Kidney tissue loading reduces the critical cooling and warming rates of VS55 and VMP cryoprotective solutions.

Critical cooling and warming rates (CCR and CWR) are two important calorimetric properties of cryoprotective agents (CPA) solutions, and achieving these rates is generally regarded as the critical criterion for successful vitrification and rewarming. In 1996, Peyridieu et al. discovered that the measured critical rates are reduced inside kidney tissue equilibrated with 30% (w/w) 2,3-butanediol compared to its free CPA solution. In general, they found a ∼5-fold reduction for CCR and a >100-fold reduction for CWR. However, to our knowledge, no follow-up studies have been conducted. We revisit this important concept, understanding that tissues never fully equilibrate with full-strength 100% CPAs during perfusion. We therefore performed measurements in a range of dilutions of two commonly employed CPA cocktails, including 75-100% VS55 (41.25-55.00% w/v) and 90-100% VMP (48.60-54.00% w/v) equilibrated with kidney tissues vs. free solution. The measured reduction in the kidney was up to 5-fold for CCR and 9-fold for CWR. After discussing possible mechanisms for this effect, curves that fit the dilution to the observed reduction in critical rate were constructed to allow extrapolation for differentially loaded tissues, which can guide the follow-up studies to find the more concentrated CPA (> 8.4 M VMP) in the M22 family to achieve human-sized kidney vitrification and rewarming.

The Fundamental Approach of the Digital Twin Application in Railway Turnouts with Innovative Monitoring of Weather Conditions.

Improving railway safety depends heavily on the reliability of railway turnouts. The realization of effective, reliable and continuous observations for the spatial analysis and evaluation of the technical condition of railway turnouts is one of the factors affecting safety in railway traffic. The mode and scope of monitoring changes in geometric parameters of railway turnouts with associated indicators needs improvement. The application of digital twins to railway turnouts requires the inclusion of fundamental data indicating their condition along with innovative monitoring of weather conditions. This paper presents an innovative solution for monitoring the status of temperature and other atmospheric conditions. A UbiBot WS1 WIFI wireless temperature logger was used, with an external DS18B20 temperature sensor integrated into an S49 (49E1)-type rail as Tszyn WS1 WIFI. Measurements were made between January and May (winter/spring) at fixed time intervals and at the same measurement point. The aim of the research is to present elements of a fundamental approach of applying digital twins to railway turnouts requiring the consideration and demonstration of rail temperature conditions as a component in the data acquisition of railway turnout condition data and other constituent atmospheric conditions through an innovative solution. The research showed that the presented innovative solution is an effective support for the application of digital twins to railway turnouts and ongoing surveying and diagnostic work of other elements of rail transport infrastructure. The applicability of the TgCWRII second temperature difference indicator in the monitoring of railway turnouts was also confirmed.

Chemical weathering and atmospheric carbon dioxide (CO2) consumption in Shanmuganadhi, South India: evidences from groundwater geochemistry.

Chemical weathering in a groundwater basin is a key to understanding global climate change for a long-term scale due to its association with carbon sequestration. The present study aims to characterize and to quantify silicate weathering rate (SWR), carbon dioxide consumption rate and carbonate weathering rate (CWR) in hard rock terrain aided by major ion chemistry. The proposed study area Shanmuganadhi is marked with superior rainfall, oscillating temperature and runoff with litho-units encompassing charnockite and hornblende-biotite gneiss. Groundwater samples (n = 60) were collected from diverse locations and analysed for major chemical constituents. Groundwater geochemistry seems to be influenced by geochemical reactions combining dissolution and precipitation of solids, cation exchange and adsorption along with minor contribution from anthropogenic activities. The SWR calculated for charnockite and hornblende-biotite gneiss was 3.07 tons km-2 year-1 and 5.12 tons km-2 year-1, respectively. The calculated CWR of charnockite and hornblende-biotite gneiss was 0.079 tons km-2 year-1 and 0.74 tons km-2 year-1, respectively. The calculated CO2 consumption rates via silicate weathering were 1.4 × 103 mol km-2 year-1 for charnockite and 5.8 × 103 mol km-2 year-1 for hornblende-biotite gneiss. Lithology, climate and relief were the key factors isolated to control weathering and CO2 consumption rates.

