Marked Activity Research Articles

Zn(II)-MOF derived N-doped carbons achieve marked ORR activity in alkaline and acidic media.

A highly efficient metal-free N-doped carbon electrocatalyst toward oxygen reduction was obtained by one-pot pyrolysis of a single Zn(II)-MOF with mixed azolate and terephthalate ligands, demonstrating E1/2 of 0.88 V (vs. RHE) in 0.1 M KOH, and 0.79 V (vs. RHE) in 0.5 M H2SO4. It represents one of the best metal-free N-doped carbon electrocatalysts for the acidic ORR.

Pathological lesions associated with Vibrio infection in Atlantic horse mackerel (Trachurus trachurus L., 1758) from the western coast of Tripoli, Libya.

Vibrios are aquatic bacteria causing Vibriosis in humans and aquatic animals. Vibriosis is a significant disease of cultured and wild fish. The present study aimed to address the impact of Vibrios on the health status of Trachurus trachurus inhabiting the coastal area of Tripoli. A total of 100 samples of (T. trachurus) were collected randomly from the Western Coast of Tripoli and Bab Al-Baher market, throughout the period from spring 2019 to summer 2019. All the sampled fish were examined externally and internally and lesions were recorded. Bacterial isolation from the liver and kidney was carried out using the appropriate culture media. Tissue samples were taken from the liver, kidney, and spleen in 10% neutral buffered formalin for a histopathology examination. The tissue sections were stained with hematoxylin and eosin to assess morphologically and Perl's Prussian blue for the demonstration of ferric iron. On average 69% of the infected fish exhibited at least one pathological lesion. Vibrios were recovered from 90% of the examined fish. The histopathological changes of the liver showed severe congestion of blood vessels (BVs), mononuclear cell infiltration surrounding a bile duct, granular degeneration and coagulative necrosis of hepatocytes in the centrilobular area, marked vacuolar degeneration of hepatocytes, activation of melano-macrophage centers (MMCs), multiple cysts of nematode in hepatic tissue (incidental finding). The histopathological alterations of the kidney showed severe congestion of BVs, vacuolar degeneration of the renal tubular epithelium, severe interstitial mononuclear cell infiltration, and marked activation of MMCs in the kidney. Moreover, congestion of BVs and marked activation of melano-macrophages (MMCs) were found in the spleen. The MMCs of most of the sampled tissues showed a strong positive reaction for ferric iron. The polluted aquatic environment by sewage at Tripoli Coast is an essential factor for triggering the pathogenicity and invasion of Vibrios to vulnerable Atlantic horse mackerel. Also, this study is a preliminary step to give a baseline for further future studies on epidemiology and control of Vibrio infection in Libyan fish.

A combined crystallographic and theoretical investigation of noncovalent interactions in 1,3,4-oxadiazole-2-thione-N-Mannich derivatives: in vitro bioactivity and molecular docking.

Two 1,3,4-oxadiazole-2-thione-N-Mannich derivatives, specifically 5-(4-chlorophenyl)-3-[(2-trifluoromethylphenylamino)methyl]-1,3,4-oxadiazole-2(3H)-thione (1) and 5-(4-chlorophenyl)-3-[(2,5-difluorophenylamino)methyl]-1,3,4-oxadiazole-2(3H)-thione (2), were synthesized and then characterized by elemental analysis and NMR (1H and 13C) spectroscopy and the single crystal X-ray diffraction method. The formed weak intermolecular interactions in the solid-state structures of these derivatives were thoroughly investigated utilizing a variety of theoretical tools such as Hirshfeld surface analysis and quantum theory of atoms in molecules (QTAIM). Furthermore, the CLP-PIXEL and density functional theory calculations were used to study the energetics of molecular dimers. Numerous weak intermolecular interactions such as C-H⋯S/Cl/F/π interactions, a directional C-Cl⋯Cl halogen bond, π-stacking, type C-F⋯F-C contact and a short F⋯O interaction, help to stabilize the crystal structure of 1. Crystal structure 2 also stabilizes with several weak intermolecular contacts, including N-H⋯S, C-H⋯N//Cl/F interactions, a highly directional C1-Cl1⋯C(π) halogen bond and C(π)⋯C(π) interaction. In vitro antimicrobial potency of compounds 1 and 2 was assessed against various Gram-positive and Gram-negative bacterial strains and the pathogenic yeast-like Candida albicans. Both compounds showed marked activity against all tested Gram-positive bacteria and weak activity against Escherichia coli and lacked inhibitory activity against Pseudomonas aeruginosa. In addition, compounds 1 and 2 displayed good in vitro anti-proliferative activity against hepatocellular carcinoma (HepG-2) and mammary gland breast cancer (MCF-7) cancer cell lines. Molecular docking studies revealed the binding modes of title compounds at the active sites of prospective therapeutic targets.

