Effects of Confluent, Roscovitine Treatment and Serum Starvation on the Cell-cycle Synchronization of Bovine Foetal Fibroblasts

The present study was designed to examine the effects of cell-cycle synchronization protocols, such as confluent, roscovitine treatment and serum starvation, in bovine foetal fibroblasts on synchronization accuracy at G0/G1, viability, apoptosis, necrosis and ploidy for use as a nuclei donor. The cells in 5-10 passages were randomly allocated into three treated groups. Cells were cultured either in Dulbecco's modified Eagle's medium (DMEM) + 10% foetal bovine serum (FBS) until 90% confluent (group 1, confluent), in DMEM + 10% FBS + 30 microM roscovitine for 12 h (group 2, roscovitine), or in DMEM + 0.5% FBS for 5 days (group 3, serum starvation). Most of the cells (>80%) in all groups were arrested at the G0/G1 stage. Although the rates did not differ, cells in group 1 showed an increased cell population arrested at the G0/G1 phase. Significantly (p < 0.05) higher rates of apoptosis occurred in group 3 than in group 1 and 2 (10% vs 6% and 6%, respectively). No differences in chromosomal abnormality were observed among groups. However, by increasing the number of cell culture passages up to 15, significantly (p < 0.05) higher chromosomal abnormality was observed than in 5 and 10 passages (39% vs 28% and 23%, respectively) in group 1. The results clearly indicated that bovine foetal fibroblasts could be effectively synchronized at G0/G1 stages by all the three different treatments, confluent, roscovitine and serum starvation. However, cells in confluent showed reduced apoptosis and necrosis when they underwent 5-10 passages, exhibiting increased percentage of cells with stable chromosome diversity. Hence, cells in confluent merit further studies before they could be used as nuclear donors.     

Screening for New Cosmeceuticals from Brown Algae Fucus vesiculosus with Antioxidant and Photo-Protecting Properties

Phlorotannins play a role in biological functions to protect the cells against UV and oxidative damage in brown algae. We hypothesized that these compounds can function as photo-protectors and antioxidants in skin care formulations. Two types of extracts (water (FV-WE) and 67% v/v ethanol (FV-EE)) from Fucus vesiculosus were obtained with a phlorotannin content between 7−14% in dry extract. Exposure to sun light during growth was included as a factor on the phlorotannin content but did not influence the phlorotannin content. However, green colored F. vesiculosus had lower total phenolic content (TPC) (FV-WE = 6.9 g GAE 100 g⁻¹ dw, FV-EE = 7.8 g GAE 100 g⁻¹ dw) compared to those with a yellow/brownish color (FV-WE = 10.4–13.7 g GAE 100 g⁻¹ dw, FV-EE = 11.2–14.0 g GAE 100 g⁻¹ dw). UVA and UVB photo protective capabilities of the extracts through different biological effective protection factors (BEPFs) were evaluated using in vitro methods; the Mansur method for sun protection factor (SPF) and calculation of effective solar absorption radiation (%ESAR) to determine SPF and UVA protection factor (UVA-PF) of the extract and in seaweed enriched lotion. The SPF was negligible, when evaluating FV-WE in lotion (10 and 20% w/w). Moreover, %ESAR of the FV-WE showed SPF and some UVA-PF, but not enough to give sufficient SPF in lotions (10% w/w). It was concluded that the concentration of UV protecting compounds in the extracts was too low to and that further fractionation and purification of phlorotannins is needed to increase the SPF.     

Effects of huanglongbing on fruit quality of sweet orange cultivars in Brazil

Citrus huanglongbing (HLB), associated with Candidatus Liberibacter asiaticus and Ca. L. americanus and transmitted by the Asian psyllid Diaphorina citri, is the most serious disease of citrus worldwide because of crop devastation and difficulty to control. Since 2004, approximately 3 million trees were eliminated in attempts to limit its spread in Brazil. Where HLB becomes endemic, the disease progression in the orchard and the increasing symptom severity throughout the tree canopy can be relatively fast, greatly reducing the economic life of affected orchards because of tree decline and yield reduction. Although the majority of the fruit from symptomatic branches drop before harvest, a significant amount of affected fruit remain attached, are available for harvest, and can affect juice quality. To quantify and compare the effects of HLB on fruit quality of the most important sweet orange cultivars grown in São Paulo State, 4–6 year-old sweet orange trees from 26 blocks (two of ‘Valencia Americana’, eight of ‘Hamlin’, four of ‘Westin’, seven of ‘Pera’, and five of ‘Valencia’) were selected prior to harvest. In each block, 14–21 HLB-symptomatic trees were chosen. In each tree, the quality of 20 fruit normal in appearance from asymptomatic branches and 20 symptomatic fruit from symptomatic branches were assessed. In general, compared to normal fruit, the symptomatic fruit were small, light, more acidic, and had lower juice percentage, Brix, total soluble solids per box, total soluble solids per fruit, and Brix/acidity ratio. These effects of fruit quality were less pronounced on early and mid season sweet orange cultivars than on late season cv. Valencia.     