Radiation treatment monitoring with DCE-US in CWR22 prostate tumor xenografts.

BACKGROUND: Longitudinal monitoring of potential radiotherapy treatment effects can be determined by dynamic contrast-enhanced ultrasound (DCE-US). PURPOSE: To assess functional parameters by means of DCE-US in a murine subcutaneous model of human prostate cancer, and their relationship to dose deposition and time-frame after treatment. A special focus has been placed to evaluate the vascular heterogeneity of the tumor and on the most suitable data analysis approach that reflects this heterogeneity. MATERIAL AND METHODS: In vivo DCE-US was acquired 24 h and 48 h after radiation treatment with a single dose of 7.5 Gy and 10 Gy, respectively. Tumor vasculature was characterized pixelwise using the Brix pharmacokinetic analysis of the time-intensity curves. RESULTS: Longitudinal changes were detected ( P < 0.001) at 24 h and 48 h after treatment. At 48 h, the eliminating rate constant of the contrast agent from the plasma, kel, was correlated ( P ≤ 0.05) positively with microvessel density (MVD; rτ = 0.7) and negatively with necrosis (rτ = -0.6) for the treated group. Furthermore, Akep, a parameter related to transcapillary transport properties, was also correlated to MVD (rτ = 0.6, P ≤ 0.05). CONCLUSION: DCE-US has been shown to detect vascular changes at a very early stage after radiotherapy, which is a great advantage since DCE-US is non-invasive, available at most hospitals, and is low in cost compared to other techniques used in clinical practice.

Concurrent regulation of LKB1 and CaMKK2 in the activation of AMPK in castrate-resistant prostate cancer by a well-defined polyherbal mixture with anticancer properties.

BACKGROUND: Zyflamend, a blend of herbal extracts, effectively inhibits tumor growth using preclinical models of castrate-resistant prostate cancer mediated in part by 5'-adenosine monophosphate-activated protein kinase (AMPK), a master energy sensor of the cell. Clinically, treatment with Zyflamend and/or metformin (activators of AMPK) had benefits in castrate-resistant prostate cancer patients who no longer responded to treatment. Two predominant upstream kinases are known to activate AMPK: liver kinase B1 (LKB1), a tumor suppressor, and calcium-calmodulin kinase kinase-2 (CaMKK2), a tumor promotor over-expressed in many cancers. The objective was to interrogate how Zyflamend activates AMPK by determining the roles of LKB1 and CaMKK2. METHODS: AMPK activation was determined in CWR22Rv1 cells treated with a variety of inhibitors of LKB1 and CaMKK2 in the presence and absence of Zyflamend, and in LKB1-null HeLa cells that constitutively express CaMKK2, following transfection with wild type LKB1 or catalytically-dead mutants. Upstream regulation by Zyflamend of LKB1 and CaMKK2 was investigated targeting protein kinase C-zeta (PKCζ) and death-associated protein kinase (DAPK), respectively. RESULTS: Zyflamend's activation of AMPK appears to be LKB1 dependent, while simultaneously inhibiting CaMKK2 activity. Zyflamend failed to rescue the activation of AMPK in the presence of pharmacological and molecular inhibitors of LKB1, an effect not observed in the presence of inhibitors of CaMKK2. Using LKB1-null and catalytically-dead LKB1-transfected HeLa cells that constitutively express CaMKK2, ionomycin (activator of CaMKK2) increased phosphorylation of AMPK, but Zyflamend only had an effect in cells transfected with wild type LKB1. Zyflamend appears to inhibit CaMKK2 by DAPK-mediated phosphorylation of CaMKK2 at Ser511, an effect prevented by a DAPK inhibitor. Alternatively, Zyflamend mediates LKB1 activation via increased phosphorylation of PKCζ, where it induced translocation of PKCζ and LKB1 to their respective active compartments in HeLa cells following treatment. Altering the catalytic activity of LKB1 did not alter this translocation. DISCUSSION: Zyflamend's activation of AMPK is mediated by LKB1, possibly via PKCζ, but independent of CaMKK2 by a mechanism that appears to involve DAPK. CONCLUSIONS: Therefore, this is the first evidence that natural products simultaneously and antithetically regulate upstream kinases, known to be involved in cancer, via the activation of AMPK.

