Anticancer Properties of 3-Dietoxyphosphorylfuroquinoline-4,9-dione.

Herein, the antitumor activity of a novel synthetic analog with 5,8-quinolinedione scaffold, diethyl (2-(2-chlorophenyl)-4,9-dioxo-4,9-dihydrofuro [3,2-g]quinolin-3-yl)phosphonate (AJ-418) was investigated on two breast cancer cell lines. This analog was selected from a small library of synthetic quinolinediones on the basis of its strong antiproliferative activity against MCF-7 and MDA-MB-231 cells and 4-5-fold lower cytotoxicity towards healthy MCF-10A cells. The morphology of MCF-7 and MDA-MB-231 cancer cells treated with AJ-418 changed drastically, while non-tumorigenic MCF-10A cells remained unaffected. In MCF-7 cells, after 24 h incubation, the increased number of apoptotic cells coincided with a decrease in proliferation and cell viability. The 24 h treatment of MDA-MB-231 cells with the tested compound reduced their cell viability and proliferation rate; however, a significant pro-apoptotic effect was visible only after longer incubation times (48 h and 72 h). Then, the maximum tolerated dose (MTD) of compound AJ-418 in C3H mice after subcutaneous administration was determined to be 160 mg/kg, showing that this analog was well tolerated and can be further evaluated to assess its potential therapeutic effect in tumor-bearing mice.

Can we detect patterns in behavioral time series of cows using cluster analysis?

Time series analysis can facilitate the detection of complex behavioral patterns and potentially provide new opportunities to assess animal welfare. The aim was to investigate whether dairy cows exhibit daily, individual patterns in activity and in area use in the barn. We predicted that behavioral patterns will be more consistent (1) within than between cows, (2) when area categorization is more specific and, thus, allows the detection of individual preferences for areas, and (3) during the night. We conducted the study at an experimental farm with 20 lactating Brown Swiss and Swiss Fleckvieh cows. The animals were housed in cubicles, and they received feed and were milked twice daily. Activity was recorded with IceTag pedometers (IceRobotics Ltd.), and area use with the SMARTBOW sensor system (Zoetis). Data were collected for 55 consecutive days and analyzed at 1-min intervals. To investigate the behavioral time series, we performed a hierarchical clustering analysis. A clustering process calculated distances between days, which were compared within and between cows based on t-tests and analyses of variance. Dendrograms of activity and area use showed that days of individual cows could not be grouped more closely together than those of different cows. A slightly better grouping was achieved with a more specific area categorization, but not during a specific time period. However, the average distances between days were always smaller within (mean ± SD; activity: 95.62 ± 76.88, lying areas: 0.14 ± 0.03, functional areas: 0.12 ± 0.01) than between cows (activity: 109.62 ± 75.33, lying areas: 0.16 ± 0.02, functional areas 0.13 ± 0.01). Considering that the time series of individual cows were slightly but always more similar compared with those between cows, and that more consistent patterns were found when the area categorization was more specific, it can be concluded that the cows exhibited weak individual preferences in area use and also weak daily individual patterns in activity and area use. Because the visual exploratory and empirical approaches used in this study do not account for variability, they do not seem to be suitable for the detection of patterns in animals that display greater plasticity in their temporal structure of activity. Thus, although determining the temporal structure of activity and area use bears the potential to assess the behavior and, in turn, for example, the physiological state and health status of cows, it does not seem to be achievable with a cluster analysis. Therefore, time series methods that account for temporal fluctuations in behavior should be further explored.

Synthesis and Anticancer Properties of New 3-Methylidene-1-sulfonyl-2,3-dihydroquinolin-4(1H)-ones.

Quinolinones have been known for a long time as broad-spectrum synthetic antibiotics. More recently, the anticancer potential of this group of compounds has been investigated. Following this direction, we obtained a small library of 3-methylidene-1-sulfonyl-2,3-dihydroquinolin-4(1H)-ones with various substituents at positions 1, 2, 6, and 7 of the quinolinone ring system. The cytotoxic activity of the synthesized analogs was tested in the MTT assay on two cancer cell lines in order to determine the structure-activity relationship. All compounds produced high cytotoxic effects in MCF-7, and even higher in HL-60 cells. 2-Ethyl-3-methylidene-1-phenylsulfonyl-2,3-dihydroquinolin-4(1H)-one, which was over 5-fold more cytotoxic for HL-60 than for normal HUVEC cells, was selected for further tests. This analog was shown to inhibit proliferation and induce DNA damage and apoptosis in HL-60 cells.

