Kriopreservasi Tanaman Purwoceng {Pimpinella Pruatjan Molk.) Dengan Teknik Vitrifikasi [Cryopreservation of Pruatjan {Pimpinella Pruatjan Molk.) by Vitrification Technique]

Pruatjan (Pimpinellapruatjan Molk.) is an Indonesian endangered medicinal plant that included in Appendix I based on CITES. Therefore it is a highly protected species. To avoid extinction of this plant, it is very important to conserve the plant. In vitro conservation is more suitable since this plant is difficult to be cultivated outside of its habitat. Cryopreservation technique may conserve this material for a long-term period. The objectives of this research were to find optimized treatments for pre culture, loading, and dehydration on cryopreservation of pruatjan. The research was conducted at Tissue Culture Laboratory in Indonesian Center for Agricultural Biotechnology and Genetic Resources Research and Development, started from May to November 2007. Pre culture was conducted using DKW basal media that added by sucrose at the level of 0.3,0.4, and 0.5M for one and three days incubation. Loading was conducted in DKW basal media containing 2M glycerol and 0.4M sucrose for 15,30, and 45 minutes duration time. Dehydration was conducted in several cryoprotectants, namely PVS1 (22% glycerol + 13% propy lene glycol + 13% etylene glycol + 6% DMSO + 3% sucrose), PVS2 (30% glycerol + 15% etylene glycol + 15% DMSO + 0,4M sucrose), PVS3 (50% glycerol + 50% sucrose), and PVS4 (35% glycerol + 20% etylene glycol + sucrose 0.6M). Result showed that pruatjan could be preserved through cryopreservation by vitrification method. The best pre culture was using 0.3 M sucrose for one day, the best loading was 30 minutes, while the best cryoprotectant was PVS2 with 90% success before freezing and 40% after freezing. The success may be improved by applying pre growth treatment, optimizing temperature of thawing, modification of recovery media and incubation condition

Karakter Anatomi Daun Kultur Purwoceng Pascakonservasi in Vitro

Leaf Anatomy Characteristic of Pruatjan Cultures Post In Vitro. Evaluation of anatomical characteristics regenerant planlet of purwoceng (Pimpinella pruatjan Molk.) cultivated on conservation medium was observed to determine the difference of characteristics performance of the planlet grown on conservation medium and those that was maintained on the normal medium. Stomata was microscopically observed on abaxial leaf paradermal section that preparated by whole mount method and leaf structure on cross section by paraffin method. The result showed that stomata density was greater on the planlets regenerated in the conservation medium but stomata length was lower than those on the normal medium. Upper epidermis, mesofil and lower epidermis length of the regenerant on the conservation medium were lower than those on the normal medium. Sorbitol and paclobutrazol applied reduced the performance of regenerated spesies than those of planlet maintained on the normal medium. Combination of both application resulted in anatomical character differences on the plants regenerated on normal medium

KRIOPRESERVASI TANAMAN PURWOCENG {Pimpinella pruatjan Molk.) DENGAN TEKNIK VITRIFIKASI

Pruatjan (Pimpinellapruatjan Molk.) is an Indonesian endangered medicinal plant that included in Appendix I based on CITES. Therefore it is a highly protected species. To avoid extinction of this plant, it is very important to conserve the plant. In vitro conservation is more suitable since this plant is difficult to be cultivated outside of its habitat. Cryopreservation technique may conserve this material for a long-term period. The objectives of this research were to find optimized treatments for pre culture, loading, and dehydration on cryopreservation of pruatjan. The research was conducted at Tissue Culture Laboratory in Indonesian Center for Agricultural Biotechnology and Genetic Resources Research and Development, started from May to November 2007. Pre culture was conducted using DKW basal media that added by sucrose at the level of 0.3,0.4, and 0.5M for one and three days incubation. Loading was conducted in DKW basal media containing 2M glycerol and 0.4M sucrose for 15,30, and 45 minutes duration time. Dehydration was conducted in several cryoprotectants, namely PVS1 (22% glycerol + 13% propy lene glycol + 13% etylene glycol + 6% DMSO + 3% sucrose), PVS2 (30% glycerol + 15% etylene glycol + 15% DMSO + 0,4M sucrose), PVS3 (50% glycerol + 50% sucrose), and PVS4 (35% glycerol + 20% etylene glycol + sucrose 0.6M). Result showed that pruatjan could be preserved through cryopreservation by vitrification method. The best pre culture was using 0.3 M sucrose for one day, the best loading was 30 minutes, while the best cryoprotectant was PVS2 with 90% success before freezing and 40% after freezing. The success may be improved by applying pre growth treatment, optimizing temperature of thawing, modification of recovery media and incubation condition

