Haem is associated with thrombosis in neonates and infants undergoing cardiac surgery for congenital heart disease.

BACKGROUND: Haem levels are associated with thrombosis in a variety of diseases, as well as being a contributing cause of thrombotic events in animal models. MATERIALS AND METHODS: We retrospectively analyzed samples from 39 children who underwent cardiac surgery with cardiopulmonary bypass, including 15 children who developed a postoperative thrombosis and 24 controls. RESULTS: Patients who developed thrombosis postoperatively had statistically significant higher average haem levels over time (presurgery to 12 h postsurgery) compared to patients who did not develop thrombosis. CONCLUSION: Higher cell-free total haem levels are associated with a higher risk of thrombosis in a paediatric cardiac surgical cohort.

Evaluating the efficacy of restoration plantings through DNA barcoding of frugivorous bird diets.

Frugivores are critical components of restoration programs because they are seed dispersers. Thus, knowledge about bird-plant trophic relationships is essential in the evaluation of the efficacy of restoration processes. Traditionally, the diet of frugivores is characterized by microscopically identifying plant residues in droppings, which is time-consuming, requires botanical knowledge, and cannot be used for fragments lacking detectable morphological characteristics (e.g., fragmented seeds and skins). We examined whether DNA barcoding can be used as a universal tool to rapidly characterize the diet of a frugivorous bird, Eurasian blackcap (Sylvia atricapilla). We used the DNA barcoding results to assess restoration efforts and monitor the diversity of potentially dispersed plants in a protected area in northern Italy. We collected 642 Eurasian Blackcap droppings at the restored site during the autumn migration over 3 years. Intact seeds and fragmented plant material were analyzed at 2 plastidial barcode loci (rbcL and trnH-psbA), and the resulting plant identifications were validated by comparison with a reference molecular data set of local flora. At least 17 plant species, including 7 of the 11 newly transplanted taxa, were found. Our results demonstrate the potential for DNA barcoding to be used to monitor the effectiveness of restoration plantings and to obtain information about fruit consumption and dispersal of invasive or unexpected plant species. Such an approach provides valuable information that could be used to study local plant biodiversity and to survey its evolution over time.

SILVER COATED PROSTHESIS IN ONCOLOGICAL LIMB SALVAGE SURGERY REDUCE THE INFECTION RATE.

Silver coatings, used in many surgical devices, have demonstrated good antimicrobial activity and low toxicity. Oncological musculoskeletal surgery have a high risk of infection, so in the last decades, silver-coated mega-prostheses have been introduced and are becoming increasingly widespread. In this study, a retrospective analysis of 158 cases of bone tumors, primary or metastatic, treated between 2005- 2015 with wide margins resection and tumor implants reconstruction, was performed. The average age was 59 years (range 11-78 years), the same surgeon with antibiotic prophylaxis according to a standard protocol treated all patients. Silver-coated prostheses were implanted in 58.5% of patients and uncoated tumor prostheses in the remaining 41.5%. Patients were re-evaluated annually and complications were recorded, focusing analysis on infective complications. The average follow-up was 39.7 months: 23.4% of patients died at a median time of 35.3 months after surgery; 18.4% developed complications that required new surgery, of which 12.6% of these were due to infection. Patients treated with silver-coated implants developed early infection in 2.2% of cases against the 10.7% of the patients treated with standard tumor prosthesis. This difference between the two groups was statistically significant. The percentage of late infections occurring from 6 months after surgery was similar in both groups. Silver blood level taken in a sample of patients at different times after surgery, always showed values well below the threshold of toxicity and no patient showed any sign of local or general toxicity secondary to silver. Our study demonstrates that tumor silver-coated implants have a rate of early infection significantly lower than traditional implants, while there were no differences in the rate of late infections as described also in the literature. We recommend the use of silver–coated prosthesis as primary implants for limb salvage surgery in primary or metastatic bone tumors, considering the absence of toxicity and the lower rate of early infection.