Algal-Derived Synthesis of Silver Nanoparticles Using the Unicellular ulvophyte sp. MBIC10591: Optimisation, Characterisation, and Biological Activities

Algal-mediated synthesis of nanoparticles (NPs) is an eco-friendly alternative for producing NPs with potent physicochemical and biological properties. Microalgae represent an ideal bio-nanofactory because they contain several biomolecules acting as passivation and stabilising agents during the biogenesis of NPs. Herein, a novel microalgae sp. was isolated, purified, and identified using light and electron microscopy and 18s rRNA sequencing. The chemical components of their watery extract were assessed using GC-MS. Their dried biomass was used to synthesise silver (Ag) NPs with different optimisation parameters. Ag-NPs were physiochemically characterised, and their anticancer and antibacterial effects were examined. The data showed that the isolated strain was 99% similar to the unicellular ulvophyte sp. MBIC10591; it was ellipsoidal to spherical and had a large cup-shaped spongiomorph chloroplast. The optimum parameters for synthesising Ag-NPs by unicellular ulvophyte sp. MBIC10591 (Uv@Ag-NPs) were as follows: mixture of 1 mM of AgNO3 with an equal volume of algal extract, 100 °C for 1 h, and pH of 7 under illumination for 24 h. TEM, HRTEM, and SEM revealed that Uv@Ag-NPs are cubic to spherical, with an average nanosize of 12.1 ± 1.2 nm. EDx and mapping analysis showed that the sample had 79% of Ag, while FTIR revealed the existence of several functional groups on the NP surface derivatives from the algal extract. The Uv@Ag-NPs had a hydrodynamic diameter of 178.1 nm and a potential charge of -26.7 mV and showed marked antiproliferative activity against PC3, MDA-MB-231, T47D, and MCF-7, with IC50 values of 27.4, 20.3, 23.8, and 40 µg/mL, respectively, and moderate toxicity against HFs (IC50 of 13.3 µg/mL). Uv@Ag-NPs also showed marked biocidal activity against Gram-negative bacteria. Escherichia coli was the most sensitive bacteria to the NPs with an inhibition zone of 18.9 ± 0.03 mm. The current study reports, for the first time, the morphological appearance of the novel unicellular ulvophyte sp., MBIC10591, and its chemical composition and potential to synthesise Uv@Ag-NPs with smaller sizes and high stability to act as anti-tumour and microbial agents.

Modulation Effect on Tubulin Polymerization, Cytotoxicity and Antioxidant Activity of 1H-Benzimidazole-2-Yl Hydrazones

1H-benzimidazol-2-yl hydrazones with varying hydroxy and methoxy phenyl moieties were designed. Their effect on tubulin polymerization was evaluated in vitro on porcine tubulin. The compounds elongated the nucleation phase and slowed down the tubulin polymerization comparably to nocodazole. The possible binding modes of the hydrazones with tubulin were explored by molecular docking at the colchicine binding site. The anticancer activity was evaluated against human malignant cell lines MCF-7 and AR-230, as well as against normal fibroblast cells 3T3 and CCL-1. The compounds demonstrated a marked antineoplastic activity in low micromolar concentrations in both screened in vitro tumor models. The most active were the trimethoxy substituted derivative 1i and the positional isomers 1j and 1k, containing hydroxy and methoxy substituents: they showed IC50 similar to the reference podophyllotoxin in both tumor cell lines, accompanied with high selectivity towards the malignantly transformed cells. The compounds exerted moderate to high ability to scavenge peroxyl radicals and certain derivatives-1l containing metha-hydroxy and para-methoxy group, and 1b-e with di/trihydroxy phenyl moiety, revealed HORAC values high or comparable to those of well-known phenolic antioxidants. Thus the 1H-benisimidazol-2-yl hydrazones with hydroxy/methoxy phenyl fragments were recognized as new agents exhibiting promising combined antioxidant and antineoplastic action.

