Cellular and biomolecular technologies for stratification of β thalassemia patients: applications in theranostics

The research described in the present PhD Thesis has been conducted in the context of a multicenter FP7 European Project called THALAMOSS (THalassemia MOdular Stratification System), having as major objective the identification of molecular markers for the development of personalized therapies for hemoglobinopathies, in particular ß-thalassemia. The ß-thalassemias are an autosomal recessive genetic disorders caused by the absence or reduction of ß-globin chains of adult hemoglobin, for which targeted and definitive treatments are, at present, not available. In order to sustain project based on stratification of ß-thalassemia patients according to clinical, genetic and molecular features, we established a systematic collection of cellular samples, the cellular Biobank, containing hematopoietic stem cells isolated from the peripheral blood, expanded, freezed and cryopreserved. To this aim, 75 subjects comprising ß-thalassemia patients and healthy donors have been recruited up to now. They were all characterized for the genotype (in order to detect pathogenic mutations) and possible fetal hemoglobin (HbF)-associated polymorphisms. More importantly, new protocols to efficiently isolate, culture, freeze and thaw hematopoietic stem cells were developed. In particular, we demonstrated that freezing, cryopreservation and thawing steps do not affect the erythroid differentiation potential of the cells and the natural erythroid differentiation process, in terms of kinetics and types of hemoglobin produced by the cells of the same patient. Moreover, we found that the cells stored in the Biobank are responsive, once thawed and sub-cultured, to treatments with known HbF inducers, including mithramycin, resveratrol, butyric acid and hydroxyurea, and are therefore suitable for the identification and development of new HbF inducers to be used for experimental therapeutic strategy for ß-thalassemia. In this context, we demonstrated that the induction effects depend on the subject’s genotype, strongly suggesting that this approach could be very useful to develop personalized therapies. In conclusion, this research activity will allow patients stratification taking into account all the phenotypic/genotypic characteristics of the single individual, in association with in vitro HbF induction under treatment with effective inducers, providing an important opportunity for the research and development of novel therapeutic strategies for ß-thalassemia

The Role of Physical Medicine and Rehabilitation in Shoulder Disorders

Shoulder pain is a common problem and it is responsible for a high proportion of patients presenting to general practice, causing work absenteeism and claims for sickness. A lot of factors and conditions can contribute to shoulder pain. The most prevalent cause is rotator cuff tendinitis; its relevance is correlated not only to its high prevalence rate but also to the fact that is disabling, causing high direct and indirect cost in industrialized country. Other causes of shoulder pain are shoulder impingement syndrome, calcific tendonitis, frozen shoulder, etc. In this context, physical medicine and rehabilitation plays a fundamental role. The conservative approach consists of several interventions. The aim is to decrease shoulder pain and to regain shoulder function, with the goal to reduce the degree of impingement, decreasing swelling and inflammation, and to minimize the risk of further injuries. The purpose of this chapter is to give an overview about shoulder disorders and their conservative treatment by means of physical therapy

Rehabilitation in Sarcopenic Elderly

Sarcopenia is a complex problem and an important emerging field in rehabilitation of the elderly. In 2010, the European working group on sarcopenia in older people (EWGSOP) described sarcopenia as a syndrome characterised by progressive and generalised loss of skeletal muscle mass and strength, associated with a risk of adverse outcomes such as physical disability, poor quality of life and death. This field of rehabilitation has been defined as ‘evaluative, diagnostic and therapeutic interventions whose purpose is to restore functional ability or enhance residual functional capability in elderly people with disabling impairments’. With growing numbers of frail older people, there is an increasing need for appropriate geriatric rehabilitation services. Definitely, sarcopenia needs a specific rehabilitation program to improve muscular mass and strength that must be integrated with a global approach with the aim to recover postural assessment, amplify sensory‐motor systems, in order to gain the necessary information for proper motor planning, to reduce risk of falls. Several physical agents in medicine permit to treat sarcopenia, like vibrations or electrical stimulation. The aim of this chapter is to give an overview about rehabilitative medicine for sarcopenia, highlighting the state of the art, presenting the most significative clinical researches and giving some inputs to set a rehabilitation protocol

