Increase In Cytotoxic Activity Research Articles

Риск срыва адаптационных перестроек и развития сердечно-сосудистых катастроф у жителей Европейского Севера и Арктики Российской Федерации

The paper presents the results on the risk criteria for disruption of adaptive restructuring and the development of cardiovascular disasters in residents of the European North and the Arctic of the Russian Federation. 980 people aged 21 to 55 years were examined, including 191 people living in the Arctic, beyond the Arctic Circle (the villages of Revda and Lovozero of the Murmansk region). The comparison group included 789 people of the specified age range living in the Arkhangelsk region. It has been established that the risk of disruption of adaptive rearrangements in Arctic residents is due to the absence of a compensatory reaction to normalize lipid metabolism with a deficiency in the content of HDL, ApoA-I ligand, the accumulation of metabolic products: cholesterol, triglycerides, LDL occurs with an increase in cytotoxic activity of lymphocytes, production of pro-inflammatory cytokines and is associated with the risk of cardiovascular disasters in Arctic residents of the Russian Federation.

In vitro effects of a natural marine algae polysaccharide (λ-carrageenan) on seabream erythrocytes, tumour cell lines and marine bacterial pathogens

AbstractCarrageenan is a high molecular weight sulphated polysaccharide used to induce experimental inflammation in mammals. In addition, it possesses a wide variety of properties that have not yet been studied in fish. This study evaluated the hemagglutinating, hemolytic, cytotoxic, and antibacterial activities of λ-carrageenan. The results showed that λ-carrageenan has hemagglutinating and hemolytic activities on gilthead seabream erythrocytes, which were dose and time-dependent during the first 6 h of incubation. No significant effects on the haemolytic activity of erythrocytes were observed after incubation for 12 or 24 h with λ-carrageenan. The PLHC-1 cell line showed significant increases in cytotoxic activity after 6 or 12 h of incubation compared with control cells, and the highest doses of λ-carrageenan caused cytotoxicity in PLHC-1 cells after 24 h of incubation. The morphology of PLHC-1 cells incubated with the highest doses of λ-carrageenan for 12 or 24 h showed obvious cell death changes compared with control cells. Interestingly, no significant variations in cytotoxic activity were observed in SAF-1 cell line after incubation with λ-carrageenan. Furthermore, λ-carrageenan showed significant dose-dependent bactericidal activity against Photobacterium damselae but had no significant effect on the bactericidal activity of Vibrio harveyi, Vibrio anguillarum, and Tenacibaculum maritimum. The study suggests that λ-carrageenan has potential applications in aquaculture and aquatic pharmaceutical industries as a hemagglutinating, hemolytic, and antibacterial agent.

Do Men and Women Differ in Hematological Adaptations to 24 Weeks of Crossfit® Training?

Regular exercise can modulate the immune system functioning through changes in the number and function of leukocytes as well as in red blood cells and other typical blood markers. High intensity exercise promotes increases in cytotoxic activity, phagocytic capacity, chemotaxis and cell apoptosis. The aim of the study was to compare the chronic effects of a 24-week training program using CrossFit® methodology on hematological variables of men vs. women. Twenty-nine CrossFit® athletes (35.3 ± 10.4 years, 175.0 ± 9.2 cm, 79.5 ± 16.4 kg) participated in the study. The blood count, the lipid profile and glucose markers were measured every two months during the study period. The erythrocyte count and hemoglobin concentrations increased in months 4 and 6 in men and women, respectively. Hematocrit levels increased in men in months 2, 4 and 6, while in women only in month 6. Red cell distribution width increased in men in month 6 when compared to the value in month 2. Segmented neutrophils increased in men in month 6 and eosinophil levels increased in women in month 6. Differences between the two sexes were observed in monocytes levels at baseline, as well as in months 2, 4 and 6. Cross-Fit® training increased red cell count indicators in both sexes, which may be related to increased erythropoiesis. Some white blood cell counts were altered and these differed between sexes. The number of lymphocytes remained stable throughout the experiment.

