The cytotoxcity activity of in vitro isolated and expanded cytotoxic T-lymphocytes and natural killer cells in bladder cancer

The expanding roles of the immune system in tumourigenesis have established immunotherapy as a potential mainstream cancer therapeutic modality. Ex vivo expanded and activated cytotoxic T-lymphocytes (CTLs) and natural killer (NK) cells have been found to be efficacious in the treatment of various types of cancers. One of the biggest limitations is the ability to generate and store cytotoxic immune cells in larger numbers without losing its cytotoxicity. Consequently, we evaluated the in vitro cytotoxic activity of freshly cultured and cryopreserved CTLs and NK cells that were expanded in vitro. We also compared the synergistic cytotoxic activity of CTLs and NK cells in combination. The cytotoxic activity was measured in bladder cancer cell lines, EJ28 (invasive) and RT112 (minimally-invasive). All experiments were run in three replicates. The cellular phenotype of the isolated and expanded effector cells was characterised using flow cytometry. MTT assay was performed to assess the dose- and time-dependent cell-mediated cytotoxic activity in the bladder cancer cells. An effector to target ratio of 1:1, 2:1, 5:1, 10:1 and 20:1 was tested after 4 h, 12 h and 24 h incubation. The fresh in vitro expanded effector cells had a high percentage of cell viability and expressed cytotoxic markers CD8+ and CD56+ in the CTL and NK cell cultures, respectively. Although the expansion capacity of the cryopreserved cells was limited, the expression of the functional markers and cytotoxic activity of these effector cells were maintained. All the effector cells exhibited significant cytotoxic activity at the effector to target ratio of 5:1 at 4 hours of co-culture. This was confirmed through the real-time observation of the morphological changes of the cells using an inverted phase contrast and time-lapse confocal microscope. The ex vivo generated CTLs and NK cells appear to retain their functionality, especially in recognizing their allogeneic target and thus, serve as a foundation to build on for future therapeutic applications

HISTORIC REVIEW ON MODERN HERBAL NANOGEL FORMULATION AND DELIVERY METHODS

Chemistry deals with herbal constituents are often coined as phytochemistry. Herbal constituents have profound improvements in drug discovery for several existing diseases. Many of these constituents are restricted from pharmaceutical discoveries due to two important reasons: pharmacodynamics and pharmacokinetics. There are many new technological strategies and comparisons have been studied to improve the herbal discoveries in the pharmaceutical market. This review paper will highlight historical evidence of nanogels which is the most important strategy applied to several herbal medicines with high patience compliance, delivery rate, and efficiency. Nanogels are nanoparticles combined with cross-linked polymer networks with desirable features to carry hydrophilic or hydrophobic drugs in a more stable condition. Nanogels are highly preferred substances for herbal medicine in terms of stability and rapid response to the external stimuli factors. Nanogel can facilitate the herbal products with higher cellular penetration than existing and hence, it proves to be the new dimension for both oral and transdermal drug delivery for several unmet diseases like cancer, diabetes, and chronic disorders. By the way, including the recent technological constituents to herbal drugs, it can possess high bioavailability, low toxicity and enhance the sustained release

Identifying clinically significant novel drug candidate for highly prevalent Alzheimer's disease

618-623Pharmacokinetics is very important in target validation and in shifting a lead compound into a drug. It is a cumbersome process in clinical research. A quantitative personation based on computed, pharmacokinetics, physicochemical properties, ILOGP, drug-likeness, medicinal chemistry friendliness, bioavailability radar and BOILED-Egg for all the synthesized, 6 novel compounds have been assessed using Swiss ADME. An effective drug can be produced from the physicochemical properties discussed in this model. The physicochemical properties of all designed Schiff bases of curcumin have been found to be optimal and so, they are perceived to have acceptable oral absorption and adequate permeability. All the monomers obeyed the rule of five by Lipinski and the oral bioavailability is accounted worldwide. The desired set of monomers have been enhanced by effective ADME screening and molecular simulation methods with Microtubuleassociated protein tau (MAPT) (PDB code: 10636) receptor could represent favourable building blocks as preferable chemotherapeutic factor in resistance to Alzheimers disease

Impaired redox environment modulates cardiogenic and ion-channel gene expression in cardiac-resident and non-resident mesenchymal stem cells

