White Lupin Drought Tolerance: Genetic Variation, Trait Genetic Architecture, and Genome-Enabled Prediction.

White lupin is a high-protein crop requiring drought tolerance improvement. This study focused on a genetically-broad population of 138 lines to investigate the phenotypic variation and genotype × environment interaction (GEI) for grain yield and other traits across drought-prone and moisture-favourable managed environments, the trait genetic architecture and relevant genomic regions by a GWAS using 9828 mapped SNP markers, and the predictive ability of genomic selection (GS) models. Water treatments across two late cropping months implied max. available soil water content of 60-80% for favourable conditions and from wilting point to 15% for severe drought. Line yield responses across environments featured a genetic correlation of 0.84. Relatively better line yield under drought was associated with an increased harvest index. Two significant QTLs emerged for yield in each condition that differed across conditions. Line yield under stress displayed an inverse linear relationship with the onset of flowering, confirmed genomically by a common major QTL. An adjusted grain yield computed as deviation from phenology-predicted yield acted as an indicator of intrinsic drought tolerance. On the whole, the yield in both conditions and the adjusted yield were polygenic, heritable, and exploitable by GS with a high predictive ability (0.62-0.78). Our results can support selection for climatically different drought-prone regions.

How I treat thrombotic thrombocytopenic purpura in pregnancy.

Thrombotic thrombocytopenic purpura (TTP) is an acute, life-threatening thrombotic microangiopathy (TMA) caused by acquired or congenital severe deficiency of ADAMTS13. Pregnancy is a recognized risk factor for precipitating acute (first or recurrent) episodes of TTP. Differential diagnosis with other TMAs is particularly difficult when the first TTP event occurs during pregnancy; a high index of suspicion and prompt recognition of TTP are essential for achieving a good maternal and fetal outcome. An accurate distinction between congenital and acquired cases of pregnancy-related TTP is mandatory for safe subsequent pregnancy planning. In this article, we summarize the current knowledge on pregnancy-associated TTP and describe how we manage TTP during pregnancy in our clinical practice.

Genome-wide association study reveals white lupin candidate gene involved in anthracnose resistance.

KEY MESSAGE: GWAS identifies candidate gene controlling resistance to anthracnose disease in white lupin. White lupin (Lupinus albus L.) is a promising grain legume to meet the growing demand for plant-based protein. Its cultivation, however, is severely threatened by anthracnose disease caused by the fungal pathogen Colletotrichum lupini. To dissect the genetic architecture for anthracnose resistance, genotyping by sequencing was performed on white lupin accessions collected from the center of domestication and traditional cultivation regions. GBS resulted in 4611 high-quality single-nucleotide polymorphisms (SNPs) for 181 accessions, which were combined with resistance data observed under controlled conditions to perform a genome-wide association study (GWAS). Obtained disease phenotypes were shown to highly correlate with overall three-year disease assessments under Swiss field conditions (r > 0.8). GWAS results identified two significant SNPs associated with anthracnose resistance on gene Lalb_Chr05_g0216161 encoding a RING zinc-finger E3 ubiquitin ligase which is potentially involved in plant immunity. Population analysis showed a remarkably fast linkage disequilibrium decay, weak population structure and grouping of commercial varieties with landraces, corresponding to the slow domestication history and scarcity of modern breeding efforts in white lupin. Together with 15 highly resistant accessions identified in the resistance assay, our findings show promise for further crop improvement. This study provides the basis for marker-assisted selection, genomic prediction and studies aimed at understanding anthracnose resistance mechanisms in white lupin and contributes to improving breeding programs worldwide.

A new standardized immunofluorescence method for potency quantification (SMPQ) of human conjunctival cell cultures.

The aim of this study is to set up a standardized and reproducible method to determine the potency (= stem cell content) of human conjunctival cell cultures by means of immunofluorescence-based analyses. This will help the development of new Advanced Therapy Medicinal Products (ATMPs) to use in future cell therapy clinical studies when fewer cells are available to perform the quality controls. To achieve this purpose, a reference standard was investigated and the expression levels of ΔNp63α (considered as a marker of conjunctival stem cells) was correlated to cell size. The limbal hTERT cells were used as reference standard to define the expression value of ΔNp63α. The mean intensity value of limbal hTERT cells ranging between 15 and 20 µm in diameter was used to distinguish between ΔNp63α bright and not bright cells. As ΔNp63α bright expression was mainly seen in the smaller cell size group (10-15 µm), we defined as conjunctival stem cells (= potency) those cells which were bright and with sizes between 10 and 15 µm. Assays on cells from clonal analyses were used to validate the method, as they do allow to observe a decrease in potency (Holoclones > Meroclones > Paraclones). The stem cell content of conjunctival grafts was found to be 11.3% ± 5.0 compared to 21.9% ± 0.6, 9.0% ± 8.1 and 0% from Holoclones, Meroclones and Paraclones, respectively. This new method, here named as Standardized Method for Potency Quantification, will allow to detect the potency in conjunctival cell cultures, thus obtaining a quality control assay responding to the GMP standards required for ATMP release.