Do we automatically detect health- or general welfare-related issues? A framework.

The early detection of health disorders is a central goal in livestock production. Thus, a great demand for technologies enabling the automated detection of such issues exists. However, despite decades of research, precision livestock farming (PLF) technologies with sufficient accuracy and ready for implementation on commercial farms are rare. A central factor impeding technological development is likely the use of non-specific indicators for various issues. On commercial farms, where animals are exposed to changing environmental conditions, where they undergo different internal states and, most importantly, where they can be challenged by more than one issue at a time, such an approach leads inevitably to errors. To improve the accuracy of PLF technologies, the presented framework proposes a categorization of the aim of detection of issues related to general welfare, disease and distress and defined disease. Each decision level provides a different degree of information and therefore requires indicators varying in specificity. Based on these considerations, it becomes apparent that while most technologies aim to detect a defined health issue, they facilitate only the identification of issues related to general welfare. To achieve detection of specific issues, new indicators such as rhythmicity patterns of behaviour or physiological processes should be examined.

Synthesis of 2,2,6-Trisubstituted 5-Methylidene-tetrahydropyran-4-ones with Anticancer Activity.

In our continuous search for new, relatively simple 2-alkylidene-1-oxoheterocycles as promising anticancer drug candidates, herein we report an efficient synthesis of 2,2,6-trisubstituted 5-methylidenetetrahydropyran-4-ones. These compounds were obtained in a four step reaction sequence, in which starting diethyl 2-oxopropylphosphonate was transformed into 2,2-disubstituted 5-diethoxyphosphoryldihydropyran-4-ones, followed by Michael addition of various Grignard reagents and Horner-Wadsworth-Emmons reaction of the obtained adducts with formaldehyde. Stereochemistry of the intermediate Michael adducts is also discussed. Final 5-methylidenetetrahydropyran-4-ones were tested for their possible antiproliferative effect against three cancer cell lines and 6-isopropyl-2,2-dimethyl-5-methylidenetetrahydropyran-4-one (11c), which showed very high cytotoxic activity against HL-60 human leukemia cells and was three times more active than known anticancer drug carboplatin, was selected for further biological evaluation, in order to disclose its mechanism of action. The obtained results indicated that 11c induced apoptosis in HL-60 cells and caused the arrest of the cell cycle in the G2/M phase, which may suggest its cytotoxic and cytostatic activity.

Phenylselanyl Group Incorporation for "Glutathione Peroxidase-Like" Activity Modulation.

The ability of organoselenium molecules to mimic the activity of the antioxidant selenoenzyme glutathione peroxidase (GPx) allows for their use as antioxidant or prooxidant modulators in several diseases associated with the disruption of the cell redox homeostasis. Current drug design in the field is partially based on specific modifications of the known Se-therapeutics aimed at achieving more selective bioactivity towards particular drug targets, accompanied by low toxicity as the therapeutic window for organoselenium compounds tends to be very narrow. Herein, we present a new group of Se-based antioxidants, structurally derived from the well-known group of GPx mimics-benzisoselenazol-3(2H)-ones. A series of N-substituted unsymmetrical phenylselenides with an o-amido function has been obtained by a newly developed procedure: a copper-catalyzed nucleophilic substitution by a Se-reagent formed in situ from diphenyl diselenide and sodium borohydride. All derivatives were tested as antioxidants and anticancer agents towards breast (MCF-7) and leukemia (HL-60) cancer cell lines. The highest H2O2-scavenging potential was observed for N-(3-methylbutyl)-2-(phenylselanyl)benzamide. The best antiproliferative activity was found for (-)-N-(1S,2R,4R)-menthyl-2-(phenylselanyl)benzamide (HL-60) and ((-)-N-(1S,2R,3S,6R)-(2-caranyl))benzamide (MCF-7). The structure-activity correlations, including the differences in reactivity of the obtained phenyl selenides and corresponding benzisoselenazol-3(2H)-ones, were performed.

A New Hybrid δ-Lactone Induces Apoptosis and Potentiates Anticancer Activity of Taxol in HL-60 Human Leukemia Cells.