Physiological and metabolic properties of a digestion-resistant maltodextrin, classified as type 3 retrograded resistant starch

There is a growing interest in highly fermentable dietary fibers having the potential to reduce risks of disease through the production of short-chain fatty acids (SCFA). Recently a digestion-resistant retrograded maltodextrin (RRM), classified as type 3 resistant starch was developed. Systematic work to determine its molecular and physiological properties was carried out to determine (1) the fraction resistant to digestion in vitro and in vivo, (2) its postconsumption effect on blood glucose in healthy volunteers, and (3) its in vitro fermentation pattern, at different ages, by use of pooled fresh human fecal inoculum. RESULTS: The digestion resistant fraction obtained in vivo from ileostomy patients (59.4%) is similar to that obtained by the AOAC method for measuring retrograded resistant starch (59.7%). The relative glycemic response after consumption of 50 g of RRM was 58.5% compared to glucose set as 100%. When exposed to colonic microbiota, in vitro obtained indigestible fractions behave similarly to those obtained in vivo in ileostomy patients. Fermentation of RRM and production of butyric acid is negligible during the first months of life but develops subsequently during weaning. In adults, RRM fermentation results in a high yield of SCFA, with butyrate representing 21-31 mol % of total SCFA. The high yield of SCFA during colonic fermentation, observed from weaning age on, as well as the potential to help reduce glycemic load may be of benefit to a number of health-related functions in the host. Further study on clear clinical end points is warranted.     

Isolation and characterization of a Mn(II)-oxidizing Bacillus strain from the demosponge Suberites domuncula

In this study we demonstrate that the demosponge Suberites domuncula harbors a Mn(II)-oxidizing bacterium, a Bacillus strain, termed BAC-SubDo-03. Our studies showed that Mn(II) stimulates bacterial growth and induces sporulation. Moreover, we show that these bacteria immobilize manganese on their cell surface. Comparison of the 16S rDNA sequence allowed the grouping of BAC-SubDo-03 to the Mn-precipitating bacteria. Analysis of the spore cell wall revealed that it contains an Mn(II)-oxidizing enzyme. Co-incubation studies of BAC-SubDo-03 with 100 μM MnCl(2) and >1 μM of CuCl(2) showed an increase in their Mn(II)-oxidizing capacity. In order to prove that a multicopper oxidase-like enzyme(s) (MCO) exists in the cell wall of the S. domuncula-associated BAC-SubDo-03 Bacillus strain, the gene encoding this enzyme was cloned (mnxG-SubDo-03). Sequence alignment of the deduced MCO protein (MnxG-SubDo-03) revealed that the sponge bacterium clusters together with known Mn(II)-oxidizing bacteria. The expression of the mnxG-SubDo-03 gene is under strong control of extracellular Mn(II). Based on these findings, we assume that BAC-SubDo-03 might serve as a Mn reserve in the sponge providing the animal with the capacity to detoxify Mn in the environment. Applying the in vitro primmorph cell culture system we could demonstrate that sponge cells, that were co-incubated with BAC-SubDo-03 in the presence of Mn(II), show an increased proliferation potential.     