Assessing mitochondria-targeted acyl hydroquinones on the mitochondrial platelet function and cytotoxic activity: Role of the linker length

IntroductionThe use of triphenylphosphonium cation (TPP+) linked to phenolic compounds by alkyl chains has a significant relevance as a mitochondrial delivery strategy in biomedicine because it affects mitochondrial bioenergetics in models of noncommunicable diseases such as cancer and cardiovascular-related conditions. Studies indicate that a long alkyl chain (10–12 carbon) increases the mitochondrial accumulation of TPP+-linked drugs. In contrast, other studies show that these compounds are consistently toxic to micromolar concentrations (as observed in platelets). In the present study, we evaluated the in vitro effect of three series of triphenylphosphonium-linked acyl hydroquinones derivates on the metabolism and function of human platelets using 3–9 carbons for the alkyl linker. Those were assessed to determine the role of the length of the alkyl chain linker on platelet toxicity. MethodsHuman platelets were exposed in vitro to different concentrations (2–40 μM) of every compound; cellular viability, phosphatidylserine exposition, mitochondrial membrane potential (ΔΨm), intracellular calcium release, and intracellular ROS generation were assessed by flow cytometry. An in silico energetic profile was generated with Umbrella sampling molecular dynamics (MD). Results and conclusionsThere was an increase in cytotoxic activity directly related to the length of the acyl chain and lipophilicity, as seen by three techniques, and this was consistent with a decrease in ΔΨm. The in silico energetic profiles point out that the permeability of the mitochondrial membrane may be involved in the cytotoxicity of phosphonium salts. This information may be relevant for the design of new TPP+ -based drugs with a safe cardiovascular profile.

Изменения иммунологических показателей у пострадавших с политравмой при применении в комплексном лечении электромагнитных волн терагерцевого диапазона

Introduction Improving the results of treatment and rehabilitation is essential for polytrauma patients. The aim of the study was to prove the effectiveness of terahertz electromagnetic waves (TEW) in polytrauma patients early after traumatic event to improve immune parameters. Material and methods The study included 20 polytrauma patients who were divided into two groups. Patients of the treatment group (n = 10) received TEW early after traumatic event in addition to standard treatment using the damage control approach. Control patients (n = 10) received no TEW therapy in the complex of therapeutic measures. Immunological studies were performed during treatment. The effectiveness of rehabilitation was assessed at a long term using an evaluation scale. Results The mean rehabilitation score was significantly higher in the treatment group compared to the controls. The treatment group showed significantly higher production of TNFα and IL-2, spontaneous production of IFNγ by lymphocytes, phagocyte activity (NBT-test) and absorption activity of monocytes after the use of TEW in comparison with controls. An increased level of IgG and IgM was also noted in patients of the treatment group. Discussion The findings showed that the use of TEW in polytrauma patients early after traumatic event resulted in the increase in cytotoxic activity of lymphocytes and phagocytic cells. The immunostimulating effect of TEW was also achieved through boosting humoral immunity. Conclusion The TEW used early after traumatic event in polytrauma patients resulted in strengthening of the immune system which contributed to more effective rehabilitation.

Synthesis and cytotoxic activity evaluation of some new 1, 3, 4-oxadiazole, 1, 3, 4-thiadiazole and 1, 2, 4- triazole derivatives attached to phthalimide.

Background and purpose:In the last few decades, nitrogen-rich heterocyclic compounds such as 1, 3, 4-thiadiazoles, 1, 2, 4-triazoles and 1, 3, 4-oxadiazoles have received considerable attention because of their notable biological properties, especially cytotoxic effects. The small molecules of mentioned azole derivatives revealed very intensive antitumor activity. In addition, phthalimide-thiadiazole and naphthalimide-triazole hybrid derivatives have shown remarkable cytotoxic effects. According to these observations, some of the hybrid derivatives containing the phthalimide-five-membered azoles were prepared in three steps in this research.Experimental approach:The thiol group of azoles was treated with ethyl chloroacetate which was followed by a reaction with hydrazine hydrate to provide acid hydrazide derivatives. Subsequently, the corresponding acid hydrazides were utilized to prepare the final derivatives through the reaction with phthalic anhydride. Cytotoxic activity of final compounds was evaluated against MCF-7 and HeLa cell lines using MTT assay.Findings/Results:Compound 3d containing two phthalimide moieties in its structure showed a significant improvement in cytotoxic activity with an IC50 value of 29 μM against HeLa cell line. Compounds 3a-3c showed less cytotoxic effects against both cell lines.Conclusion and implications:The combination of the thiadiazole nucleus with two phthalimide structures increased the cytotoxic activity against the HeLa cell line. This increase in cytotoxic activity is probably due to its being more lipophilic characteristic and interaction of this derivative with the biological targets of two directions.

