In vitro testing for genotoxicity of indigo naturalis assessed by micronucleus test.

In the field of cosmetic dyes, used for coloring the hair and skin, there is a clear tendency to replace the widely used synthetic dyes by natural colorants, such as henna and mixtures of henna with indigo. The aim of this study was to estimate the genotoxicity of water and DMSO solutions of indigo naturalis (prepared from Indigofera tinctoria leaves) using the cytokinesis-blocked micronucleus (CBMN) assay in the human metabolically active HepG2 cell line. The cytotoxic effects of indigo solutions were first assessed by propidium iodide and fluorescein-diacetate simultaneous staining. For both solutions, cytotoxicity was always under 10%. Data obtained in the CBMN assay (for all concentrations tested) indicated that the frequency of MN (micronuclei) in exposed cells was no higher than the control. Both the water and DMSO solutions showed the same behavior. These results indicate that indigo naturalis exhibits neither cytotoxicity, nor genotoxicity for all concentrations tested, which may justify excluding indigofera and its components from the list of carcinogenic agents

Evaluation of Bone Mineral Density in a Cohort of Children with Growth Hormone Deficiency

Background: Growth Hormone (GH) plays an important role in linear growth and in bone turnover during childhood. GH deficiency (GHD) may cause secondary osteoporosis associated to low bone mineral density (BMD), impairment of bone turnover and increased fracture rate. The effects of treatment with recombinant human Growth Hormone (rhGH) on bone metabolism are controversial. We aimed to assess BMD using dual energy x-ray absorptiometry (DEXA) among a cohort of children with GHD before rhGH therapy. Furthermore, we aimed to evaluate the association between BMD and auxological, biochemical and therapeutic data at baseline and during rhGH therapy. Methods: We enrolled 193 patients (9.68 ± 3.27 years, 58% males, 75% in a pre-pubertal age) with diagnosed GHD. DEXA was performed before treatment. Anamnestic, anthropometric, biochemical and radiological data were evaluated at baseline and during rhGH treatment (6, 12 and 24 months). Results: The median value of BMD Z-score before rhGH therapy was -1.15 ± 0.97. Analyzing BMD values at baseline, we found differences between pubertal and pre-pubertal patients (BMD SDS -1.54± 0.95 vs. -0.97±0.93; p < 0.001) and between patients with a normal brain magnetic resonance imaging (MRI) and subjects with a pathologic MRI (BMD SDS -1.09±0.99 vs.-1.48±0.82 p 0.03, respectively). The absolute value of BMD (g/cm²) was positively correlated with height SDS (r 0.20, p <0.05), BMI SDS (r 0.24, p <0.05) and IGF-1 values (r 0.33, p <0.05); BMD SDS value was positively correlated with target height SDS (r 0.28, p <0.05) and BMI SDS (r 0.36, p <0.05) whereas there was a negative correlation between BMD SDS and the age at GHD diagnosis (r² 0.40, p <0.05). There was no association between BMD values and biochemical and therapeutic data. Conclusions: Our study shows that pubertal patients have a lower BMD than pre-pubertal ones as a consequence of the mild bone demineralization (BMD Z-score -1.15±0.97) secondary to GHD. Therefore, our data suggest starting rhGH therapy early as to promote an optimal growth. DEXA might represent a valid mean to complete the diagnosis in GHD patients and to optimally orient the therapeutic decisions; it should be repeated at the end of treatment in order to evaluate its effect on bone metabolism