An experimental evaluation of anticonvulsant activity of aqueous extract of Caralluma dalzielii N.E. Brown

Ethnopharmacological RelevanceCaralluma dalzielii belongs to the family of Asclepiadiaceae. It is a succulent perennial plant widely distributed across the Sahel region from Senegal to Northwest Nigeria. Extracts from Caralluma dalzielii are used traditionally in the treatment of pain, infertility, epilepsy and other neurological diseases. Aim of the studyThe aim of the present study was to evaluate the anticonvulsant effects of aqueous extracts of aerial parts of Caralluma dalzielii using various experimental models. Materials and methodsThe anticonvulsant effect of the plant extract (250, 500 and 1000 mg/kg p.o) were assessed using maximal electroshock (MES) model in chicks, pentylenetetrazole (PTZ) and strychnine (STN)-induced seizures models in mice. Diazepam-induced sleep in chicks was used to study the sedative property of the plant. In the anticonvulsant and sedative property studies, the onset and duration of seizures and sleep were recorded respectively. The oral acute toxicity study was assessed. The extract was screened preliminarily for its phytochemicals and was further analysed using Gas chromatography-mass spectrometry (GC–MS) to identify its major chemical components. ResultsThe oral acute toxicity studies showed that the LD50 was greater than 5000 mg/kg in mice. Pre-treatment of mice with the aqueous extract of Caralluma dalzielii produced a dose-dependent protection against PTZ-induced seizures and mortality with 100% protection at the highest dose of the extract. The extract also significantly increased the latency/onset of myoclonic jerks and reduced the duration of tonic seizures dose dependently. In STN-induced seizure, the extract had no protection against seizure induction similar to diazepam the positive control. However, it significantly (p < 0.05) prolonged the latency of seizure and increased the survival time before death in a dose dependent manner. Caralluma dalzielii had zero protection against maximal electroshock. Phytochemical screening showed the presence of flavonoids, tannins, saponins, steroids, glycosides, cardiac glycosides and phenols. GC–MS analysis revealed the presence of some compounds. ConclusionThe extract of Caralluma dalzielii possesses anticonvulsant activity with less marked sedative activity. It also contains several phytochemicals which may be responsible for the activity. This study serves to validate the use of this plant in folkloric medicine for the treatment of epilepsy and convulsions.

Manipulation of Amino Acid Levels with Artificial Diets Induces a Marked Anticancer Activity in Mice with Renal Cell Carcinoma.

Targeted therapies with antiangiogenic drugs (e.g., sunitinib) and immune checkpoint inhibitors (e.g., anti-PD-1 antibodies) are the standard of care for patients with metastatic renal cell carcinoma. Although these treatments improve patient survival, they are rarely curative. We previously hypothesized that advanced cancers might be treated without drugs by using artificial diets in which the levels of specific amino acids (AAs) are manipulated. In this work, after showing that AA manipulation induces selective anticancer activity in renal cell carcinoma cells in vitro, we screened 18 artificial diets for anticancer activity in a challenging animal model of renal cell carcinoma. The model was established by injecting murine renal cell carcinoma (Renca) cells into the peritoneum of immunocompetent BALB/cAnNRj mice. Mice survival was markedly improved when their normal diet was replaced with our artificial diets. Mice fed a diet lacking six AAs (diet T2) lived longer than mice treated with sunitinib or anti-PD-1 immunotherapy; several animals lived very long or were cured. Controlling the levels of several AAs (e.g., cysteine, methionine, and leucine) and lipids was important for the anticancer activity of the diets. Additional studies are needed to further evaluate the therapeutic potential and mechanism of action of this simple and inexpensive anticancer strategy.

Notch pathway regulates osimertinib drug-tolerant persistence in EGFR-mutated non-small-cell lung cancer.