A Sensitive and Simple Method to Assess NK Cell Activity by RT-qPCR for Granzyme B Using Spleen and Blood

A new assay method for natural killer (NK) cell activity was established using quantitative RT-PCR (RT-qPCR) to determine the gene expression of granzyme B (GzmB) and perforin 1 (Prf1). The RT-qPCR method was compared to a conventional cytotoxic assay. Upregulated expression of GzmB and Prf1 mRNA and enhanced cytotoxic activity were observed in splenocytes from lipopolysaccharide (LPS)-treated rats. A high correlation, R2 = 0.71, was observed between the gene expression of GzmB and the cytotoxic activity of splenocytes from these rats, indicating that GzmB RT-qPCR is a reliable alternative method to assess NK cell activity/activation. Remarkably, 12.6- to 59.7-fold upregulation of GzmB mRNA expression was observed in leukocytes, the spleen, and splenocytes from LPS-injected rats. Its upregulation appeared to be dose-dependent on the LPS concentration in the range of 0.01 - 0.1 mg/kg. Whereas, only 1.3- to 1.9-fold increase of cytotoxic activity was detected in splenocytes from the rats treated with LPS in the same range. From these, it is evident that, to assess NK cell activity/activation, the GzmB RT-qPCR method is highly sensitive compared with the conventional cytological assay. Furthermore, this GzmB RT-qPCR method is advantageous, as it does not require freshly prepared splenocytes and cell culture procedures. The convenience of GzmB RT-qPCR enables the use of whole blood, leukocytes, the spleen, and/or their frozen samples to evaluate NK cell activity/activation.

Cellular immune response following head injury

The incidence of head injuries is increasing particularly in cities. The aim was to study cellular immune response during the first 24 hours following head injuries among men aged 41-60 years and older. Methods: Altogether, 90 men aged 41-60 years were enrolled within 24 hours after head injury. Phenotypes of lymphocytes CD3+, CD4+, CD5+, CD71+, CD8+, CD16+, CD25+, CD95+, HLA-DR+, CD10+ were studied. Results. Proportion of mature T-lymphocytes increased in parallel with the increase of severity of trauma (11.1 % in mild traumas vs. 37.5 % in severe cases). In cases with average degree of trauma low concentrations of activators were detected (CD25+ = 0,23 ± 0,03 • 109 c/l; CD71+ = 0,24 ± 0,01 • 109 c/l and HLA-DR+ = 0,26 ± 0,02 • 109 c/l) combined with T-cells deficiency. Concentrations of CD8+ and CD95+ increase with the severity of the trauma: (0,64 ± 0,04) - (0,76 ± 0,01) • 109 c/l; (0,51 ± 0,02) - (0,56 ± 0,01) • 109 c/l, respectively. Conclusions: An increase in helper-suppressor ratio (k > 2) due to T-helpers as a compensation for low concentrations of CD5+, CD3+, CD25+, CD71+ and HLA-DR+ seems to be associated with favorable prognosis. In severe head injuries a decrease in helpersuppressor ratio (k < 2) accompanied by CD5+, CD3+ deficiency can be explained by an increase in CD8+. An increase in cytotoxic activity of lymphocytes during the first 24 hours after trauma may be associated with unfavorable prognosis.

Intratumoral Distribution and pH-Dependent Drug Release of High Molecular Weight HPMA Copolymer Drug Conjugates Strongly Depend on Specific Tumor Substructure and Microenvironment.