Effect of raw material on cooking quality and nutritional composition of durum wheat spaghetti.

In this study the effect of semolina and wholemeal flour from six durum wheat cultivars on the pasta cooking and nutritional quality was evaluated. The wholemeal spaghetti samples showed an improvement in the chemical composition (high protein and insoluble dietary fibre content) but they have a decline in the cooking quality (high cooking loss) with respect to the semolina spaghetti. In particular, the wholemeal spaghetti Cappelli and Core samples recorded the highest protein and insoluble dietary fibre content, respectively. As compared to the other samples, the wholemeal spaghetti Iride recorded a higher cooking loss. Moreover, the wholemeal spaghetti showed the lowest overall quality due to the low score of elasticity, firmness and colour. Specifically, the wholemeal Cappelli recorded a slight rise of the overall quality with respect to other wholemeal samples. In conclusion, the wholemeal spaghetti Cappelli was found to be an optimum compromise between the sensory and nutritional quality.

Sweet liking predicts liking and familiarity of some alcoholic beverages, but not alcohol intake: A population study using a split-sample approach.

Sweetness has been proposed to be an important quality in the decision to consume alcohol, and strong preferences for sweet tastes have been associated with alcohol abuse. However, alcohol is characterized by a number of other sensory properties, including astringency and bitterness that may drive preference and consumption. Spinelli et al. (2021) classified individuals into three sweet-sensory liking clusters (High Sweet-Liking, Moderate Sweet-Liking, and Inverted-U) that differed in their sweetness optima and sensory-liking patterns (relationship between liking and sweetness, bitterness and astringency perception in a food model). The current paper replicates the sweet sensory-liking clusters in a new set of participants (n = 1976), and extends the predicted value of these clusters examining their relationship to wine and other types of alcoholic beverages by gender using a split-sample approach on a total of over 3000 adults. The sweet sensory-liking clusters had a predictive relationship for the familiarity and liking of some alcoholic beverages characterized by stronger tastes, but not weekly alcohol intake levels. Thus, although sweet sensory-liking clusters may be associated with the type of beverages and frequency with which a person will drink and enjoy a type of alcoholic beverage, they are poor predictors of the quantity of alcohol that a person ingests over the course of a week.

Idiopathic erythrocytosis: a germline disease?

Polycythemia Vera (PV) is typically caused by V617F or exon 12 JAK2 mutations. Little is known about Polycythemia cases where no JAK2 variants can be detected, and no other causes identified. This condition is defined as idiopathic erythrocytosis (IE). We evaluated clinical-laboratory parameters of a cohort of 56 IE patients and we determined their molecular profile at diagnosis with paired blood/buccal-DNA exome-sequencing coupled with a high-depth targeted OncoPanel to identify a possible underling germline or somatic cause. We demonstrated that most of our cohort (40/56: 71.4%) showed no evidence of clonal hematopoiesis, suggesting that IE is, in large part, a germline disorder. We identified 20 low mutation burden somatic variants (Variant allelic fraction, VAF, < 10%) in only 14 (25%) patients, principally involving DNMT3A and TET2. Only 2 patients presented high mutation burden somatic variants, involving DNMT3A, TET2, ASXL1 and WT1. We identified recurrent germline variants in 42 (75%) patients occurring mainly in JAK/STAT, Hypoxia and Iron metabolism pathways, among them: JAK3-V722I and HIF1A-P582S; a high fraction of patients (48.2%) resulted also mutated in homeostatic iron regulatory gene HFE-H63D or C282Y. By generating cellular models, we showed that JAK3-V722I causes activation of the JAK-STAT5 axis and upregulation of EPAS1/HIF2A, while HIF1A-P582S causes suppression of hepcidin mRNA synthesis, suggesting a major role for these variants in the onset of IE.

Responsiveness to warning sensations and anxiety-related psychological traits modulate individual differences in preference for vegetable foods with varied sensory properties.