Osimertinib is a third-generation epidermal growth factor receptor (EGFR)-tyrosine kinase inhibitor (TKI) that has shown marked antitumor activity in patients with EGFR-mutated non-small-cell lung cancer (NSCLC). However, these effects are transient and most patients develop resistance. Reversible drug-tolerant persister (DTP) cells are defined as a small subpopulation of cells with markedly reduced sensitivity and non-genetic acquired resistance to EGFR-TKIs. Notch is a transmembrane receptor that plays an important role in tumorigenesis. We previously reported that there is significant crosstalk between the Notch and EGFR pathways in NSCLC. Moreover, the Notch pathway is associated with resistance to previous-generation EGFR-TKIs. However, the role of Notch in osimertinib resistance is not fully understood. In this study, we evaluated whether Notch is involved in osimertinib resistance. We show that NOTCH1 and Notch target genes are upregulated in osimertinib DTP cells, and that the addition of a γ-secretase inhibitor (GSI), a Notch inhibitor, impairs drug-tolerant persistence in vitro and in vivo. Compared with osimertinib, combined GSI and osimertinib suppress phospho-ERK partly by enhancing DUSP1 expression. Furthermore, Notch1 and HES1 were upregulated after EGFR-TKI treatment in half of human EGFR-mutated NSCLC tumor tissues. These results suggest that the combination of GSI and osimertinib may be a potential therapy for EGFR-mutated NSCLC.

Disparate Regions of the Human Chemokine CXCL10 Exhibit Broad-Spectrum Antimicrobial Activity against Biodefense and Antibiotic-Resistant Bacterial Pathogens.

CXCL10 is a pro-inflammatory chemokine produced by the host in response to microbial infection. In addition to canonical, receptor-dependent actions affecting immune-cell migration and activation, CXCL10 has also been found to directly kill a broad range of pathogenic bacteria. Prior investigations suggest that the bactericidal effects of CXCL10 occur through two distinct pathways that compromise the cell envelope. These observations raise the intriguing notion that CXCL10 features a separable pair of antimicrobial domains. Herein, we affirm this possibility through peptide-based mapping and structure/function analyses, which demonstrate that discrete peptides derived from the N- and C-terminal regions of CXCL10 mediate bacterial killing. The N-terminal derivative, peptide P1, exhibited marked antimicrobial activity against Bacillus anthracis vegetative bacilli and spores, as well as antibiotic-resistant clinical isolates of Klebsiella pneumoniae, Acinetobacter baumannii, Enterococcus faecium, and Staphylococcus aureus, among others. At bactericidal concentrations, peptide P1 had a minimal degree of chemotactic activity, but did not cause red blood cell hemolysis or cytotoxic effects against primary human cells. The C-terminal derivative, peptide P9, exhibited antimicrobial effects, but only against Gram-negative bacteria in low-salt medium─conditions under which the peptide can adopt an α-helical conformation. The introduction of a hydrocarbon staple induced and stabilized α-helicity; accordingly, stapled peptide P9 displayed significantly improved bactericidal effects against both Gram-positive and Gram-negative bacteria in media containing physiologic levels of salt. Together, our findings identify and characterize the antimicrobial regions of CXCL10 and functionalize these novel determinants as discrete peptides with potential therapeutic utility against difficult-to-treat pathogens.

Discovery of Novel N-Heterocyclic-Fused Deoxypodophyllotoxin Analogues as Tubulin Polymerization Inhibitors Targeting the Colchicine-Binding Site for Cancer Treatment.

Natural products are a major source of anticancer agents and play critical roles in anticancer drug development. Inspired by the complexity-to-diversity strategy, novel deoxypodophyllotoxin (DPT) analogues were designed and synthesized. Among them, compound C3 exhibited the potent antiproliferative activity against four human cancer cell lines with IC50 values in the low nanomolar range. Additionally, it showed marked activity against paclitaxel-resistant MCF-7 cells and A549 cells. Moreover, compound C3 can inhibit tubulin polymerization by targeting the colchicine-binding site of tubulin. Further study revealed that compound C3 could arrest cancer cells in the G2/M phase and disrupt the angiogenesis in human umbilical vein endothelial cells. Meanwhile, C3 remarkably inhibited cancer cell motility and migration, as well as considerably inhibited tumor growth in MCF-7 and MCF-7/TxR xenograft model without obvious toxicity. Collectively, these results indicated that compound C3 may be a promising tubulin polymerization inhibitor development for cancer treatment.

