Phytocompounds-based therapeutic approach: Investigating curcumin and green tea extracts on MCF-7 breast cancer cell line.

BACKGROUND: Breast cancer (BC) has transcended lung cancer as the most common cancer in the world. Due to the disease's aggressiveness, rapid growth, and heterogeneity, it is crucial to investigate different therapeutic approaches for treatment. According to the World Health Organization (WHO), Plant-based therapeutics continue to be utilized as safe/non-toxic complementary or alternative treatments for cancer, even in developed countries, regardless of how cutting-edge conventional therapies are. Despite their low bioavailability, curcumin (CUR) and green tea (GT) represent safer therapeutic options. Due to their potent molecular-modulating properties on various cancer-related molecules and signaling pathways, they are considered gold-standard therapeutic agents and have been incorporated into the development of one or more therapeutic strategies of BC treatment. METHODS: We investigated the modulatory role of CUR and GT extracts on significant multi molecular targets in MCF-7 BC cell line to assess their potential as BC multi-targeting agents. We analyzed the phytocompounds in GT leaves using High-performance liquid chromatography (HPLC) and Gas chromatography-mass spectrometry (GC-MS) techniques. The mRNA expression levels of Raf-1, Telomerase, Tumor necrosis factor alpha (TNF-α) and Interleukin-8 (IL-8) genes in MCF-7 cells were quantified using quantitative real-time PCR (qRT-PCR). The cytotoxicity of the extracts was assessed by the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay and the released Lactate dehydrogenase (LDH), a valuable marker for identifying the programmed necrosis (necroptosis). Additionally, the concentrations of the necroptosis-related proinflammatory cytokines (TNF-α and IL-8) were measured using enzyme-linked immunosorbent assay (ELISA). RESULTS: In contrast to the GT, the results showed the anticancer and cytotoxic properties of CUR against MCF-7 cells, with a relatively higher level of released LDH. The CUR extract downregulated the oncogenic Raf-1, suppressed the Telomerase and upregulated the TNF-α and IL-8 genes. Results from the ELISA showed a notable increase in IL-8 and TNF-α cytokines levels after CUR treatment, which culminated after 72 h. CONCLUSIONS: Among both extracts, only CUR effectively modulated the understudy molecular targets, achieving multi-targeting anticancer activity against MCF-7 cells. Moreover, the applied dosage significantly increased levels of the proinflammatory cytokines, which represent a component of the cytokines-targeting-based therapeutic strategy. However, further investigations are recommended to validate this therapeutic approach.

Engineering of fluorescent or photoactive Trojan probes for detection and eradication of ß-Amyloids.

Trojan horse technology institutes a potentially promising strategy to bring together a diagnostic or cell-based drug design and a delivery platform. It provides the opportunity to re-engineer a novel multimodal, neurovascular detection probe, or medicine to fuse with blood-brain barrier (BBB) molecular Trojan horse. In Alzheimer's disease (AD) this could allow the targeted delivery of detection or therapeutic probes across the BBB to the sites of plaques and tangles development to image or decrease amyloid load, enhance perivascular Aß clearance, and improve cerebral blood flow, owing principally to the significantly improved cerebral permeation. A Trojan horse can also be equipped with photosensitizers, nanoparticles, quantum dots, or fluorescent molecules to function as multiple targeting theranostic compounds that could be activated following changes in disease-specific processes of the diseased tissue such as pH and protease activity, or exogenous stimuli such as, light. This concept review theorizes the use of receptor-mediated transport-based platforms to transform such novel ideas to engineer systemic and smart Trojan detection or therapeutic probes to advance the neurodegenerative field.

Developing Trojan horses to induce, diagnose and suppress Alzheimer's pathology.

There are many obstacles impeding the Alzheimer's disease (AD) research. For instance, its early diagnosis to identify individuals at risk has not been successful so far. AD animal models cannot be created without genetic pre-disposition or surgical manipulation. Single gene/protein delivery has so far failed to achieve significant clinical improvements in multifactorial AD. We hypothesize that the blood-brain barrier (BBB) penetration issues are the major obstacle in the development of current Alzheimer's causative, diagnostic, and multi-targeted therapeutic probes, and partly the cause of the failure of more than 99% of intervention trials. To overcome this problem, shuttle peptides or monoclonal antibodies for receptors on BBB can act as molecular Trojan horses to transport the fused novel classes of re-engineered AD causative agents, diagnostic probes, or multiple function neurovascular medicines across the BBB via receptor-mediated transport to cause, diagnose, or improve the AD phenotype, respectively. Here, we propose the design of such Trojan horses, comprising three essential components that could (i) reverse Aß amyloidosis, (ii) clear liberated Aß, and (iii) improve angiogenesis or endothelial metabolic dysfunction, besides alleviating the inflammation, to eventually enhance neuronal health, cerebral blood flow, and cognitive function. Such Trojan horses can aid in AD research by diagnosing Aß-oligomers at earlier stages, creating improved animal models by exposing transgenic animals to amyloid-inducing agents, and allowing treatment by novel neurovascular medicines.