Prevalence and risk factors of postpartum depression and adjustment disorder during puerperium - a retrospective research.

OBJECTIVE: This observational study was designed to evaluate risk factors and distribution of Adjustment Disorder (AD) and Postnatal Depression (PPD), after delivery and a positive screening at Camposampiero Hospital (2012-2017). BACKGROUND: After childbirth, women are vulnerable to develop psychological disorders. Since the effects of psychopathology are relevant, prevention and early intervention are of paramount importance. Recognising risk factors is therefore necessary. METHODS: Women were screened after delivery, between the 6th and 8th week, using EPDS. Depressive symptoms were examined by psychological assessment. Socio-demographic, clinical and obstetric variables were analysed to identify women at greater risk for developing perinatal psychological disorders. RESULTS: Of the 3102 women screened, 14.6% resulted positive: 23.6% of them suffered from AD and 5.5% from PPD. Regarding AD, previous induced abortion, unwanted pregnancy, unemployment and family history of mental disorders were the most relevant risk factors. Higher risks for PPD were: not attending antenatal classes, unwanted pregnancy, previous psychopathology and family history of mental disorders. CONCLUSION: In our study, distribution and risk factors for PPD and AD are in line with those reported in literature. Paying attention to subclinical symptoms co-related to items of EPDS helps healthcare professionals to be more sensitive in detecting suffering women.

Long-term neuropsychological sequelae, emotional wellbeing and quality of life in patients with acquired thrombotic thrombocytopenic purpura.

Neurological symptoms related to microthrombosis are the hallmark of acute manifestations of acquired thrombotic thrombocytopenic purpura (TTP). Despite the achievement of hematological remission, patients may report persisting neurological impairment that affects their quality of life. To assess the long-term neuropsychological consequences of acute TTP, we recruited 35 acquired TTP patients (77% females, median age at onset 41 years, interquartile range: 35-48) regularly followed at our out-patient clinic of thrombotic microangiopathies in Milan (Italy) from December 2015 to October 2016. Patients underwent a psychological evaluation of memory and attentional functions, emotional wellbeing and health-related quality of life at least three months after their last acute TTP event (median 36 months, interquartile range: 17-54). During the psychological consultation, 17 patients (49%) referred persisting subjective neurological impairment in the frame of a remission phase, with at least one symptom as disorientation, loss of concentration, dizziness, lack of balance, headache and diplopia. Neuropsychological assessment revealed lower scores than the Italian general population pertaining to direct, indirect and deferred memory. A higher degree of impairment of memory domains was found in patients with neurological involvement at the time of presentation of the first acute TTP episode. Anxiety and depression were detected in seven (20%) and 15 (43%) patients, respectively. Health-related quality of life was lower than the Italian general population, with mental domains more impacted than physical domains (mean difference 58.43, 95% confidence interval: 71.49-45.37). Our study demonstrates compromised memory and attention functions, persisting anxiety/depression symptoms and a generally reduced quality of life in patients recovering from acute acquired TTP. New clinical strategies should be considered to improve these symptoms.

Clinical Factors Influencing the Outcomes of an Acellular Dermal Matrix for Skin Cancer Treatment: A Retrospective Study.

OBJECTIVE: To evaluate the efficacy and safety of a biologically engineered dermal matrix used in reconstructive surgery after skin tumor resection, focusing on the frequency of successful grafting and identifying potential factors influencing treatment outcomes. DESIGN AND PARTICIPANTS: This retrospective analysis involved consecutive patients diagnosed with skin cancer in any area of the body and for which treatment with a dermal skin template was recommended as alternative to traditional surgery. MAIN OUTCOME MEASURES: Percentage of successful grafting and the patient and tumor characteristics influencing treatment outcome via univariate analysis. MAIN RESULTS: A total of 302 patients were included. Surgical reconstruction with the matrix was effective in 88.9% of the patients within 21 days of surgery. Notably, the matrix was successful regardless of tumor location, type, or size. Infection was the only variable significantly associated with graft failure (P < .001). CONCLUSIONS: The studied dermal matrix provides an efficient alternative to traditional reconstructive surgery in patients who present specific comorbidities or risk factors. The only variable significantly associated with graft failure was infection, which should be properly controlled through appropriate treatment.