Potential role of dietary parsley and/or parsley nanoparticles against zinc oxide nanoparticles toxicity induced physiological, and histological alterations in Nile tilapia, Oreochromis niloticus

In the recent study, Nile tilapia, Oreochromis niloticus was exposed to sublethal concentration of Zinc Oxide nanoparticles (ZnO-NPs) (8 mg/L) to study the toxic influence on biochemical and oxidative stress biomarkers. Also, the protective role of dietary parsley leaves and parsley nanoparticles was evaluated. Fish were divided into 6 groups of 3 replicates as follows: group I: was served as control, group II: was fed free basal diet containing parsley leaves (4 g/kg of diet). group III: was fed diet containing parsley nanoparticles (4 g/kg of diet). group IV: exposed to (8 mg/L) ZnO-NPs and fed on basel diet for 6 weeks. group V: exposed to (8 mg/L) ZnO-NPs and fed on parsley leaves (4 g/kg of diet). group VI: exposed to (8 mg/L) ZnO-NPs and fed on parsley nanoparticles (4 g/kg diet) for 6 weeks. At the end of experiment, blood and liver tissues were collected for biochemical analysis and histopathological investigation. Results exhibited significant (p < 0.05) increase in the levels of urea, creatinine, total protein, albumin, globulin, glucose, cortisol, cholesterol, and IgM during ZnO-NPs exposure for 6 weeks. Concurrent with those changes, marked decrease (p < 0.05) in serum triglycerides and IgG levels were observed. On the other hand, the histopathological alterations of the examined liver of tilapia intoxicated with ZnO-NPs showed severe congestion, necrosis and apoptosis of hepatic cells, and marked activation of kupffer cells with mononuclear inflammatory cellular infiltration mainly lymphocytes were prevalent. Intake of parsley and parsley nanoparticles in groups (V & VI) after exposure to ZnO –NPs ameliorated hyperglycemia, total protein and lipid profiles, and blunted the increase in cortisol and modulated the levels of urea, creatinine, and IgM of ZnO –NPs treated fish, and enhanced the hepatic architecture of tilapia fish fed on parsley and nano parsley. The obtained results revealed that the dietary parsley leaves and parsley nanoparticles reduced the influence of ZnO-NPs toxicity on Nile tilapia which was more evidence in fish fed on parsley nanoparticles. It could be concluded that parsley has a hepatoprotective effect in tilapia fish against ZnO-NPs toxicity.

An exopolysaccharide from Leuconostoc mesenteroides showing interesting bioactivities versus foodborne microbial targets

An exopolysaccharide (EPS_B3) produced by a Leuconostoc mesenteroides strain isolated from a semi-hard Italian cheese was chemically and biologically characterized. HPLC-SEC, NMR, FT-IR and monosaccharide composition experiments were performed. Antimicrobial, antibiofilm, bifidogenic, antioxidant, and DNA-protective activity of EPS_B3 were also studied. Results revealed that EPS_B3 was a mixture of two high-molecular-weight dextran with low branching degree. Moreover, EPS_B3 displayed significant antibacterial activity against eight foodborne pathogens and inhibited biofilm formation by Listeria monocytogenes. EPS_B3 also evidenced bifidogenic activity, stimulating the growth of three probiotic bifidobacteria, and improving the tolerance of Bifidobacterium animalis subsp. lactis to oxygen stress. It also protected plasmid DNA from hydrogen peroxide damage. Only limited antioxidant capacity was observed.In conclusion, data suggest that EPS_B3 might be exploited in the context of functional foods especially for its marked antimicrobial activity as well as for the ability to improve the viability of bifidobacteria in probiotic foods. However, further studies should be carried out to assess the ability of EPS_B3 to reach intact the target site (i.e., gastrointestinal tract) to consider the possibility of use it as a new functional ingredient in foods.