Redox homeostasis plays a crucial role in the regulation of self-renewal and differentiation of stem cells. However, the behavioral actions of mesenchymal stem cells in redox imbalance state remain elusive. In the present study, the effect of redox imbalance that was induced by either hydrogen peroxide (H2O2) or ascorbic acid on human cardiac-resident (hC-MSCs) and non-resident (umbilical cord) mesenchymal stem cells (hUC-MSCs) was evaluated. Both cells were sensitive and responsive when exposed to either H2O2 or ascorbic acid at a concentration of 400 µmol/L. Ascorbic acid pre-treated cells remarkably ameliorated the reactive oxygen species level when treated with H2O2. The endogenous antioxidative enzyme gene (Sod1, Sod2, TRXR1 and Gpx1) expressions were escalated in both MSCs in response to reactive oxygen species elevation. In contrast, ascorbic acid pre-treated hUC-MSCs attenuated considerable anti-oxidative gene (TRXR1 and Gpx1) expressions, but not the hC-MSCs. Similarly, the cardiogenic gene (Nkx 2.5, Gata4, Mlc2a and β-MHC) and ion-channel gene (IKDR, IKCa, Ito and INa.TTX) expressions were significantly increased in both MSCs on the oxidative state. On the contrary, reduced environment could not alter the ion-channel gene expression and negatively regulated the cardiogenic gene expressions except for troponin-1 in both cells. In conclusion, redox imbalance potently alters the cardiac-resident and non-resident MSCs stemness, cardiogenic, and ion-channel gene expressions. In comparison with cardiac-resident MSC, non-resident umbilical cord-MSC has great potential to tolerate the redox imbalance and positively respond to cardiac regeneration

Identifying clinically significant novel drug candidate for highly prevalent Alzheimer's disease

Pharmacokinetics is very important in target validation and in shifting a lead compound into a drug. It is a cumbersome process in clinical research. A quantitative personation based on computed, pharmacokinetics, physicochemical properties, ILOGP, drug-likeness, medicinal chemistry friendliness, bioavailability radar and BOILED-Egg for all the synthesized, 6 novel compounds have been assessed using Swiss ADME. An effective drug can be produced from the physicochemical properties discussed in this model. The physicochemical properties of all designed Schiff bases of curcumin have been found to be optimal and so, they are perceived to have acceptable oral absorption and adequate permeability. All the monomers obeyed the rule of five by Lipinski and the oral bioavailability is accounted worldwide. The desired set of monomers have been enhanced by effective ADME screening and molecular simulation methods with Microtubuleassociated protein tau (MAPT) (PDB code: 10636) receptor could represent favourable building blocks as preferable chemotherapeutic factor in resistance to Alzheimers disease

Autologous Immune Enhancement Therapy in Recurrent Ovarian Cancer with Metastases: A Case Report

Current therapeutic modalities for ovarian cancer such as chemotherapy, radiotherapy and surgery have been reported to yield only marginal success in improving survival rates of patients and have associated adverse effects. We report here a case of recurrent stage IV ovarian cancer, treated with cell-based autologous immune enhancement therapy (AIET) along with chemotherapy and followed up for 18 months. A 54-year-old female was diagnosed with a recurrence of ovarian carcinoma 1 year after initial surgical removal followed by chemotherapy for stage IIIC ovarian carcinoma. When diagnosed in 2010 with recurrence, she had liver and spleen metastases with a CA-125 level of 243 U/ml and a stage IV clinical status. Six infusions of AIET using autologous in vitro expanded and activated natural killer (NK) cells (CD3–CD56+) and activated T lymphocytes (CD3+CD56+) were administered in combination with 6 cycles of chemotherapy with carboplatin and doxorubicin. Following this treatment, CA-125 decreased to 4.7 U/ml along with regression of the metastatic lesions and an improved quality of life. No adverse reactions were reported after the AIET transfusions. Eighteen months of follow-up revealed a static nonprogressive disease. Combining AIET with chemotherapy and other conventional treatments has been found to be effective in our experience, as reported earlier, even in patients with advanced ovarian cancer, and we recommend this strategy be considered in treating similar cases

A safety evaluation of intravenous administration of ex vivo expanded human peripheral blood-derived NK cells : a preclinical study