Sarcopenia in Chronic Illness and Rehabilitative Approaches

Primary sarcopenia is considered to be age-related when no other cause is evident, other than aging itself. Secondary sarcopenia should be considered when one or more other causes are evident, such as multiple chronic conditions. Previous studies have reported that low muscle strength and impaired physical performance can be found in chronic diseases, including metabolic disease (diabetes, hypertension, and obesity), arthritis, osteoporosis, cancer, chronic kidney disease, chronic obstructive pulmonary disease, neuromuscular disease, and chronic infection. The development of preventive and therapeutic strategies against secondary sarcopenia and wasting disorders in general is an epidemiological need. The planning of a complex rehabilitation program in sarcopenia associated to chronic conditions, in the context of a comprehensive treatment, is made up of a nutritional support, exercise, correction of lifestyles, and the use of advanced physical energies. Therefore, for the purposes of the optimal management, it is essential to identify the pathogenesis and clinical characteristics that can affect the different rehabilitative treatment

Nonsteroidal Anti-inflammatory Drugs: Integrated Approach to Physical Medicine and Rehabilitation

Inflammation is an immediate response to damage; in acute phase, it is a form of defense for body and it aims to restitutio ad integrum, in the chronic form itself becomes disease. This mechanism determines inflammatory diseases that are a group of clinical disorders which are characterized by abnormal inflammatory responses such as osteoarthritis, in myalgic syndromes (like fibromyalgia or miofascial sindrome), in some forms of headache, in peripheral vascular disease, in many malignancies. In Physical and Rehabilitation Medicine, the use of analgesic drugs (including NSAIDs) is a crucial resource inside a complex bioprogressive rehabilitative project. A part of the classic use per os is characterized by a serious and systemic side effect and there is also a possibility to administer drugs through other routes. Antalgic and rehabilitative mesotherapy (ARM) is a minimally invasive technique consisting of subcutaneous injections of bioactive substances. Other alternatives are represented by iontophoresis, phonophoresis, phytotherapy, and topical application. The purpose of this chapter is to give an overview about the state of the art regarding the use of NSAIDs in physical medicine and rehabilitation

A Rational Approach to Drug Repositioning in β-thalassemia: Induction of Fetal Hemoglobin by Established Drugs

Drug repositioning and the relevance of orphan drug designation for β-thalassemia is reviewed. Drug repositioning and similar terms ('drug repurposing', 'drug reprofiling', 'drug redirecting', 'drug rescue', 'drug re-tasking' and/or 'drug rediscovery') have gained great attention, especially in the field or rare diseases (RDs), and represent relevant novel drug development strategies to be considered together with the 'off-label' use of pharmaceutical products under clinical trial regimen. The most significant advantage of drug repositioning over traditional drug development is that the repositioned drug has already passed a significant number of short- and long-term toxicity tests, as well as it has already undergone pharmacokinetic and pharmacodynamic (PK/PD) studies. The established safety of repositioned drugs is known to significantly reduce the probability of project failure. Furthermore, development of repurposed drugs can shorten much of the time needed to bring a drug to market. Finally, patent filing of repurposed drugs is expected to catch the attention of pharmaceutical industries interested in the development of therapeutic protocols for RDs. Repurposed molecules that could be proposed as potential drugs for β-thalassemia, will be reported, with some of the most solid examples, including sirolimus (rapamycin) that recently has been tested in a pilot clinical trial

Postnatal and non-invasive prenatal detection of β-thalassemia mutations based on Taqman genotyping assays

The β-thalassemias are genetic disorder caused by more than 200 mutations in the β-globin gene, resulting in a total (β0) or partial (β+) deficit of the globin chain synthesis. The most frequent Mediterranean mutations for β-thalassemia are: β039, β+ VSI-110, β+IVSI-6 and β0IVSI-1. Several molecular techniques for the detection of point mutations have been developed based on the amplification of the DNA target by polymerase chain reaction (PCR), but they could be labor-intensive and technically demanding. On the contrary, Taq- Man® genotyping assays are a simple, sensitive and versatile method suitable for the single nucleotide polymorphism (SNP) genotyping affecting the human β-globin gene. Four Taq- Man® genotyping assays for the most common β-thalassemia mutations present in the Mediterranean area were designed and validated for the genotype characterization of genomic DNA extracted from 94 subjects comprising 25 healthy donors, 33 healthy carriers and 36 β- thalassemia patients. In addition, 15 specimens at late gestation (21-39 gestational weeks) and 11 at early gestation (5-18 gestational weeks) were collected from pregnant women, and circulating cell-free fetal DNAs were extracted and analyzed with these four genotyping assays. We developed four simple, inexpensive and versatile genotyping assays for the postnatal and prenatal identification of the thalassemia mutations β039, β+IVSI-110, β+IVSI-6, β0IVSI-1. These genotyping assays are able to detect paternally inherited point mutations in the fetus and could be efficiently employed for non-invasive prenatal diagnosis of β-globin gene mutations, starting from the 9th gestational week

