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SummaryIn patients with common variable immune deficiencies, primary vaccination followed by two booster doses is recommended for protection against COVID-19. Seroconversion has been shown in 60% of patients. We have no information on whether serum antibodies reflect the generation of durable immune memory.In a longitudinal study on 47 common variable immune deficiencies patients who received the third and fourth vaccine dose, we show that the measurement of specific antibodies is not sufficient to predict the establishment of immune memory and the ability to respond to antigen re-exposure.Our results indicate that the combination of antibodies and memory B cells responses represents a more reliable read-out of vaccine immune efficacy in vulnerable patients.This analysis may not only identify individuals remaining unprotected after vaccination and unable to respond to additional booster doses, but also address the search for the underlying immune defect and suggest patient-tailored management strategies.

Digital ischemia due to arterial defects need urgent surgical management. The traditional treatment consists of vascular reconstruction using a reversed autologous venous graft as a bypass. Very few studies have described the use of arterial grafts for digital artery reconstruction. This cadaver study characterized the forearm perforator arteries to assess the potential feasibility of using them as donor grafts for digital artery reconstruction. Eleven forearms and twenty hands were dissected from freshly injected cadavers. All clinically significant perforators (>0.5 mm) derived from radial or ulnar arteries and digital arteries were evaluated. The digital palmar arteries were measured at three points: metacarpophalangeal (MCP) joint, proximal interphalangeal (PIP) joint, and distal interphalangeal (PIP) joint. In the 11 forearms analyzed, 5.5 ± 1.3 perforators from radial or ulnar arteries with a diameter of at least 0.5 mm were found per dissection. The mean diameters were 0.9 ± 0.18 mm proximally and 0.8 ± 0.15 mm distally; the mean length was 35.6 ± 11.35 mm. The mean diameters for the dominant and non-dominant arteries were 1.5 and 1.3 mm at the MCP, 1.3 and 1.0 mm at the PIP, 0.8 and 0.7 mm at the DIP, respectively. The forearms are good donor sites as they have large-diameter arteries of suitable length for arterial grafting. These new arterial grafts may be suitable for vascular reconstruction of digital arteries starting from the PIP joint.

Growing evidence report that non-genetic-driven events such as enhancer reprogramming promote neoplastic transformation and strongly contribute to the phenotypical heterogeneity of cancers as much as genetic variation. In this context, we investigated the role of enhancers in sustaining oncogenic transformation in B-Cell Acute Lymphoblastic leukemia in children (BCP-ALL), a type of cancer caused by the accumulation of lymphoid progenitor cells in the bone marrow and a leading cause of cancer-related mortality in children. Using next-generation sequencing (ATAC-seq), we built the most up-to-date map of chromatin accessibility in pediatric BCP-ALL. We observed that enhancer activity dynamically changes during cancer progression and represents principal phenomena underlying phenotypic–functional characteristics of BCP-ALL progression. BCP-ALL patients are dominated by a regulatory repertoire (N=∼11k) originally represented at diagnosis that shrinks under treatments and subsequently re-expands, driving the relapse. We then deployed a wide range of in-vivo, in-vitro assays, and in-silico analyses to demonstrate the impact of enhancer activity in determining the phenotypical complexity. CRISPR-Cas-9-mediated validation of selected productive enhancers demonstrated a high capability of these regions to control MYB and DCTD oncogenic activities. Taken together, these findings provide direct support to the notion that enhancer plasticity is a crucial determinant of the BCP-ALL phenotype.

The fact that honey bees have a relatively simple nervous system that allows complex behaviors has made them an outstanding model for studying neurobiological processes. Studies on learning and memory routinely use appetitive and aversive learning paradigms that involve recording of the proboscis or the sting extension. However, these protocols are based on all-or-none responses, which has the disadvantage of occluding intermediate and more elaborated behaviors. Nowadays, the great advances in tracking software and data analysis, combined with affordable video recording systems, have made it possible to extract very detailed information about animal behavior. Here we describe antennal movements that are elicited by odor that have no, positive or negative valence. We show that animals orient their antennae towards the source of the odor when it is positive, and orient them in the opposite direction when the odor is negative. Moreover, we found that this behavior was modified between animals that had been trained based on protocols of different strength. Since this procedure allows a more accurate description of the behavioral outcome using a relatively small number of animals, it represents a great tool for studying different cognitive processes and olfactory perception.

