoazepines by alkyne-carbonyl-metathesis.

Imidazo[1,2-a]benzoazepines were prepared by Brønsted acid-mediated intramolecular alkyne-carbonyl metathesis (ACM). The starting materials, imidazole and benzimidazole derivatives, were prepared by N-alkylation, formylation and Sonogashira cross-coupling reaction. The final intramolecular ACM delivered the final products in good to excellent yields and with a wide tolerance towards functional groups.

Regioselective Synthesis of Naphthothiophenes by Pd Catalyzed Cross-Coupling Reactions and Alkyne-Carbonyl Metathesis.

Naphthothiophenes were prepared from commercially available 2,3-dibromothiophenes in two steps by one-pot Suzuki/Sonogashira or Sonogashira/Suzuki coupling reactions, followed by intramolecular alkyne-carbonyl-metathesis reactions. The final cyclization reaction proceeds in the presence of p-toluenesulfonic acid and provides a rapid access to two series of isomeric naphthothiophenes.

Synthesis of Dibenzotropones by Alkyne-Carbonyl Metathesis.

Dibenzocycloheptanones (dibenzotropones) were prepared by Brønsted acid mediated intramolecular alkyne-carbonyl metathesis (ACM) reactions. The cyclization precursors are readily available by Sonogashira reaction of 2-bromobenzoyl chloride with terminal alkynes, followed by Suzuki reactions with benzaldehydes. The ACM reactions are highly modular and atom economic and allow for the construction of two regioisomeric series of dibenzotropones.

Relationship between endocrine changes and bone markers in pediatric thalassemic patients after hematopoietic stem cell transplantation.

BACKGROUND: Beta thalassemia major and its treatment by hematopoietic stem cell transplantation can have deleterious effects on bone integrity and a main part of such effects is due to their deleterious effects on endocrine systems. So, we assessed the effects of endocrine changes during HSCT (Hematopoietic Stem Cell Transplantation) on growing bones of pediatric thalassemic patients. METHODS: Bone-specific alkaline phosphatase and osteocalcin (bone formation markers), N-terminal telopeptide (NTX, bone resorption marker), calcium (Ca), phosphorus (P), alkaline phosphatase (Alk ph), parathyroid hormone (PTH), vitamin D (vit D), prolactin, luteinizing hormone (LH), follicle-stimulating hormone (FSH), thyroxine (T4), triiodothyronine (T3), thyroid-stimulating hormone (TSH), insulin-like growth factor 1 (IGF-1), testosterone (in males) or estradiol (in females), measured in 20 major thalassemic patients with mean age of 10.8±3.9 years. Parameters at the baseline (before HSCT), and 1 month and 3 months after HSCT. RESULTS: After stem cell transplantation, changes of mean serum levels of NTX, osteocalcin, prolactin, LH, T4, IGF-1, testosterone (in males), Ca, Alk ph, PTH, and vit D were not significant, but bone specific Alk ph, P, T3, TSH, FSH and estradiol changed significantly (P=0.013, P=0.001, P=0.48, P=0.02, P=0.04 and P=0.001, respectively). After one month, there was a significant positive relationship between osteocalcine and T3 (p= 0.009). After 3 months, also, there was a significant positive relationship between osteocalcine and T3 and T4 as well as a negative one with IGF-1 (P<0.001, P<0.02 and P<0.03, respectively). CONCLUSIONS: Endocrine disorders do not appear to have an overall positive or negative effect on bone metabolism (anabolism or catabolism) in HSCT pediatric thalassemic patients in short term (three months).

Correction to: Clinical and genetic analysis of two wolfram syndrome families with high occurrence of wolfram syndrome and diabetes type II: a case report.

Following publication of the original article [1], the authors flagged that the name of 'Asal Hojjat' was misspelled; the name had been spelled as 'Asal Hojat'.

Clinical and genetic analysis of two wolfram syndrome families with high occurrence of wolfram syndrome and diabetes type II: a case report.