Rectus Femoris Characteristics in Post Stroke Spasticity: Clinical Implications from Ultrasonographic Evaluation

In stroke survivors, rectus femoris (RF) spasticity is often implicated in gait pattern alterations such as stiff knee gait (SKG). Botulinum toxin type A (BoNT-A) is considered the gold standard for focal spasticity treatment. However—even if the accuracy of injection is crucial for BoNT-A efficacy—instrumented guidance for BoNT-A injection is not routinely applied in clinical settings. In order to investigate the possible implications of an inadequate BoNT-A injection on patients' clinical outcome, we evaluated the ultrasound-derived RF characteristics (muscle depth, muscle thickness, cross-sectional area and mean echo intensity) in 47 stroke survivors. In our sample, we observed wide variability of RF depth in both hemiparetic and unaffected side of included patients (0.44 and 3.54 cm and between 0.25 and 3.16 cm, respectively). Moreover, our analysis did not show significant differences between treated and non-treated RF in stroke survivors. These results suggest that considering the inter-individual variability in RF muscle depth and thickness, injection guidance should be considered for BoNT-A treatment in order to optimize the clinical outcome of treated patients. In particular, ultrasound guidance may help the clinicians in the long-term follow-up of muscle quality

Estado nutricional de adultos y niños menores a 5 años en dos zonas de Guatemala: Resultados del proyecto “Él y Ella, tiempo e ingreso: dinámicas intra-hogar e impacto sobre la nutrición de hogares agrícolas"

Proyecto IMMANAEn esta nota informativa, explicamos los cálculos del índice de masa corporal (IMC) para adultos en hogares agrícolas de Guatemala, así como también la implementación de indicadores para evaluar el estado nutricional de los niños menores a 5 años (talla por edad, peso por edad y peso por altura). Este trabajo es parte del proyecto “Él y Ella, tiempo e ingreso: dinámicas intra-hogar e impacto sobre la nutrición de hogares agrícolas”, liderado por el Centro Internacional de Agricultura Tropical (CIAT) y la Universidad de Florida1 con fondos de la iniciativa de investigación competitiva para desarrollar métodos y métricas innovadores para las acciones de agricultura y nutrición (IMMANA Grants)2 La prevalencia de desnutrición crónica en niños guatemaltecos menores a 5 años es de 46% según el Ministerio de Salud Pública de Guatemala (INE, 2015). Desnutrición crónica se refiere a la talla insuficiente respecto a la edad y es consecuencia de condiciones socioeconómicas inadecuadas que están asociadas con una deficiente nutrición y salud de la madre, las cuales se reflejan en el cuidado inadecuado de los niños. La desnutrición crónica impide que los niños desarrollen plenamente su potencial físico y cognitivo (WHO, 2018). Al mismo tiempo y paradójicamente, Guatemala también muestra una presencia significante de sobrepeso y obesidad entre su población. La prevalencia de sobrepeso en adultos es de 56% y de obesidad 21% (WHO, 2017)

Aplicação do teste de elegibilidade multidimensional na definição do público-alvo beneficiário de políticas públicas

Este artigo apresenta o Teste de Elegibilidade Multidimensional como um método alternativo de seleção de beneficiários para políticas sociais no caso brasileiro. Como a aplicação do referido método requer a utilização de modelos estatísticos com dados coletados em pesquisas domiciliares por amostragem, elabora-se um modelo de regressão logística que relaciona a probabilidade de um domicílio estar em condição de pobreza com suas características físicas e com variáveis socioeconômicas de seus moradores. As probabilidades previstas pelo modelo constituem os escores de propensão (propensity scores) à pobreza que podem ser utilizados como critério de inclusão de domicílios em programas de transferência de renda. As estimativas dos escores de propensão e uma medida de desempenho da focalização foram calculadas com base nas informações da PNAD 2003