The innate aversion to warning sensations is an important barrier to the acceptance of vegetable food often characterized by bitter and sour tastes, and astringency. Large individual variations exist in preference for this food category. The present study aimed at exploring differences in demographics, anthropometrics, taste responsiveness, personality traits and attitudes in consumers differing in their preference for vegetable food with varied levels of warning sensations. A panel of Italian consumers (n = 718; 53.6% women, age 18-74 years) self-reported familiarity with, preference for and choice of vegetables with high and low levels of warning sensations. Two clusters were identified: High Warning-Vegetable Consumers (HWVC, n = 464) and Low-Warning Vegetable Consumers (LWVC, n = 254). HWVC showed higher familiarity with and preference for vegetables as a whole and higher choice of vegetables characterized by warning sensations than LWVC. HWVC were more represented by older and normal weight individuals as compared to LWVC. Differences among clusters in liking for and perception of a phenol-enriched plant-based food model specifically developed to induce different levels of bitterness, sourness and astringency were found. HWVC rated bitterness, sourness, and astringency lower and liking higher than LWVC. Scores in anxiety-related psychological traits were lower while attitudes to healthy and high-quality food choice were higher in HWVC than in LWVC. The results of the present study depicted a coherent interplay among several person-related dimensions in modulating preference for vegetable foods. Higher responsiveness to warning sensations, higher level of anxiety-related traits, lower importance assigned to food healthy/quality aspects and younger age all acted as barriers to exposure and acceptance of vegetable food and call for a multidimensional approach to promote the consumption of this food category.

Bioplasty for vertebral fractures: preliminary results of a pre-clinical study on goats using autologous modified skin fibroblasts.

The debate is still ongoing about the long term effects of the mininvasive vertebral augmentation techniques and their usefulness in treating more complex cases where a bone inducing effect more than a merely bone substitution would be suitable, such as the vertebral fractures in young patients. We previously developed a clinically relevant gene therapy approach using modified dermal fibroblasts for inducing bone healing and bone formation in different animal models. The aim of this study is to show the feasibility of a minimally invasive percutaneous intrasomatic ex vivo gene therapy approach to treat thoracolumbar vertebral fractures and anterior column bone defects in a goat model.

A biochemical genomics approach for identifying genes by the activity of their products.

For the identification of yeast genes specifying biochemical activities, a genomic strategy that is rapid, sensitive, and widely applicable was developed with an array of 6144 individual yeast strains, each containing a different yeast open reading frame (ORF) fused to glutathione S-transferase (GST). For the identification of ORF-associated activities, strains were grown in defined pools, and GST-ORFs were purified. Then, pools were assayed for activities, and active pools were deconvoluted to identify the source strains. Three previously unknown ORF-associated activities were identified with this strategy: a cyclic phosphodiesterase that acts on adenosine diphosphate-ribose 1"-2" cyclic phosphate (Appr>p), an Appr-1"-p-processing activity, and a cytochrome c methyltransferase.

Global Profile: Going beyond liking to better understand product experience.

Over the last few years, interest in collecting an increasing variety of information in order to acquire a deeper and more comprehensive understanding of the consumer experience with products has steadily grown. The present study was designed to develop a methodological approach to explore the Global Profile of products within a category. A product-specific questionnaire was developed using one-on-one online interviews conducted with a modified version of the Repertory Grid Method (RGM) combined with semiotic analysis, EmoSemio, extended to cover all the main dimensions of product experience of the category of processed tomato. A 96 item questionnaire - including liking, sensory properties, emotions, emotional and functional conceptualisations and contextual appropriateness measures - was developed and employed in a home use test with 196 consumers who evaluated 9 products (one per day). All the statements were found to discriminate between products with the exception of one. A Multiple Factor Analysis showed that emotions were highly correlated with functional/emotional conceptualisations, while sensory properties were mainly related to emotional conceptualisations (memories) and uses in the recipes. The information provided by the simultaneous collection of these different dimensions allows to go beyond liking and may be used in product development and innovation in order to better understand the consumer experience of a product.