Metastatic phenotype in CWR22 prostate cancer xenograft following castration.

BACKGROUND: CWR22 is a human xenograft model of primary prostate cancer (PCa) that is often utilized to study castration recurrent (CR) PCa. CWR22 recapitulates clinical response to androgen deprivation therapy (ADT), in that tumors regress in response to castration, but can recur after a period of time. METHODS: Two cohorts of mice, totaling 117 mice were implanted with CWR22, allowed to develop tumors, castrated by pellet removal and followed for a period of 32 and 50 weeks. Mice presenting with tumors >2.0 cm(3) at the primary site, moribund appearance, or palpable masses other than the primary tumor were sacrificed prior to the endpoint of the study. Tumor tissue, serum, and abnormal lesions were collected upon necropsy and analyzed by IHC, H&E, and PCR for presence of metastatic lesions arising from CWR22. RESULTS: Herein, we report that CWR22 progresses after castration from a primary, hormonal therapy-naïve tumor to metastatic disease in 20% of castrated nude mice. Histological examination of CWR22 primary tumors revealed distinct pathologies that correlated with metastatic outcome after castration. CONCLUSION: This is the first report and characterization of spontaneous metastasis in the CWR22 model, thus, CWR22 is a bona-fide model of clinical PCa representing the full progression from androgen-sensitive, primary PCa to metastatic CR-PCa.

Characterization of fibroblast-free CWR-R1ca castration-recurrent prostate cancer cell line.

BACKGROUND: The previously established CWR-R1 cell line has been used as an in vitro model representing castration-recurrent prostate cancer. Microscopic observation of subconfluent cells demonstrated two distinct cellular morphologies: polygonal closely aggregated epithelial cells surrounded by bipolar fibroblastic cells with long processes. This study sought to establish and characterize a fibroblast-free derivative of the CWR-R1 cell line. METHODS: The CWR-R1ca cell line was established from CWR-R1 cells by removing fibroblasts using multiple cycles of short-term trypsinization, cloning, and pooling single-cell colonies. Authentication of fibroblast-free CWR-R1ca cells was demonstrated by analyzing the expression of cytodifferentiation and prostate-associated markers, DNA and cytogenetic profiling, and growth pattern in the absence or presence of androgen. RESULTS: CWR-R1ca is an androgen-sensitive cell line that expresses the androgen receptor (AR) and its splice variant 7 and the luminal epithelia markers, CK-8, CK-18, and c-Met. CWR-R1fb fibroblasts isolated from CWR-R1 cells express AR, hepatocyte growth factor-α, and mouse ß-actin but not AR-V7 or epithelial markers. Cytogenetic analysis of CWR-R1ca cells revealed a hyperdiploid male with numerical gains in chromosomes 1, 7, 8, 10, 11, and 12, deletion of one chromosome 2 allele, structural abnormalities that include der(1)t(1:4), der(4)t(2:4), der(10)t(4:10), and an unbalanced reciprocal translocation between chromosome 6 and 14. DNA-profiling revealed that CWR-R1ca cells had significant short-tandem repeat marker homology with CWR22Pc and CWR22Rv1 cell lines, which indicated lineage derivation from CWR22 prostate cancer xenografts. CWR-R1ca cells were responsive to the growth stimulatory effects of dihydrotestosterone (DHT) in the femtomolar range. CONCLUSION: This study establishes CWR-R1ca cells as a fibroblast-free derivative of the castration-recurrent CWR-R1 cell line. Prostate 76:1067-1077, 2016. © 2016 Wiley Periodicals, Inc.

Origin of year-long bean (Phaseolus dumosus Macfady, Fabaceae) from reticulated hybridization events between multiple Phaseolus species.