Design, synthesis and antitumor evaluation of novel pyrazolo[3,4-d]pyrimidines incorporating different amino acid conjugates as potential DHFR inhibitors

The present study aimed to investigate the antitumor effect of simultaneous inhibition of dihydrofolate reductase (DHFR) enzyme. We designed some novel pyrazolo[3,4-d]pyrimidines bearing different amino acid conjugates as efficient antifolate agents attributable to their structural similarity with methotrexate (MTX) and MTX-related antifolates. All compounds were tested to screen their enzymatic inhibition against DHFR compared with the reference drug MTX and for their in vitro antitumor cytotoxicity against six MTX-resistant cancer cell lines. The flow cytometry indicated that the most potent compound 7f arrested MCF-7 cells in the S-phase and induced apoptosis. Western blot for visualisation proved the ability of compound 7f to induce the expression of proapoptotic caspases and Bax proteins in MCF-7 breast cancer cell line beside its ability to diminish the expression of antiapoptotic Bcl-2 protein. Molecular modelling studies concluded that compound 7f displayed better binding energy than that of the normal ligand MTX.

Sub-lethal doses of imidacloprid alter food selection in the invasive garden ant Lasius neglectus

Despite several restrictions to their use, neonicotinoid insecticides are still widely employed worldwide. Residual sub-lethal amounts of these chemicals can have detrimental effects on the behavior of non-target insects. Toxic effects on economically important species such as bees have been widely documented, but less is known about their toxic action on other social insects, such as ants. In this study, we assessed the effect of different sub-lethal doses of the neonicotinoid imidacloprid on the ability of colonies of the invasive ant Lasius neglectus to select the most profitable resource. We used Y-shaped mazes having an imidacloprid-polluted or an unpolluted sucrose solution on the two branches. Two sucrose (0.1 M, 0.5 M) and two imidacloprid (1 μg/ml, 10 μg/ml) concentrations were used. In parallel, we evaluated the marking activity of foragers who fed on the same solutions. We found that the 0.1 M sugar solution polluted with 1 μg/ml imidacloprid was significantly more frequently selected in binary choices experiments than the unpolluted resource. Moreover, the ingestion of the same combination of sugar and imidacloprid significantly increased the marking rate of foragers. The higher concentration of the pollutant had lower effects, probably because of the hormesis phenomenon. Results suggest that the lower sub-lethal dose of imidacloprid can lead ants to select again the polluted resource. This “active” selection of the pollutant may magnify the negative effects on the colonies. Due to their ecological role, any impairment of ant survival or behavior may have detrimental cascade effects on the whole ecosystem.

A Novel Spirocyclic Dimer (SpiD3) Resensitizes CLL Cells to Venetoclax

Chronic lymphocytic leukemia (CLL) is an incurable, heterogeneous B-cell malignancy with a dependence on B-cell receptor signaling and subsequent activation of nuclear factor-kappa B (NF-κB) for growth and survival. Almost all CLL cells evade apoptosis primarily through overexpressing the B-cell lymphoma/leukemia 2 (BCL2) protein, making it an attractive therapeutic target. Venetoclax (VEN), a BH3-mimetic that selectively inhibits BCL2, is highly cytotoxic to CLL cells and has shown high response rates in refractory/relapsed (R/R) CLL. Despite the clinical success of VEN in CLL, a subset of patients (~25%) fail to achieve minimal residual disease and succumb to progressive disease and/or drug resistance. Factors contributing to VEN resistance have been reported such as mutations in BCL2 (e.g., BCL2G101V), marked NF-κB pathway activation, and increased expression of other anti-apoptotic BCL2 family proteins (e.g., MCL1, BCL-xL and BFL-1) that are not targeted by VEN. This indicates the need for novel therapeutic approaches for R/R CLL that bypass resistance mechanisms and/or target alternative B-cell survival pathways. We therefore hypothesize that targeting NF-κB signaling would augment the efficacy of VEN-based regimens and/or mitigate resistance to VEN in CLL. To investigate this, we used a novel spirocyclic dimer, SpiD3, that covalently binds to NF-κB proteins, IKKβ and RELA/P65 (Rana S, etal, 2022). We previously established the anti-leukemic properties of SpiD3, reporting the IC50 below 1 μM in CLL cell lines (Eiken A, et al, 2021). To evaluate if SpiD3 can potentially enhance VEN cytotoxicity in CLL, we conducted synergy assays in CLL cell lines (HG3 and OSUCLL). Cell proliferation was measured using the MTS assay following a 72-hour treatment with increasing concentrations of SpiD3 and/or VEN. Consistent with our hypothesis, 0.5 μM SpiD3 showed strong synergy with 0.25 μM VEN in HG3 cells and with 0.125 μM VEN in OSUCLL , which is less than the reported plasma concentration of VEN in CLL patients (0.79 - 1.14 μM). Since, NF-κB activation is implicated in VEN-resistance, we next evaluated SpiD3 in a preclinical CLL model of acquired VEN resistance. We developed VEN resistance in OSUCLL cells through prolonged exposure to increasing concentrations of VEN until the cells remained viable in the presence of 22 μM VEN. Remarkably, SpiD3 elicited robust cytotoxicity in the VEN-resistant OSUCLL cells with an IC50 of ~1 μM (Figure 1). Treatment of VEN-resistant OSUCLL cells with SpiD3 for 4 hours resulted in reduced expression of anti-apoptotic proteins (MCL1 and BFL-1) and increased levels of the pro-apoptotic protein, PUMA. This was coupled with marked PARP cleavage. In contrast to VEN, SpiD3 reduced P65 and MYC expression and attenuated phosphorylation of ERK (Thr202/Tyr204) and PRAS (Thr246), all factors known to contribute to VEN resistance. Further interrogation of differential gene expression following SpiD3 treatment in VEN-resistant cells is ongoing to delineate the molecular mechanism underlying SpiD3's anti-tumor properties in VEN-resistant CLL. In summary, our data (a) supports the efficacious combination of SpiD3 with VEN as a viable therapeutic approach in CLL to bypass NF-κB activation underlying VEN resistance and (b) introduces SpiD3 as an alternative therapeutic approach for VEN-resistant CLL patients. Figure 1View largeDownload PPTFigure 1View largeDownload PPT Close modal

