ASPETTI ETICI RELATIVI ALLA CRIOCONSERVAZIONE DI CELLULE STAMINALI DA SANGUE DI CORDONE OMBELICALE

ABSTRACT Il dibattito sulle cellule staminali, nell’ultimo decennio è stato oggetto di privilegiato interesse sia nell’ambito della ricerca scientifica e bioetica che nella divulgazione a mezzo stampa nazionale ed internazionale; inoltre coinvolge a diverso titolo una pluralità di problematiche. Le applicazioni terapeutiche delle cellule staminali, in corso già da un decennio, per la cura di diverse patologie, risultano essere altamente promettenti ed utilizzabili anche nell’ambito della medicina rigenerativa. Gli interrogativi etici differiscono profondamente a seconda della fonte da cui le cellule staminali sono prelevate. Pertanto è necessario definire quale sia l’origine delle staminali. Assume una valenza assai diversa effettuare ricerche e sperimentazioni su staminali adulte o su staminali embrionali in quanto l’utilizzo di queste ultime comporta sempre la distruzione dell’embrione da cui sono prelevate. E’ proprio intorno a tale differenza, o meglio intorno allo status dell’embrione, che si accendono i dibattiti bioetici più roventi. Fino a pochi anni fa, il cordone ombelicale era considerato, una volta eliminato dal corpo del neonato, uno scarto biologico. La scoperta della presenza di cellule staminali emopoietiche, utilizzabili ai fini trapiantologici ha incrementato l’interesse scientifico nei confronti di esso, suscitando numerose aspettative tra i ricercatori e i clinici. Inoltre la notevole capacità delle cellule cordonali di sopportare le procedure di crioconservazione a lungo termine, ha reso concreto e possibile il loro utilizzo sia in pazienti che necessitavano di una ricostituzione emopoietica sia nell’ambito della medicina rigenerativa. Il prelievo e l’utilizzo di cellule staminali isolate da cordone ombelicale non suscitano interrogativi etici rilevanti, ma l’attenzione si riversa sulle possibilità di donazione (autologa o allogenica) e sulle caratteristiche delle banche (pubbliche o private). Pertanto anche la tematica delle cellule staminali cordonali e delle biobanche coinvolgono a pieno titolo i diritti fondamentali della persona. E’ proprio sul dibattito circa il bancaggio di sangue cordonale che si incentra questa attività di dottorato, focalizzando l’attenzione sugli aspetti etici relativi ai benefici della donazione e della conservazione di sangue cordonale, e sulla sperimentazione di nuove modalità di crioconservazione che ne migliorino l’efficienza ed incrementino la vitalità delle cellule staminali cordonali dopo lo scongelo

Making the Communication of CCS more "human"

CCS communication has proven a tough challenge, particularly for the difficulty in raising interest for the technology, which is still unknown to the majority of the population, and for the complexity of conveying information about its potential for reducing emissions. In this paper we present a research based effort for bringing CCS nearer to people, through visual material developed taking into account emotional needs related to the technology. The production of a short introductory film on CCS is illustrated and its testing with a sample of 700 high school students

Peptide-mediated Interference of TIR Domain Dimerization in MyD88 Inhibits Interleukin-1-dependent Activation of NF-κB *

Myeloid differentiation factor 88 (MyD88) plays a crucial role in the signaling pathways triggered by interleukin (IL)-1 and Toll-like receptors in several steps of innate host defense. A crucial event in this signaling pathway is represented by dimerization of MyD88, which allows the recruitment of downstream kinases like IRAK-1 and IRAK-4. Herein, we have investigated the function of the Toll/IL-1 receptor (TIR) domain in MyD88 homodimerization in cell-free and in vitro experimental settings by using epta-peptides that mimic the BB-loop region of the conserved TIR domain of different proteins. By using a pull-down assay with purified glutathione S-transferase-MyD88 TIR or co-immunoprecipitation experiments, we found that epta-peptides derived from the TIR domain of MyD88 and IL-18R are the most effective in inhibiting homodimerization with either the isolated TIR or full-length MyD88. Moreover, we demonstrated that a cell permeable analog of MyD88 epta-peptide inhibits homodimerization of MyD88 TIR domains in an in vitro cell system and significantly reduces IL-1 signaling, as assayed by activation of the downstream transcription factor NF-kappaB. Our results indicate that the BB-loop in TIR domain of MyD88 is a good target for specific inhibition of MyD88-mediated signaling in vivo

