The giant diploid faba genome unlocks variation in a global protein crop

Publisher Copyright: © 2023, The Author(s).Increasing the proportion of locally produced plant protein in currently meat-rich diets could substantially reduce greenhouse gas emissions and loss of biodiversity1. However, plant protein production is hampered by the lack of a cool-season legume equivalent to soybean in agronomic value2. Faba bean (Vicia faba L.) has a high yield potential and is well suited for cultivation in temperate regions, but genomic resources are scarce. Here, we report a high-quality chromosome-scale assembly of the faba bean genome and show that it has expanded to a massive 13 Gb in size through an imbalance between the rates of amplification and elimination of retrotransposons and satellite repeats. Genes and recombination events are evenly dispersed across chromosomes and the gene space is remarkably compact considering the genome size, although with substantial copy number variation driven by tandem duplication. Demonstrating practical application of the genome sequence, we develop a targeted genotyping assay and use high-resolution genome-wide association analysis to dissect the genetic basis of seed size and hilum colour. The resources presented constitute a genomics-based breeding platform for faba bean, enabling breeders and geneticists to accelerate the improvement of sustainable protein production across the Mediterranean, subtropical and northern temperate agroecological zones.Peer reviewe

Effect of bisphosphonates on cartilage turnover assessed with a newly developed assay for collagen type II degradation products

Background: Animal studies of arthritis have suggested that bisphosphonates may have chondroprotective abilities. Objective: To evaluate the effect of bisphosphonate treatment on cartilage degradation. Methods: Type II collagen is almost exclusively localised in cartilage, where it is the major structural component of the tissue. Hence fragments derived from this protein should represent a specific index for cartilage degradation. The urinary concentration of collagen type II C-telopeptide degradation products (CTX-II) was measured by a new immunoassay (enzyme linked immunosorbent assay (ELISA)). The serum concentration of collagen type I C-telopeptide degradation products (CTX-I), a marker of bone degradation, was also measured by ELISA. Participants: Two groups were studied. The alendronate group included 63 healthy postmenopausal women aged 45–54 randomly allocated to receive three years' treatment with 1 mg, 5 mg, 10 mg, or 20 mg alendronate daily or placebo. In the third year the women receiving 20 mg were switched to placebo. The ibandronate group included 119 women at least 10 years after the menopause aged <75 randomly allocated to receive 12 months' treatment with 0.25 mg, 0.5 mg, 1.0 mg, 2.5 mg, or 5 mg ibandronate daily or placebo followed by 12 months without treatment. Results: 20 mg of alendronate and 2.5 and 5 mg of ibandronate treatment produced significant decreases in urinary CTX-II to about 50% of baseline. The level reached after three months of treatment remained practically constant during the following 12–36 treatment months. When treatment was withdrawn CTX-II values returned towards baseline. Serum CTX-I also decreased rapidly within three months, but to a level of about 30% of baseline. Conclusions: The urinary excretion of CTX-II, a new marker of cartilage degradation, decreases significantly in response to bisphosphonate. This suggests that bisphosphonates may have chondroprotective effects in humans. By measurement of CTX-II it should be possible to monitor the effects of drugs that potentially inhibit cartilage destruction

Cartilage turnover assessed with a newly developed assay measuring collagen type II degradation products: influence of age, sex, menopause, hormone replacement therapy, and body mass index

Background: Cartilage normally has a slow turnover but in arthritis increased metabolism results in degradation of the tissue. Objective: To assess cartilage turnover in a sample of the general population by an assay measuring cartilage derived urinary collagen type II (CTX-II) C-telopeptide degradation products. Methods: CTX-II concentrations were measured in urine samples from 615 healthy men and women aged 20–87 years, and the influence of age, sex, menopause, hormone replacement therapy (HRT), and body mass index (BMI) was assessed. Results: CTX-II concentrations showed age dependent variations, with notable differences between men and women. Mean (SD) CTX-II concentration in postmenopausal women (220 (118) ng/mmol, n=25) was significantly higher than in an age matched group of premenopausal women (112 (79) ng/mmol, n=26, p<0.001). CTX-II concentration in women using HRT (118 (57) ng/mmol, n=50) was significantly lower than in an age and BMI matched group of women not receiving HRT (215 (99) ng/mmol, n=50, p<0.001). In subjects with a BMI ≥25 kg/m(2), CTX-II concentrations were significantly higher than in those with a BMI <25 kg/m(2) (185 (114) v 148 (91) ng/mmol, p<0.001). Conclusions: Cartilage turnover, as assessed by measuring urinary degradation products of CTX-II varies considerably with age, and significant differences between CTX-II levels in men and women as well as in pre- and postmenopausal women are found. Further studies are required to validate the marker for assessing cartilage degradation in arthritis

