Design,Synthesis and in Vitro Tumor Cytotoxicity Evaluation of 3,5‐Diamino‐N‐substituted Benzamide Derivatives as Novel GSK‐3β Small Molecule Inhibitors

Glycogen synthase kinase‐3 (GSK‐3) plays an important regulatory role in various signaling pathways; such as PI3 K/AKT, which is closely related to the occurrence and development of tumors. At present, the most reported active GSK‐3 inhibitors have the same structure: lactam ring or amide structure. To find out the GSK‐3β small molecule inhibitor with novel, safe, efficient and more uncomplicated synthesis method, we analyzed in‐depth reported crystal‐binding patterns of GSK‐3β small molecule inhibitor with GSK‐3β protein, and designed and synthesized 17 non‐reported 3,5‐diamino‐N‐substituted benzamide compounds. Their structures were confirmed by ¹H‐NMR, ¹³C‐NMR, and HR‐MS. The preliminary screening of tumor cytotoxicity of compounds in vitro was detected by MTT, and their structure–activity relationships were illustrated. The results have shown that 3,5‐diamino‐N‐[3‐(trifluoromethyl)phenyl]benzamide ( 4d ) exhibited significant tumor cytotoxicity against human colon cancer cells (HCT‐116) with IC₅₀ of 8.3 μm and showed commendable selectivity to GSK‐3β. In addition, Compound 4d induced apoptosis to some extent and possessed modest PK properties.     

Clinanthus microstephium,an Amaryllidaceae Species with Cholinesterase Inhibitor Alkaloids: Structure−Activity Analysis of Haemanthamine Skeleton Derivatives

Plants of the Amaryllidaceae family are well‐known (not only) for their ornamental value but also for the alkaloids that they produce. In this report, the first phytochemical study of Clinanthus genus was carried out. The chemical composition of alkaloid fractions from Clinanthus microstephium was analyzed by GC/MS and NMR. Seven known compounds belonging to three structural types of Amaryllidaceae alkaloids were identified. An epimeric mixture of a haemanthamine‐type compound (6‐hydroxymaritidine) was tested as an inhibitor against acetyl‐ and butyrylcholinesterase enzymes (AChE and BChE, respectively), two enzymes relevant in the treatment of Alzheimer's disease, with good results. Structure–activity relationships through molecular docking studies with this alkaloid and other structurally related compounds were discussed.     

Prediction of structural consequences for disease causing variants in C21orf2 protein using computational approaches

Amyotrophic lateral sclerosis (ALS), a progressive motor-neurone disease, affects individuals usually aged between 50 and 70 years. C21orf2, recently identified as the new ALS susceptibility gene, harbours rare missense mutations that cause this fatal disease. We used bioinformatics and molecular modelling approaches to study specific ALS-associated mutations in C21orf2. Both native and mutant structures of the protein obtained from homology modelling were analysed in detail to gain insights into the potential impact of these mutations on the protein structure and its function. Our analyses reveal that more than 75% of the mutations are likely to be deleterious. These effects seem to carry through to mouse C21orf2 as well, indicating that mouse would make a viable animal model to study this ALS gene in detail.     

Insight derived from molecular dynamics simulation into dynamics and molecular motions of cuticle-degrading serine protease Ver112

Cuticle-degrading serine protease Ver112, which derived from a nematophagous fungus Lecanicillium psalliotae, has been exhibited to have high cuticle-degrading and nematicidal activities. We have performed molecular dynamics (MD) simulation based on the crystal structure of Ver112 to investigate its dynamic properties and large-scale concerted motions. The results indicate that the structural core of Ver112 shows a small fluctuation amplitude, whereas the substrate binding sites, and the regions close to and opposite the substrate binding sites experience significant conformational fluctuations. The large concerted motions obtained from essential dynamics (ED) analysis of MD trajectory can lead to open or close of the substrate binding sites, which are proposed to be linked to the functional properties of Ver112, such as substrate binding, orientation, catalytic, and release. The significant motion in the loop regions that is located opposite the binding sites are considered to play an important role in modulating the dynamics of the substrate binding sites. Furthermore, the bottom of free energy landscape (FEL) of Ver112 are rugged, which is mainly caused by the fluctuations of substrate binding regions and loops located opposite the binding site. In addition, the mechanism underlying the high flexibility and catalytic activity of Ver112 was also discussed. Our simulation study complements the biochemical and structural studies, and provides insight into the dynamics-function relationship of cuticle-degrading serine protease Ver112.     