Advanced cellular models for rare disease study: exploring neural, muscle and skeletal organoids

Organoids are self-organized, three-dimensional structures derived from stem cells that can mimic the structure and physiology of human organs. Patient-specific induced pluripotent stem cells (iPSCs) and 3D organoid model systems allow cells to be analyzed in a controlled environment to simulate the characteristics of a given disease by modeling the underlying pathophysiology. The recent development of 3D cell models has offered the scientific community an exceptionally valuable tool in the study of rare diseases, overcoming the limited availability of biological samples and the limitations of animal models. This review provides an overview of iPSC models and genetic engineering techniques used to develop organoids. In particular, some of the models applied to the study of rare neuronal, muscular and skeletal diseases are described. Furthermore, the limitations and potential of developing new therapeutic approaches are discussed

effects of a new human recombinant mnsod in the treatment of photoaging and actinic keratosis

Physiological processes, as aerobic metabolism and inflammatory response, generate reactive oxygen species (ROS) that may induce cellular injury when their amount is increased and antioxidant defense mechanisms are overwhelmed. Also, ROS are generated following UV skin irradiation able to deplete the natural antioxidant defenses in the skin. The increase in exposure to UV may lead to photoaging and precancerous skin lesions (actinic keratosis). New antioxidant strategies in the prevention and therapy of skin lesions are urgently needed. In this study, we evaluated the antioxidant efficacy of a recombinant form of human manganese superoxide dismutase able to inhibit reactive oxygen species production in some patients affected by severe photoaging and actinic keratosis

Continuous monitoring of natural CO2 emissions near Rome: lessons for low-level CO2 leakage detection

Continuous monitoring has been carried out at a fluvial flood-plain site near Rome for over a year. There is a mix of biogenic CO2 and deep geogenic CO2 at the site at relatively low concentrations and fluxes compared with other natural CO2 seepage sites studied previously. Factors such as temperature and soil moisture clearly affect the CO2 concentration and flux and seasonal and diurnal influences are apparent. Statistical approaches are being used to try to define these relationships and separate out the two gas components, which would be necessary in any quantification of leakage from CO2 storage

Clinical expression of facioscapulohumeral muscular dystrophy in carriers of 1-3 D4Z4 reduced alleles: Experience of the FSHD Italian National Registry

OBJECTIVES: Facioscapulohumeral muscular dystrophy type 1 (FSHD1) has been genetically linked to reduced numbers ( 64 8) of D4Z4 repeats at 4q35. Particularly severe FSHD cases, characterised by an infantile onset and presence of additional extra-muscular features, have been associated with the shortest D4Z4 reduced alleles with 1-3 repeats (1-3 DRA). We searched for signs of perinatal onset and evaluated disease outcome through the systematic collection of clinical and anamnestic records of de novo and familial index cases and their relatives, carrying 1-3 DRA. SETTING: Italy. PARTICIPANTS: 66 index cases and 33 relatives carrying 1-3 DRA. OUTCOMES: The clinical examination was performed using the standardised FSHD evaluation form with validated inter-rater reliability. To investigate the earliest signs of disease, we designed the Infantile Anamnestic Questionnaire (IAQ). Comparison of age at onset was performed using the non-parametric Wilcoxon rank-sum or Kruskal-Wallis test. Comparison of the FSHD score was performed using a general linear model and Wald test. Kaplan-Meier survival analysis was used to estimate the age-specific cumulative motor impairment risk. RESULTS: No patients had perinatal onset. Among index cases, 36 (54.5%) showed the first signs by 10 years of age. The large majority of patients with early disease onset (26 out of 36, 72.2%) were de novo; whereas the majority of patients with disease onset after 10 years of age were familial (16, 53.3%). Comparison of the disease severity outcome between index cases with age at onset before and over 10 years of age, failed to detect statistical significance (Wald test p value=0.064). Of 61 index cases, only 17 (27.9%) presented extra-muscular conditions. Relatives carrying 1-3 DRA showed a large clinical variability ranging from healthy subjects, to patients with severe motor impairment. CONCLUSIONS: The size of the D4Z4 allele is not always predictive of severe clinical outcome. The high degree of clinical variability suggests that additional factors contribute to the phenotype complexity