In the search for new drug candidates, researchers turn to natural substances isolated from plants which may be either used directly or may serve as a source for chemical modifications. An interesting strategy in the design of novel anticancer agents is based on the conjugation of two or more biologically active structural motifs into one hybrid compound. In this study, we investigated the anticancer potential of 4-benzyl-5,7-dimethoxy-4-methyl-3-methylidene-3,4-dihydro-2H-chroman-2-one (DL-247), a new hybrid molecule combining a chroman-2-one skeleton with an exo-methylidene bond conjugated with a carbonyl group, in human myeloid leukemia HL-60 cell line. The cytotoxicity of the new compound was tested using MTT assay. The effect of DL-247 on cell proliferation and apoptosis induction were studied by flow cytometry, fluorometric assay and ELISA analysis. DL-247 displayed high cytotoxic activity (IC50 = 1.15 µM, after 24 h incubation), significantly inhibited cell proliferation and induced apoptosis by both, the intrinsic and extrinsic pathways. A combination of DL-247 with taxol exhibited a strong synergistic effect on DNA damage generation, apoptosis induction and inhibition of cell growth.

Under temperate weather conditions, dairy goats use an outdoor run more with increasing warmth and avoid light wind or rain.

Access to an outdoor run might provide some benefits for the social and activity behavior of dairy goats. The aim of this study was to investigate the influence of weather parameters on outdoor run use by dairy goats under temperate weather conditions. Data were collected from February to April and in October, 2014, on 14 commercial dairy goat farms in Switzerland and Germany for 14 d per farm. Temperature, humidity, solar radiation, wind speed, and rain amount were measured continuously using a weather station. The outcome variables, average proportion of the herd in the outdoor run (proportion of herd in run) and average frequency of goats entering the outdoor run per hour in relation to group size (frequency of entries to run), were assessed by video data. Temperature, humidity, and solar radiation were combined to create the parameter "warmth" by means of a principal component analysis, whereas wind speed and rain (rain events and rain amount) were used in their original form as explanatory variables. Additionally, the effect of the comprehensive climate index on outdoor run use was investigated. Data were analyzed using linear mixed effects models. Increasing "warmth" increased, whereas increasing wind speed reduced, the proportion of herd in run. With increasing comprehensive climate index, the proportion of herd in run and the frequency of entries to run increased. On days without rain, the proportion of herd in run and the frequency of entries to run were markedly higher than during comparable daytime hours with rain. In the hour before the rain started and after it had stopped, more goats were in the outdoor run than during rain hours, but still fewer than during comparable daytime hours on days without rain. Furthermore, the proportion of herd in run and the frequency of entries to run decreased to a very low level when the rain amount exceeded 1 mm/h. To conclude, in the measured ranges of temperature (-0.8 to +16.5°C), humidity (34.8 to 99.2%), solar radiation (25.9 to 519.8 W/m2), and wind speed (0 to 4 m/s), warmer conditions increased the outdoor run use, whereas the goats clearly avoided light wind and rain. Therefore, in the outdoor runs, protection against rain and wind could promote their use under temperate weather conditions. Furthermore, ensuring optimized access to solar radiation by considering the exposure of the outdoor run might be of advantage, as direct sun exposure induces warmer conditions.

Stress indicators in dairy cows adapting to virtual fencing.

Virtual fencing (VF) enables livestock grazing without physical fences by conditioning animals to a virtual boundary delimited with an audio tone (AT) and an electric pulse (EP). The present study followed the adaptation process of lactating dairy cows to a VF system with changing virtual boundaries and investigated its impact on animal welfare. Twenty cows were divided into stratified groups (2× VF; 2× electric fencing, EF) of five individuals. Each group grazed half-days in a separate EF paddock of comparable size during 3 d of acclimation (P0), followed by 21, 14, 14, and 7 d of experimental treatment (P1 to P4). At the start of the trial, all cows were equipped with an IceQube pedometer (Peacock Technology Ltd, Stirling, UK) and a VF collar (Nofence AS, Batnfjordsøra, Norway). During P0, cows were accustomed to their first paddock with a deactivated virtual boundary and wearing the sensors. In P1 to P4, an active virtual boundary for the VF groups, and a second EF for the EF groups was set up parallel to an outer EF within their paddock. Throughout the trial, the sensors continuously tracked cow positions and activity behavior at 15-min intervals. From P1 onwards, the VF collars additionally recorded each AT and EP per cow with a georeferenced time stamp. During P0 to P4, daily feed intake, body weight, and milk yield were recorded in the barn. A total of 26 milk samples were collected per cow to determine milk cortisol levels. Behavioral observations were conducted for 2 h on day 23 to record agonistic behaviors, vocalizations, and excretions. The total number of stimuli per cow ranged from 37 to 225 ATs (mean ±â€…SD: 1.9 ±â€…3.3 per day) and 3 to 11 EPs (mean ±â€…SD: 0.1 ±â€…0.7 per day) throughout the trial. The maximum number of EPs per day was 8 for an individual cow and occurred once on D1. Mean EP/AT decreased by 55% during the first three half-days of grazing and with each paddock change from 0.2 EP/AT in week 1 to 0.03, 0.02, and 0 EP/AT in weeks 4, 6, and 8, respectively. Linear and generalized mixed effects models revealed that milk yield and cortisol, feed intake, body weight, and activity and lying behavior did not significantly differ between VF and EF groups. A higher number of agonistic behaviors were observed in the VF groups when the VF system was activated. However, due to the short observation periods only few contacts were observed in total. Overall, all cows adapted to the VF system without evidence of lasting adverse effects on animal welfare.