Click Chemistry with Polymers, Dendrimers, and Hydrogels for Drug Delivery

This is a post-peer-review, pre-copyedit version of an article published in Pharmaceutical Research. The final authenticated version is available online at: https://doi.org/10.1007/s11095-012-0683-yDuring the last decades, great efforts have been devoted to design polymers for reducing the toxicity, increasing the absorption, and improving the release profile of drugs. Advantage has been also taken from the inherent multivalency of polymers and dendrimers for the incorporation of diverse functional molecules of interest in targeting and diagnosis. In addition, polymeric hydrogels with the ability to encapsulate drugs and cells have been developed for drug delivery and tissue engineering applications. In the long road to this successful story, pharmaceutical sciences have been accompanied by parallel advances in synthetic methodologies allowing the preparation of precise polymeric materials with enhanced properties. In this context, the introduction of the click concept by Sharpless and coworkers in 2001 focusing the attention on modularity and orthogonality has greatly benefited polymer synthesis, an area where reaction efficiency and product purity are significantly challenged. The purpose of this Expert Review is to discuss the impact of click chemistry in the preparation and functionalization of polymers, dendrimers, and hydrogels of interest in drug deliveryThis work was financially supported by the Spanish Ministry of Science and Innovation (CTQ2009-10963 and CTQ2009-14146-C02-02) and the Xunta de Galicia (10CSA209021PR and CN2011/037)S

Functional gallic acid-based dendrimers as synthetic nanotools to remodel amyloid-β-42 into noncytotoxic forms

The self-assembly of amyloid-β (Aβ) generates cytotoxic oligomers linked to the onset and progression of Alzheimer’s disease (AD). As many fundamental molecular pathways that control Aβ aggregation are yet to be unraveled, an important strategy to control Aβ cytotoxicity is the development of bioactive synthetic nanotools capable of interacting with the heterogeneous ensemble of Aβ species and remodel them into noncytotoxic forms. Herein, the synthesis of nanosized, functional gallic acid (Ga)-based dendrimers with a precise number of Ga at their surface is described. It is shown that these Ga-terminated dendrimers interact by H-bonding with monomeric/oligomeric Aβ species at their Glu, Ala, and Asp residues, promoting their remodeling into noncytotoxic aggregates in a process controlled by the Ga units. The multivalent presentation of Ga on the dendrimer surface enhances their ability to interact with Aβ, inhibiting the primary and secondary nucleation of Aβ fibrillization and disrupting the Aβ preformed fibrils.The authors acknowledge the financial support from the EC (FORECAST-668983), “Programa Operacional Regional do Norte”, “Fundo Social Europeu”, Norte2020 TERM&SC, for the PhD grant NORTE-08-5369-FSE-000044, the Spanish Ministry of Science and Innovation (RTI2018-102212-B-I00), the Xunta de Galicia (ED431C 2018/30; Centro singular de investigación de Galicia accreditation 2019−2022, ED431G 2019/03), European Regional Development Fund-ERDF, and the Galician Supercomputing Centre (CESGA) and the MAT2016-80266-R of the Spanish Ministry of Science and Innovation

Fibroblast Activation Protein specific optical imaging in Non-Small Cell Lung Cancer

Fibroblast activation protein (FAP) is a cell surface propyl-specific serine protease involved in the regulation of extracellular matrix. Whilst expressed at low levels in healthy tissue, upregulation of FAP on fibroblasts can be found in several solid organ malignancies, including non-small cell lung cancer, and chronic inflammatory conditions such as pulmonary fibrosis and rheumatoid arthritis. Their full role remains unclear, but FAP expressing cancer associated fibroblasts (CAFs) have been found to relate to a poor prognosis with worse survival rates in breast, colorectal, pancreatic, and non-small cell lung cancer (NSCLC). Optical imaging using a FAP specific chemical probe, when combined with clinically compatible imaging systems, can provide a readout of FAP activity which could allow disease monitoring, prognostication and potentially stratify therapy. However, to derive a specific signal for FAP any sequence must retain specificity over closely related endopeptidases, such as prolyl endopeptidase (PREP), and be resistant to degradation in areas of active inflammation. We describe the iterative development of a FAP optical reporter sequence which retains FAP specificity, confers resistance to degradation in the presence of activated neutrophil proteases and demonstrates clinical tractability ex vivo in NSCLC samples with an imaging platform

Functional responses in a lizard along a 3.5-km altitudinal gradient

Aim: Physiological and metabolic performance are key mediators of the functional response of species to environmental change. Few environments offer such a multifaceted array of stressors as high-altitude habitats, which differ markedly in temperature, water availability, UV radiation and oxygen pressure compared to low-altitude habitats. Species that inhabit large altitudinal gradients are thus excellent models to study how organisms respond to environmental variation. Location: Tenerife island, Canary Islands archipelago (Spain). Taxon: Tenerife lizard (Gallotia galloti, Lacertidae). Methods: We integrated data on age structure, thermal and hydric regulatory behaviour and four metabolic and stress-related biomarkers for an insular lizard that inhabits an extreme altitudinal range (sea level to 3700 m a.s.l.), to understand how an ectotherms' age, ecophysiology and metabolism can be affected by extreme environmental variation. Results: We found marked differences in metabolic stress markers associated with altitude (particularly in the abundance of carbonyl metabolites and relative telomere length), but without a linear pattern along the altitudinal cline. Contrary to expectations, longer telomeres and lower carbonyl content were detected at the highest altitude, suggesting reduced stress in these populations. Evaporative water loss differed between populations but did not follow a linear altitudinal gradient. Lizard age structure or thermal physiological performance did not markedly change across different altitudes. Mixed signals in life-history and thermal ecology across populations and altitude suggest complex responses to variable conditions across altitude in this species. Main Conclusions: Our integrative study of multiple functional traits demonstrated that adaptation to highly divergent environmental conditions in this lizard is potentially linked to an interplay between plasticity and local adaptation variably associated with different functional traits