In recent years, high-intensity focused ultrasound (HIFU) has emerged as a new and promising non-invasive and non-ionizing ablative technique for the treatment of localized solid tumors. Extensive pre-clinical and clinical studies have evidenced that, in addition to direct destruction of the primary tumor, HIFU-thermoablation may elicit long-term systemic host anti-tumor immunity. In particular, an important consequence of HIFU treatment includes the release of tumor-associated antigens (TAAs), the secretion of immuno-suppressing factors by cancer cells and the induction of cytotoxic T lymphocyte (CTL) activity. Radiation therapy (RT) is the main treatment modality used for many types of tumors and about 50% of all cancer patients receive RT, often used in combination with surgery and chemotherapy. It is well known that RT can modulate anti-tumor immune responses, modifying micro-environment and stimulating inflammatory factors that can greatly affect cell invasion, bystander effects, radiation tissue complications (such as fibrosis), genomic instability and thus, intrinsic cellular radio-sensitivity. To date, various combined therapeutic strategies (such as immuno-therapy) have been performed in order to enhance RT success in treating locally advanced and recurrent tumors. Recent works suggested the combined use of HIFU and RT treatments to increase the tumor cell radio-sensitivity, in order to synergize the effects reaching the maximum results with minimal doses of ionizing radiation (IR). Here, we highlight the opposite immuno-modulation roles of RT and HIFU, providing scientific reasons to test, by experimental approaches, the use of HIFU immune-stimulatory capacity to improve tumor radio-sensitivity, to reduce the RT induced inflammatory response and to decrease the dose-correlated side effects in normal tissues.