Virtual fences are commercially available but face restrictions in some countries due to animal welfare concerns. For virtual fencing (VF), animals are equipped with collars that emit audio tones (ATs) followed by electric pulses (EPs) when they cross a virtual boundary tracked by global navigation. Existing studies have so far not covered the aspect of longer-term learning, impacting possibly VF suitability. The present study followed therefore the learning process of dairy cows with changing virtual boundaries and examined behavior and stress indicators in dairy cows during an 8-wk adaptation to VF across four experimental periods. Four control and treatment groups of five cows each were investigated. EPs occurred most frequently on days 1 to 3 and remained low for the remaining experiment. In the latter two experimental periods, almost no EPs were recorded while ATs were still triggered, indicating that it took the animals two introductions to a new fence line to respond to the ATs only. Animal welfare was assessed by monitoring cow activity and lying behavior, milk yield, milk cortisol, feed intake, body weight, and frequencies of agonistic interactions, vocalizations, and excretions. All cows adapted to the VF system without compromising animal welfare during the study period.

Historical Perspective and Current Trends in Anticancer Drug Development.

Cancer is considered one of the leading causes of death in the 21st century. The intensive search for new anticancer drugs has been actively pursued by chemists and pharmacologists for decades, focusing either on the isolation of compounds with cytotoxic properties from plants or on screening thousands of synthetic molecules. Compounds that could potentially become candidates for new anticancer drugs must have the ability to inhibit proliferation and/or induce apoptosis in cancer cells without causing too much damage to normal cells. Some anticancer compounds were discovered by accident, others as a result of long-term research. In this review, we have presented a brief history of the development of the most important groups of anticancer drugs, pointing to the fact that they all have many side effects.

Stereoselective Synthesis and Anticancer Activity of 2,6-Disubstituted trans-3-Methylidenetetrahydropyran-4-ones.

In this report, we present efficient and stereoselective syntheses of 2,6-disubstituted trans-3-methylidenetetrahydropyran-4-ones and 2-(4-methoxyphenyl)-5-methylidenetetrahydropyran-4-one that significantly broaden the spectrum of the available methylidenetetrahydropyran-4-ones with various substitution patterns. Target compounds were obtained using Horner-Wadsworth-Emmons methodology for the introduction of methylidene group onto the pyranone ring. 3-Diethoxyphosphoryltetrahydropyran-4-ones, which were key intermediates in this synthesis, were prepared by fully or highly stereoselective addition of Gilman or Grignard reagents to 3-diethoxyphosphoryldihydropyran-4-ones. Addition occurred preferentially by axial attack of the Michael donors on the dihydropyranone ring. Relative configurations and conformations of the obtained adducts were assigned using a detailed analysis of the NMR spectra. The obtained methylidenepyran-4-ones were evaluated for cytotoxic activity against two cancer cell lines (HL-60 and MCF-7). 2,6-Disubstituted 3-methylidenetetrahydropyran-4-ones with isopropyl and phenyl substituents in position 2 were more cytotoxic than analogs with n-butyl substituent. Two of the most cytotoxic analogs were then selected for further investigation on the HL-60 cell line. Both analogs induced morphological changes characteristic of apoptosis in cancer cells, significantly inhibited proliferation and induced apoptotic cell death. Both compounds also generated DNA damage, and one of the analogs arrested the cell cycle of HL-60 cells in the G2/M phase. In addition, both analogs were able to inhibit the activity of topoisomerase IIα. Based on these findings, the investigated analogs may be further optimized for the development of new and effective topoisomerase II inhibitors.

Synthesis and structure-activity relationship study of novel 3-diethoxyphosphorylfuroquinoline-4,9-diones with potent antitumor efficacy.