A phase I study of recombinant human interleukin-21 (rIL-21) in combination with sunitinib in patients with metastatic renal cell carcinoma (RCC)

BACKGROUND: sunitinib induces partial responses in 47% of patients with metastatic renal cell carcinoma (mRCC). However, the achievement of complete responses remains scarce and all patients will eventually develop progressive disease. Recombinant interleukin-21 (rIL-21) is a novel cytokine, which is believed to deliver sustained cellular anti-tumor response and the combination of both agents may work synergistically. MATERIAL AND METHOD: from July 2007 to July 2008 in this phase I trial nine therapy-naive patients with metastatic RCC in five European centers were enrolled. Patients with either good or intermediate risk according to Memorial Sloan-Kettering Cancer Center (MSKCC) were eligible without restrictions to histology subtype nor measurable disease. Patients were treated with increasing doses of rIL-21 administered subcutaneously (s.c.) in combination with sunitinib 50 mg once daily (OD) orally at the '4 weeks on/2 weeks off' schedule. Dose-escalation was applied by a conventional '3+3 design'. Planned dose levels (DL) for rIL-21 were 3, 10, 30 and 100 microg/kg s.c. The primary endpoint was to determine the maximum tolerated dose (MTD) and recommended dose (rd). secondary objectives included pharmacokinetics of sunitinib and ril-21, and the induction of ril-21 antibodies. RESULTS: at 10 microg/kg two dose-limiting toxicities (DLT) occurred in four patients, consisting of grade 4 neutropenia and grade 3 thrombocytopenia. The MTD was 3 microg/kg rIL-21 combined with sunitinib 50 mg OD at the '4 weeks on/2 weeks off' schedule. Frequent occurring adverse events were injection site reaction, stomatitis, fatigue and dysgeusia. CONCLUSIONS: the combination of sunitinib 50 mg at the '4 weeks on/2 weeks off' schedule and 10 microg/kg IL-21 was not tolerated due to hematological DLTs. The dose level of 3 microg/kg rIL-21 was considered too low to be therapeutically relevant for further evaluation and therefore the study was discontinued

Activity of recombinant human interleukin-21 (rIL-21) in patients (pts) with stage IV malignant melanoma (MM) without prior treatment : clinical data from a phase IIa study

Background: IL-21 has a key role in cancer immunology. We have previously described a phase 1 study of rIL-21 in pts with advanced MM. We now report final clinical results from a phase 2a study in treatment-naïve pts with advanced MM. Methods: Open-label, two-stage phase 2a trial. Primary objective: antitumor efficacy as determined by response rate. Secondary objectives: safety, effects on blood biomarkers, measurement of anti-rIL-21 antibodies. Eligibility: unresectable MM, measurable disease by RECIST, no prior systemic therapy (adjuvant interferon permitted), adequate major organ function, good performance status, no significant autoimmune disease, life expectancy ≥4 months. rIL-21 was administered at 30 µg/kg/dose in cycles of 5 dosing days followed by 9 days of rest for 6 weeks (wks), followed by 2 wks off treatment. Further cycles were offered to pts without tumor progression causing symptoms and requiring other therapy. Only pts with PR or CR by RECIST at wk 16 were offered further treatment. Results: Stage 1 of the study comprised 14 pts. One confirmed CR was observed and as per protocol 10 more pts were accrued to stage 2 (total n=24: 10 female, 14 male). Best tumor response any time on study included 1 confirmed CR and 1 confirmed PR (both with lung mets) with overall RR 8.3%, 8 SD (33.3%) and 14 PD (58.3%). The pt with CR by wk 8 had PD 11 wks later. The pt with PR had SD at wk 8 but confirmed PR at wk 16. 6/8 pts with SD at wk 8 had progressed by wk 16. Another pt had CR in target lesions and SD in non-target, but PD in brain (overall PD). One pt had 9 cycles of treatment, 10 had 6, and 13 had 3–4. Treatment was well tolerated and safety was similar to the phase 1 trial. 8 pts had SAEs: hypersensitivity (1), hepatitis (1), fever (2), rigor (1), cellulitis (1), sinus tachycardia (1), cancer related pain (4). One pt withdrew due to adverse events at wk 8 and 3 pts had the dose reduced to 10 µg/kg. Conclusions: rIL-21 administered at 30 µg/kg/day in 5-day cycles every 2nd wk was well tolerated by pts with metastatic MM and confirmed responses by RECIST including one CR were observed. Future studies will evaluate potential for further anti-tumor activity at higher doses of rIL-21.1 page(s