A Longitudinal Study of Decision Making in the Process of Acquiring a Dog

To prevent potential problems in the relationship between people and their dogs, it is important to engage in a thoughtful decision-making process with regard to acquiring a dog. To map the most important elements in the decision-making process, a social cognitive model was applied using seven psychological constructs: perceived advantages of having a dog; perceived disadvantages; the social norm; self-efficacy; optimism; expected commitment; and the intention to acquire a dog. People who were planning to acquire a dog within one year were asked to fill in an online questionnaire and another one 14 months later. The social cognitive constructs were operationalized in the baseline questionnaire, and in the follow-up participants were asked to report whether they had actually acquired a dog during the 14-month interval. The sample (n?=?588) largely consisted of females (88%) and those who had a high level of education (64%). The mean age of the participants was 40.9 years. The data showed significant relations among the decision-making variables, and significant differences between demographic variables related to these. In univariate longitudinal analyses, several social cognitive variables significantly predicted acquiring a dog in the 14-month period. When intention to acquire a dog was entered into the multivariate model, it remained the only predictor (OR = 2.16, p?<?0.001); the model explained 33% of the variance in acquiring a dog. It was possible to assess the main constructs that play a role in decision making regarding acquiring a dog: Most baseline measures were related to actual later behavior, also when taking into account all changes in other internal and external factors that may have taken place. Insight into the decision-making process makes it possible to intervene in it for the good of people, dogs, and their relationships.     

Siberian spruce tree ring anatomy: imprint of development processes and their high-temporal environmental regulation

Wood anatomy was offered as spatiotemporal proxy record for tracheid differentiation kinetics due to its advantages in terms of much longer cover period and less demanding measurements. In this study, external and internal regulation of earlywood-to-latewood transition and properties of latewood of Picea obovata Ledeb were considered. The values and interrelations between cell number, tree ring width, maximal and mean radial cell diameter, maximal cell wall thickness and position of the transition to thick-walled tracheids were investigated within site and along the altitudinal gradient. Correlations with moving 21-day climatic series were used to estimate high-resolutional external influences. Relationships between tree ring traits are spatially stable and close within one stage of differentiation and between cells production and expansion. Relationships between sites differ in upper and lower parts of the gradient. Most of traits respond to the primary limiting factors near summer solstice; however, maximal cell wall thickness responds positively to the temperatures at the + 10 °C threshold. Altitudinal anatomical patterns revealed interaction of intrinsic and external factors in the regulation of tracheid differentiation. Timing of climatic response highlighted role of photoperiod as a trigger in the earlywood-to-latewood transition, and crucial role of the growth season ending for latewood development.     

Ensifer fredii symbiovar vachelliae nodulates endemic Vachellia gummifera in semiarid Moroccan areas

The purpose of this work was to study the genetic diversity of the nodule-forming bacteria associated with native populations of Vachellia gummifera growing wild in Morocco. The nearly complete 16S rRNA gene sequences from three selected strains, following ARDRA and REP-PCR results, revealed they were members of the genus Ensifer and the sequencing of the housekeeping genes recA, gyrB, dnaK and rpoB, and their concatenated phylogenetic analysis, showed that the 3 strains belong to the species E. fredii. Based on the nodC and nodA phylogenies, the 3 strains clearly diverged from the type and other reference strains of E. fredii and formed a clearly separated cluster. The strains AGA1, AGA2 and AGB23 did not form nodules on Glycine max, Phaseolus vulgaris and Medicago truncatula, and effectively nodulated V. gummifera, Acacia cyanophylla, Prosopis chilensis and Leucaena leucocephala. Based on similarities of the nodC and nodA symbiotic genes and differences in the host range, the strains isolated from Moroccan endemic V. gummifera may form a different symbiovar within Ensifer species, for which the name “vachelliae” is proposed.     

Schiff bases of tryptamine as potent inhibitors of nucleoside triphosphate diphosphohydrolases (NTPDases): Structure-activity relationship