Dear Sir, The RH genes RHD and RHCE encode two proteins that represent the clinically most important blood group system defined by the sequences of red cell membrane proteins. RHD and RHCE, encoding the Rh proteins (D and Cc/Ee, respectively), are organised in tandem on chromosome 1p34-p36 and probably derived from duplication of a common ancestral gene. Many RH genes carry point mutations, or have rearrangements and exchanges between RHD and RHCE which result from gene conversion events. RHCE encode hybrid proteins that have RhCE-specific amino acids in RhD, or RhD-specific residues in RhCE. These might generate new antigens in the Rh blood group system, and alter or weaken expression of the conventional antigens1,2. Reduced expression of D antigen occurs in an estimated 0.2%–1% of Caucasians. Historically, red blood cell antigens that react with anti-D only after extended testing with the indirect antiglobulin test are called weak D. Weak D expression primarily results from single point mutations in RHD which encode amino acid changes predicted to be intracellular or in the transmembrane regions of RhD. These affect the efficiency of insertion, and, therefore, the quantity of RhD protein in the membrane, reflected in the reduced number of D antigen sites on the red blood cells. Red blood cells with partial D antigen type as D-positive, but individuals often produce anti-D when stimulated by transfusion or pregnancy. Some partial D, similar to weak D, result from point mutations in RHD that cause single amino acid changes. But, in contrast to weak D, these changes are located on the extracellular regions and alter or create new epitopes1,2. Molecular methods for blood group genotyping became available more than 10 years ago and are useful methods to help to clarify immunogenetic doubts or to verify results3. Molecular RHD blood group typing is very efficient for managing donors and patients carrying any of the various molecular types of weak D and partial D. Weak D and partial D expression are caused by a large number of RHD alleles and variations of the antigen structure of RhD result either in a partial (partial D) or a weak D phenotype (weak D). Weak and partial D result in quantitative and qualitative changes in Rh protein expression respectively. The clinical relevance of these changes are, according to Flegel1,2, that weak D subjects belonging to weak type 1, 2, 3, 4.0, 4.1 and 5 can be treated as Rh-positive and be transfused by Rh-positive red blood cells, while subjects with weak type 4.2–11 and 15 should be treated as Rh-negative and transfused with Rh-negative red blood cells. Partial D can produce different protein epitope expression and, therefore, induce specific antibody production. In this situation, partial D subjects should be considered Rh-negative and transfused with Rh-negative red blood cells1,2. In our daily practice, D antigens are determined serological agglutination tests according to the guidelines of Italian Society for Transfusion Medicine and Immunohaematology4. In particular, during routine Rhesus tests, a microtitre plate-based assay employing two different anti-D (D-Rapid, clone RUM-1 IgM and D-Fast, clone IgM; Immucor Gamma, Immucor, Inc. Norcross, GA, USA) is used. Thereafter, the D negative samples are tested for Du with two different methods: a microtitre plate employing Anti-D Duo IgG-IgM (clone IgG/IgM clone Th28+MS26; Galileo Capture R ImmucorGamma) using solid phase capture and a gel matrix test employing one anti-D (Id-Dia Clone Anti-D; DiaMed GmbH, Switzerland). If the result is positive, the samples are tested with a gel matrix direct antiglobulin assay. The serological analysis for allelic D variant is based on “Partial Rh Typing” (ImmucorGamma) using six monoclonal IgG antisera and Capture-R Select (ImmucorGamma) in a solid phase method. Molecular biology analysis is performed using commercial kits from BAGene Health Care GmbH (Weak D-TYPE; Partial D-TYPE; BAG Health Care GmbH, Germany). The basic material for typing with BAGene DNA-SSP kits is purified DNA from peripheral blood mononuclear cells. The test is based on sequence-specific primers (SSP) - polymerase chain reaction (PCR). BAGene Partial D-TYPE allows for the molecular genetic determination of partial D such as DII, DIII, DIV, DV, DVI, DVII, DAU, DBT, DFR, DHMi, DHMii, DNB and DHAR (Rh33)5, whereas BAGene Weak D-TYPE allows the molecular genetic determination of weak D types including 1, 2, 3, 4.0/4.1, 4.2, 5, 11, 15 and 175. Both methods are based on the fact that primer extension, and hence successful PCR, relies on an exact match at the 3′-end of both primers. Therefore, only if the primers entirely match the target sequence is amplification obtained; this is subsequently visualised by agarose gel electrophoresis. In 2010, a survey performed at our Unit, the Immunohaematology and Transfusion Medicine Unit of the “Paolo Giaccone” University Hospital in Palermo, revealed that out of 11 samples (from 8 males and 3 females) analysed and regarded by preliminary analysis as weak D, only eight were confirmed by complete serological analysis as weak D, whereas the other three samples did not give satisfactory results. A genetic protocol was, therefore, used, which gave the results of D weak type 1/DCS, type 11/DCS, and 5/DAR respectively. On the basis of this outcome, we re-evaluated the eight patients assessed as weak D by serological analysis getting D weak type 5/DCS as the most frequent result. These results were not influenced by the patients’ gender or age. Given the strong immunogenicity of the D antigen and the high rate of immunisation of D-negative individuals after the transfusion of D-positive red blood cells, the determination of RHD alleles is of special significance1,2,5. Immunisation of D-negative individuals can occur following transfusions of D-positive red blood cells and in D-negative pregnant women carrying D-positive foetuses5. The aim of this study was to use a genetic protocol to confirm and/or clarify D antigen doubts in order to prevent immunisation of patients. At first glance, serological analysis for D weak appeared to be trustworthy regarding common D weak phenotypes whereas analysis of non-common phenotypes was less satisfactory. In particular, D weak type 5 was easily identifiable by serological analysis whereas D weak types 1 and 11 seemed to display lower antibody affinity and was, therefore, less well identified. With respect to this problem, the use of the genetic protocol was decisive for obtaining correct results. The evidence of D partial results was confirmed by the presence of DCS and DAR variants. The genetic resolution of these variants is, so far, limited, and these two results are, therefore, taken in the context of D-positive results. However, given the low number of samples screened, this study cannot be decisive and other samples need to be analysed.

Centenarians are the best example of extreme human longevity, and they represent a selected population in which the appearance of major age-related diseases, such as cancer, and cardiovascular diseases among others, has been consistently delayed or escaped. The study of the long-lived individual genetic profile has the purpose to possibly identify the genes and the allelic variations influencing extended life expectancy, hence considering them as biomarkers of age-related diseases onset and development. The present study shows no significant differences between allelic variations of ABO blood groups among a group of centenarians from Western Sicily.

Alzheimer disease (AD) is a heterogeneous and progressive neurodegenerative disease, which in Western society mainly accounts for senile dementia. Today many countries have rising aging populations and are facing an increased prevalence of age-related diseases, such as AD, with increasing health-care costs. Understanding the pathophysiology process of AD plays a prominent role in new strategies for extending the health of the elderly population. Considering the future epidemic of AD, prevention and treatment are important goals of ongoing research. However, a better understanding of AD pathophysiology must be accomplished to make this objective feasible. In this paper, we review some hot topics concerning AD pathophysiology that have an important impact on therapeutic perspectives. Hence, we have focused our attention on inflammation, cytokines, immune response, apolipoprotein E (APOE), cholesterol, oxidative stress, as well as exploring the related therapeutic possibilities, i.e., nonsteroidal antiinflammatory drugs, cytokine blocking antibodies, immunotherapy, diet, and curcumin.