A new hexapeptide from the leader peptide of rMnSOD enters cells through the oestrogen receptor to deliver therapeutic molecules

A 24-amino acid leader peptide of a new human recombinant manganese superoxide dismutase can enter cells and carry molecules. Here, we demonstrated that six of the 24 amino acids penetrate cells through a particular gate represented by a specific amino acid sequence of the oestrogen receptor (ER). We analysed the internalization of the synthetic hexapeptide and the cytotoxic activity of the hexapeptide conjugated to cisplatin on a cell line panel. In most cell lines, the hexapeptide delivered an amount of cisplatin that was 2 to 8 times greater than that released by cisplatin when the drug was used alone. This increased delivery increases the therapeutic index of cisplatin and reduces side effects caused by a high dosage or long-term treatment times. We may consider this hexapeptide a new molecular carrier to deliver molecules with therapeutic activity into ER+ cells for diagnostic purposes and clinical or immune therapy

Increased prevalence of unstable HLA-C variants in HIV-1 rapid-progressor patients

HIV-1 infection in the absence of treatment results in progression toward AIDS. Host genetic factors play a role in HIV-1 pathogenesis, but complete knowledge is not yet available. Since less-expressed HLA-C variants are associated with poor HIV-1 control and unstable HLA-C variants are associated with higher HIV-1 infectivity, we investigated whether there was a correlation between the different stages of HIV-1 progression and the presence of specific HLA-C allotypes. HLA-C genotyping was performed using allele-specific PCR by analyzing a treatment-naïve cohort of 96 HIV-1-infected patients from multicentric cohorts in the USA, Canada, and Brazil. HIV-1-positive subjects were classified according to their different disease progression status as progressors (Ps, n = 48), long-term non-progressors (LTNPs, n = 37), and elite controllers (ECs, n = 11). HLA-C variants were classified as stable or unstable according to their binding stability to β2-microglobulin/peptide complex. Our results showed a significant correlation between rapid progression to AIDS and the presence of two or one unstable HLA-C variants (p-value: 0.0078, p-value: 0.0143, respectively). These findings strongly suggest a link between unstable HLA-C variants both at genotype and at allele levels and rapid progression to AIDS. This work provides further insights into the impact of host genetic factors on AIDS progression

Physical activity practiced at a young age is associated with a less severe subsequent clinical presentation in facioscapulohumeral muscular dystrophy

Background: In facioscapulohumeral muscular dystrophy (FSHD), it is not known whether physical activity (PA) practiced at young age is associated with the clinical presentation of disease. To assess this issue, we performed a retrospective cohort study concerning the previous practice of sports and, among them, those with medium-high cardiovascular commitment in clinically categorized carriers of a D4Z4 reduced allele (DRA). Methods: People aged between 18 and 60 were recruited as being DRA carriers. Subcategory (classical phenotype, A; incomplete phenotype, B; asymptomatic carriers, C; complex phenotype, D) and FSHD score, which measures muscle functional impairment, were assessed for all participants. Information on PAs was retrieved by using an online survey dealing with the practice of sports at a young age. Results: 368 participants were included in the study, average age 36.6 years (SD = 9.4), 47.6% male. The FSHD subcategory A was observed in 157 (42.7%) participants with average (± SD) FSHD score of 5.8 ± 3.0; the incomplete phenotype (category B) in 46 (12.5%) participants (average score 2.2 ± 1.7) and the D phenotype in 61 (16.6%, average score 6.5 ± 3.8). Asymptomatic carriers were 104 (subcategory C, 28.3%, score 0.0 ± 0.2). Time from symptoms onset was higher for patients with A (15.8 ± 11.1 years) and D phenotype (13.3 ± 11.9) than for patients with B phenotype (7.3 ± 9.0). The practice of sports was associated with lower FSHD score (-17%) in participants with A phenotype (MR = 0.83, 95% CI = 0.73-0.95, p = 0.007) and by 33% in participants with D phenotype (MR = 0.67, 95% CI = 0.51-0.89, p = 0.006). Conversely, no improvement was observed in participants with incomplete phenotype with mild severity (B). Conclusions: PAs at a young age are associated with a lower clinical score in the adult A and D FSHD subcategories. These results corroborate the need to consider PAs at the young age as a fundamental indicator for the correct clinical stratification of the disease and its possible evolution