Background and Aims Improved understanding of the secondary gene pools of crops is essential for advancing genetic gain in breeding programmes. Common bean, Phaseolus vulgaris, is a staple crop with several wild relatives in its secondary gene pool. The year-long bean, P. dumosus, an important crop in Guatemala, is considered particularly closely related to P. vulgaris and a potential source of novel variation. However, the genetic diversity and relationship to other Phaseolus species of P. dumosus remain unclear. Methods We conducted the first comprehensive investigation of P. dumosus genetic diversity using both nuclear and chloroplast genome markers. Our nuclear marker set included over 700 markers present within the Phaseolus DArT (Diversity Arrays Technology) array, which we applied to P. dumosus and other relatives of P. vulgaris (including every secondary gene pool species: P. acutifolius, P. albescens, P. coccineus and P. costaricensis). Key Results Phaseolus dumosus arose from hybridization of P. vulgaris and P. coccineus, followed by at least two later hybridizations with sympatric congener populations. Existing P. dumosus collections have low genetic diversity. Conclusions The under-utilized crop P. dumosus has a complex hybrid origin. Further sampling in the region in which it arose may uncover additional germplasm for introgressing favourable traits into crops within the P. vulgaris gene pool.

Surveying and mapping cereals and legumes wild relatives in Mount Hermon (Bekaa, Lebanon).

Crop Wild Relatives (CWR) should be highly prioritized, monitored, and conserved as they have an immense effect on sustainability and livelihood. In this study we aim to survey and map cereal and legume wild relatives of Fabaceae and Poaceae families. Mount Hermon, Bekaa side, Lebanon. A set of 46 CWR species were targeted based on desk selection analysis and prioritization by the International Center for Agricultural Research in Dry Areas genebank for their potential importance in breeding programs. A botanical survey of 17 sites of the various habitats of Mount Hermon was performed during April-June 2021 using a systematic transect/quadrate sampling method. Recorded genera and species were accurately georeferenced and then mapped with the DIVA-GIS program. In total, 854 occurrences were observed belonging to 34 species of Fabaceae and 12 species of Poaceae. High H' Shannon diversity values were recorded in three sites (Al Fakiaa, Sham El Hafour and Ain Ata- al Berke) of the Mount with values ranking between 2.45 and 2.83. This was confirmed by the richness distribution maps of genera and species. Richness distribution maps provide relevant clues on candidate sites for high concentrations of each of the species under study. At least the three sites, hosting 87% of the surveyed CWR's species, can be considered for further in situ conservation actions.

Enhanced recovery after chest wall resection and reconstruction: a clinical practice review.

Since the late 1990s, and Henrik Kehlet's hypothesis that a reduction of the body's stress response to major surgeries could decrease postoperative morbidity, "Enhanced Recovery After Surgery" (ERAS) care pathways have been streamlined. They are now well accepted and considered standard in many surgical disciplines. Yet, to this day, there is no specific ERAS protocol for chest wall resections (CWRs), the removal of a full-thickness portion of the chest wall, including muscle, bone and possibly skin. This is most unfortunate because these are high-risk surgeries, which carry high morbidity rates. In this review, we propose an overview of the current key elements of the ERAS guidelines for thoracic surgery that might apply to CWRs. A successful ERAS pathway for CWR patients would entail, as is the standard approach, three parts: pre-, peri- and postoperative elements. Preoperative items would include specific information, targeted patient education, involvement of all members of the team, including the plastic surgeons, smoking cessation, dedicated nutrition and carbohydrate loading. Perioperative items would likely be standard for thoracotomy patients, namely carefully selective pre-anesthesia sedative medication only in some rare instances, low-molecular-weight heparin throughout, antibiotic prophylaxis, minimization of postoperative nausea and vomiting, avoidance of fluid overload and of urinary drainage. Postoperative elements would include early mobilization and feeding, swift discontinuation of intravenous fluid supply and chest tube removal as soon as safe. Optimal pain management throughout also appears to be critical to minimize the risk of respiratory complications. Together, all these items are achievable and may hold the key to successful introduction of ERAS pathways to the benefit of CWR patients.

Review of Crop Wild Relative Conservation and Use in West Asia and North Africa.

