System Integration and Certification. The Market Demand for Clarity and Transparency/Part 2

Rosana G. Moreira, Editor-in-Chief; Texas A&M UniversityThis is an Invited Paper from International Commission of Agricultural Engineering (CIGR, Commission Internationale du Genie Rural) E-Journal Volume 5 (2003): L. Bodria. System Integration and Certification. The Market Demand for Clarity and Transparency/Part 2. Club of Bologna. Nov. 16, 2002. Vol. V. February 2003

Studio di fattibilit\ue0 di un sistema ottico portatile semplificato per la valutazione del grado di maturaione di mirtilli

Le fasi di maturazione dei mirtilli fino alla raccolta vengono oggi seguite senza l\u2019aiuto di strumenti in grado di fornire dati oggettivi e anche il momento della raccolta viene deciso solamente sulla base dell\u2019analisi visiva dei frutti e dell\u2019esperienza del produttore. I produttori potrebbero quindi essere supportati nelle loro decisioni da strumenti semplici e portatili, utilizzabili direttamente in campo per la valutazione del grado di maturazione dei frutti. Pertanto \ue8 stata studiata la possibilit\ue0 di realizzare uno strumento ottico portatile ed economico, basato su tecnologia a LED e sulla misura e l\u2019elaborazione del valore di riflettanza diffusa in corrispondenza di un numero limitato di lunghezze d\u2019onda opportunamente selezionate. La selezione delle lunghezze d\u2019onda pi\uf9 significative per l\u2019individuazione dei frutti a maturazione commerciale adatti alla raccolta, \ue8 stata effettuata partendo da spettri Vis/NIR raccolti nell\u2019intervallo 445-970 nm di campioni della variet\ue0 Brigitta. Le bacche raccolte sono state suddivise in quattro classi di maturazione in funzione del colore. Gli spettri acquisiti sono stati sottoposti a un\u2019Analisi delle Componenti Principali (PCA, The Unscrambler\uae 9.6) che ha permesso di evidenziare in particolare due bande (attorno a 650 nm e a 740 nm) in corrispondenza delle quali risultano massimizzate le differenze tra gli spettri dei campioni completamente maturi e quelli delle altre classi. I valori di riflettanza registrati a queste lunghezze d\u2019onda sono stati utilizzati per creare dei rapporti normalizzati rispetto alla lunghezza d\u2019onda di 850 nm e sulla base di tali rapporti \ue8 stato elaborato un algoritmo semplificato per la classificazione dei frutti. La classificazione dei campioni in base all\u2019indice di maturazione elaborato ha mostrato un\u2019elevata capacit\ue0 di discriminare i frutti completamente maturi da quelli parzialmente o non maturi (93% di campioni correttamente classificati). I risultati di questo studio indicano pertanto la fattibilit\ue0 di uno strumento semplificato ed economico, basato sull\u2019impiego di sole tre lunghezze d\u2019onda, capace di valutare rapidamente e in condizioni di campo il grado di maturazione dei mirtilli

Feasibility of a simplified handheld optical system for blue-berries ripeness field evalutation

During last years, small fruits farming and marketing have increased widely in Italy in response to consumer\u2019s interest in bioactive and health protecting compounds fruits rich. At present, the ripening stage and harvest time is normally estimated by a visual analysis based on grower\u2019s experience. Farm growers could be supported in their decisions by simple and portable devices, used to directly assess the berries ripeness in the field and accordingly to plan the best harvest time. The aim of this work was to study the feasibility of a simplified handheld and inexpensive optical device, based on measurements and processing of diffuse reflectance at a few wavelengths appropriately selected. This study is focused on selecting the most significant wavelengths able to identify in the field the blueberries ready to be harvested. To this aim Vis/NIR spectra in the range 445-970 nm were acquired for Vaccinium corymbosum (\u2018Brigitta\u2019 cultivar) during two different growing seasons (2005 and 2006), harvested in Valtellina area (Lombardy, Italy). Spectra measurements were taken in the field on individual berries along their equator region. Fruits samplings were performed weekly, and picked fruits were divided in four ripeness classes according to external colour. The PCA of 634 fruits spectra highlighted two principal spectral bands (around 680 nm and 740 nm) in which differences among fully (class IV) and not completely ripe (classes IIII)samples spectra are maximized. Reflectance values at these wavelengths were used to obtain spectral ratios normalised to 850 nm reflectance. Based on these relations, a simple fruits classification algorithm was proposed. Berries ripeness grading based on the proposed index showed a high ability in discriminating fully ripe fruits from partially ripe fruits and unripe fruits. More then the 93% of samples were correctly classified in validation phase. The results of this study demonstrate the feasibility of a simplified and low-cost handheld device, based on the use of only three wavelengths, able to quickly estimate blueberry ripeness in the field, with special reference to the last and most sensitive stages of ripening process

