Rehabilitative treatment of patients with covid-19 infection: The p.a.r.m.a. evidence based clinical practice protocol

Background: The impact of the SARS-CoV-2 on the National Health System (NHS) required a reorganization of the various levels of care, which also involved the rehabilitation reality. Aim of the work: A clinical practice review of the literature was conducted to provide operational-rehabilitation guidelines adapt-ed to the local reality and to the recent corporate reorganization in the context of the COVID-19 emergency. Methods: A practice review of the available scientific evidence was regularly conducted from the start of the COVID-19 pandemic to periodically update the clinical practice guidelines. Articles that met the following inclusion criteria were included: studies conducted on human adult subjects with COVID-19 infection, un-dergoing rehabilitation in any hospitalization setting. Results: The results of this clinical practice update were periodically discussed with colleagues and collaborators in a multi-professional team, in order to guarantee a good clinical practice protocol, named P.A.R.M.A. Conclusions: The P.A.R.M.A. protocol is the result of a periodic review literature update, which has allowed us to take charge of patients affected by COVID-19 ac-cording to the most up-to-date clinical evidences, guaranteeing a shared and uniform treatment within a local reality in an era of health emergency. (www.actabiomedica.it)

Combining gene therapy with fetal hemoglobin induction on erythroid precursor cells from β039 and IVS-I-110 thalassemia patients

Gene therapy is one of the strategy to correct the lack of β-globin synthesis in human hematopoietic stem cells (HSC) derived from β-thalassemia patients. However, the use of lentiviral vector carrying a therapeutic β-globin gene might fall short in achieving a complete reversion of the β-thalassemic phenotype due to current limitations in vector design and myeloablative regimen. Following gene transfer all or a large proportion of erythroid cells might express suboptimal levels of β-globin, impairing the therapeutic potential of the treatment. The aim of the present study was to evaluate whether, in absence of complete reversion of the β-globin phenotype upon gene transfer, it is possible to use fetal hemoglobin induction to eliminate the residual -globin aggregates and achieve normal levels of hemoglobin. Gene therapy was performed with the lentiviral vector T9W. Induction of fetal hemoglobin was obtained using mithramycin (MTH). Levels of mRNA and hemoglobins were determined by qRT-PCR and HPLC. Erythroid progenitor cells from five β039/β039 and three β039/IVS-I-110 thalassemia patients were treated with T9W, which expresses the human -globin gene, and mithramycin, separately or in combination. When transduction with the T9W lentiviral vector is insufficient to completely eliminate the unpaired -globin chains, combination of -globin gene transfer therapy together with fetal hemoglobin induction was found very efficacious to remove the excess of -globin-proteins in thalassemic erythroid progenitor cells, independently from the genotype. The real clinical in vivo benefit, in our opinion, might be assayed in humanized transgenic mice mimicking thalassemia, in which HbF can be induced, the gene therapy is possible and the data obtained are informative

Increase in gamma-globin mRNA content in human erythroid cells treated with angelicin analogs

The aim of the present study was to identify molecular analogs of angelicin (ANG) able to increase erythroid differentiation of K562 cells and expression of gamma-globin genes in human erythroid precursor cells, with low effects on apoptosis. ANG-like molecules are well-known photosensitizers largely used for their antiproliferative activity in the treatment of different skin diseases (i.e., psoriasis, vitiligo, eczema, and mycosis fungoides). To verify the activity of these derivatives, we employed three experimental cell systems: (1) the human leukemic K562 cell line, (2) K562 cell clones stably transfected with a pCCL construct carrying green-EGFP under the gamma-globin gene promoter, and (3) the two-phase liquid culture of human erythroid progenitors isolated from normal donors and beta-thalassemia patients. The results of our study suggest that trimethyl ANG is a powerful inducer of erythroid differentiation, compared with known inducers, such as ANG, cytosine arabinoside, mithramycin, and cisplatin. These data could have practical relevance, because pharmacologically mediated regulation of human gamma-globin gene expression, with the consequent induction of fetal hemoglobin, is considered a potential therapeutic approach in hematological disorders including beta-thalassemia and sickle cell anemia

Bergamot (Citrus bergamia Risso) fruit extracts as gamma-globin gene expression inducers: Phyto-chemical and functional perspectives

Epicarps of Citrus bergamia fruits from organic farming were extracted with the object to obtain derived products differently rich in coumarins and psoralens. The extracts were chemically characterized by 1H-NMR (nuclear magnetic resonance), GC-FID (gas chromatography-flame ionization detector), GC-MS (gas chromatography-mass spectrometry) and HPLC (high pressure liquid chromatography) for detecting and quantifying the main constituents. Both bergamot extracts and chemical standards corresponding to the main constituents detected were then assayed for their capacity to increase erythroid differentiation of K562 cells and expression of gamma-globin genes in human erythroid precursor cells. Three experimental cell systems were employed: (a) the human leukemic K562 cell line, (b) K562 cell clones stably transfected with a pCCL construct carrying green-EGFP (enhanced green fluorescence protein) under the gamma-globin gene promoter and (c) the two-phase liquid culture of human erythroid progenitors isolated from healthy donors. The results suggest that citropten and bergapten are powerful inducers of differentiation and gamma-globin gene expression in human erythroid cells. These data could have practical relevance, since pharmacologically-mediated regulation of human gamma-globin gene expression, with the consequent induction of fetal hemoglobin, is considered a potential therapeutic approach in hematological disorders, including beta-thalassemia and sickle cell anemia

Increase in gamma-globin mRNA content in human erythroid cells treated with angelicin analogs

The aim of the present study was to identify molecular analogs of angelicin (ANG) able to increase erythroid differentiation of K562 cells and expression of gamma-globin genes in human erythroid precursor cells, with low effects on apoptosis. ANG-like molecules are well-known photosensitizers largely used for their antiproliferative activity in the treatment of different skin diseases (i.e., psoriasis, vitiligo, eczema, and mycosis fungoides). To verify the activity of these derivatives, we employed three experimental cell systems: (1) the human leukemic K562 cell line, (2) K562 cell clones stably transfected with a pCCL construct carrying green-EGFP under the gamma-globin gene promoter, and (3) the two-phase liquid culture of human erythroid progenitors isolated from normal donors and beta-thalassemia patients. The results of our study suggest that trimethyl ANG is a powerful inducer of erythroid differentiation, compared with known inducers, such as ANG, cytosine arabinoside, mithramycin, and cisplatin. These data could have practical relevance, because pharmacologically mediated regulation of human gamma-globin gene expression, with the consequent induction of fetal hemoglobin, is considered a potential therapeutic approach in hematological disorders including beta-thalassemia and sickle cell anemia