Ensuring global food security in the face of climate change is critical to human survival. With a predicted human population of 9.6 billion in 2050 and the demand for food supplies expected to increase by 60% globally, but with a parallel potential reduction in crop production for wheat by 6.0%, rice by 3.2%, maize by 7.4%, and soybean by 3.1% by the end of the century, maintaining future food security will be a challenge. One potential solution is new climate-smart varieties created using the breadth of diversity inherent in crop wild relatives (CWRs). Yet CWRs are threatened, with 16-35% regarded as threatened and a significantly higher percentage suffering genetic erosion. Additionally, they are under-conserved, 95% requiring additional ex situ collections and less than 1% being actively conserved in situ; they also often grow naturally in disturbed habitats limiting standard conservation measures. The urgent requirement for active CWR conservation is widely recognized in the global policy context (Convention on Biological Diversity post-2020 Global Biodiversity Framework, UN Sustainable Development Goals, the FAO Second Global Plan of Action for PGRFA, and the FAO Framework for Action on Biodiversity for Food and Agriculture) and breeders highlight that the lack of CWR diversity is unnecessarily limiting crop improvement. CWRs are not spread evenly across the globe; they are focused in hotspots and the hottest region for CWR diversity is in West Asia and North Africa (WANA). The region has about 40% of global priority taxa and the top 17 countries with maximum numbers of CWR taxa per unit area are all in WANA. Therefore, improved CWR active conservation in WANA is not only a regional but a critical global priority. To assist in the achievement of this goal, we will review the following topics for CWRs in the WANA region: (1) conservation status, (2) community-based conservation, (3) threat status, (4) diversity use, (5) CURE-CWR hub: (ICARDA Centre of Excellence), and (6) recommendations for research priorities. The implementation of the recommendations is likely to significantly improve CWRs in situ and ex situ conservation and will potentially at least double the availability of the full breadth of CWR diversity found in WANA to breeders, and so enhance regional and global food and nutritional security.

Thoracic cavity remodeling and pulmonary function change after chest wall resection.

Background: Chest wall resection (CWR) is an essential procedure for treating malignancies and infectious conditions of the chest wall. However, there are few studies on the pulmonary function and changes in thoracic cavity volume (TCV) related to CWR. This study aims to investigate the effects of CWR on long-term changes in TCV and pulmonary function. Methods: Data of patients who underwent CWR between 2001 and 2021 were retrospectively reviewed. Patients who underwent single rib or lung resection rather than wedge resection were excluded. TCV (liter) was defined as the sum of the right and left TCVs (RCV and LCV) and was measured using computed tomography image reconstruction software. Changes in pulmonary function and TCV 1 year postoperatively were analyzed. Results: A total of 45 patients were included. The number of resected ribs was 2 in 16 (35.6%) and ≥3 in 29 (64.4%) patients. Thirty patients underwent reconstruction. Long-term post-CWR decreased in forced vital capacity (FVC) (-7.9%, P=0.004) and forced expiratory volume in 1 second (FEV1) (-7.0%, P=0.002) were significant. There was no significant decrease in FEV1/FVC ratio (-3.0%, P=0.06), diffusing capacity of the lung for carbon monoxide (DLCO) (-5.9%, P=0.18) and TCV (-3.1%, P=0.10). There was no correlation between changes in TCV and decreases in FVC (r=0.12, P=0.56) or FEV1 (r=0.15, P=0.45). After right-side CWR (n=27), RCV (-7.8%, P=0.01) decreased significantly, whereas LCV (+2.1%, P=0.58) did not. The left-side CWR exhibited an identical pattern. (LCV: -8.5%, P=0.004; RCV: +1.3%, P=0.85). In the ≥3 rib-resection group, FVC (-9.5%, P=0.02), FEV1 (-7.9%, P=0.02) and TCV (-6.4%, P=0.04) decreased significantly. No significant changes were noted in the 2 rib-resection group. There were no significant differences in the changes in pulmonary function nor TCV between the reconstruction and no-reconstruction groups. Conclusions: The long-term decrease in pulmonary function after CWR was significant, especially after ≥3-rib resection.

Screening of Comprehensive Panel of Cultivated and Wild Vigna Species for Resistance to Pulse Beetle, Callosobruchus chinensis L.