[Sexuality and physical disorders]. / Sexualité et maladie.

In every kind of pathology each patient is also conditioned by one's sexual identity. For this reason a physician should not avoid considering the possible presence for each patient of some sexual problem, even if it is not directly linked to the mean disease. The same doctor could be moreover surprised by a positive reaction from the patient if sexuality is mentioned even within severe physical disorders.

Release of biologically active CD154 during collection and storage of platelet concentrates prepared for transfusion.

BACKGROUND: Millions of platelet transfusions are given each year. Transfusion reactions occur in as many as 30% of patients receiving unmodified platelet transfusions. The cause of some transfusion reactions remains unclear. The current paradigm suggests that platelet concentrates (PC) contain proinflammatory mediators that are released by white blood cells during collection, processing and storage. CD154 (CD40 ligand, CD40L) is a potent inflammatory mediator, normally sequestered inside the resting platelet, that is known to translocate to the platelet membrane and be shed into plasma in response to agonist activation. We hypothesized that platelet-soluble CD154 (sCD154) is 'spontaneously' released by transfused platelets and plays a major role in transfusion reactions. OBJECTIVES: To determine the time course and biological properties of CD154 translocation and release during collection and storage of platelets for transfusion. METHODS: We measured surface and sCD154 in platelets prepared by the platelet-rich plasma method or apheresis by fluorescence-activated cell sorting and enzyme-linked immunosorbent assay, respectively. The specific biological activity of platelet sCD154 was assayed by stimulation of the CD154/CD40 pathway in known CD40-positive cells with PC-derived supernatants. RESULTS AND CONCLUSIONS: We demonstrate that PCs prepared for transfusion have high levels of membrane-bound CD154 and sCD154, with maximum levels being seen 72 h after platelet collection. Importantly, we show that platelet-derived sCD154 potently stimulates CD40-positive cells. We propose that platelet-derived CD154 is a key 'cytokine' responsible for adverse reactions associated with platelet transfusions. Improved methods of platelet collection and/or storage, which limit CD154 expression, could reduce the risks of transfusion reaction.

Maresin 1 induces a novel pro-resolving phenotype in human platelets.

Essentials Specialized proresolving mediators (SPMs) promote the resolution of inflammation. This study sought to investigate the effects of SPMs on human platelet function. The SPM, Maresin 1, enhanced hemostatic, but suppressed inflammatory functions of platelets. SPMs uniquely regulate platelet function and may represent a new class of antiplatelet agents. SUMMARY: Background Antiplatelet therapy is a cornerstone of modern medical practice and is routinely employed to reduce the likelihood of myocardial infarction, thrombosis and stroke. However, current antiplatelet therapies, such as aspirin, often have adverse side-effects, including increased risk of bleeding, and some patients are relatively 'aspirin-resistant'. Platelets are intimately involved in hemostasis and inflammation, and clinical consequences are associated with excessive or insufficient platelet activation. Objectives A major unmet need in the field of hematology is the development of new agents that safely prevent unwanted platelet activation in patients with underlying cardiovascular disease, while minimizing the risk of bleeding. Here, we investigate the potential of endogenously produced, specialized pro-resolving mediators (SPMs) as novel antiplatelet agents. SPMs are a recently discovered class of lipid-derived molecules that drive the resolution of inflammation without being overtly immunosuppressive. Methods Human platelets were treated with lipoxin A4, resolvin D1, resolvin D2, 17-HDHA or maresin 1 for 15 min, then were subjected to platelet function tests, including spreading, aggregation and inflammatory mediator release. Results We show for the first time that human platelets express the SPM receptors, GPR32 and ALX. Furthermore, our data demonstrate that maresin 1 differentially regulates platelet hemostatic function by enhancing platelet aggregation and spreading, while suppressing release of proinflammatory and prothrombotic mediators. Conclusions These data support the concept that SPMs differentially regulate platelet function and may represent a novel class of antiplatelet agents. SPMs also may play an important role in the resolution of inflammation in cardiovascular diseases.

