Current Pharmaceutical Biotechnology - Volume 19, Issue 12, 2018

Thymoquinone (TQ), derived from the seeds of Nigella sativa, has lately been shown as a miracle drug because of its wide range of therapeutic effects against various diseases, including cancer, asthma, diabetes, colitis and infectious diseases. In the present review, we aimed to decipher the molecular mechanisms of therapeutic action of TQ by modulating the cell signaling pathways. Many in vivo and in vitro studies have demonstrated the therapeutic efficacy of TQ against a wide range of ailments. TQ possesses potent anti-inflammatory and immunomodulatory effects by specifically targeting the NF-kB, IL-1β and TNF-α signaling pathways. The anticancer activity of TQ has been primarily shown by altering the expression of signal transducers and activator transcription (STAT3), PTEN and p53 genes. TQ alleviates the hyperglycemia-associated complications, the hepatic or renal ailments through its potent antioxidant and anti-inflammatory properties. Interestingly, the liposome- or nanoparticle-based TQ formulations have shown greater effectiveness against various diseases in animal models. Thus, the understanding of the molecular mechanisms of TQ action may lead to the development of its therapeutic formulations to cure a wide variety of diseases.

This review article presents the concepts of aggregates, coaggregates, and a liquid droplet of proteins and compares the concentrated states in the presence of small additives to control their formation and dissociation. The aggregates composed of single protein molecules result mainly from hydrophobic interactions between unfolded protein molecules. Thus, the aggregation of protein can be effectively suppressed by small additives that increase the solubility of hydrophobic solutes, typically arginine (Arg) and chaotropes. In contrast, coaggregation that is composed of two or more types of proteins results from both hydrophobic and attractive electrostatic interactions between even partially unfolded protein molecules. Accordingly, coaggregation is more controllable than simple aggregation using various types of small additives, such as ions and osmolytes, as well as Arg and chaotropes. The liquid droplets of proteins observed in living cells and a protein-polyelectrolyte complex (PPC) undergo liquid-liquid phase separation driven only by electrostatic interactions. Thus, the liquid droplets and PPCs are redissolved when the concentration of ions is increased. The properties of electrostatic or hydrophobic interactions, solid-like or liquid-like states, and apparently spherical and amorphous structures are simple but valuable criteria that can be used to control protein aggregation and condensation using small additives.

Objective: To identify the clinical and adverse effects of intravenous chemotherapy combined with intraperitoneal chemotherapy in the treatment of gastric cancer with abdominal peritoneal carcinomatosis. Methods: Retrospective analysis was performed on 49 gastric cancer patients with abdominal peritoneal carcinomatosis. Patients were divided into two groups: patients subjected to intravenous chemotherapy (n = 27) were defined as the Control group and patients subjected to combined intravenous and intraperitoneal chemotherapy (n = 22) were defined as the Combination Therapy group. Then the objective efficacy, survival, and adverse effects of two groups were evaluated. Results: The clinical characteristics of two groups were equally distributed. Compared with the Control group, the Objective Response Rate (ORR) and Disease Control Rate (DCR) of the Combination Therapy group were significantly higher (all P<0.05). The Median Survival Time (MST) of the Control group was (6.57 ± 0.75) months, which was significantly shorter than in the Combination Therapy group (15.03 ± 2.31) months (P < 0.05). In addition, the 12-, 18-, 24-, 30-, 36-, and 60-month survival rates of the Control group were all significantly lower than those of the Combination Therapy group. Furthermore, the incidence of adverse reactions in the two groups was not statistically significant; neither of the treatments resulted in severe complications. Conclusion: Compared with intravenous chemotherapy alone, the combined intravenous and intraperitoneal chemotherapy was more effective in improving quality of life and extending survival time in patients with abdominal peritoneal carcinomatosis from gastric cancer. In addition, the combination treatment is tolerable and therefore worth further clinical study.