BACKGROUND: Mutations of the WFS1 gene are responsible for most cases of Wolfram syndrome (WS), a rare, recessively inherited neurodegenerative disorder characterized by juvenile-onset non-autoimmune diabetes mellitus and optic atrophy. Variants of WFS1 are also associated with non-syndromic hearing loss and type-2 diabetes mellitus (T2DM). Our study adds to literature significant associations between WS and T2DM. CASE PRESENTATION: In this study, we analyzed the clinical and genetic data of two families with high prevalence of WS and T2DM. Genetic linkage analysis and DNA sequencing were exploited to identify pathogenic variants. One novel pathogenic variant (c.2243-2244insC) and one known pathogenic (c.1232_1233delCT) (frameshift) variant were identified in exon eight of WFS1 gene. CONCLUSIONS: The mutational and phenotypic spectrum of WS is broadened by our report of novel WFS1 mutation. Our results reveal the value of molecular analysis of WFS1 in the improvement of clinical diagnostics for WS. This study also confirms the role of WFS1 in T2DM.

Homozygosity mapping and direct sequencing identify a novel pathogenic variant in the CISD2 gene in an Iranian Wolfram syndrome family.

AIMS: Wolfram syndrome (WS) is a rare recessive neurodegenerative disorder characterized by diabetes mellitus and optic atrophy. Mortality and morbidity rate of the disease is high in adulthood due to neurological and respiratory defects. So far, two WS genes, WFS1 (more than 90% of cases) and CISD2, have been identified. In the present study, we aimed to determine the role of WFS2 in a group of Iranian WS families. METHODS: We recruited 27 families with the clinical diagnosis of WS. Homozygosity mapping was implemented using short tandem repeat polymorphic markers and bi-directional sequencing of the CISD2 gene in families negative for WFS1 mutations. The candidate variant was checked among family members. In silico analysis and protein modeling were applied to assess the pathogenic effect of the variant. Tetra-primers ARMS PCR was set up for checking the variant in 50 ethnic-matched controls. RESULTS: One family showed homozygosity by descent at WFS2. A novel missense variant, c.310T > C (p.S104P), was found in exon 2 of the CISD2 gene. Computational predictions revealed its pathogenic effect on protein structure, function, and stability. Parents and his healthy brother were heterozygous for the variant. The variant was not observed in the control group. CONCLUSIONS: This is the first study that elucidates the role of the CISD2 gene among Iranian WS families with a novel disease-causing missense variant. Next-generation sequencing could unravel disease-causing genes in remained families to expand genetic heterogeneity of WS.

A Patient with Trisomy 4p and Monosomy 10q

Translocations are the most common structural abnormality in the human genome. Carriers of balanced chromosome rearrangements exhibit increased risk of abortion or a chromosomally-unbalanced child. The present study was carried out in 2017 at the Iranian Blood Transfusion Research Center. This study reported a rare chromosomal disorder with 4p duplication and 10q distal deletion syndrome which is associated with various complications at birth. Defects included the following characteristics: dysmorphic facial characteristic, hand or foot anomalies, growth retardation, developmental delay, strabismus, heart defects and renal anomalies. Cytogenetic analysis and array CGH were performed and, for the first time, we reported a patient with trisomy 4p16.3p12 and monosomy 10q26.3. The patient was found to have: arr 4p16.3p12 (37,152-45,490,207) x3, 10q26.3 (134,872,562-135,434,149) x1 genomic imbalances.

Clinical and molecular assessment of 13 Iranian families with Wolfram syndrome.

PURPOSE: Wolfram syndrome (WS) is a rare genetic disorder described by a pattern of clinical manifestations such as diabetes mellitus, diabetes insipidus, optic nerve atrophy, sensorineural hearing loss, urinary tract abnormalities, and psychiatric disorders. WFS1 and WFS2 loci are the main genetic loci associated with this disorder. METHODS: In the current study, we investigated associations between these loci and WS via STR markers and homozygosity mapping in 13 Iranian families with WS. All families were linked to WFS1 locus. RESULTS: Mutation analysis revealed four novel mutations (Q215X, E89X, S168Del, and E391Sfs*51) in the assessed families. Bioinformatics tools confirmed the pathogenicity of the novel mutations. Other identified mutations were previously reported in other populations for their pathogenicity. CONCLUSIONS: The current study adds to the mutation repository of WS and shows a panel of mutations in Iranian population. Such panel would facilitate genetic counseling and prenatal diagnosis in families with WS cases.

The activity and tissue distribution of thioredoxin reductase in basal cell carcinoma.