A farm-scale pilot plant for biohydrogen and biomethane production by two-stage fermentation

Hydrogen is considered one of the possible main energy carriers for the future, thanks to its unique environmental properties. Indeed, its energy content (120 MJ/kg) can be exploited virtually without emitting any exhaust in the atmosphere except for water. Renewable production of hydrogen can be obtained through common biological processes on which relies anaerobic digestion, a well-established technology in use at farm-scale for treating different biomass and residues. Despite two-stage hydrogen and methane producing fermentation is a simple variant of the traditional anaerobic digestion, it is a relatively new approach mainly studied at laboratory scale. It is based on biomass fermentation in two separate, seuqential stages, each maintaining conditions optimized to promote specific bacterial consortia: in the first acidophilic reactorhydrogen is produced production, while volatile fatty acids-rich effluent is sent to the second reactor where traditional methane rich biogas production is accomplished. A two-stage pilot-scale plant was designed, manufactured and installed at the experimental farm of the University of Milano and operated using a biomass mixture of livestock effluents mixed with sugar/starch-rich residues (rotten fruits and potatoes and expired fruit juices), afeedstock mixture based on waste biomasses directly available in the rural area where plant is installed. The hydrogenic and the methanogenic reactors, both CSTR type, had a total volume of 0.7m3 and 3.8 m3 respectively, and were operated in thermophilic conditions (55 2 °C) without any external pH control, and were fully automated. After a brief description of the requirements of the system, this contribution gives a detailed description of its components and of engineering solutions to the problems encountered during the plant realization and start-up. The paper also discusses the results obtained in a first experimental run which lead to production in the range of previous laboratory results, with a typical hydrogen and methane specific productivity of 2.2 and 0.5 Nm3/m3reactor per day, in the first and second stage of the plant respectively. At our best knowledge, this plant is one of the very first prototypes producing biohydrogen at farm scale, and it represents a distributed, small scale demonstration to obtain hydrogen from renewable waste-sources

PCSK9 ablation attenuates Aβ pathology, neuroinflammation and cognitive dysfunctions in 5XFAD mice

Background: Increasing evidence highlights the importance of novel players in Alzheimer's disease (AD) pathophysiology, including alterations of lipid metabolism and neuroinflammation. Indeed, a potential involvement of Proprotein convertase subtilisin/kexin type 9 (PCSK9) in AD has been recently postulated. Here, we first investigated the effects of PCSK9 on neuroinflammation in vitro. Then, we examined the impact of a genetic ablation of PCSK9 on cognitive performance in a severe mouse model of AD. Finally, in the same animals we evaluated the effect of PCSK9 loss on Aβ pathology, neuroinflammation, and brain lipids. Methods: For in vitro studies, U373 human astrocytoma cells were treated with Aβ fibrils and human recombinant PCSK9. mRNA expression of the proinflammatory cytokines and inflammasome-related genes were evaluated by q-PCR, while MCP-1 secretion was measured by ELISA. For in vivo studies, the cognitive performance of a newly generated mouse line - obtained by crossing 5XFADHet with PCSK9KO mice – was tested by the Morris water maze test. After sacrifice, immunohistochemical analyses were performed to evaluate Aβ plaque deposition, distribution and composition, BACE1 immunoreactivity, as well as microglia and astrocyte reactivity. Cholesterol and hydroxysterols levels in mouse brains were quantified by fluorometric and LC-MS/MS analyses, respectively. Statistical comparisons were performed according to one- or two-way ANOVA, two-way repeated measure ANOVA or Chi-square test. Results: In vitro, PCSK9 significantly increased IL6, IL1B and TNFΑ mRNA levels in Aβ fibrils-treated U373 cells, without influencing inflammasome gene expression, except for an increase in NLRC4 mRNA levels. In vivo, PCSK9 ablation in 5XFAD mice significantly improved the performance at the Morris water maze test; these changes were accompanied by a reduced corticohippocampal Aβ burden without affecting plaque spatial/regional distribution and composition or global BACE1 expression. Furthermore, PCSK9 loss in 5XFAD mice induced decreased microgliosis and astrocyte reactivity in several brain regions. Conversely, knocking out PCSK9 had minimal impact on brain cholesterol and hydroxysterol levels. Conclusions: In vitro studies showed a pro-inflammatory effect of PCSK9. Consistently, in vivo data indicated a protective role of PCSK9 ablation against cognitive impairments, associated with improved Aβ pathology and attenuated neuroinflammation in a severe mouse model of AD. PCSK9 may thus be considered a novel pharmacological target for the treatment of AD