Microbial Proteins as Novel Industrial Biotechnology Hosts to Treat Epilepsy.

Epilepsy is characterized by the hyperexcitability of various neuronal circuits that results due to the imbalance between glutamate-mediated excitation of voltage-gated cation channels and γ-amino butyric acid (GABA)-mediated inhibition of anion channels leading to aberrant, sporadic oscillations or fluctuations in neuronal electrical activity. Epilepsy with a risk of mortality and around 65 million sufferers of all ages all over the world is limited therapeutically with high rates of adverse reactions, lack of complete seizure control, and over 30% patients with refractory epilepsy. The only alternative to medicines is to identify and surgically remove the seizure foci in the brain or to abort the seizures just as they begin using an implanted cerebral electrode. However, these alternatives are unable to precisely aim aberrant neuronal circuits while leaving others unaltered. Epilepsy animal models also constitute the identical constraint. Thus, a better target-specific approach is needed to study and treat epilepsy. Unicellular green algae Chlamydomonas reinhardtii expresses a channelrhodopsin-2 (ChR2) sodium ion channel protein that controls the phototaxis movement of algae in response to blue light. Similarly, archaeon Natronomonas pharaonis (NpHR) expresses a monovalent Cl- channel protein halorhodopsin that responds to yellow light. These features of ChR2 and NpHR proteins can be used in optogenetic techniques to manipulate the bi-directional firing pattern of neuronal circuits in an attempt to better understand the pathophysiology of epileptic seizures as well as to discover novel potential drugs to treat epilepsy.

Hepatitis C Virus Associations with Non Hodgkin's Lymphoma: Insights on Inflammation/Angiogenesis and CD Markers.

We aimed to investigate any association between hepatitis C virus (HCV) infection and non-Hodgkin's lymphoma (NHL) in the view of cytokines that control inflammation/angiogenesis and their correlation with certain CD markers. NHL patients with or without HCV infection were studied. CD5, CD30, CD3, CD20 and CD45 were immunohistochemically evaluated. Plasma levels of vascular endothelial and platelet derived growth factors (VEGF, and PDGF), tumor necrosis factor (TNF-α), transforming growth factor (TGF-ß), interleukin-6 (IL-6), IL-8, IL-4, IL-12 and interferon gamma (IFN-γ) were detected by enzyme-linked immunosorbent assay (ELISA). HCV+ve NHL patients showed a significant reduction in VEGF, PDGF, IFN-γ, CD5 and CD45 and a significant increase in IL-12 and IL-8. In conclusion, there was a significant change in cytokine secretion and expression of CD markers in HCV+ve NHL patients. Based on our results, HCV infection in NHL patients requires more in-depth investigations to explore any role in lymphoma progression.

Seminal soluble fas relationship with oxidative stress in infertile men with varicocele.

OBJECTIVE: To assess seminal plasma soluble Fas (sFas) relationship with oxidative stress and varicocele (Vx) grade in infertile men. METHODS: In all, 230 men were prospectively investigated: fertile men without Vx, fertile men with Vx, infertile men without Vx, and infertile men with Vx. In their semen, seminal oxidant (malondialdehyde [MDA]), antioxidants (ascorbic acid, glutathione peroxidase [GPx], catalase [CAT], and superoxide dismutase [SOD]), and seminal sFas were assessed. RESULTS: Either fertile or infertile men with Vx demonstrated significantly higher seminal oxidants (MDA) and significantly lower seminal antioxidants (SOD, GPx, CAT, and ascorbic acid), sFas compared with fertile or infertile men without Vx. Infertile men with or without Vx had significantly higher seminal MDA and significantly lower seminal antioxidants, sFas compared with fertile men with or without Vx. Men with Vx grade III had significantly higher seminal MDA and significantly lower antioxidants, sFas compared with Vx grade II and I, respectively. Seminal sFas demonstrated significant positive correlation with sperm count, sperm motility, sperm normal forms, seminal ascorbic acid, SOD, GPx, and CAT and significant negative correlation with seminal MDA. CONCLUSION: Down regulation of seminal sFas in Vx associated men is related to increased oxidative stress and is correlated with Vx grade.

RANTES, TNF-α, oxidative stress, and hematological abnormalities in hepatitis C virus infection.