PURPOSE: Basal cell carcinoma (BCC) is the most prevalent cancer worldwide. Different mechanisms are proposed to be involved in its pathogenesis such as oxidative stress. Oxidative stress, which is the consequence of the disruption of redox balance in favor of oxidants, is involved in the initiation or progression of many tumors. Thioredoxin reductase (TrxR) is a key enzyme of the thioredoxin (Trx) system, containing Trx and TrxR and NADPH, which is one of the main cellular oxidoreductases with an essential role in cellular health and survival through providing and maintaining redox balance. Therefore, we aimed to study and compare the activity and tissue distribution of TrxR in tumoral tissue and its healthy margin in patients with BCC. METHODS: After biopsy and taking samples from 18 patients, TrxR activity was measured using a commercial kit and its tissue distribution was assessed immunohistochemically. RESULTS: Both the activity and tissue distribution of TrxR in tumoral tissues were significantly higher compared to their healthy margins. Regarding the tissue distribution, this significant increase in TrxR in tumoral tissues was documented based on both staining intensity and abundance of positive cells in immunohistochemistry. CONCLUSIONS: Based on these results, it is concluded that TrxR is involved in the pathogenesis of BCC; however, more investigations are required to clarify whether this increase is a consequence of BCC or it is an initiating mechanism.

Significant expressivity of Wolfram syndrome: phenotypic assessment of two known and one novel mutation in the WFS1 gene in three Iranian families.

Wolfram syndrome also known as DIDMOAD (Diabetes Insipidus, Diabetes Mellitus, Optic Atrophy, and Deafness) is a rare neurodegenerative autosomal recessive disorder. There is evidence of variable expressivity both in patients and heterozygous carriers. In this study, we describe three Persian Wolfram syndrome families with differences in the age of onset, signs and symptoms of the disease. We clinically evaluated affected families for verifying WS clinical diagnosis. After linkage analysis via 5 STR markers, molecular analysis for WFS1 was performed by direct sequencing for patients and available family members. Three homozygous mutations were identified including c.1885 C>T, c.2205C>A both in exon 8 and c.460+1G>A in intron 4. The mutation c.2205C>A was found to be novel. We report interesting phenotype-genotype correlations: homozygous c.1885C>T and c.2205C>A variants were correlated with quite different disease severity and onset in the siblings. We report a rare case of WS with homozygous c.1885C>T who is married and has a healthy child. c.460+1G>A showed a possible partial dominant inheritance put forth by a heterozygous parent showing partial WS symptoms while her daughter displayed typical WS symptoms. Due to variable expressivity, detailed clinical examination and molecular diagnostics should be used to confirm WS and a more exact recurrence risk data.

Molecular characterization of WFS1 in an Iranian family with Wolfram syndrome reveals a novel frameshift mutation associated with early symptoms.

Wolfram syndrome (WS) is a rare autosomal recessive neurodegenerative disorder that represents a likely source of childhood diabetes especially among countries in the consanguinity belt. The main responsible gene is WFS1 for which over one hundred mutations have been reported from different ethnic groups. The aim of this study was to identify the molecular etiology of WS and to perform a possible genotype-phenotype correlation in Iranian kindred. An Iranian family with two patients was clinically studied and WS was suspected. Genetic linkage analysis via 5 STR markers was carried out. For identification of mutations, DNA sequencing of WFS1 including all the exons, exon-intron boundaries and the promoter was performed. Linkage analysis indicated linkage to the WFS1 region. After DNA sequencing of WFS1, one novel pathogenic mutation, which causes frameshift alteration c.2177_2178insTCTTC (or c.2173_2177dupTCTTC) in exon eight, was found. The genotype-phenotype correlation analysis suggests that the presence of the homozygous mutation may be associated with early onset of disease symptoms. This study stresses the necessity of considering the molecular analysis of WFS1 in childhood diabetes with some symptoms of WS.

Cytokine gene polymorphism and graft-versus-host disease: a survey in Iranian bone marrow transplanted patients.

Graft versus host disease (GVHD) is a major complication of bone marrow transplantation (BMT). Numerous studies have shown the potential role of cytokine genotypes in the occurrence of GVHD. In this retrospective, case-control study we aimed to investigate the association between 13 cytokine genes and acute GVHD (aGVHD) after HLA-identical sibling BMT in 91 Iranian subjects. Negative association was found between aGVHD and donor IL-10/GCC haplotype or donor IL-4Ra-A allele in the population study. When compared within the leukemia subgroup, we observed positive association between recipient IL-1α -889/C allele and aGVHD. Also there were negative association between recipient IL-10/CAA haplotype and donor IL-4Ra/A allele and development of aGVHD. Among the different genotypes only donor IL-4Ra and donor IL-12 showed significant association. We conclude that several cytokine polymorphisms are positively and negatively associated with aGVHD in Iranian HLA matched siblings, of which IL-4Ra and IL-12 may play important roles.