Stimulus-sensitive polymer drug conjugates based on high molecular weight N-(2-hydroxypropyl) methacrylamide (HPMA) copolymers carrying doxorubicin via a pH-dependent cleavable bond (pHPMA-Dox) were previously shown to be able to overcome multi-drug resistance. Nevertheless, a tumor type dependent differential response was observed. Although an improved and more selective tumor accumulation of pHPMA-Dox is generally achieved due to the enhanced permeability and retention (EPR) effect, little is known about the fate of these conjugates upon entering the tumor tissue, which could explain the different responses. In this study, we compared in vitro and in vivo accumulation and Dox-activation of pHPMA-Dox in three cancer cell line models (1411HP, A2780cis, HT29) and derived xenograft tumors using a near-infrared fluorescence-labeled pHPMA-Dox conjugate. Firstly, cytotoxicity assays using different pH conditions proved a stepwise, pH-dependent increase in cytotoxic activity and revealed comparable sensitivity among the cell lines. Using multispectral fluorescence microscopy, we were able to track the distribution of drug and polymeric carrier simultaneously on cellular and histological levels. Microscopic analyses of cell monolayers confirmed the assumed mechanism of cell internalization of the whole conjugate followed by intracellular cleavage and nuclear accumulation of Dox in all three cell lines. In contrast, intratumoral distribution and drug release in xenograft tumors were completely different and were associated with different tissue substructures and microenvironments analyzed by Azan- and Hypoxisense®-staining. In 1411HP tumors, large vessels and less hypoxic/acidic microenvironments were associated with a pattern resulting from consistent tissue distribution and cellular uptake as whole conjugate followed by intracellular drug release. In A2780cis tumors, an inconsistent pattern of distribution partly resulting from premature drug release was associated with a more hypoxic/acidic microenvironment, compacted tumor tissue with compressed vessels and specific pre-damaged tissue structures. A completely different distribution pattern was observed in HT29 tumors, resulting from high accumulation of polymer in abundant fibrotic structures, with small embedded vessels featuring this tumor type together with pronounced premature drug release due to the strongly hypoxic/acidic microenvironment. In conclusion, the pattern of intratumoral distribution and drug release strongly depends on the tumor substructure and microenvironment and may result in different degrees of therapeutic efficacy. This reflects the pronounced heterogeneity observed in the clinical application of nanomedicines and can be exploited for the future design of such conjugates.

Abstract 017: Cytolytic Natural Killer Cells Play a Direct Role in Causing Hypertension and Intrauterine Growth Restriction in Pregnant Rats

The R educed U terine P erfusion P ressure (RUPP) rat model of placental ischemia mirrors many of the hallmark features of preeclampsia (PE) such as hypertension, intrauterine growth restriction (IUGR), and immune activation, which includes increased cytolytic natural killer cells (cNKs). We have previously demonstrated that depletion of NK cells in RUPP rats improves PE pathophysiology. In this study we tested the hypothesis that RUPP-stimulated NKs have increased cytotoxicity and play a direct role in inflammation, oxidative stress, hypertension, and intrauterine growth restriction in pregnant rats. NK cells were isolated from the placentas of normal pregnant (NP) and RUPP rats on gestation day (GD) 19 and assessed for cytotoxic activity. RUPP NK cells demonstrated a 5-fold increase in cytotoxic activity vs NP NK cells. On GD 12, 5x10 6 NK cells from RUPP rats were infused into a subset of NP rats. On GD 19, circulating and placental total NK cells and cNKs were quantified via flow cytometry in NP, RUPP, and NP+RUPP NK rats. MAP, fetal and placental weights, placental reactive oxygen species (ROS), and placental cytokines were also measured. Placental cNK cells (%gated) were significantly increased in RUPP (18.5±3.6%) and NP+RUPP NK (16.1±3.6%) compared to NP (4.1±0.8%) controls. Circulating cNKs followed a similar trend. MAP increased from 100 mmHg in NP to 126 mmHg in RUPP, and 119 mmHg in NP+RUPP NK (p<0.05 vs NP). Fetal weight decreased from 2.4±0.03 g in NP to 2.0±0.04 g in RUPP and to 2.1±0.02 g in NP+RUPP NK (p<0.05 vs NP). Placental weights followed a similar trend. Placental cNK cytokines were significantly increased in RUPP and in NP recipients of RUPP NK cells: TNF-α: NP- 23±3.1, RUPP- 62.3±7.8 pg/mg, NP+RUPP NK-43.9±5.7 pg/mg (p<0.05 vs NP); IFNγ: NP- 3.3±0.7, RUPP- 7.6±1.2 pg/mg, NP+RUPP NK-6.6±1.9 pg/mg (p<0.05 vs NP). Placental VEGF decreased from 6.5±1.3 pg/mg in NP to 2±0.8pg/mg in RUPP and 2.9±0.6 pg/mg in NP+RUPP NK. Circulating levels followed a similar trend. Placental ROS significantly increased from 27.3±4.6 RLUs/min/mg in NP to 63.1±5.7 RLUs/min/mg in RUPP and 84±17.6 RLUs/min/mg in NP+RUPP NK. These data demonstrate a direct role for cNKs in causing PE pathophysiology and identifies cNKs as a novel therapeutic target for treatment of PE.