Background: The local therapy is a particularly proper approach in treatment of cancer diseases, at which the drugs are highly toxic. Unfortunately, antitumor drugs diffusion is quite often limited by hyaluronic acid layers abundantly found in a cancer tissue. Methods: The undertaken approach assumed relaxation of hyaluronan layer by using a hydrolytic enzyme - hyaluronidase. The considered process corresponds to the heterogenic catalysis with substrate occurred as a layer and a catalyst in a native form. It was shown that enzyme concentration around 4 mg/L kept for 10 minutes in body fluid surrounding HA layers is enough to allow the drug to diffuse. Results: Cyanocobalamin, as a model drug, was applied in diffusion processes testing. The possibility of hyaluronidase releasing in the place close to hyaluronic acid layers was also considered. Hydrogel carriers promoting fast release of hyaluronidase were characterized. The profile of releasing Hase from the tested carriers based on Κ-carrageenan or sodium alginate with polyvinyl alcohol corresponded to Higuchi model. Conclusion: An expected rate value (the mass transport coefficient app. 4.0.10-7 m/s) was obtained for blends 0.5% Κ-carrageenan, 4% polyvinyl alcohol and 0.5% sodium alginate, 6% polyvinyl alcohol.

Background: Trichilia silvatica, popularly known as “catiguá-branco”, is distributed in Brazil (Mato Grosso do Sul), and members of this genus are commonly used for the treatment of rheumatism (arthritis). The aim of this research was to investigate the anti-inflammatory, antioxidant and antiproliferative activities of the methanolic extract of the leaves (MEL-TS) and bark (MEB-TS) of T. silvatica. We also evaluated the concentration of phenolic compounds, flavonoids, flavonol, and condensed tannins by liquid chromatography - photodiode array (LC/PDA) analysis. Methods: The MEL-TS and MEB-TS revealed the presence of caffeic acid in both extracts by LC/PDA. The samples were evaluated for antioxidant activity using free-radical scavenging and lipoperoxidation assays. The anti-inflammatory effects were studied in carrageenan-induced paw edema, pleurisy and zymosan-induced arthritis. Results: The MEL-TS and MEB-TS showed the total phenolic concentration (270.8 ± 17.10 mg gallic acid equivalents GAE/g extract and 278.8 ± 25.13 mg GAE/ g extract, respectively), and flavonoids in MEL-TS (209.30 ± 2.91 mg quercetin equivalents QE/ g extract). In the lipoperoxidation assay, exhibited moderate antioxidant activity with IC50 values ≤ 35.32 μg/mL. Both extracts inhibited oedema induced by carrageenan at 2 h and 4 h, inhibited leukocyte migration at 6 h post administration, and did not impact zymosan-induced arthritis. Finally, MEL-TS was particularly effective against prostate cell line (GI50 ≤ 0.22 μg/mL). Conclusion: Overall, the results indicated that T. silvatica reduce migration leukocytes activity, edema formation in these models of experimental arthritis could explain the popular use for treatment of inflammatory processes (rheumatism).

Background: Over the last years, a new generation of oral anticoagulants (NOACs), including apixaban, rivaroxaban and dabigatran, has been developed for the control of thrombosis and related disorders. The presence of food within the gastrointestinal (GI) tract can potentially affect the oral bioavailability of drugs. Objective: In the present paper, we evaluated the stability of these drugs in in vitro GI digestion, with and without the main macronutrients such as proteins, lipids, carbohydrates, and fibers, and their ability to enter into the systemic circulation. In addition, we examined the percentage of the drug binding to plasma proteins, such as human serum albumin, high density- and low density lipoproteins. Methods: The NOACs bioaccessibility was evaluated by an in vitro procedure simulating the gastrointestinal enzymatic system, while their bioavailability was studied by cell culture of Caco-2 cells and in vitro study of transepithelial transport. The in vitro transepithelial permeated NOACs were added to plasma protein solutions simulating the average fasting plasma protein concentrations. The NOACs detection was carried out by HPLC-DAD/ESI-MS analysis. Results: GI digestion significantly reduces intestinal bioaccessibility and bioavailability of NOACs, especially as regards apixaban and dabigatran. Interestingly, the co-digestion of fibers led to a strong decrease of NOAC intestinal bioaccessibility and bioavailability, while the effects of the other macronutrients, as well as a low fiber standard meal, had no significant influence in this sense. Conclusion: Dabigatran, rivaroxaban and apixaban may be administered independently of a standard meal, provided that it does not include a high amount of dietary fibers.