The management of severe vaginal obstruction from genital chronic graft-versus-host disease: diagnosis, surgical technique and follow-up.

AIM: Chronic graft-versus-host disease (GVHD) is one of the most important systemic late-onset complications of haematopoietic stem cell transplantation. Gynaecological manifestations are considered relatively rare, and involve lower genital tract skin and mucosa, causing vulvar scarring, vaginal stenosis, affecting the patients' sexual life, and leading to more serious complications as haematocolpos, haematometra and abscesses. Genital GVHD can be treated with topical therapy when mild to moderate disease is present. Surgery is indicated in advanced and complicated cases to restore normal anatomy. The aim of this study is to propose a standard approach for the management of such condition when medical therapy is not effective. METHODS: From May 2000 to January 2002, 8 patients suffering from genital chronic GVHD were operated in our institution. We describe clinical and sonographic presurgical assessment, simple or ultrasonographic guided surgical technique, postsurgical treatment and follow-up. RESULTS: Surgery was completely successful in restoring genital anatomy in all the cases. Mean follow-up was of 17 months (6 to 38 months). Two patients early discontinued the postsurgical treatment. At 1 month complete vaginal patency was found in 6 cases, weak partial adhesions in 1 case, while 1 patient refused follow-up. Eventually, complete vaginal patency was maintained in 2 cases, and partial adhesions were found in 5 cases. CONCLUSIONS: The combined use of clinical examination and endosonography provides a precise assessment of the level of the obstruction. The surgical technique here described is feasible and successful in restoring normal anatomy, while long term results seem related to the compliance towards the postsurgical treatment.

[Sexual problems after allogenic stem cell transplantation]. / Problèmes d'ordre sexuet après greffe allogénique de cellules souches.

Allogenic stem cell transplantation is an effective procedure for malignant haematological diseases, for these reasons the number of long-term survivors has increased as well as the concern for their quality of life. The late effects of transplantation can highly influence the patient's sexuality and relationships through psychological and physical factors. Psychological factors: regression due to sickness, staff-dependency, emotional and physical partner-dependence, distress related to sterility. Physical factors: hypo-oestrogenism, genital mucosae dryness and sometimes vulval or vaginal chronic graft-versus-host disease. Addressing the topic of intimacy may cause anxiety and embarrassment among nurses, haematological staff and practitioners with the risk of leaving patients in a sort of "conspiracy of silences".

Mammalian odorant binding proteins.

Odorant binding proteins (OBPs) pertain to one of the most abundant classes of proteins found in the olfactory apparatus. OBPs are a sub-class of lipocalins, defined by their property of reversibly binding volatile chemicals, that we call 'odorants'. Numerous sequences of OBPs are now available, derived from protein sequencing from nasal mucus material, or from DNA sequences. The structural knowledge of OBPs has been improved too in recent years, with the availability of two X-ray structures. The physiological role of OBPs remains, however, essentially hypothetical, and most probably, not linked to a function of odor transport. The present knowledge on OBP biochemistry, sequence and structure will be examined here in relation to the different functional hypotheses proposed for OBPs.

In vivo cytoreduction studies and cell sorting--enhanced tumor-cell detection in high-risk neuroblastoma patients: implications for leukapheresis strategies.