Kidney international 102(1), 160-172 (2022). doi:10.1016/j.kint.2022.02.042, Published by Elsevier, New York, NY

Summary Background Presenting symptoms of childhood cancers might mimic those of rheumatic diseases. However, the evidence available to guide differential diagnosis remains scarce. Preventing wrong or delayed diagnosis is therefore important to avoid incorrect administration of glucocorticoid or immunosuppressive therapy and worsening of prognosis. As such, we aimed to assess the prevalence and characteristics of presenting musculoskeletal manifestations in patients at cancer onset and to identify the factors that differentiate childhood malignancies with arthropathy from juvenile idiopathic arthritis. Methods We did a multicentre, cross-sectional study at 25 paediatric haemato-oncology centres and 22 paediatric rheumatology centres in Italy. We prospectively recruited patients who were younger than 16 years that were newly diagnosed with cancer or juvenile idiopathic arthritis. We excluded patients with glucocorticoid pre-treatment (>1 mg/kg per day of oral prednisone or equivalent for ≥2 consecutive weeks). We collected data for patients with a new diagnosis of cancer or juvenile idiopathic arthritis using an electronic case report form on a web-based platform powered by the Cineca Interuniversity Consortium. The primary outcome was to describe the frequency and characteristics of musculoskeletal manifestations at cancer onset; and the secondary outcome was to identify factors that could discriminate malignancies presenting with arthropathy, with or without other musculoskeletal symptoms, from juvenile idiopathic arthritis using multivariable logistic regression analysis. Findings Between May 1, 2015, and May 31, 2018, 1957 patients were eligible, of which 1277 (65%) had cancer and 680 (35%) had juvenile idiopathic arthritis. Musculoskeletal symptoms occurred in 324 (25% [95% CI 23·0–27·8]) of 1277 patients with cancer, of whom 207 had arthropathy. Patients with malignant bone tumours had the highest frequency of musculoskeletal symptoms (53 [80%] of 66), followed by patients with Langerhans histiocytosis (16 [47%] of 34), leukaemia (189 [32%] of 582), soft-tissue sarcomas (16 [24%] of 68), and neuroblastoma (21 [19%] of 109). In the 324 patients with cancer and musculoskeletal symptoms, the most common complaints were joint pain (199 [61%]), followed by limb bone pain (112 [35%]). Joint involvement had a prevalent monoarticular pattern (100 [48%] of 207) and oligoarticular pattern (86 [42%] had 2–4 joints involved and 20 [10%] had >4 joints involved), with the most frequently involved joints being the hip (88 [43%] of 207) and knee (81 [39%]). On multivariable analysis, limb bone pain was the independent variable most strongly associated with cancer (odds ratio [OR] 87·80 [95% CI 18·89–408·12]), followed by weight loss (59·88 [6·34–565·53]), thrombocytopenia (12·67 [2·40–66·92]), monoarticular involvement (11·30 [4·09–31·19]), hip involvement (3·30 [1·13–9·61]), and male sex (2·40 [1·03–5·58]). Factors independently associated with juvenile idiopathic arthritis were morning stiffness (OR 0·04 [95% CI 0·01–0·20]), joint swelling (0·03 [0·01–0·09]), and involvement of the small hand joints (0·02 [0–1·05]). Interpretation Our study provides detailed information about presenting musculoskeletal manifestations of childhood cancers and highlights the clinical and laboratory features that are most helpful in the differential diagnosis with juvenile idiopathic arthritis. Funding Associazione Lorenzo Risolo.