TGF-Beta codon 25 polymorphism and the risk of graft-versus-host disease after allogenic hematopoietic stem cell transplantation.

Some of the genotypes of cytokines are associated with acute graft versus host disease after bone marrow transplantation. The purpose of the present investigation was to find out the possible association between transforming growth factor beta-1 (TGF-beta1) codon 25 polymorphism (rs:1800471) and acute graft versus host disease (aGVHD) after bone marrow transplantation from the sibling with the similar HLA among the Iranian population. In this retrospective case-control investigation, 172 subjects including 86 Iranian patients and their siblings with the similar HLA as donor/recipient pairs were recruited. All of the patients were diagnosed with one group of blood disorder consisting of Acute Myeloid Leukemia (AML)=40, Acute Lymphoblastic Leukemia (ALL)=25 and Chronic Myeloid Leukemia (CML)=21. PCR-SSP method was carried out to ascertain TGF- beta1 codon 25 G/C polymorphism genotypes. The frequency of TGF- beta1 codon 25 GG, GC and CC genotypes among all cases were 77.3%, 21.5% and 1.2%, respectively. Recipients with the GG genotype developed severe aGVHD significantly more than those with CC or GC genotypes (Odds Ratio =12.133, P=0.015). Genetic background of TGF-beta1 may be involved in aGVHD development and/or severity in the patients who received Bone Marrow Transplantation (BMT) from their siblings with the similar HLA among the Iranian population.

Frequencies of HLA-DRB1 in Iranian normal population and in patients with acute lymphoblastic leukemia.

BACKGROUND: Recognition of HLA alleles is useful in transplantation and in anthropological and disease studies. Acute lymphoblastic leukemia (ALL) is the most common blood cancer. It is now generally agreed that both genetic and environmental factors play an interactive role in the development of ALL disease. It is unknown whether there exists a restriction to certain MHC genotypes in leukemia like ALL. METHODS: Genetic construct of HLA DRB1 was studied in Iranian normal populations and in patients with acute lymphoblastic leukemia using PCR-SSP method. RESULTS: It was shown that the most common allele in DRB1 locus in normal population was DRB1*11 (20%), whereas DRB1*09 was the least frequent allele (0.9%). Additionally, this study presented the results of HLA-DRB1 typing in 106 ALL patients and compared them with normal individuals. Comparison of the results between the normal population and the patient group revealed that there was allelic association between the DRB1*13 and the disease. Results showed that the difference between the frequencies of DRB1*13 in patients and normal individuals was significant (p=0.04), but there was a moderate difference among the frequencies of DRB1*04, *07, and *09 in childhood (0-15 years) ALL. The frequencies of DRB1*13, *04, and *07 in patients were 2.5, 16, 4.5% and, in normal individuals, were 11.4, 10, and 8.3%, respectively. CONCLUSIONS: It should be concluded that DRB1*13, which showed a decrease in patients, should be protective against acute lymphoblastic leukemia (ALL), whereas DRB1*04, which was moderately increased in patients, could be considered a susceptible allele for childhood ALL.

Transforming growth factor-beta1 codon 10 polymorphism is associated with acute GVHD after allogenic BMT in Iranian population.

BACKGROUND: Certain cytokine genotypes are associated with acute graft versus host disease (aGVHD) after bone marrow transplantation (BMT). The present study aimed to determine existing association between TGF-beta1 codon 10 polymorphism and aGVHD after HLA-identical sibling BMT in the Iranian population. MATERIAL/METHODS: In a retrospective case-control study, 168 subjects including 84 Iranian HLA-identical sibling BMT donor/recipient pairs were recruited. All of the patients were affected by hematological malignancies (AML=39, ALL=23 and CML=22). PCR-SSP method was performed to determine TGF-beta1 codon 10 T/C polymorphism genotypes. RESULTS: The frequency of TGF-beta1 codon 10 TT, TC and CC genotypes among all subjects were 26.8%, 33.3% and 39.9% respectively. Recipients with the T allele developed aGVHD significantly less than those without the T allele (odds ratio =0.334, P=0.026). CONCLUSIONS: Genetic background of TGF-beta1 may be involved as a protective factor in the development of aGVHD in HLA-matched sibling BMT in Iranian population. Moreover, this finding may indicate that the genetic markers in Iranians are, at least to some extent, linked to distinct genetic event from Japanese.