Phenolic content and antioxidant activity of hydromethanolic and aqueous extracts of aerial parts of Phlomis crinita.

Phlomis crinita is a plant species of the family Lamiaceae including more than 100 perennial herbs, shrubs, and sub-shrubs species native to the Mediterranean, Central Asia, and India. This species is commonly a good natural source of various secondary metabolites. Therefore, the present study was conducted to determine phenolic content and antioxidant activity of hydromethanolic (PC ME) and aqueous extracts (PC AQE) of aerial parts of P. crinita. Total polyphenols, flavonoids and tannins were quantified, respectively by the methods of Folin-Ciocalteu reagent, aluminum trichloride (AlCl3) and Bate-Smith method. The in vitro antioxidant activity was assessed using DPPH and ABTS•+ radical scavenging, β-carotene-linoleic acid, reducing power and ferrous ion chelating activity assays. PC ME showed high level of tannins (132,13 ±0.68 µg TAE/mg extract) and total phenolic content (82.71±0.79 µg GAE/mg extract), in addition a marked inhibiting oxidation activity of β-carotene/ linoleic acid (74.10%) was observed. Results showed also a higher iron-chelating activity of PC ME (0.20 mg/mL) compared to PC AQE (0.046 mg/mL). The plant extracts revealed a significant antioxidant activity as evidenced by the DPPH and ABTS radical scavenging activity (IC50 = 0.103 mg/mL for PC ME and 0.144 mg/mL for PC AQE) for DPPH assay and (IC50 = 0.0130 mg/mL for PC ME and 0.0187 mg/mL for PC AQE) , as well as the PC ME exhibits higher reducing power (IC50 =0.288mg/mL) than PC AQE (0.296 mg/mL). As a result, P.crinita is suggested as a promising and effective therapeutic medicinal plant for the treatment of several diseases.

The total phenol, flavonoid content and antioxidant activity of Zehneria maysorensis stem and leaf extracts

Zehneria species have enormous ethno-botanical value and are used by different tribes for food as wild edible plants and treatment of various ailments. The present study was designed to carry out the preliminary phytochemical screening, total phenol, flavonoid quantification followed by an investigation of the in vitro antioxidant activities of Zehneria maysorensis. Phytochemical analysis showed the presence of saponins, steroids, alkaloids, phenols and tannins. Ethanol extracts of stem and leaf showed maximum phenol and flavonoid content. The extract also confirmed DPPH radical scavenging activity. A marked dose dependent antioxidant activity was observed in the trials.

Determining the potential use of biosurfactants in preventing endodontic infections.

