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Progress on the study of the molecular and cellular biology of septins would be greatly accelerated by the development of small molecules that directly inhibit higher-order septin assembly in vivo. By comparison, molecules like latrunculin, paclitaxil, benomyl, etc. allow researchers to acutely perturb the actin or tubulin cytoskeletal networks. Two small molecules, forchlorfenuron (FCF; N-(2-chloro-4pyridyl)-N-phenylurea) and 1-ethyl-3-(4-methoxyphenyl)-6-methylpyrimido[5,4-e][1,2,4]triazine-5,7-dione (PubChem CID 906558), have documented effects on septin localization and/or function, although for each molecule there is also strong evidence for off-target effects. In this chapter we provide a summary of ways to utilize FCF to alter higher-order septin assembly properties in living cells., (Copyright © 2016 Elsevier Inc. All rights reserved.)

Identification and evaluation of safety of forchlorfenuron ((1-(2-chloro-4-pyridyl)-3-phenylurea)), 1, metabolites after biotransformation in kiwifruit is the objective of this study. To elucidate properties of these metabolites, liquid chromatography hybrid ion trap time-of-flight mass spectrometry (LC-IT-TOF-MS) was applied, with MetID Solution and Formula Predictor Software in positive mode. Cytotoxicity of forchlorfenuron and its metabolites were tested through sulforhodamine B assays against normal Chinese hamster ovary cells (CHO). As deduced from characteristic fragment ions of forchlorfenuron, then confirmed by comparison with synthetic standards, as well as characterized by NMR and mass spectrometry techniques, results indicate the presence of 4-hydroxyphenyl-forchlorfenuron, 2, 3-hydroxyphenyl-forchlorfenuron, 3, and forchlorfenuron-4-O-β-D-glucoside, 5. Forchlorfenuron (IC50 = 12.12 ± 2.14 μM) and 4-hydroxyphenyl-forchlorfenuron (IC50 = 36.15 ± 1.59 μM), exhibits significant cytotoxicity against CHO, while 3-hydroxyphenyl-forchlorfenuron and forchlorfenuron-4-O-β-D-glucoside show no cytotoxicity.

An enzymatic procedure based on a catalase biosensor for the detection of forchlorfenuron (CPPU) has been reported in this work. Catalase was immobilized on boron nitride (BN) sheets dispersed in chitosan by adsorption. The immobilized catalase exhibited direct electron transfer character and excellent electrocatalytic activity towards H2O2 reduction. After introducing CPPU into the H2O2 containing phosphate buffer solution, the catalase-catalyzed H2O2 reduction current decreased. By measuring the current decrease, CPPU can be determined in the range of 0.5-10.0 µM with the detection limit of 0.07 μM. The non-competitive inhibition behavior of CPPU towards catalase was verified by the Lineweaver-Burk plots. Long stability character has been ascribed to this biosensor. Possible use of this biosensor in flow systems is illustrated. The proposed biosensor has been successfully applied to CPPU determination in fruits samples with satisfactory results., (Copyright © 2014 Elsevier B.V. All rights reserved.)

In 2008, in Central Italy, a low dosage of CPPU solution, 4 μL L(-1) (6 hL/ha), was sprayed on the canopy of vines of 'Hayward' kiwifruit, at the "break of sepals", about one week before anthesis, to study its effects on fruit weight/size and on qualitative and nutritional characteristics. At harvest, CPPU, with respect to control, significantly increased the fresh weight by about 12% (+12.6 g fruit(-1)) and consequently the yield per vine, without affecting fruit shape, firmness, dry matter (%), total soluble solids, glucose, fructose, sucrose, starch, citrate, malate, vitamin C and soluble and insoluble oxalic acid. After 3 months of storage, CPPU-treated kiwifruits and the control fruit showed no difference in dry matter content, fruit firmness and total soluble solids. The results indicate that a low dosage of CPPU applied in pre-anthesis can improve fruit weight/size without any negative effect on fruit qualitative and nutritional characteristics., (Copyright © 2014 Elsevier Ltd. All rights reserved.)

