Luigi Inguglia

Curcumin as a Possible Lead Compound against Hormone‐Independent, Multidrug‐Resistant Breast Cancer

We examine the possible evidence that the phytochemical curcumin may overcome resistance to hormonal and cytotoxic agents in breast cancer. We present our observations on MCF‐7R, a multidrug‐resistant (MDR) variant of the MCF‐7 breast cancer cell line. In contrast to MCF‐7, MCF‐7R lacks aromatase and estrogen receptor α (ERα) and overexpresses the multidrug transporter ABCB1 and the products of different genes implicated in cell proliferation and survival, like c‐IAP‐1, NAIP, survivin, and COX‐2. Nevertheless, in cytotoxicity and cell death induction assays, we found that the antitumor activity of curcumin is substantial both in MCF‐7 and in MCF‐7R. We elaborated the diketone system of curcumin into different analogues; the benzyloxime and the isoxazole and pyrazole heterocycles showed remarkable increases in the antitumor potency both in the parental and in the MDR MCF‐7 cells. Furthermore, curcumin or, more potently, the isoxazole analogue, produced early reductions in the amounts of relevant gene transcripts that were diverse (i.e., they were relative to Bcl‐2 and Bcl‐XL in MCF‐7 and the inhibitory of apoptosis proteins and COX‐2 in MCF‐7R) in the two cell lines. Thus, the two compounds exhibited the remarkable property of being able to modify their molecular activities according to the distinct characteristics of the parental and MDR cells. We discuss also how curcumin may (1) exert antitumor effects in breast cancer through ER‐dependent and ER‐independent mechanisms; and (2) act as a drug transporter‐mediated MDR reversal agent. Overall, the structure of curcumin may represent the basis for the development of new, effective anticancer agents in hormone‐independent MDR breast cancer.

Antimicrobial and Antibiofilm Activity of a Recombinant Fragment of β-Thymosin of Sea Urchin Paracentrotus lividus

With the aim to obtain new antimicrobials against important pathogens such as Staphylococcus aureus and Pseudomonas aeruginosa, we focused on antimicrobial peptides (AMPs) from Echinoderms. An example of such peptides is Paracentrin 1 (SP1), a chemically synthesised peptide fragment of a sea urchin thymosin. In the present paper, we report on the biological activity of a Paracentrin 1 derivative obtained by recombination. The recombinant paracentrin RP1, in comparison to the synthetic SP1, is 22 amino acids longer and it was considerably more active against the planktonic forms of S. aureus ATCC 25923 and P. aeruginosa ATCC 15442 at concentrations of 50 µg/mL. Moreover, it was able to inhibit biofilm formation of staphylococcal and P. aeruginosa strains at concentrations equal to 5.0 and 10.7 µg/mL, respectively. Molecular dynamics (MD) simulations allowed to rationalise the results of the experimental investigations, providing atomistic insights on the binding of RP1 toward models of mammalian and bacterial cell membranes. Overall, the results obtained point out that RP1 shows a remarkable preference for bacterial membranes, in excellent agreement with the antibacterial activity, highlighting the promising potential of using the tested peptide as a template for the development of novel antimicrobial agents.

Cytotoxic activity of Holothuria tubulosa (Echinodermata) coelomocytes

ABSTRACT The immune system of marine invertebrates, in particular that of holothurians, still requires further study. Our research showed that coelomocyte cells contained in the coelomic fluid of the sea cucumber, Holothuria tubulosa, are able to lyse, in vitro, red blood cells in rabbits and sheep. A plaque‐forming assay showed spherule cells to be the effector cells, able to release cytotoxic molecules after xenogenic cell contact. The coelomocyte lysate supernatant, analysed by polyacrylamide gel electrophoresis overlay technique, using rabbit and sheep erythrocytes, showed two different haemolytic protein patterns: one calcium dependent and the other calcium independent. The fractions of each pattern were resolved on a polyacrylamide gel and calcium‐dependent and independent coelomocyte lysate patterns were compared. HighlightsCoelomocytes of Holothuria tubulosa, are able to lyse, in vitro, red blood cells in rabbits and sheep.Coelomocytes are able to release cytotoxic molecules after xenogenic cell contact causing plaque lysis.Cytotoxic molecules are proteins showing a different lysis pattern in presence or in absence of calcium ions.