Rituximab in Children with Steroid-Dependent Nephrotic Syndrome: A Multicenter, Open-Label, Noninferiority, Randomized Controlled Trial.

Steroid-dependent nephrotic syndrome (SDNS) carries a high risk of toxicity from steroids or steroid-sparing agents. This open-label, noninferiority, randomized controlled trial at four sites in Italy tested whether rituximab is noninferior to steroids in maintaining remission in juvenile SDNS. We enrolled children age 1-16 years who had developed SDNS in the previous 6-12 months and were maintained in remission with high prednisone doses ( 650.7 mg/kg per day). We randomly assigned participants to continue prednisone alone for 1 month (control) or to add a single intravenous infusion of rituximab (375 mg/m(2); intervention). Prednisone was tapered in both groups after 1 month. For noninferiority, rituximab had to permit steroid withdrawal and maintain 3-month proteinuria (mg/m(2) per day) within a prespecified noninferiority margin of three times the levels among controls (primary outcome). We followed participants for 651 year to compare risk of relapse (secondary outcome). Fifteen children per group (21 boys; mean age, 7 years [range, 2.6-13.5 years]) were enrolled and followed for 6460 months (median, 22 months). Three-month proteinuria was 42% lower in the rituximab group (geometric mean ratio, 0.58; 95% confidence interval, 0.18 to 1.95 [i.e., within the noninferiority margin of three times the levels in controls]). All but one child in the control group relapsed within 6 months; median time to relapse in the rituximab group was 18 months (95% confidence interval, 9 to 32 months). In the rituximab group, nausea and skin rash during infusion were common; transient acute arthritis occurred in one child. In conclusion, rituximab was noninferior to steroids for the treatment of juvenile SDNS

Mutations in DSTYK and dominant urinary tract malformations.

ABSTRACT Introduction Congenital abnormalities of the kidney of the urinary tract are the most common cause of pediatric kidney failure. These disorders are highly heterogeneous, and their etiology is poorly understood. Methods We performed genome-wide linkage analysis and whole-exome sequencing in a family with autosomal dominant congenital abnormalities of the kidney of the urinary tract (7 affected family members). We also performed sequence analysis in 311 unrelated patients, as well as histologic and functional studies. Results Linkage analysis identified five regions of the genome that were shared among all affected family members. Exome sequencing identified a single rare deleterious variant within these linkage intervals, a heterozygous splice-site mutation in dual serine/threonine and tyrosine protein kinase (DSTYK). This variant, which resulted in aberrant gene product splicing, was present in all affected family members. Additional independent DSTYK mutations, including nonsense and splice-site mutations, were detected among 7/311 unrelated patients. DSTYK is highly expressed in the maturing epithelia of all major organs, localizing to cell membranes. Knockdown in zebrafish resulted in multi-organ developmental defects, resembling loss of fibroblast growth factor (FGF) signaling. Consistent with this finding, DSTYK colocalizes with FGF receptors in the ureteric bud and metanephric mesenchyme. Finally, DSTYK knockdown in human embryonic kidney cells inhibited FGF-stimulated ERK-phosphorylation, the principal signal downstream of receptor tyrosine kinases. Conclusions We detected DSTYK mutations in 2.2% of patients with congenital abnormalities of the kidney and urinary tract whom we studied, suggesting that DSTYK is a major determinant of human urinary tract development, downstream of FGF signaling

Strong protective effect of the APOL1 p.N264K variant against G2-associated focal segmental glomerulosclerosis and kidney disease