The main strategic objective of the sPATIALS3 project is the creation and strengthening of a Lombardy hub of transversal skills for agrifood production and human nutrition, to provide technologically advanced solutions to the agrifood sector. In synergy with private enterprises, sPATIALS3 aims at developing food products of higher nutritional quality, safety, traceability and packaging aimed at a wide range of consumers, with particular attention to vulnerable groups of patients affected by specific diseases. Work Package 1 of the project is devoted to the characterization and selection of raw materials of plant origin, primarily seeds. Three categories of seeds are considered: pulses (Phaseolus vulgaris), cereals (Triticum aestivum and Zea mays) and oil seeds (Camelina sativa and different species of Linum). The common strategy consists in the identification of genotypes characterized by improved nutritional and nutraceutical profile, achieved through the selection of different materials based on their increased content in beneficial compounds, the reduced content in antinutritional factors, or reduced allergenicity. Depending on the specific features of each plant, various seed quality traits are under investigation. A key role is played by the new high resolution NMR spectrometer, acquired within the project, operating at 600 MHz and equipped with a cryoprobe (Prodigy). This last probe delivers a sensitivity enhancement of a factor of 3, thus allowing the identification and quantification of metabolites and bioactive compounds present even in small amount in the various extracts. The main results obtained so far include the identification of seeds with the following characteristics: oPhaseolus vulgaris genotypes with reduced content in antinutritional compounds (lectins, digestive enzyme inhibitors, phytate, and galactooligosaccharides) or with increased iron concentration; oTriticum aestivum genotypes with reduced content of free asparagine, responsible for the formation of acrylamide, a neurotoxic compound suspected to be carcinogenic which seeps during the baking of bakery products; oLombard Zea mays varieties with reduced content of specific members of the ?-zein storage protein family, responsible for allergenic reactions in humans and pigs; oCamelina sativa genotypes with a healthy omega6/omega3 ratio and increased glucosinolate content, trait that correlates with the presence of isothiocyanates (anticancer compounds); oLinum species and varieties showing a genetic variability in the content of oil and different phenolic compounds.

Common bean (Phaseolus vulgaris L.) is the most important grain legume for human consumption providing up to 15% of total daily calories and 36% of total daily protein in parts of Africa and the Americas. As a legume, it also has a role in sustainable agriculture owing to its ability to fix atmospheric nitrogen. Wild common bean is organized in two geographically isolated and genetically differentiated wild gene pools (Mesoamerican and Andean) that diverged from a common ancestral wild population more than 100,000 years ago. From these wild gene pools, common bean was independently domesticated in Mexico and in South America nearly 8,000 years ago, and these domestication events were followed by local adaptations resulting in landraces with distinct characteristics (Schmutz et al. 2014). Domestication led to morphological changes in seed and leaf sizes, in the growth habit and photoperiod responses, variation in seed coat color and pattern that distinguish culturally adapted classes of beans. This unique example of parallel domestication is the subject of the PARDOM project that, starting from the Phaseolus replicated experiment, aims at understanding common bean genome evolution and adaptation. In the framework of the PARDOM project, we are developing TILLING-by-sequencing and genome editing technological platforms for the functional validation of candidate domestication genes in common bean. For the development of the TILLING-by-seq platform, DNA from seeds of a P. vulgaris TILLING population developed in the Mesoamerican genotype BAT93 (Porch et al. 2009; Cominelli et al. 2018) was extracted. A three-dimensional pooling system of 54 pools, each of 96 samples on average, at resolution of a population of 1728 individuals was used for NGS targeted sequencing based on custom capture probes. For the genotyping, a total of 719 genes of interest were chosen, based on the presence of one or more signals of domestication, differential expression between the Andean genotype and Mesoamerican genotype, known involvement in the phenomenon of shattering, seed development and in the cytokinin hormonal pathway. Among these genes, 27 had a complete CDS sequence coverage, whereas for the others the first 1-3 exons were covered, for a total of approximately 491Mb. The validation of candidate genes for domestication is currently in progress also via forward genetics, following the identification of target regions in coding sequences for genome editing based on CRISPR/Cas9 technology. Fifteen target candidate domestication genes have been selected, based on the presence of one or more signals of domestication. Current editing approach is directed toward MYB26, encoding a transcription factor involved in pod shattering phenotype. Given the challenges posed by common bean transformation (biolistic transgenesis), the genome editing approach is being simultaneously carried out also on soybean ( Glycine max) homologous genes.