Metabolic Syndrome, Mild Cognitive Impairment, and Dementia: A Meta-Analysis of Longitudinal Studies.

OBJECTIVE: A systematic review and a meta-analysis of both clinical and population-based studies was undertaken according to the Preferred Reporting Items for Systematic Reviews and Meta-Analyses statement to clarify whether Metabolic Syndrome (MetS) is a risk or a protective factor for incident dementia, Alzheimer disease (AD), and vascular dementia (VaD), and whether it's involved in progression to dementia in patients affected by mild cognitive impairment (MCI). METHODS: Search terms included ("metabolic syndrome" OR "syndrome x" OR "plurimetabolic syndrome") AND ("dementia" OR "Alzheimer disease" OR "vascular dementia" OR "mild cognitive impairment" OR "MCI"). Research was restricted to articles published in English between January 1, 2000 and August 31, 2018. No age limit was set. RESULTS: At the end of the selection procedure, nine longitudinal studies were selected for the meta-analysis: six studies enrolled cognitively well-functioning participants and three studies involved MCI patients. A total of 18,313 participants aged older than 40 years with mean MetS prevalence of 22.7% were followed on average for 9.41years. A fixed model was used to estimate pooled hazard ratios and 95% confidence intervals. CONCLUSION: No statistically significant pooled association emerged between MetS and incident dementia and AD. MetS increased the incidence of pure VaD. MetS increased the risk of progression from MCI to dementia. Follow-up length might be a key factor in investigating these associations further. Because MetS is constituted by a set of potentially modifiable factors, further studies with longer follow-up and repeated assessment of both MetS and cognitive status are desirable to draw definite conclusions.

Towards xeno-free cultures of human limbal stem cells for ocular surface reconstruction.

Isolated limbal epithelial stem cells (LESCs) were cultured with or without a 3T3 murine fibroblast feeder-layer (FL) in 4 different culture media on culture plates or on denuded human amniotic membrane (AM) support and fibrin gel support: (1) control medium supplemented with fetal bovine serum; (2) control medium supplemented with the synthetic serum "XerumFree™ XF205" (XF); (3) CnT-20 medium supplemented with "XerumFree™ XF205" (CnT-XF) and (4) CnT-20 medium supplemented with human AB serum (CnT-AB). The three xenogeneic media were compared to standard condition (control + FL) and parameters assessed included cell morphology, proliferative potential, number of passages, assessment of clonogenic and abortive colonies, life span, ∆Np63α expression and epithelial morphology on AM. During serial cultivation of LESCs, most of the tested xeno-free media supported similar numbers of cell passages, total colony number, cumulative cell doublings (CCD) rates and expression of ∆Np63α compared to control. The conditions cultivated with a FL showed a non-statistically significant higher number of cell passages and CCD rates before senescence when compared to the same conditions cultured without FL. Except for the control medium, only XF medium enabled the growth of cells on AM. The expression of ∆Np63α was comparable in all the cultures grown onto AM, when compared to the controls on fibrin gel. In conclusion, the xeno-free media enabled LESC culture both on plastic and on denuded human AM. Despite the analyses were carried out in a statistically low number of samples and need re-assessment in a larger cohort, our results suggest that the production of a completely xeno-free LESC graft could be beneficial for future clinical applications.

Preservation of Ocular Epithelial Limbal Stem Cells: The New Frontier in Regenerative Medicine.

Significant advances have been made in the field of ocular regenerative medicine. Promising stem cell-based therapeutic strategies have been translated into the clinical practice over the last few decades. These new stem cell-based therapies offer the possibility of permanently restoring corneal epithelium in patients with severe disabling and blinding ocular surface disease. The European Union has already classified stem cell-based therapies as "medicinal products". Therefore, manipulation is strictly regulated according to the defined conditions of good manufacturing practice, with the production of stem cell therapeutics at only accredited production sites authorized by the national regulatory agencies. In this regard, as first medical products are licensed for commercial use in Europe enabling a more widespread access to a stem cell-based therapy, the need for safe, validated and reproducible techniques for ex vivo cultured tissue preservation and distribution are coming to the forefront of research. However, these provide various new challenges for biobanking industry such as the retention of viability, good functionality of stem cells and sterility issues. This chapter provides an overview of the current advances in the field of corneal/limbal epithelial stem cell culture preservation techniques using either hypothermic storage or cryopreservation methods, that were used in different culturing steps (from stem cell isolation to the ex vivo epithelial graft preparation), with the reported impact on the post-thawing product recovery.