Pulses are a key source of dietary proteins in human nutrition. Despite several efforts to increase the production, various constraints, such as biotic and abiotic factors, threaten pulse production by various means. Bruchids (Callosobruchus spp.) are the serious issue of concern, particularly in storage conditions. Understanding host-plant resistance at morphological, biochemical and molecular levels is the best way to minimize yield losses. The 117 mungbean (Vigna radiata L. Wilczek) genotypes, including endemic wild relatives, were screened for resistance against Callosobruchus chinensis; among them, two genotypes, PRR 2008-2 and PRR 2008-2-sel, which belong to V. umbellata (Thumb.), were identified as highly resistant. The expression of antioxidants in susceptible and resistant genotypes revealed that the activity of phenylalanine ammonia lyase (PAL) was upregulated in the highly resistant wild Vigna species and lower in the cultivated susceptible genotypes, along with other biomarkers. Further, the SCoT-based genotyping revealed SCoT-30 (200 bp), SCoT-31 (1200 bp) and SCoT-32 (300 bp) as unique amplicons, which might be useful for developing the novel ricebean-based SCAR markers to accelerate the molecular breeding programme.

Solving the mystery of Obake rice in Africa: population structure analyses of Oryza longistaminata reveal three genetic groups and evidence of both recent and ancient introgression with O. sativa.

The undomesticated rice relative Oryza longistaminata is a valuable genetic resource for the improvement of the domesticated Asian rice, Oryza sativa. To facilitate the conservation, management, and use of O. longistaminata germplasm, we sought to quantify the population structure and diversity of this species across its geographic range, which includes most of sub-Saharan Africa, and to determine phylogenetic relationships to other AA-genome species of rice present in Africa, including the prevalence of interspecific hybridization between O. longistaminata and O. sativa. Though past plant breeding efforts to introgress genes from O. longistaminata have improved biotic stress resistance, ratooning ability, and yield in O. sativa, progress has been limited by substantial breeding barriers. Nevertheless, despite the strong breeding barriers observed by plant breeders who have attempted this interspecific cross, there have been multiple reports of spontaneous hybrids of O. sativa and O. longistaminata (aka "Obake") obtained from natural populations in Africa. However, the frequency and extent of such natural introgressions and their effect on the evolution of O. longistaminata had not been previously investigated. We studied 190 O. longistaminata accessions, primarily from the International Rice Research Institute genebank collection, along with 309 O. sativa, 25 Oryza barthii, and 83 Oryza glaberrima control outgroups, and 17 control interspecific O. sativa/O. longistaminata hybrids. We analyzed the materials using 178,651 single-nucleotide polymorphisms (SNPs) and seven plastid microsatellite markers. This study identified three genetic subpopulations of O. longistaminata, which correspond geographically to Northwestern Africa, Pan-Africa, and Southern Africa. We confirmed that O. longistaminata is, perhaps counterintuitively, more closely related to the Asian species, O. sativa, than the African species O. barthii and O. glaberrima. We identified 19 recent spontaneous interspecific hybrid individuals between O. sativa and O. longistaminata in the germplasm sampled. Notably, the recent introgression between O. sativa and O. longistaminata has been bidirectional. Moreover, low levels of O. sativa alleles admixed in many predominantly O. longistaminata accessions suggest that introgression also occurred in the distant past, but only in Southern Africa.

CmWRKY15-1 Promotes Resistance to Chrysanthemum White Rust by Regulating CmNPR1 Expression.

Chrysanthemum white rust (CWR), a disease caused by the fungus Puccinia horiana Henn., seriously impairs the production and ornamental value of chrysanthemums. We previously isolated the disease-resistance gene CmWRKY15-1 from the chrysanthemum and generated CmWRKY15-1 transgenic plants. Here, we determined that CmWRKY15-1-overexpressing lines of the susceptible cultivar 'Jinba' show higher defensive enzyme activity and lower H2O2 levels than a wild type after inoculation with P. horiana, indicating that CmWRKY15-1 positively regulates plant responses to P. horiana. To further explore the mechanism underlying this effect, we performed RNA sequencing using the leaves of wild-type and CmWRKY15-1-RNA interference lines of the resistant cultivar 'C029' after treatment with P. horiana. We identified seven differentially expressed genes in the salicylic acid (SA) pathway, including CmNPR1 (Non-expressor of pathogenesis-related genes 1), encoding an important regulator of this pathway. We isolated the CmNPR1 promoter by hiTAIL-PCR and predicted that it contains pathogen-induced W-box elements. The promoter region of CmNPR1 was activated by P. horiana in a ß-glucuronidase activity assay. Yeast one-hybrid assays showed that CmWRKY15-1 binds to the CmNPR1 promoter region to regulate its expression. Finally, we confirmed the interaction between CmWRKY15-1 and CmNPR1 in a bimolecular fluorescence complementation assay. We propose that CmWRKY15-1 interacts with CmNPR1 to activate the expression of downstream pathogenesis-related genes that enhance resistance to P. horiana through the SA pathway. These findings shed light on the mechanism underlying resistance to CWR.