The use of natural killer (NK) cells in the treatment of various cancers is emerging as a promising approach in adoptive immunotherapy. However, the safety of ex vivo activated and expanded cells in in vivo conditions remain unknown. In this study, the toxicity of NK cells was evaluated at different doses, with 5 × 106, 20 × 106 and 50 × 106 cells injected intravenously into pre-irradiated (30Gy) immunodeficient mice twice a week for three weeks and the mice were followed-up on for 90 days. Throughout the study, no mortality, abnormal clinical signs, or behavioural changes related to the testing material were observed in either the treated or control groups of mice. There were no significant variations in food and water consumption between both genders in the NK cell treated and control groups. However, certain significant changes were observed between the groups in the clinical biochemistry and urine analysis reports. As autopsy showed no significant variations in absolute and relative organ weights between the groups, except for the livers of the treated mice. The histopathological analysis also demonstrated that there were no significant abnormalities in most of the organs of both genders, except for the liver. Some necrotic lesions were observed in the livers of both the treated and control mice, and these lesions may be due to the effects of irradiation or could be common in NOD.SCID mice. The findings of this study indicate that intravenous administration of NK cells is safe and does not cause any adverse effects up to the dose of 50 × 106 cells/mouse

Generation and characterization of human cardiac resident and non-resident mesenchymal stem cell

Despite the surgical and other insertional interventions, the complete recuperation of myocardial disorders is still elusive due to the insufficiency of functioning myocardiocytes. Thus, the use of stem cells to regenerate the affected region of heart becomes a prime important. In line with this human umbilical cord-derived mesenchymal stem cells (hUC-MSCs) have gained considerable interest due to their potential use for mesodermal cell based replacement therapy and tissue engineering. Since MSCs are harvested from various organs and anatomical locations of same organism, thus the cardiac regenerative potential of human cardiac-derived MSCs (hC-MSCs) and human umbilical cord Wharton’s Jelly derived MSC (hUC-MSCs) were tested concurrently. At in vitro culture, both hUC-MSCs and hC-MSCs assumed spindle shape morphology with expression of typical MSC markers namely CD105, CD73, CD90 and CD44. Although, hUC-MSCs and hC-MSCs are identical in term of morphology and immunophenotype, yet hUC-MSCs harbored a higher cell growth as compared to the hC-MSCs. The inherent cardiac regenerative potential of both cells were further investigated with mRNA expression of ion channels. The RT-PCR results demonstrated that both MSCs were expressing a notable level of delayed rectifier-like K+ current (I KDR ) ion channel, yet the relative expression level was considerably varied between hUC-MSCs and hC-MSCs that Kv1.1(39 ± 0.6 vs 31 ± 0.8), Kv2.1 (6 ± 0.2 vs 21 ± 0.12), Kv1.5 (7.4 ± 0.1 vs 6.8 ± 0.06) and Kv7.3 (27 ± 0.8 vs 13.8 ± 0.6). Similarly, the Ca2+-activated K+ current (I KCa ) channel encoding gene, transient outward K+ current (I to ) and TTX-sensitive transient inward sodium current (I Na.TTX ) encoding gene (Kv4.2, Kv4.3 and hNE-Na) expressions were detected in both groups as well. Despite the morphological and phenotypical similarity, the present study also confirms the existence of multiple functional ion channel currents IKDR, IKCa, Ito, and INa.TTX in undifferentiated hUC-MSCs as of hC-MSCs. Thus, the hUC-MSCs can be exploited as a potential candidate for future cardiac regeneration

Synthesis and Electrochemical Performances of gamma-KCoPO4 Nanocrystals as Promising Electrode for Aqueous Supercapatteries

Herein, discrete gamma-KCoPO4 nanocrystals were prepared by a facile, green and fast sol-gel route. The lattice parameters as well as positional and displacement parameters of atoms in the average gamma-KCoPO4 structure with a space group of P2(1)/n were calculated using full profile Rietveld refinement. Monodispersed quadrangular gamma-KCoPO4 nanocrystals with improved phase purity and crystallinity were found through XRD patterns and HRTEM images. Subsequently, the prepared gamma-KCoPO4 nanocrystals were tested as electrode material for supercapatteries in aqueous electrolytes. The gamma-KCoPO4 electrode shows superior specific charge capacity of 309 C g(-1) at 1 mV s(-1) in 1 M KOH, compared to 1 M NaOH (222 C g(-1)) and 1 M LiOH (77 C g(-1)). Further, it exhibits improved electrochemical activity by delivering an impressive specific charge capacity of 100 C g(-1) at a current of 0.6 mA cm(-2) in an aqueous electrolyte medium with acceptable capacity retention. A lab-scale supercapattery was assembled resembling the commercial device. The fabricated device delivered an enhanced specific energy of 28 W h kg(-1) and a specific power of 1600 W kg(-1) and prolonged cycle life of about 5000 cycles