Proteome alterations in erythrocytes with PIEZO1 gain-of-function mutations

: Gain-of-function (GoF) mutations in PIEZO1 cause dehydrated hereditary stomatocytosis (DHS) or hereditary xerocytosis, an autosomal dominant hemolytic anemia characterized by high reticulocyte count, a tendency to macrocytosis, and mild jaundice, as well as by other variably penetrant clinical features, such as perinatal edema, severe thromboembolic complications after splenectomy, and hepatic iron overload. PIEZO1 mutations in DHS lead to slowing inactivation kinetics of the ion channel and/or facilitating channel opening in response to physiological stimuli. To characterize the alterations of red blood cell proteome in PIEZO1 mutated patients, we used a differential approach to compare the proteome of DHS patients (16 patients from 13 unrelated pedigrees) versus healthy subjects. We identified new actors in the regulation of the complex landscape of ion and volume balance of erythrocytes mediated by PIEZO1. Particularly, the main impaired processes in DHS patients were: ion homeostasis, transmembrane transport, regulation of vesicle-mediated transport, and the proteasomal catabolic process. Functional assays demonstrated a co-expression of PIEZO1 and Band3 when PIEZO1 was activated. Moreover, the alteration of the vesicle-mediated transport was functionally demonstrated by the increased vesiculation rate in DHS patients compared to healthy controls. This finding provides also an explanation of the pathogenetic mechanism underlying the increased thrombotic rate observed in these patients. Finally, the newly identified proteins, involved in the intracellular signaling pathways altered by PIEZO1 mutations, could be used in the future as potential druggable targets in DHS

The HLA Variant rs6903608 Is Associated with Disease Onset and Relapse of Immune-Mediated Thrombotic Thrombocytopenic Purpura in Caucasians

Immune-mediated thrombotic thrombocytopenic purpura (iTTP) is a rare, life-threatening thrombotic microangiopathy caused by severe ADAMTS13 (a disintegrin and metalloproteinase with thrombospondin motifs 13) deficiency, recurring in 30-50% of patients. The common human leukocyte antigen (HLA) variant rs6903608 was found to be associated with prevalent iTTP, but whether this variant is associated with disease relapse is unknown. To estimate the impact of rs6903608 on iTTP onset and relapse, we performed a case-control and cohort study in 161 Italian patients with a first iTTP episode between 2002 and 2018, and in 456 Italian controls. Variation in rs6903608 was strongly associated with iTTP onset (homozygotes odds ratio (OR) 4.68 (95% confidence interval (CI) 2.67 to 8.23); heterozygotes OR 1.64 (95%CI 0.95 to 2.83)), which occurred over three years earlier for each extra risk allele (β -3.34, 95%CI -6.69 to 0.02). Of 153 survivors (median follow-up 4.9 years (95%CI 3.7 to 6.1)), 44 (29%) relapsed. The risk allele homozygotes had a 46% (95%CI 36 to 57%) absolute risk of relapse by year 6, which was significantly higher than both heterozygotes (22% (95%CI 16 to 29%)) and reference allele homozygotes (30% (95%CI 23 to 39%)). In conclusion, HLA variant rs6903608 is a risk factor for both iTTP onset and relapse. This newly identified biomarker may help with recognizing patients at high risk of relapse, who would benefit from close monitoring or intensified immunosuppressive therapy

Heterogeneous clinical presentation in ICF syndrome: correlation with underlying gene defects.

Immunodeficiency with centromeric instability and facial anomalies (ICF) syndrome is a primary immunodeficiency, predominantly characterized by agammaglobulinemia or hypoimmunoglobulinemia, centromere instability and facial anomalies. Mutations in two genes have been discovered to cause ICF syndrome: DNMT3B and ZBTB24. To characterize the clinical features of this syndrome, as well as genotype-phenotype correlations, we compared clinical and genetic data of 44 ICF patients. Of them, 23 had mutations in DNMT3B (ICF1), 13 patients had mutations in ZBTB24 (ICF2), whereas for 8 patients, the gene defect has not yet been identified (ICFX). While at first sight these patients share the same immunological, morphological and epigenetic hallmarks of the disease, systematic evaluation of all reported informative cases shows that: (1) the humoral immunodeficiency is generally more pronounced in ICF1 patients, (2) B- and T-cell compartments are both involved in ICF1 and ICF2, (3) ICF2 patients have a significantly higher incidence of intellectual disability and (4) congenital malformations can be observed in some ICF1 and ICF2 cases. It is expected that these observations on prevalence and clinical presentation will facilitate mutation-screening strategies and help in diagnostic counseling.Journal ArticleResearch Support, N.I.H. ExtramuralSCOPUS: ar.jinfo:eu-repo/semantics/publishe