African Americans have a significantly higher risk of developing chronic kidney disease, especially focal segmental glomerulosclerosis -, than European Americans. Two coding variants (G1 and G2) in the APOL1 gene play a major role in this disparity. While 13% of African Americans carry the high-risk recessive genotypes, only a fraction of these individuals develops FSGS or kidney failure, indicating the involvement of additional disease modifiers. Here, we show that the presence of the APOL1 p.N264K missense variant, when co-inherited with the G2 APOL1 risk allele, substantially reduces the penetrance of the G1G2 and G2G2 high-risk genotypes by rendering these genotypes low-risk. These results align with prior functional evidence showing that the p.N264K variant reduces the toxicity of the APOL1 high-risk alleles. These findings have important implications for our understanding of the mechanisms of APOL1-associated nephropathy, as well as for the clinical management of individuals with high-risk genotypes that include the G2 allele

De novo TRIM8 variants impair its protein localization to nuclear bodies and cause developmental delay, epilepsy, and focal segmental glomerulosclerosis

Focal segmental glomerulosclerosis (FSGS) is the main pathology underlying steroid-resistant nephrotic syndrome (SRNS) and a leading cause of chronic kidney disease. Monogenic forms of pediatric SRNS are predominantly caused by recessive mutations, while the contribution of de novo variants (DNVs) to this trait is poorly understood. Using exome sequencing (ES) in a proband with FSGS/SRNS, developmental delay, and epilepsy, we discovered a nonsense DNV in TRIM8, which encodes the E3 ubiquitin ligase tripartite motif containing 8. To establish whether TRIM8 variants represent a cause of FSGS, we aggregated exome/genome-sequencing data for 2,501 pediatric FSGS/SRNS-affected individuals and 48,556 control subjects, detecting eight heterozygous TRIM8 truncating variants in affected subjects but none in control subjects (p = 3.28 × 10−11). In all six cases with available parental DNA, we demonstrated de novo inheritance (p = 2.21 × 10−15). Reverse phenotyping revealed neurodevelopmental disease in all eight families. We next analyzed ES from 9,067 individuals with epilepsy, yielding three additional families with truncating TRIM8 variants. Clinical review revealed FSGS in all. All TRIM8 variants cause protein truncation clustering within the last exon between residues 390 and 487 of the 551 amino acid protein, indicating a correlation between this syndrome and loss of the TRIM8 C-terminal region. Wild-type TRIM8 overexpressed in immortalized human podocytes and neuronal cells localized to nuclear bodies, while constructs harboring patient-specific variants mislocalized diffusely to the nucleoplasm. Co-localization studies demonstrated that Gemini and Cajal bodies frequently abut a TRIM8 nuclear body. Truncating TRIM8 DNVs cause a neuro-renal syndrome via aberrant TRIM8 localization, implicating nuclear bodies in FSGS and developmental brain disease

Genetic drivers of kidney defects in the digeorge syndrome

BACKGROUND The DiGeorge syndrome, the most common of the microdeletion syndromes, affects multiple organs, including the heart, the nervous system, and the kidney. It is caused by deletions on chromosome 22q11.2; the genetic driver of the kidney defects is unknown. METHODS We conducted a genomewide search for structural variants in two cohorts: 2080 patients with congenital kidney and urinary tract anomalies and 22,094 controls. We performed exome and targeted resequencing in samples obtained from 586 additional patients with congenital kidney anomalies. We also carried out functional studies using zebrafish and mice. RESULTS We identified heterozygous deletions of 22q11.2 in 1.1% of the patients with congenital kidney anomalies and in 0.01% of population controls (odds ratio, 81.5; P = 4.5×1014). We localized the main drivers of renal disease in the DiGeorge syndrome to a 370-kb region containing nine genes. In zebrafish embryos, an induced loss of function in snap29, aifm3, and crkl resulted in renal defects; the loss of crkl alone was sufficient to induce defects. Five of 586 patients with congenital urinary anomalies had newly identified, heterozygous protein-Altering variants, including a premature termination codon, in CRKL. The inactivation of Crkl in the mouse model induced developmental defects similar to those observed in patients with congenital urinary anomalies. CONCLUSIONS We identified a recurrent 370-kb deletion at the 22q11.2 locus as a driver of kidney defects in the DiGeorge syndrome and in sporadic congenital kidney and urinary tract anomalies. Of the nine genes at this locus, SNAP29, AIFM3, and CRKL appear to be critical to the phenotype, with haploinsufficiency of CRKL emerging as the main genetic driver