Microbial biofilms play a dominant role in the failure of endodontic therapies. Bacterial adhesion is the first step in the establishment of biofilms, activating the host immune response leading to tissue damage. Biosurfactants are microbe-derived tensioactive molecules with latent anti-adhesive and anti-microbial activity. This study reports the extraction and characterization of a biosurfactant from Lactobacillus (L.) plantarum (Lp-BS) and investigates its anti-microbial and anti-adhesive properties compared to rhamnolipid, a commercially available biosurfactant. Lp-BS, extracted from L. plantarum during the growth phase, was characterized as a glycoprotein, able to reduce surface tension and emulsify non-polar liquids. Proteomic analysis of Lp-BS identified three bacterial adhesin-like proteins, suggesting roles in hindering bacterial adhesion. Lp-BS did not show significant anti-microbial activity against endodontic pathogens from the Streptococcus (Strep.) anginosus group or Enterococcus (Ent.) faecalis at 50mg/ml. However, anti-adhesive activity on abiotic surfaces was observed against both Strep. anginosus and Strep. intermedius. Rhamnolipid exhibited strong anti-microbial activity, with minimum inhibitory concentrations of 0.097mg/ml against Strep. anginosus, and 0.048mg/ml against Strep. constellatus and Strep. intermedius, in addition to a marked anti-adhesive activity. These findings offer preliminary evidence for the potential application of biosurfactants as an anti-microbial and/or anti-adhesive pharmacotherapy in endodontics.

Spatio-temporal patterns of the European wildcat in a Mediterranean coastal area

Like most small felids, the European wildcat Felis silvestris is a rather elusive species, poorly detectable in the wild, due to several aspects of its biology. Camera trapping can represent a suitable tool to understand temporal activity patterns and habitat preferences of such elusive species. We used intensive camera trapping over two full years to investigate spatio-temporal patterns of the wildcat in a Mediterranean coastal protected area (Maremma Regional Park, central Italy). At the seasonal scale, the wildcat showed a marked twilight activity in summer and winter (mating period), and nocturnal activity in autumn. Conversely, a peak of activity during the day, i.e., in broad daylight, was reported in spring. Reproductive females may limit their nocturnal movements in spring to increase protection from predators to their kittens at the den, although further data are required to support this conclusion. At seasonal, semestral, and yearly temporal scales, the frequency of wildcat detections increased along with availability of shrubwood. These results emphasise the importance for this small felid of areas with dense vegetation cover (Mediterranean maquis and shrubwood, in our study area). Areas densely covered with shrubby vegetation are expected to provide benefits to this elusive small cat in terms of reduced human disturbance (included tourists), availability of prey (e.g., small mammals), as well as shelter, essential to ensure protection towards potential larger predators.

The antibacterial and antibiofilm activity of Granudacyn in vitro in a 3D collagen wound infection model.

It is widely agreed that infection and the formation of biofilms play a major role in increasing inflammation and delaying wound healing. The aim of this study was to evaluate, in vitro, the antimicrobial activity of the wound irrigation solution, Granudacyn (Mölnlycke Health Care AB, Sweden) against planktonic bacteria and mature biofilms of clinically relevant bacterial species. Quantitative evaluation of bacterial numbers and confocal and/or scanning electron microscopy were used to evaluate the wound irrigation solution's antimicrobial/antibiofilm activity in standard laboratory conditions as well as in a three-dimensional (3D) collagen wound infection model. The wound irrigation solution exhibited a rapid and strong antibacterial activity against both Gram-positive and Gram-negative strains isolated from infected wounds in planktonic form, with a reduction in bacterial number of >4 Logs after as little as one minute of treatment. The wound irrigation solution also exerted an evident activity against preformed biofilms of Pseudomonas aeruginosa and Staphylococcus aureus (>3 Log and >1 Log reduction in colony forming unit number, respectively, after 15 minutes of incubation). Although the wound irrigation solution was partially inhibited in the presence of simulated wound fluid, it maintained a marked antibiofilm activity in in vivo-like conditions (ie. in a 3D collagen wound infection model) with a strong killing and a mild debridement effect, which was superior to standard saline. The results obtained in this study suggest that although the wound irrigation solution used might be partially inhibited by wound exudate, it has the potential to effectively kill wound infecting planktonic as well as biofilm bacteria.