Congenital and acquired ADAMTS13 deficiency: Two mechanisms, one patient.

Thrombotic thrombocytopenic purpura (TTP) is a life-threatening microangiopathy with a heterogeneous and largely unpredictable course. It is caused by ADAMTS13 deficiency, that can be either congenital or due to anti-ADAMTS13 autoantibodies development. ADAMTS13 deficiency is necessary but not always sufficient to cause acute clinical manifestations and trigger factors may be needed. We report the case of a woman diagnosed with congenital TTP in her adulthood, presenting with anti-ADAMTS13 autoantibodies in acute phase during ticlopidine consumption. Noteworthy, the two ADAMTS13 mutations identified in this patient are novel: one is a splice-site mutation located in intron 11 (c.1308+2_5delTAGG) and the other is a point missense mutation in exon 29 (c.4184T>C leading to p.Leu1395Pro substitution). Since congenital TTP is an extremely rare disease and drug-induced TTP is an uncommon side effect of treatment with ticlopidine, the simultaneous occurrence of both mechanisms of disease in one patient is exceptional. This case represents TTP as a multifactorial disease, with ADAMTS13 genetic abnormality and environmental exposures acting together in determining individual clinical phenotype.

Conversion of Mountain Beech Coppices into High Forest: An Example for Ecological Intensification.

Converting beech coppices into high forest stands has been promoted in the last decades as a management goal to attenuate the negative effects that frequent clearcutting may have on soil, landscape, and biodiversity conservation. The silvicultural tool usually adopted is the gradual thinning of shoots during the long span of time required to complete the conversion, that also allows the owner to keep harvesting some wood. This research reports and discusses, in the light of the ecological intensification approach, the results achieved from an experimental test started more than 25 years ago in a 42-year-old beech (Fagus sylvatica L.) coppice with standards in central Italy. The effects of various thinning intensities (three treatments plus a control) on the stand growth and structure are assessed by successive forest inventories. Analyses are integrated by spatial indices to assess stem density and canopy cover. Converting beech coppices into high forest through gradual thinning of shoots proves to be an effective step down the road to silvicultural systems characterized by continuous forest cover, as a tool of ecological intensification suitable to guarantee both public and private interests. Thinning has led to stands with fewer but larger stems, thus accelerating the long conversion process while maintaining both wood harvesting capability and environmental services.

Prevalence and gender differences in late-life depression: a population-based study.

OBJECTIVE: The worldwide phenomenon of an aging population combined with the increasing prevalence of depression in late life are issues that need to be addressed. This study aims to estimate the frequency of depression and subthreshold depression occurring in a sample of cognitively well-functioning, community-dwelling, older Italian adults and to investigate sociodemographic and clinical correlates of depression, exploring gender differences. METHODS: We used a cross-sectional analyses of survivors in a population-based study (the Faenza Project) that included 359 subjects aged 74 years and older (49.3% women). A modified version of Cambridge Examination for Mental Disorders of Elderly Persons-Revised was administered to all participants. Prevalence rates of depression and 95% confidence intervals (CIs) were estimated according to International Classification of Diseases, Tenth Revision criteria. Statistical analyses were implemented to describe sociodemographic and clinical features associated with depression. Odds ratios were estimated by multivariate logistic regression, and the dependant variable was any type of depression. RESULTS: Overall prevalence of depression was 25.1% (95% CI: 20.6-29.6), with no evidence of gender difference. Prevalence of mild, moderate, and severe depression was 16.4% (95% CI: 12.6-20.2), 7.5% (95% CI: 4.8-10.2), and 1.1 (95% CI: -0.4-2.6), respectively. A rate of 5.6% of the population complained of subthreshold depressive symptoms. After age 81, depression occurrence decreased as age increased. The association between depression and functional measures, such as primary activity, mobility, and disability in performing household chores, were stronger in men than in women. Similarly, severely disabling conditions like stroke were more strongly associated with depression in men than in women. CONCLUSION: Our data suggest a disparity between men and women regarding the impact of depression on everyday life. Specific gender differences need to be taken into account for the evaluation of the depression-related burden in late life.