Mycobacterium smegmatis alters the production of secondary metabolites by marine-derived Aspergillus niger.

It is generally accepted that fungi have a number of dormant gene clusters for the synthesis of secondary metabolites, and the activation of these gene clusters can expand the diversity of secondary metabolites in culture. Recent studies have revealed that the mycolic acid-containing bacterium Tsukamurella pulmonis activates dormant gene clusters in the bacterial genus Streptomyces. However, it is not clear whether the mycolic acid-containing bacteria activate dormant gene clusters of fungi. We performed co-culture experiments using marine-derived Aspergillus niger with Mycobacterium smegmatis, a mycolic acid-containing bacteria. The co-cultivation resulted in the production of a pigment by A. niger and increased cytotoxic activity of the extract against human prostate cancer DU145 cells. An analysis of secondary metabolites in the extract of the co-culture broth revealed that the increase in cytotoxic activity was caused by the production of malformin C (1), and that TMC-256A1 (2), desmethylkotanin (3), and aurasperone C (4) were selectively produced under co-culture conditions. In addition, further study suggested that direct interaction between the two microorganisms was necessary for the production of the pigment and the cytotoxic compound malformin C (1) from A. niger. Given the biological activities of malformin C, including cytotoxic activity, our approach for increasing the production of bioactive secondary metabolites has important practical applications and may facilitate structural analyses of novel bioactive compounds.

Korean mistletoe lectin enhances natural killer cell cytotoxicity via upregulation of perforin expression.

Natural killer (NK) cells are crucial components of the innate immune system, providing the first line of defense against pathogens. In a previous study, we demonstrated prophylactic activity of water extract of Korean mistletoe (Viscum album coloratum) on tumor metastasis. However, the leading compound from water extract of Korean mistletoe was not clearly addressed. The purpose of this research was mainly focused on addressing the effect of Korean mistletoe lectin (KMLC) on NK cell cytotoxicity, and the ability of cytokine secretion as well as its signal transduction, mitogen-activated protein kinase (MAPK) pathway. KMLC was used to test NK cell-mediated cytotoxicity in vitro and in vivo. Non-isotope cytotoxicity assay (bis-N,N,N',N'-tetraacetic acid (BATDA) release assay) was performed to test the cytotoxicity of NK cells against target tumor cells. Receptor expression was checked by flow cytometry analysis and MAPK signal molecules were analyzed by immunoblotting. KMLC at 200 ng/mL increased the cytotoxicity of NK92 cells by 35% compared with untreated cells. KMLC-treated (at 100 ng/mL) mice splenocytes showed a 20% increase in cytotoxic activity. Also, the B chain, one of the subchains of KMLC, increases perforin expression. We demonstrated that the signal transduction controlling NK cell cytotoxicity was mediated by upregulation of the NKG2D receptor and expression of a cytotoxic effector molecule. These results suggested that KMLC possessed immunological activity, mediated by NK cell activation.

Immune function in arctic mammals: Natural killer (NK) cell-like activity in polar bear, muskox and reindeer

Natural killer (NK) cells are a vital part of the rapid and non-specific immune defense against invading pathogens and tumor cells. This study evaluated NK cell-like activity by flow cytometry for the first time in three ecologically and culturally important Arctic mammal species: polar bear (Ursus maritimus), muskox (Ovibos moschatus) and reindeer (Rangifer tarandus). NK cell-like activity for all three species was most effective against the mouse lymphoma cell line YAC-1, compared to the human leukemia cell line K562; NK cell response displayed the characteristic increase in cytotoxic activity when the effector:target cell ratio increased. Comparing NK activity between fresh and cryopreserved mouse lymphocytes revealed little to no difference in function, highlighting the applicability of cryopreserving cells in field studies. The evaluation of this important innate immune function in Arctic mammals can contribute to future population health assessments, especially as pollution-induced suppression of immune function may increase infectious disease susceptibility.