BACKGROUND: Chronic infection with hepatitis C virus (HCV) is associated with failures of T-cell-mediated immune clearance and with abnormal B-cell growth and activation. Hepatitis C virus infection is characterized by a systemic oxidative stress that is most likely caused by a combination of chronic inflammation, iron overload, liver damage, and proteins encoded by HCV. After a viral infection, multiple proinflammatory mediators contribute to recruitment of immune cells to the liver and to the generation of an antiviral immune response. Recent publications mark chemokines and their receptors as key players in leukocyte recirculation through the inflamed liver. MATERIALS AND METHODS: The present study involved 75 male subjects, divided into 2 groups: group 1 (n = 30), control group; group 2 (n = 45), patients with chronic HCV. For all subjects, the following investigations were performed: estimation of the levels of bilirubin, albumin, prothrombin concentration, glycosylated hemoglobin, creatinine, α-fetoprotein, HCV RNA, and activities of alanine and aspartate transaminases as well as alkaline phosphatase. In addition, regulated on activation normal T cell expressed and secreted (RANTES), tumor necrosis factor alpha, malondialdehyde (MDA) and nitric oxide (NO) were assessed. Plasma HCV-RNA concentration (viral load) was determined by real-time polymerase chain reaction (PCR) StepOne system using Applied Biosystem. Complete blood picture was assayed using Abbott Cell-Dyn 3700 hematology analyzer. RESULTS: There were significant increases of the levels of RANTES, tumor necrosis factor alpha, MDA, and NO in HCV-infected patients compared with the control group (P <0.05); and in these patients, these levels showed significant positive correlation with the HCV RNA viral load. Also, mild leukopenia, thrombocytopenia, neutropenia, and lymphocytosis, with consequent significant increase in the lymphocytes/neutrophils ratio, were detected in these patients. CONCLUSION: The data support the concept of chemokines (RANTES) as mediators of liver cell injury in HCV infection. In addition, MDA and NO levels might be used as monitoring markers for oxidative stress in hepatitis C infection.

Effect of smoking status on seminal parameters and apoptotic markers in infertile men.

PURPOSE: We assessed semen parameters, sperm apoptotic markers and seminal plasma cotinine in infertile smokers. MATERIALS AND METHODS: A total of 160 men were divided into 4 equal groups, including fertile smokers, fertile nonsmokers, infertile smokers and infertile nonsmokers. Smoking was classified as mild--fewer than 10, moderate--10 to 20 or heavy--more than 20 cigarettes daily. All men underwent semen analysis, and assessment of sperm caspase-9, Smac/DIABLO, DNA fragmentation and seminal plasma cotinine. RESULTS: Infertile men, particularly smokers, have significantly lower semen variables and significantly higher sperm Smac/DIABLO, caspase-9 activity, the percent of DNA fragmentation and seminal plasma cotinine. The mean number of cigarettes smoked daily and smoking duration significantly correlated positively with sperm Smac/DIABLO, caspase-9 activity, the percent of DNA fragmentation and seminal plasma cotinine, and significantly correlated negatively with tested semen variables. Heavy smoking was associated with a significant increase in sperm Smac/DIABLO, caspase-9 activity and seminal plasma cotinine, and with a significant decrease in tested semen variables compared with those in moderate or mild smokers. CONCLUSIONS: Smoking has a negative impact on semen variables. It is associated with increased sperm caspase-9, Smac/DIABLO and the percent of DNA fragmentation, especially in infertile heavy smokers.

Assessment of seminal plasma laminin in fertile and infertile men.

AIM: To assess laminin levels in the seminal plasma of infertile and fertile men, and to analyze the correlation of laminin levels with sperm count, age, sperm motility and semen volume. METHODS: One hundred and twenty-five recruited men were equally divided into five groups according to their sperm concentration and clinical examination: fertile normozoospermia, oligoasthenozoospermia, non-obstructive azoospermia (NOA), obstructive azoospermia (OA) and congenital bilateral absent vas deferens (CBAVD). The patients' medical history was investigated and patients underwent clinical examination, conventional semen analysis and estimation of seminal plasma laminin by radioimmunoassay. RESULTS: Seminal plasma laminin levels of successive groups were: 2.82 +/- 0.62, 2.49 +/- 0.44, 1.77 +/- 0.56, 1.72 +/- 0.76, 1.35 +/- 0.63 U/mL, respectively. The fertile normozoospermic group showed the highest concentration compared to all infertile groups with significant differences compared to azoospermic groups (P<0.05). Testicular contribution was estimated to be approximately one-third of the seminal laminin. Seminal plasma laminin demonstrated significant correlation with sperm concentration (r = 0.460, P < 0.001) and nonsignificant correlation with age (r = 0.021, P = 0.940), sperm motility percentage (r = 0.142, P = 0.615) and semen volume (r = 0.035, P = 0.087). CONCLUSION: Seminal plasma laminin is derived mostly from prostatic and testicular portions and minimally from the seminal vesicle and vas deferens. Estimating seminal laminin alone is not conclusive in diagnosing different cases of male infertility.