Background: Endoglin is a marker of active, proliferating endothelial cells of blood vessels. In many cancers, it is present in both peripheral vessels and vessels located inside the tumor. Endoglin is more specific and sensitive compared to other tumor angiogenesis markers. It is suggested that endoglin can be considered a reliable marker of disease outcome. Objective: The aim of the study was to assess the expression of endoglin and to determine its potential usefulness as a complementary molecular marker of endometrial cancer. Method: The study included 60 women who underwent hysterectomy: 45 with endometrioid endometrial cancer (study group) and 15 without neoplastic changes (control group). The study group was further divided according to the degree of histological differentiation: G1, 17; G2, 15; and G3, 13. The expression of endoglin was determined immunohistochemically with mouse anti-Endoglin monoclonal antibody. The obtained reactions were evaluated using light microscopy. Results: Analysis of endoglin expression in endothelium showed that it reached 145% of the control. In G2, we observed that the endoglin level decreased and was similar to the control, while in G3 it increased and was even higher than in G1. In cancer cells, endoglin expression increased with the grade of endometrial cancer. Conclusion: Endoglin can be considered a valuable complementary molecular marker, allowing to visualize the advancement of the cancer process, including endometrial cancer.

Background: Endostatin (ES) is a promising anti-angiogenesis protein and has been approved for the treatment of non-small cell lung cancer, but short half-life, poor stability and nonspecific delivery caused great pain to patients and produced unsatisfactory treatment effectiveness. Objective: In this work, in order to overcome these disadvantages, ES was covalently modified by polysulfated heparin (PSH) with the expectancy of longer half-life, higher anti-angiogenesis activity and better cellular uptake. Methods: To characterize the cellular uptake, flow cytometry and confocal laser scanning microscopy were used to study the intracellular localization of fluorescein isothiocyanate-labeled ES and PSH-ES in EAhy926 endothelial cells. Zebrafish model was used to study the anti-angiogenesis activities of ES and its derivatives in vivo. The 125I-radiolabeled ES and PSH-ES were administered to healthy BALC/c mice for the pharmacokinetics study. Results: Compared with ES, better cellular uptake effects were detected in PSH-ES group. Both ES and PSH-ES showed inhibition on the intersegmental vessels formation, while PSH-ES displayed a higher one. The half-life of PSH-ES was lengthened and area under the curve (AUC) was increased. At the same time, ES and PSH-ES were both widely and rapidly distributed in the lungs, livers, kidneys and hearts with little difference. Conclusion: The results indicated that PSH displayed good properties as a novel glyco-modifier for protein and peptide. The results also showed that PSH-ES displayed better cellular uptake, higher antiangiogenesis activity and prolonged half-life, which would lead to better anti-tumour effects.

Background: The further functionalization of natural existing biomaterials is a very efficient method to introduce additional advanced characteristics on a unique structural composition and architecture. Objective: As an example, different animal sources, if properly treated, can be used to develop bone xenograft active in hard tissues regeneration. In this sense, it is also important to consider that the selected process has to take into consideration the intrinsic variability of the base material itself and possibly being able to compensate for it. Methods: In this work we characterize cancellous bovine bone treated by deposition of polymer and collagen and we show that the added components not only lead to a more resistant and more hydrophilic material, but also reduce the conventional correlation between apparent density and elastic modulus, which, in general, is a major source of uncertainty and risk in xenografts usage. Results: Moreover, though intrinsically reinforcing the material, the deposition process leaves the specific open-porous structure, that allows cells proliferation and vessels ingrowth, basically unaltered. Conclusion: The final material combines in a single piece and at the same time, mechanical resistance, homogeneous mechanical response and proper structural characteristics that allow further integration within the patient autochthonous tissues.