Objective: To develop a protocol for breaking of seed dormancy and increasing the seed germination rate of Bunium persicum., Methods: The seeds were treated with 3.1, 6.3, 12.5, 25, 50 and 100 µmol/L of benzyl aminopurine, gibberellic acid (GA3), thidiazuron (TDZ) and forchlorfenuron. Then, seeds were transferred to two different temperature conditions including room temperature (25 °C) and chilling temperature (2-5 °C)., Results: The treatment of moist seeds with chilling temperature (2-5 °C) broke seed dormancy and showed maximum germination, which was 54.7% after 60 d treatment. Also, the treatment of dry seeds with chilling temperature broke seed dormancy with 9.3% germination rate after 120 d. Treatment of seeds with different level of plant growth regulators showed that under moist-room condition, there was evidence of higher and lower seed germination rate: GA3 (100 µmol/L) with 46.7% and TDZ (50 µmol/L) with 6.67% respectively. In addition, the results showed that under moist-chilling condition, TDZ (6.3 µmol/L) with 53.3% seed germination rate had higher influence on breaking seed dormancy. Treatment of seeds with combination of TDZ and GA3 under moist-chilling condition revealed higher rate of breaking of seed dormancy when 6.3 µmol/L TDZ was combined with 100 µmol/L GA3, showing 93.7% germination rate., Conclusions: The effect of plant growth regulators coupled with chilling temperature on breaking of seed dormancy could provide a large number of seedlings while the long juvenile time which is the next restricting factor of plantation still remained. Thus, the subsequent growth of seedlings to provide a large number of corms is necessary for successful plantation.

Septins are guanosine-5'-triphosphate-binding proteins involved in wide-ranging cellular processes including cytokinesis, vesicle trafficking, membrane remodelling and scaffolds, and with diverse binding partners. Precise roles for these structural proteins in most processes often remain elusive. Identification of small molecules that inhibit septins could aid in elucidating the functions of septins and has become increasingly important, including the description of roles for septins in pathogenic phenomena such as tumorigenesis. The plant growth regulator forchlorfenuron, a synthetic cytokinin known to inhibit septin dynamics, likely represents an informative probe for septin function. This report deals with septins of the human blood fluke Schistosoma mansoni and their interactions with forchlorfenuron. Recombinant forms of three schistosome septins, SmSEPT5, SmSEPT7.2 and SmSEPT10, interacted with forchlorfenuron, leading to rapid polymerization of filaments. Culturing developmental stages (miracidia, cercariae, adult males) of schistosomes in FCF at 50-500 μM rapidly led to paralysis, which was reversible upon removal of the cytokinin. The reversible paralysis was concentration-, time- and developmental stage-dependent. Effects of forchlorfenuron on the cultured schistosomes were monitored by video and/or by an xCELLigence-based assay of motility, which quantified the effect of forchlorfenuron on fluke motility. The findings implicated a mechanism targeting a molecular system controlling movement in these developmental stages: a direct effect on muscle contraction due to septin stabilization might be responsible for the reversible paralysis, since enrichment of septins has been described within the muscles of schistosomes. This study revealed the reversible effect of forchlorfenuron on both schistosome motility and its striking impact in hastening polymerization of septins. These novel findings suggested routes to elucidate roles for septins in this pathogen, and exploitation of derivatives of forchlorfenuron for anti-schistosomal drugs., (Copyright © 2014 Australian Society for Parasitology Inc. Published by Elsevier Ltd. All rights reserved.)

The paper presents a novel approach for the determination of three cytokinin compounds, thidiazuron (TDZ), 1,3-diphenylurea (1,3-DPU) and forchlorfenuron (CPPU), in fruit and vegetables samples using liquid chromatography with electrospray ionization and time-of-flight mass spectrometry (LC-ESI-TOFMS). Analytes were extracted from the sample matrix with ethanol, and the extract, after dilution with water, was submitted to dispersive liquid-liquid microextraction (DLLME). Once acetonitrile and 1,2-dichloroethane had been selected as extraction and disperser solvents, respectively, the influence of the following experimental parameters was studied using a Plackett-Burman design: volume of extraction and disperser solvents, sample mass and time and speed of centrifugation. The best analytical conditions were 250 µL 1,2-dichloroethane, 1.5 mL acetonitrile, 5 g sample mass, and centrifugation at 3000 rpm for 3 min. The optimized method provided DLs in the range 0.02-0.05 ng g(-1), depending on the compound. Satisfactory recovery values between 89 and 106% were obtained for spiked samples (kiwifruit, watermelon, grape and tomato) in the 0.2-1.0 ng g(-1) concentration range, depending on the compound. None of the target analytes was detected in any of the samples analyzed., (Copyright © 2013 Elsevier B.V. All rights reserved.)