Antibacterial Activity of Desert Truffles from Saudi Arabia Against Staphylococcus aureus and Pseudomonas aeruginosa

Medicinal mushrooms represent an unlimited source of polysaccharides with nutritional, antitumoral, antibacterial, and immune-stimulating properties. Traditional studies of epigeous higher Basidiomycetes have recently been joined by studies of hypogeous fungi and, in particular, of so-called desert truffles. With the aim to obtain novel agents against bacteria of clinical importance, we focused on the edible desert truffle mushrooms Tirmania pinoyi, Terfezia claveryi, and Picoa juniperi as sources of new antimicrobial agents. In particular, we investigated the in vitro antibacterial activity of acid-soluble protein extracts (aqueous extracts) of these 3 species against the Gram-positive human pathogenic reference strain Staphylococcus aureus ATCC 29213 and the Gram-negative strain Pseudomonas aeruginosa ATCC 15442. The acid-soluble protein extracts of T. pinoyi and T. claveryi showed minimum inhibitory concentrations of 50 μg/mL against tested pathogens. We believe that such preliminary results are promising to obtain a valuable antibiotic alternative to fight antibiotic-resistant pathogens.

Anesthetic efficacy of ketamine–diazepam, ketamine–xylazine, and ketamine–acepromazine in Caspian Pond turtles (Mauremys caspica)

Objectives: The objective of this study was to assess the efficacy of different anesthetic drug combinations on the Caspian Pond turtles (Mauremys caspica). Subjects and Methods: Three groups of the Caspian Pond turtles (n = 6) were anesthetized with three different drug combinations. Initially, a pilot study was conducted to determine the best drug doses for the anesthetization of the turtles, and according to these results, ketamine–diazepam (120 mg/kg ketamine hydrochloride [5%] and 2 mg/kg diazepam [5%]), ketamine–acepromazine (120 mg/kg ketamine hydrochloride [5%] and 1 mg/kg acepromazine [1%]), and ketamine–xylazine (120 mg/kg ketamine hydrochloride [5%] and 1 mg/kg xylazine [2%]) were injected intramuscularly. The onset times of anesthetization and the recovery time were measured. Statistical analysis of the data was performed using one-way analysis of variance followed by t-tests, and P < 0.05 was considered statistically significant. Results: There were statistically significant differences in the mean of the onset times of anesthesia and recovery time among the three drug combinations depending on the treatment used. The onset of anesthesia of the animals treated with the ketamine–diazepam combination was 60% and 42% shorter, for male and female turtles, respectively, compared to that obtained with the ketamine–acepromazine combination and 64% (male turtles) and 50% (female turtles) shorter than that obtained with the ketamine–xylazine combination. Further, the recovery time, in male turtles, was 17% shorter in animals treated with the first drug combination than those treated with the ketamine–acepromazine combination and 37% shorter than those treated with the ketamine–xylazine combination. The recovery time, in female turtles, did not seem to be significantly different among treatments. Conclusions: The study showed that the ketamine–diazepam drug combination is the anesthetic combination with the fastest onset time and shortest recovery time.

Antimicrobial Activity of the Extracts of Terfezia Claveryi and Tirmania Pinoyi Against Gram-positive and Gram-negative Bacteria Causal Agent of Diseases in Tomato

Antimicrobial Activity of the Extracts of Terfezia claveryi and Tirmania pinoyi Against Gram-positive and Gram-negative Bacteria Causal Agent of Diseases in Tomato Maria Letizia Gargano , Patrizia Bella , Stefano Panno, Vincenzo Arizza, Luigi Inguglia, Vittoria Catara, Giuseppe Venturella, Salvatore Davino* Department of Agricultural and Forest Science (SAF), University of Palermo, Viale delle Scienze Bld. 5, 90123 Palermo (Italy) Department of Biological, Biochemistry and Pharmacological Science and Technology (STEBICEF), Via Archirafi 18-28, 90122 Palermo, (Italy) Department of Agricultural, Food and Environment (Di3A), Via Santa Sofia 100, 91123 Catania, (Italy) salvatore.davino@unipa.it # These authors contributed equally to the study