Bone marrow (BM) microenvironment contributes to the regulation of normal hematopoiesis through a finely tuned balance of self-renewal and differentiation processes, cell-cell interaction and secretion of cytokines that during leukemogenesis are altered and favor tumor cell growth. In pediatric acute myeloid leukemia (AML), chemotherapy is the standard of care, but still >30% of patients relapse. The need to accelerate the evaluation of innovative medicines prompted us to investigate the mesenchymal stromal cells (MSCs) role in the leukemic niche to define its contribution to the mechanisms of leukemia escape. We generated humanized three-dimensional (3D) niche with AML cells and MSCs derived from either patients (AML-MSCs) or healthy donors. We observed that AML cells establish physical connections with MSCs, mediating a reprogrammed transcriptome inducing aberrant cell proliferation and differentiation, and severely compromising their immunomodulatory capability. We confirmed that AML cells modulate h-MSCs transcriptional profile promoting functions similar to the AML-MSCs when co-cultured in vitro, thus facilitating leukemia progression. Conversely, MSCs derived from BM of patients at time of disease remission showed recovered healthy features, at transcriptional and functional levels, including the secretome. We proved that AML blasts alter MSCs activities in the BM niche, favoring disease development and progression. We discovered that a novel AML-MSCs selective CaV1.2 channel blocker drug, Lercanidipine, is able to impair leukemia progression in 3D niche both in vitro and when implanted in vivo, if used in combination with chemotherapy, supporting the hypothesis that synergistic effects can be obtained by dual targeting approaches.

Contains fulltext : 225968.pdf (Publisher’s version ) (Closed access) Immunosuppression in IgA nephropathy (IgAN) should be reserved for patients at high-risk of disease progression, which KDIGO guidelines determine based solely on proteinuria 1g or more/day. To investigate if treatment decisions can be more accurately accomplished using individualized risk from the International IgAN Prediction Tool, we simulated allocation of a hypothetical immunosuppression therapy in an international cohort of adults with IgAN. Two decision rules for treatment were applied based on proteinuria of 1g or more/day or predicted risk from the Prediction Tool above a threshold probability. An appropriate decision was defined as immunosuppression allocated to patients experiencing the primary outcome (50% decline in eGFR or ESKD) and withheld otherwise. The net benefit and net reduction in treatment are the proportion of patients appropriately allocated to receive or withhold immunosuppression, adjusted for the harm from inappropriate decisions, calculated for all threshold probabilities from 0-100%. Of 3299 patients followed for 5.1 years, 522 (15.8%) experienced the primary outcome. Treatment allocation based solely on proteinuria of 1g or more/day had a negative net benefit (was harmful) because immunosuppression was increasingly allocated to patients without progressive disease. Compared to using proteinuria, treatment allocation using the Prediction Tool had a larger net benefit up to 23.4% (95% confidence interval 21.5-25.2%) and a larger net reduction in treatment up to 35.1% (32.3-37.8%). Thus, allocation of immunosuppression to high-risk patients with IgAN can be substantially improved using the Prediction Tool compared to using proteinuria.

Background: Genitourinary syndrome of menopause (GSM) is the new term for vulvovaginal atrophy (VVA). The condition is relevant in more than 50% of women, having an adverse impact on quality of life and sexual relationships. Objective: To assess the efficacy and safety of a new type of non-ablative laser, Solid State Vaginal Laser (SSVL), for vaginal tissue regeneration and rejuvenation. Method: Eighty participants with GSM symptoms were treated with a total of 4 treatments in about two months (every 15 - 20 days) of a non-ablative SSVL (LASEmaR 1500TM-EUFOTON). A cumulative intensity of GSM symptoms using a 10-cm VAS (dryness and/or burning and/or dyspareunia), the vaginal health index (VHI), the Female Sexual Function Index (FSFI) were evaluated. Urinary Incontinence Short Form (ICIQ-UI SF) and vaginal bioptic samples were also collected. Results: Improvement following the SSVL was observed on VHIS, VVA symptoms and sexual female function. This finding was also ratified by the improvement of vaginal histological features. After the SSVL treatment, almost all patients (91%) affected by urinary incontinence obtained the complete remission of symptoms. Conclusion: The objective evaluation of VHIS, FSFI and ICIQ-UI SF scores and the histological results indicates a real favorable effect of SSVL on GSM and on urinary incontinence.