Overexpression of NTPDases leads to a number of pathological situations such as thrombosis, and cancer. Thus, effective inhibitors are required to combat these pathological situations. Different classes of NTPDase inhibitors are reported so far including nucleotides and their derivatives, sulfonated dyes such as reactive blue 2, suramin and its derivatives, and polyoxomatalates (POMs). Suramin is a well-known and potent NTPDase inhibitor, nonetheless, a range of side effects are also associated with it. Reactive blue 2 also had non-specific side effects that become apparent at high concentrations. In addition, most of the NTPDase inhibitors are high molecular weight compounds, always required tedious chemical steps to synthesize. Hence, there is still need to explore novel, low molecular weight, easy to synthesize, and potent NTPDase inhibitors.Keeping in mind the known NTPDase inhibitors with imine functionality and nitrogen heterocycles, Schiff bases of tryptamine, 1–26, were synthesized and characterized by spectroscopic techniques such as EI-MS, HREI-MS, ¹H-, and ¹³C NMR. All the synthetic compounds were evaluated for the inhibitory avidity against activities of three major isoforms of NTPDases: NTPDase-1, NTPDase-3, and NTPDase-8. Cumulatively, eighteen compounds were found to show potent inhibition (Ki = 0.0200–0.350 μM) of NTPDase-1, twelve (Ki = 0.071–1.060 μM) of NTPDase-3, and fifteen compounds inhibited (Ki = 0.0700–4.03 μM) NTPDase-8 activity. As a comparison, the Kis of the standard inhibitor suramin were 1.260 ± 0.007, 6.39 ± 0.89 and 1.180 ± 0.002 μM, respectively. Kinetic studies were performed on lead compounds (6, 5, and 21) with human (h-) NTPDase-1, -3, and -8, and Lineweaver-Burk plot analysis showed that they were all competitive inhibitors. In silico study was conducted on compound 6 that showed the highest level of inhibition of NTPDase-1 to understand the binding mode in the active site of the enzyme.     

Non‐verbal mate retention behaviour in women and its relation to couple's relationship adjustment and satisfaction

The function of mate retention strategies is to preserve the bond between romantic partners and to prevent desertion, which is also why such behaviours are often elicited by the presence of a potential rival. Most existing studies on mate retention are based on self‐reports, which are prone to various biases. In this study, we observed non‐verbal behaviour of 47 long‐term romantic heterosexual couples during their interview with a female experimenter, whom the women in the couples may have perceived as a potential rival. We measured non‐verbal displays of mate retention tactics (direct guarding, intimacy inducement, appearance enhancement, and love and care display), as well as relationship maintenance strategies of openness and positivity. We expected that relationship quality as reported by both partners in the couple would be positively associated with ‘benefit‐provisioning’ mate retention behaviours and negatively associated with a ‘cost‐inflicting’ mate retention tactic (direct guarding). Relationship quality was assessed by Spanier's Dyadic Adjustment Scale using both the total score of dyadic adjustment and a Dyadic Satisfaction subscale. We found a significant association between both partners' dyadic adjustment and satisfaction and woman's appearance enhancement behaviours (such as self‐touch, hair flip and primp) in the presence of her partner and a potential rival. We also found a negative association between partner's dyadic adjustment and woman's direct guarding of partner using short looks, and a positive association between woman's intimacy‐inducing behaviour using long gaze and her own dyadic satisfaction and her partner's dyadic adjustment. Our results highlight the specific role of female attractiveness in later stages of romantic relationships within the context of mate retention and draw attention to the significance of the use of observational techniques in addition to self‐reports.     

Probable origin and toxin profile of Alexandrium tamarense (Lebour) Balech from southern Brazil

The distribution of the toxic dinoflagellate Alexandrium tamarense Lebour has apparently expanded within the southern hemisphere during the last 2 decades. Toxic blooms of A. tamarense were recorded in Argentinean coastal waters since 1980; however, the first documented bloom in southern Brazil was in 1996. In this study, 13 strains of A. tamarense from southern Brazil were isolated and kept in culture. Phylogenetic analysis using RFLP and DNA sequences of the D1–D2 region of large subunit ribosomal DNA (rDNA) clearly indicates that Brazilian strains are most closely related to other South American strains. The strains from South America are placed firmly within a phylogenetic clade which contains strains from North America, northern Europe and northern Asia, previously called the North American clade. Possible dispersal hypotheses are discussed. The cultures were also analyzed for saxitoxin and its derivatives by high performance liquid chromatography (HPLC). The main saxitoxin groups found were the low toxicity N-sulfocarbamoyl group, C1, 2 (30–84%), followed by the high potency carbamate toxins, gonyautoxins 1, 4 (6.6–55%), gonyautoxins 2, 3 (0.3–29%), neosaxitoxin (1.4–24%) and saxitoxin (0–4.4%). The toxin composition is similar to that of other strains from South America, supporting a close relationship between A. tamarense from southern Brazil and other areas of South America. Toxicity values were variable (7.07–65.92 pg STX cell⁻¹), with the higher range falling among the most toxic values recorded for cultures of A. tamarense, indicating the significant risk for shellfish contamination and human intoxication during blooms of this species along the southern Brazilian coast.