Herein we report an efficient synthesis of a series of regioisomeric N,O-syn and N,O-anti 3-diethoxyphosphorylfuroquinoline-4,9-diones combining furoquinoline-5,8-dione skeleton, present in several highly cytotoxic compounds, with diethoxyphosphoryl moiety. The cytotoxic activity of the obtained analogs was tested against two human cancer cell lines: promyelocytic leukemia HL-60 and breast cancer adenocarcinoma MCF-7 and for comparison on human umbilical vein endothelial cells HUVEC and mammary gland/breast MCF-10 A cells. Several diethoxyphosphorylfuroquinoline-4,9-diones proved to be highly cytotoxic for cancer cells with IC50 values even below 0.1 µM. Interestingly, N,O-syn 3-diethoxyphosphorylfuroquinoline-4,9-diones were 3- to 7-fold more active against HL-60 cells than the respective N,O-anti regioisomers. The most promising analogs 9c and 9i, with the highest cancer/healthy cells cytotoxicity ratio, were further evaluated to establish their mode of action. In HL-60 cells these analogs enhanced intracellular ROS generation and NAD(P)H:quinone oxidoreductase 1 (NQO1) depletion which led to the cell cycle arrest in the S-phase, reduced cell proliferation, DNA damage and apoptosis.

Intramolecular C-N Bond Formation via Thermal Arene C-H Bond Activation Supported by Au(III) Complexes.

One of the main tactics to access C-N bonds from inactivated C-H functionalities is direct transition metal-supported aminations. Due to the often harsh reaction conditions, the current goal in the field is the search for more mild and sustainable transformations. Herein, we present the first solvent-free thermally induced C-N bond formation driven by Au(III) salts. The general structure of the products was confirmed by 1H, 13C, 15N NMR, TGA-DTA and ATR/FT-IR analysis. Additionally, all derivatives were tested as catalysts in a three-component coupling reaction between phenylacetylene, benzaldehyde and piperidine and as anticancer agents on HL-60 and MCF-7 cell lines.

Crystal Growth, Single Crystal Structure, and Biological Activity of Thiazolo-Pyridine Dicarboxylic Acid Derivatives.

Four novel TPDCA derivatives were prepared via a supersaturation method combining TPDCA with water, N-methyl-2-pyrrolidone (NMP), Na(PO2H2), and ammonia solution: 2(C9H7NO5S)H2O (1), (C9H7NO5S)C5H9NO (2), (C9H7NO5S)Na(PO2H2) (3), and (C9H5NO5S)(NH4)2(H2O) (4). Their crystal structures were determined by single-crystal X-ray diffraction. Compounds (1) and (2) crystallize in the monoclinic space groups P21 and P21/c, respectively, whereas compounds (3) and (4) crystallize in the triclinic space group P1̅. Weak and moderate hydrogen bonds were detected in the four compounds. In the biological tests, (1) and (3) exhibited significant antibacterial activity against Escherichia coli and Staphylococcus aureus; in addition, (1) was cytotoxic against leukemia HL-60 cells with the IC50 value of 158.5 ± 12.5 µM.

Molecular mechanisms of apoptosis induced by a novel synthetic quinolinone derivative in HL-60 human leukemia cells.

The mortality rates for acute myeloid leukemia are very high, necessitating the search for novel chemotherapeutic candidates. Herein, we investigated the anticancer potential of a new synthetic compound, 2-ethyl-3-methyliden-1-tosyl-2,3-dihydroquinolin-4-(1H)-one (AJ-374) against myeloid leukemia HL-60 cell line. This analog was selected from the small library of synthetic dihydroquinolinones on the basis of its strong antiproliferative activity against HL-60 cells and 30-fold lower cytotoxicity towards healthy HUVEC cells. AJ-374 promoted the arrest of the cells in the subG0/G1 phase of the cell cycle in the first 24 h. Treatment of HL-60 cells with AJ-374 caused an increase in annexin-V positive cells, activation of caspase-8, -9 and -3, dissipation of the mitochondrial membrane potential and enhancement of FAS protein level. Apoptosis induction triggered by this quinolinone was blocked by the pre-treatment of the cells with caspase-8, -9 and -3 inhibitors. The obtained results indicated that AJ-374-induced apoptosis was executed by both, the extrinsic and intrinsic pathways. The cytotoxic activity of AJ-374 was also associated with down-regulation of the mitogen-activated protein kinase (MAPK) pathway and was independent of reactive oxygen species generation. Taken together, these results suggest that AJ-374 exerts a potent anticancer effect on leukemia cells, with a wide safety margin, which makes this analog an attractive drug candidate for further testing.