A review of the indigenous coffee resources of Uganda and their potential for coffee sector sustainability and development.

Uganda is a major global coffee exporter and home to key indigenous (wild) coffee resources. A comprehensive survey of Uganda's wild coffee species was undertaken more than 80 years ago (in 1938) and thus a contemporary evaluation is required, which is provided here. We enumerate four indigenous coffee species for Uganda: Coffea canephora, C. eugenioides, C. liberica (var. dewevrei) and C. neoleroyi. Based on ground point data from various sources, survey of natural forests, and literature reviews we summarise taxonomy, geographical distribution, ecology, conservation, and basic climate characteristics, for each species. Using literature review and farm survey we also provide information on the prior and exiting uses of Uganda's wild coffee resources for coffee production. Three of the indigenous species (excluding C. neoleroyi) represent useful genetic resources for coffee crop development (e.g. via breeding, or selection), including: adaptation to a changing climate, pest and disease resistance, improved agronomic performance, and market differentiation. Indigenous C. canephora has already been pivotal in the establishment and sustainability of the robusta coffee sector in Uganda and worldwide, and has further potential for the development of this crop species. Coffea liberica var. dewevrei (excelsa coffee) is emerging as a commercially viable coffee crop plant in its own right, and may offer substantial potential for lowland coffee farmers, i.e. in robusta coffee growing areas. It may also provide useful stock material for the grafting of robusta and Arabica coffee, and possibly other species. Preliminary conservation assessments indicate that C. liberica var. dewevrei and C. neoleroyi are at risk of extinction at the country-level (Uganda). Adequate protection of Uganda's humid forests, and thus its coffee natural capital, is identified as a conservation priority for Uganda and the coffee sector in general.

In situ occurrence and protection of crop wild relatives in Italian sites of natura 2000 network: Insights from a data-driven approach.

Aim of this work is to evaluate the in situ status of different crop wild relative species in Italy by analysing the geographic distribution of their populations and to suggests possible strategies to improve their future conservation. The work has been focused on different species of the Allium, Beta, Brassica, Secale and Triticum genera that are of priority at European and global levels for the economic importance of the related crops, the level of threat, as well as the potential for use. Using information available in the Italian National Geoportal, geographical distribution and the overall percentage of populations occurring in Natura 2000 sites was initially analysed. In addition, due to the economic importance of the genus and species distribution in Italy, Brassica glabrescens, B. insularis, B. macrocarpa, B. montana, B. procumbens, B. rupestris, B. villosa were the object of additional analyses based on more detailed occurrence data, retrieved from multiple databases, and including land cover/land use and in situ and ex situ density analyses. Geographical distribution data were retrieved for 1,996 in situ populations belonging to 60 crop wild relative species: Allium (43), Brassica (11), Triticum (4), Beta (1) and Secale (1). Percentages of population occurring in Natura 2000 sites are quite different when the different species are considered; this also applies to Brassica species in most need of protection. Results of land cover/land use analysis showed that Brassica populations outside Natura 2000 areas mainly occur in anthropized sites while those within Natura 2000 mainly in sites characterised by natural and seminatural conditions. Areas where genetic reserves could be instituted and that could be the target of future Brassica resources collection missions are also suggested. Based on a large dataset of punctual geographical distribution data of population occurrences across the territory, this research shows that, in Italy, crop wild relatives in situ are in a quite precarious condition especially when species in most need of protection are considered. Our data also highlight the role of Natura 2000 Network in favouring in situ protection of these precious resources in Europe.