城市微型公共空间景观设计

随着城市的发展,城市公共空间对城市生活的重要性不断凸显,城市微型公共空间作为其层次结构中不可或缺的一环,在世界范围内也得到了长足发展。但对于我国城市微型公共空间景观设计领域而言,却一直未能得到足够的重视。以深圳市罗湖区嘉宾公园为研究对象,阐述城市微型公共景观设计对于城市公共空间层次结构的重要意义,并展现其为城市所带来的独特魅力。深圳市罗胡区嘉实公园的建成为我国城市微型公共空间提供了一个极佳的参考样本。     

Updated Trends on the Biodiscovery of New Marine Natural Products from Invertebrates

From 1990–2019, a total of 15,442 New Marine Natural Products from Invertebrates (NMNPIs) were reported. The 2010s saw the most prolific decade of biodiscovery, with 5630 NMNPIs recorded. The phyla that contributed most biomolecules were the Porifera (sponges) (47.2%, 2659 NMNPIs) and the Cnidaria (35.3%, 1989 NMNPIs). The prevalence of these two phyla as the main sources of NMNPIs became more pronounced in the 2010s. The tropical areas of the Pacific Ocean yielded more NMNPIs, most likely due to the remarkable biodiversity of coral reefs. The Indo-Burma biodiversity hotspot (BH) was the most relevant area for the biodiscovery of NMNPIs in the 2010s, accounting for nearly one-third (1819 NMNPIs) of the total and surpassing the top BH from the 1990s and the 2000s (the Sea of Japan and the Caribbean Islands, respectively). The Chinese exclusive economic zone (EEZ) alone contributed nearly one-quarter (24.7%) of all NMNPIs recorded during the 2010s, displacing Japan’s leading role from the 1990s and the 2000s. With the biodiscovery of these biomolecules steadily decreasing since 2012, it is uncertain whether this decline has been caused by lower bioprospecting efforts or the potential exhaustion of chemodiversity from traditional marine invertebrate sources.     

Socio-Economic and Scientific Impact Created by Whitefly-Transmitted,Plant-Virus Disease Resistant Tomato Varieties in Southern India

Research carried out to assess the impact of open-pollinated Tomato leaf curl virus (ToLCV)-resistant tomatoes and hybrids on the livelihoods of resource-poor farmers in Southern India is described and discussed. Three high-yielding ToLCV-resistant tomato varieties were developed initially using conventional breeding and screening techniques involving inoculation by ToLCV-viruliferous whitefly, Bemisia tabaci. In 2003 and 2004, respectively, these varieties were released officially by the Karnataka State Seed Committee and the Indian Ministry of Agriculture through notification in the Gazette of India. From 2003 to 2005, eleven seed companies bought breeder seed of the ToLCV-resistant varieties and used them to begin breeding F₁ hybrids from them. Socio-economic studies carried out to assess the benefits obtained from growing the ToLCV-resistant varieties found that farmers could gain up to 10 times the profit by growing the ToLCV-resistant varieties compared to the pre-existing ToLCV-susceptible varieties. Adoption of ToLCV-resistant tomatoes was also associated with reduced pesticide use. Extra income from tomato sales was prioritised by farmers to pay for children's education, better nutrition and medicines. In a joint effort with the commercial seed sector in India, a promotional field day was organised in 2007. As well as the three ToLCV-resistant varieties, 62 ToLCV-resistant hybrid tomatoes were exhibited during a farmer-field day by 17 commercial seed companies and several public institutes. Tomatoes with ToLCV-resistance are now grown widely in South India and seeds of the three open-pollinated varieties have been distributed to more than 12 countries. In 2007, a conservative estimate of the financial-benefit to cost of the research ratio was already more than 837:1.     

DRIS norms and limiting nutrients in banana cultivation in the South of Ecuador

Abstract

Foliar analysis is an effective method to diagnose the nutritional status of plants. However, the mineral concentration in foliar tissue has traditionally been evaluated by assessing the activity of each element, without considering the interactions between them. To address this, dual interactions were calculated using the Diagnosis and Recommendation Integrated System (DRIS) to identify which crop nutrients are most influential in nutrient imbalances and which are the most limiting nutrients for the nutritional status of banana crops in Ecuador. To achieve this, a regional survey of the nutritional status and its productivity levels was conducted for 188 different sites during the crop season in 2017–2018, involving banana cultivars ‘Vallery’ and ‘Williams’, from the Cavendish subgroup. The DRIS calculation method was combined with Beaufils and Jones functions. From the initial 188 foliar samples, 83 samples (representing 44% of the population) were considered to represent the high-yield reference population, with yields of 38–60 t ha^?1. The DRIS method defined the mean nutritional balance index, which was not found to be statistically correlated (p?>?0.05) with productivity, revealing that there was no significant association with the nutritional status of the plants. Specific DRIS norms were obtained and indicated that deficiencies in K, N, Ca, and Fe, and excesses in Mn, B, Cl, Zn, S, Cu, and Mg were the most limiting nutrients for banana cultivars in the south of Ecuador.