PURPOSE: To improve autologous leukapheresis strategies in high-risk neuroblastoma (NB) patients with extensive bone marrow involvement at diagnosis. PATIENTS AND METHODS: Anti-G(D2) immunocytochemistry (sensitivity, 1 in 10(5) to 10(6) leukocytes) was used to evaluate blood and bone marrow disease at diagnosis and during the recovery phase of the first six chemotherapy cycles in 57 patients with stage 4 NB and bone marrow disease at diagnosis. A total of 42 leukapheresis samples from the same patients were evaluated with immunocytology, and in 24 of these patients, an anti-G(D2) immunomagnetic enrichment step was used to enhance tumor-cell detection. RESULTS: Tumor cytoreduction was much faster in blood compared with bone marrow (3.2 logs after the first cycle and 2.1 logs after the first two cycles, respectively). Bone marrow disease was often detectable throughout induction, with a trend to plateau after the fourth cycle. By direct anti-G(D2) immunocytology, a positive leukapheresis sample was obtained in 7% of patients after either the fifth or sixth cycle; when NB cell immunomagnetic enrichment was applied, 25% of patients had a positive leukapheresis sample (sensitivity, 1 in 10(7) to 10(8) leukocytes). CONCLUSION: Standard chemotherapy seems to deliver most of its in vivo purging effect within the first four cycles. In patients with overt marrow disease at diagnosis, postponing hematopoietic stem-cell collection beyond this point may not be justified. Tumor-cell clearance in blood seems to be quite rapid, and earlier collections via peripheral-blood leukapheresis might be feasible. Immunomagnetically enhanced NB cell detection can be highly sensitive and can indicate whether ex vivo purging should be considered.

Lateral recognition of a dye hapten by a llama VHH domain.

Camelids, camels and llamas, have a unique immune system able to produce heavy-chain only antibodies. Their VH domains (VHHs) are the smallest binding units produced by immune systems, and therefore suitable for biotechnological applications through heterologous expression. The recognition of protein antigens by these VHHs is rather well documented, while less is known about the VHH/hapten interactions. The recently reported X-ray structure of a VHH in complex with a copper-containing azo-dye settled the ability of VHH to recognize haptens by forming a cavity between the three complementarity-determining regions (CDR). Here we report the structures of a VHH (VHH A52) free or complexed with an azo-dye, RR1, without metal ion. The structure of the complex illustrates the involvement of CDR2, CDR3 and a framework residue in a lateral interaction with the hapten. Such a lateral combining site is comparable to that found in classical antibodies, although in the absence of the VL.

Crystallization and preliminary diffraction analysis of the catalytic domain of xylanase Z from Clostridium thermocellum.

The catalytic domain of a thermostable xylanase from Clostridium thermocellum has been expressed in Escherichia coli and crystallized from a polyethylene glycol 2000 solution by the hanging drop method. Crystals belong to the triclinic space group P1 with cell dimensions a = 46.8 A, b = 50.8 A, c = 70.3 A, alpha = 100.7 degrees, beta = 83.8 degrees, gamma = 101.6 degrees, and two molecules in the unit cell. These crystals diffract X-rays to at least 1.8 A resolution and are suitable for high-resolution X-ray analysis.

Crystal structure of a ternary complex between human chorionic gonadotropin (hCG) and two Fv fragments specific for the alpha and beta-subunits.

Human chorionic gonadotropin (hCG), is a placental hormone which exerts its major effect by stimulating progesterone production, crucially sustaining the early weeks of pregnancy. Detection of hCG with specific monoclonal antibodies (mAbs) has become the chosen means for pregnancy diagnosis. We have used antibody Fv fragments derived from two high-affinity mAbs, one against the alpha and the other against the beta-hCG subunit to enable the crystallisation of intact or desialylated hCG. Crystals of a ternary complex composed of Fv anti-alpha/hCG/Fv anti-beta were found to diffract to 3.5 A resolution, and the structure was solved by molecular replacement. In the crystal, the two Fvs keep hCG as in a molecular cage, providing good protein-protein contacts and leaving enough space for the saccharides to be accommodated in the cell solvent. The two Fvs were found not to interact directly through their complementary-determining regions with the hCG saccharides, but only with the protein. The hCG structure in the ternary complex was very close to that of the HF partially deglycosylated hormone, thus indicating that neither the saccharides nor the Fvs had any substantial influence on hormone structure.