The electrical pulse application enhances intra-cellular localization and potentiates cytotoxicity of curcumin in breast cancer cells.

Breast cancer is the most common cancer of women, and fifth leading cause of mortality worldwide. Existing breast cancer regimens are costly and produce severe side effects. This highlights a need for the development of efficient novel therapies, which are cost effective and limit side effects. An electrical pulse (EP)-based chemo therapy, known as electrochemotherapy (ECT) using the natural compound curcumin could be an effective alternative. ECT is a non-surgical modality, which produces excellent anti-tumor efficacy at small drug concentrations due to increased uptake of drugs. In clinics, ECT is shown to be effective in treating advanced, recurrent, and metastatic breast cancers, which are refractory to multiple modalities. ECT with curcumin triggers apoptotic cell death in breast cancer cells and could be an effective alternative, due to curcumin's low cost and reduced side-effects. However, there is a lack of studies quantifying the uptake of curcumin in response to EP application. Towards this, we determined the uptake of different curcuminoids (curcumin, desmethoxycurcumin, and bisdemethoxycurcumin) upon EP application and their impact on cell cytotoxicity. Additionally, we studied the combined effect of calcium chloride (CaCl2) and a curcuminoids (Cur) mixture, based on initial studies suggesting calcium electroporation as a potential inexpensive anti-cancer treatment. Our results indicate EP with Cur increases cellular uptake, cell shrinkage, and cytotoxicity. The EP + Cur resulted in the highest uptake of the bisdemethoxycurcumin. Further, EP also potentiated the cytotoxicity of CaCl2 and of the Cur and CaCl2 combination against breast cancer cells and caused apoptosis. Our preliminary data pave the way to further studies on Cur and CaCl2 combination treating breast cancer.

Efficacy of auditory training using the auditory brainstem response to complex sounds: auditory processing disorder and specific language impairment.

OBJECTIVES: This study verified the efficacy of a formal auditory training (AT) program in children with hearing and language disorders using the auditory brainstem response to complex sounds (c-ABR) with and without background noise. PATIENTS AND METHODS: Thirty children (7-13 years) were distributed into four groups: typical development (TD: n = 7), auditory processing disorder (APD: n = 9) and specific language disorder (SLIa: n = 6, and SLIb: n = 8). All children underwent behavioral assessment of auditory processing and c-ABR with and without background noise. The APD and SLIa groups underwent 8 weeks of formal AT, but all children were reevaluated 12 weeks after the initial assessment. RESULTS: The TD group presented better behavioral performance than the other groups. For c-ABR in silence, no significant differences were observed among groups or assessments, except regarding VA complex measures, which were altered in the APD group. For c-ABR with background noise, however, the APD, SLIa and SLIb groups presented delayed latencies. Groups that underwent formal AT improved behavioral performance and decreased latencies to c-ABR in background noise at final assessment. CONCLUSIONS: These data suggest that efficacy of formal AT can be demonstrated by c-ABR with background noise.

Activity of sap from Croton lechleri on rat vascular and gastric smooth muscles.

The effects of red sap from Croton lechleri (SdD), Euphorbiaceae, on vascular and gastric smooth muscles were investigated. SdD, from 10 to 1000 microg/ml, induced concentration-dependent vasoconstriction in rat caudal arteries, which was endothelium-independent. In arterial preparations pre-constricted by phenylephrine (0.1 microM) or KCl (30 mM), SdD also produced concentration-dependent vasoconstriction. To study the mechanisms implicated in this effect we used selective inhibitors such as prazosin (0.1 microM), an antagonist of alpha(1)-adrenoceptors, atropine (0.1 microM), an antagonist of muscarinic receptors, and ritanserin (50 nM), a 5-HT(2A) antagonist; none of these influenced vasoconstriction caused by SdD. Likewise, nifedipine (50 nM), an inhibitor of L-type calcium channels, did not modify the action of SdD. Capsaicin (100 nM), an agonist of vanilloid receptors, also did not affect vasoconstriction by SdD. We also investigated the action of SdD (10-1000 microg/ml) on rat gastric fundus; per se the sap slightly increased contractile tension. When the gastric fundus was pre-treated with SdD (100 microg/ml) the contraction induced by carbachol (1 microM) was increased, whereas that by KCl (60mM) or capsaicin (100 nM) were unchanged. The data shows that SdD increased contractile tension in a concentration-dependent way, both on vascular and gastric smooth muscles. The vasoconstriction is unrelated to alpha(1), M, 5-HT(2A) and vanilloid receptors as well as L-type calcium channels. SdD increased also contraction by carbachol on rat gastric fundus. Thus for the first time, experimental data provides evidence that sap from C. lechleri owns constricting activity on smooth muscles.

Production of anthocyanins by Catharanthus roseus.

A stable cell suspension line of Catharanthus roseus producing anthocyanin was obtained. In this strain it was found that approximately 30% of cells regularly accumulated these metabolites and that anthocyanin accumulation occurred between the second half of log phase and the stationary phase of the culture growth cycle. The anthocyanins in the suspension cultures were compared with those biosynthetized in the flowers both of regenerated by somatic embryogenesis and field-grown plants. Six anthocyanins were identified in all the examined samples, three 3-O-glucosides and three 3-O-(6-O-p-coumaroyl) glucosides of petunidin, malvidin and hirsutidin. The hirsutidin coumaroyl glucoside has not been reported previously, and was predominat in all samples. The anthocyanin relative content was similar for cell suspensions and flowers from regenerated plants but different from field-grown plant flowers; instead, the total content was almost the same for the two flower types and higher compared to suspension culture content.

In vivo and in vitro production of alkaloids by Haplophyllum patavinum.

A protocol for shoot induction from callus of Haplophyllum patavinum was established. Two known furoquinoline (skimmianine and haplopine), and three quinolone (edulinine, ribalinine and isoplatydesmine) alkaloids were isolated for the first time from plant material, callus and shoot cultures of this species. The structures of these compounds have been characterised on the basis of spectroscopic evidence.

Monitoring the production yields of vincristine and vinblastine in Catharanthus roseus from somatic embryogenesis. Semiquantitative determination by flow-injection electrospray ionization mass spectrometry.

A semiquantitative determination of two bis-indole antitumor alkaloids, vincristine and vinblastine, has been performed by flow-injection electrospray ionization mass spectrometric analysis of the extracts of Catharanthus roseus. Leaves and flowers of two different phenotypes (pink flower and white flower) obtained from somatic embryogenesis were thus examined and compared with the field-grown mother plant. Different amounts of vincristine and vinblastine were detected depending on the examined samples.

Traditional antileukodermic herbal remedies in the Mediterranean area.

More than 100 antileukodermic recipes, involving about 80 plant species used in Italy through the centuries, were collected from Renaissance texts, folk medicine and contemporary herbal usage. The possible efficacy of the recorded plants in stimulating physiological skin pigmentation is discussed in the light of present phytochemical and pharmacological knowledge. Many plants contain erythemogenic substances, some of which (furocoumarins, hypericin) induce light-mediated hypermelanogenesis. Other plants can colour the epidermal keratin (tannins, anthraquinones) or the subcutaneous fat depot (beta-carotene). The usefulness of other plants remains obscure.