The Effect of Genome Parametrization and SNP Marker Subsetting on Genomic Selection in Autotetraploid Alfalfa.

BACKGROUND: Alfalfa, the most economically important forage legume worldwide, features modest genetic progress due to long selection cycles and the extent of the non-additive genetic variance associated with its autotetraploid genome. METHODS: To improve the efficiency of genomic selection in alfalfa, we explored the effects of genome parametrization (as tetraploid and diploid dosages, plus allele ratios) and SNP marker subsetting (all available SNPs, only genic regions, and only non-genic regions) on genomic regressions, together with various levels of filtering on reading depth and missing rates. We used genotyping by sequencing-generated data and focused on traits of different genetic complexity, i.e., dry biomass yield in moisture-favorable (FE) and drought stress (SE) environments, leaf size, and the onset of flowering, which were assessed in 143 genotyped plants from a genetically broad European reference population and their phenotyped half-sib progenies. RESULTS: On average, the allele ratio improved the predictive ability compared with other genome parametrizations (+7.9% vs. tetraploid dosage, +12.6% vs. diploid dosage), while using all the SNPs offered an advantage compared with any specific SNP subsetting (+3.7% vs. genic regions, +7.6% vs. non-genic regions). However, when focusing on specific traits, different combinations of genome parametrization and subsetting achieved better performances. We also released Legpipe2, an SNP calling pipeline tailored for reduced representation (GBS, RAD) in medium-sized genotyping experiments.

Retinal Microvascular Alterations in Hidradenitis Suppurativa Patients: A Pilot Study Using Optical Coherence Tomography Angiography.

Background: Hidradenitis suppurativa (HS) is a relapsing-remitting inflammatory disease characterized by the progression of asymptomatic nodules to deep-seated lesions and fistula formation that leads to suppuration and scarring. Optical coherence tomography angiography (OCTA) is a new non-invasive imaging technique that carefully analyzes retinal microvasculature networks with high-resolution imaging. Recent studies have demonstrated that retinal vessel density and retinal perfusion reflect systemic inflammatory responses. This study's aim was to analyze OCTA-derived retinal microvasculature parameters to understand if patients affected by HS and without any relevant ocular or systemic comorbidities showed impaired retinal vascular function and morphology. Method: We performed a case-control study of HS patients and age- and sex-matched control cohort. A total of 20 eyes from 10 HS patients and 30 eyes from 15 healthy controls were analyzed, and OCTA-derived microvasculature parameters were compared between groups. Results: OCTA images showed that HS patients, compared to healthy controls, were typically characterized by higher values of the foveal avascular zone (FAZ) both in the superficial capillary plexus (SCP) and in the deep capillary plexus (DCP), and by lower values of vessel density (VD)-SCP, VD-DCP, and vessel length density (VLD)-SCP in the foveal region. These findings partially reflect changes that have been demonstrated in diabetic patients that could be induced by a protracted metabolic or systemic inflammatory dysregulation. Conclusions: In conclusion, OCTA enables large-scale, non-invasive visual screening and follow-up of the retinal vasculature features, providing a new strategy for the prevention and monitoring of visual changes in HS patients.

From MASH to HCC: the role of Gas6/TAM receptors.

Metabolic dysfunction-associated steatohepatitis (MASH) is the replacement term for what used to be called nonalcoholic steatohepatitis (NASH). It is characterized by inflammation and injury of the liver in the presence of cardiometabolic risk factors and may eventually result in the development of hepatocellular carcinoma (HCC), the most common form of primary liver cancer. Several pathogenic mechanisms are involved in the transition from MASH to HCC, encompassing metabolic injury, inflammation, immune dysregulation and fibrosis. In this context, Gas6 (Growth Arrest-Specific 6) and TAM (Tyro3, Axl, and MerTK) receptors may play important roles. The Gas6/TAM family is involved in the modulation of inflammation, lipid metabolism, fibrosis, tumor progression and metastasis, processes which play an important role in the pathophysiology of acute and chronic liver diseases. In this review, we discuss MASH-associated HCC and the potential involvement of the Gas6/TAM system in disease development and progression. In addition, since therapeutic strategies for MASH and HCC are limited, we also speculate regarding possible future treatments involving the targeting of Gas6 or TAM receptors.