Chest wall reconstruction with implantable cross-linked porcine dermal collagen matrix: Evaluation of clinical outcomes.

Objectives: The aim of the study is to evaluate clinical applications, safety, and effectiveness of a porcine-derived acellular cross-linked dermal matrix biological mesh in chest wall reconstruction. Methods: We retrospectively analyzed a prospective multicenter database of chest wall reconstructions using a biological mesh in adult patients undergoing operation between October 2013 and December 2020. We evaluated preoperative data, type of resection and reconstruction, hospitalization, 30-day morbidity and mortality, and overall survival. Results: A total of 105 patients (36 women [34.2%]; mean age, 57.0 ± 16.1 years; range, 18-90 years) were included, they have admitted for: primary chest wall tumor (n = 52; 49.5%), secondary chest wall tumor (n = 29; 27.6%), lung hernia (n = 12; 11.4%), trauma (n = 10; 9.6%), and infections (n = 2; 1.9%). The surgical sites were preoperatively defined as at high risk of infection in 28 patients (26.7%) or as infected in 16 (15.2%) patients. Thirty-days morbidity was 30.5% (n = 32 patients); 14 patients (13.3%) had postoperative complications directly related to chest wall surgical resection and/or reconstruction. We experienced no 30-day mortality; 1-year and 2-year mortality was 8.4% and 16.8%, respectively. Conclusions: Biological mesh represents a valuable option in chest wall reconstruction even when surgical sites are infected or at high-risk of infections. This mesh shows low early and late postoperative complication rates and excellent long-term stability.

Engineering sorghum for higher 4-hydroxybenzoic acid content.

Engineering bioenergy crops to accumulate coproducts in planta can increase the value of lignocellulosic biomass and enable a sustainable bioeconomy. In this study, we engineered sorghum with a bacterial gene encoding a chorismate pyruvate-lyase (ubiC) to reroute the plastidial pool of chorismate from the shikimate pathway into the valuable compound 4-hydroxybenzoic acid (4-HBA). A gene encoding a feedback-resistant version of 3-deoxy-d-arabino-heptulonate-7-phosphate synthase (aroG) was also introduced in an attempt to increase the carbon flux through the shikimate pathway. At the full maturity and senesced stage, two independent lines that co-express ubiC and aroG produced 1.5 and 1.7 dw% of 4-HBA in biomass, which represents 36- and 40-fold increases compared to the titer measured in wildtype. The two transgenic lines showed no obvious phenotypes, growth defects, nor alteration of cell wall polysaccharide content when cultivated under controlled conditions. In the field, when harvested before grain maturity, transgenic lines contained 0.8 and 1.2 dw% of 4-HBA, which represent economically relevant titers based on recent technoeconomic analysis. Only a slight reduction (11-15%) in biomass yield was observed in transgenics grown under natural environment. This work provides the first metabolic engineering steps toward 4-HBA overproduction in the bioenergy crop sorghum to improve the economics of biorefineries by accumulating a value-added coproduct that can be recovered from biomass and provide an additional revenue stream.

Crop Adaptation: Weedy and Crop Wild Relatives as an Untapped Resource to Utilize Recent Increases in Atmospheric CO2.

Adaptation measures are necessary to ensure the stability and performance of the food supply relative to anthropogenic climate change. Although a wide range of measures have been proposed (e.g., planting dates, crop choices, drought resistance), there may be a ubiquitous means to increase productivity relatively quickly. Numerous studies have shown that the projected increase in atmospheric CO2 can stimulate crop growth and seed yield with noted intra-specific differences within crop cultivars, suggesting potential differences to CO2 that could be exploited to enhance seed yield in the future. However, it is worth emphasizing that atmospheric CO2 has already risen substantially (≈27% since 1970) and that, at present, no active effort by breeders has been made to select for the CO2 increase that has already occurred. In contrast, for weedy or crop wild relatives (CWR), there are indications of evolutionary adaptation to these recent increases. While additional steps are needed, the identification and introgression of these CO2-sensitive traits into modern crop cultivars may be a simple and direct means to increase crop growth and seed yield.