Fabry disease is an X‐linked disorder resulting from a deficiency of the lysosomal enzyme alpha‐galactosidase leading to the accumulation of glycolipids in the cells. It has been demonstrated that polymorphisms of alfa‐galactosidase might influence the onset and development of inflammatory cardiovascular age related disease.Familiar Mediterranean fever (FMF) is a autosomal recessive inflammatory based disease, characterized by recurrent fever, peritonitis, arthritis, pleuritis, and cardiac problems. MEFV encodes the pyrin protein it has been suggested to play a role in the innate immunity regulating the inflammatory response. Several data suggest a role of FMF polymorphisms in worsening recurrent inflammatory diseases. The aim of this study is to analyze the interaction between polymorphisms of this two diseases in order to find possible association. We have analyzed 46 patients with cardiac and neurological diseases holding Fabry's disease polymorphisms. Surprisingly, we have found that out of the 46 patients 9 individuals were bearing also Pyrin allelic mutations (more that 19.6%). Since the percentage of Pyrin mutation in a healthy population is more or less 5%, we can conclude that the presence of these mutations might load of further inflammatory burden the status on Fabry's patients.

Ageing is a complex process that negatively impacts the development of the different systems and its ability to function. On the other hand, the rate of ageing in humans is not uniform, due to genetic heterogeneity and the influence of environmental factors. Thus, the ageing rate, measured as the decline of functional capacity and stress resistance, seems to be different in every individual. Therefore, attempts have been made to analyse this individual age, the so-called biological age, in comparison to chronological age. Age-related changes in body function or composition that could serve as a measure of biological age and predict the onset of age-related diseases and/or residual lifetime are termed biomarkers of ageing. Such biomarkers of ageing should help on the one hand to characterise this biological age and, as age is a major risk factor in many degenerative diseases, could be subsequently used on the other hand to identify individuals at high risk of developing age-associated diseases or disabilities. Unfortunately, most of the markers under discussion are related to age-related diseases rather than to age, so none of these markers discussed in literature is a true biomarker of ageing. Hence, we discuss some disease-related biomarkers useful for a better understanding of ageing and the development of new strategies to counteract it, essential for improving the quality of life of the elderly population. Biomarkers discussed are based on immunosenescence, inflammatory responses and oxidative stress, since the review is based on data from author laboratories rather than on an extensive review of the literature. However, this kind of knowledge is useful to anti-ageing strategies aimed to slow ageing and to postpone death by preventing infectious diseases and delaying the onset of age-related diseases.

The concept of Vascular Dementia (VaD) has been recognized for over a century, but its definition and diagnostic criteria remain unclear. Conventional definitions identify the patients too late, miss subjects with cognitive impairment short of dementia, and emphasize consequences rather than causes, the true bases for treatment and prevention. We should throw out current diagnostic categories and describe cognitive impairment clinically and according to commonly agreed instruments that document the demographic data in a standardized manner and undertake a systematic effort to identify the underlying aetiology in each case. Increased effort should be targeted towards the concept of and criteria for Vascular Cognitive Impairment and Post-Stroke Dementia as well as for genetic factors involved, especially as these categories hold promise for early prevention and treatment.

Alzheimer's disease (AD) is a heterogeneous and progressive neurodegenerative disease which in Western society mainly accounts for clinical dementia. Inflammation plays a key role in AD and dissecting the genetics of inflammation may provide an answer to the possible treatment. Hence, the better understanding of different molecular and cellular inflammatory mechanisms is crucial for complete knowledge of AD pathophysiology, and for its prevention and drug therapy. Accordingly, in the present study we evaluated whether the pro-inflammatory polymorphisms of lipopolysaccaride-receptors, +896A/G Toll-Like Receptor (TLR4) and -260C/T CD14, are risk factors for AD. The study included both 626 AD patients (427 women and 199 men; age range: 53-98 years; mean age: 74.88+/-8.44) from Northern Italy and age and gender matched controls. Our results demonstrate that the +896A/G TLR4 single nucleotide polymorphism (SNP) is associated with AD, whereas no association has been observed with -260C/T CD14 SNP. Furthermore, no differences have been observed evaluating the combined presence of +896A+TLR4/-260T+CD14 "high responder"(proinflammatory-profile). However, our results showing the involvement of TLR4 in AD pathophysiology, strengthen the suggestion that systemic inflammation plays a key role in AD. Carriers of high responder SNP, affected by mild cognitive impairment might, be the ideal target for a preventive treatment with biologics as monoclonal antibodies directed against the pro-inflammatory cytokines to decrease the level of systemic inflammation involved in AD pathophysiology.