Improvement the production of cytotoxic metabolites by Streptomyces griseus KJ623766

The cell-free culture supernatant (CFCS) obtained from Streptomyces (S.) grisues (accession codeKJ623766) fermentation, locally isolated Streptomyces strains from Egyptian soil sample, showed potent cytotoxic activity against Caco2 cell line. Fermentation was carried out in a 14 L laboratory fermenter, under optimum conditions of 28 °C, 200 rpm, 5 standard liters per minute (SLPM) aeration, 2 bar airflow pressure and uncontrolled pH. After 72 h of incubation, the cell-free culture supernatant (CFCS) was collected and extracted using ethyl acetate (1:1, v/v) at pH 7.0. Using 3-(4,5-dimethylthazol-2-yl)-2,5-diphenyl tetrazolium-bromide (MTT) assay, the ethyl acetate extract showed potential cytotoxic activity against Caco2 with CD50of 14 µg/mL. This showed an increase in cytotoxic activity by about 1.6 folds when compared to results obtained from shake flask (CD5022 µg/mL). Production improvement of cytotoxic activity was carried out also by genetic manipulation using a dose of 4 KiloGray (KGy) of gamma radiation. Fifteen out of forty-seven mutants showed higher potential cytotoxic activities when compared to that of the wild-type strain of S.griseus KJ623766. Mutants G31, G44, and G45 showed the most potent cytotoxic activities where they exhibited about 7 folds increase in potential cytotoxic activity with CD50 of 3.2 ±0.2, 2.9 ±0.1 and 3.25 ±0.43 µg/mL, respectively.

B7-H3 as a promising target for cytotoxicity T cell in human cancer therapy.

Targeting B7-H3 over-expressed tumor cells with anti-B7-H3 monoclonal antibodies inhibits tumor growth. Here we demonstrated the expression of B7 family homologue 3 (B7-H3) in a wide range of human tumor cells and further investigated whether B7-H3 could be served as a target for T-cell mediated immunotherapy against human cancers. The specific cytotoxic activity of activated T cell (ATC) armed with a novel anti-CD3 x anti-B7-H3 bispecific antibody (B7-H3Bi-Ab) against tumor cell was evaluated in vitro and in vivo. In contrast with unarmed ATC, an increase in cytotoxic activity of B7-H3Bi-armed ATC against tumor cells was observed at effector/target (E/T) ratios of 5:1, 10:1, and 20:1. Moreover, B7-H3Bi-armed ATC secreted more IFN-γ, TNF-α and IL-2 than unarmed ATC. Infusion of B7-H3Bi-armed ATC inhibited tumor growth in severe combined immunodeficiency (SCID) xenograft models, along with a significant survival benefit. Therefore, treatment with novel B7-H3Bi-armed ATC will be a promising strategy for current cancer immunotherapy.

Physico-chemical characterisation, cytotoxic activity, and biocompatibility studies of tamoxifen-loaded solid lipid nanoparticles prepared via a temperature-modulated solidification method

Context: Solid lipid nanoparticles (SLNs) can efficiently and efficaciously incorporate anti-cancer agents. Objective: To prepare and characterise tamoxifen (TAM)-loaded SLNs. Materials and methods: Glyceryl monostearate, Tween-80, and trehalose were used in SLNs. SLNs were tested via dynamic light scattering (DLS), transmission electron microscopy (TEM), differential scanning calorimetry (DSC), X-ray diffraction (XRD), and Fourier transform infrared spectroscopy (FTIR). Results: Characterisation studies revealed SLNs of about 540 nm with a negative surface charge and confirmed the entrapment of TAM in the SLNs. The entrapment efficiency was estimated to be 60%. Discussion: The in vitro drug release profile demonstrated a gradual increase followed by a release plateau for several days. A drug concentration-dependent increase in cytotoxic activity was observed when the SLNs were evaluated in cell cultures. Conclusion: Biocompatible and stable lyophilised SLNs were successfully prepared and found to possess properties that may be utilised in an anti-cancer drug delivery system.

An approach to C-glycosidic conjugates of isoflavones

Abstract A novel approach to isoflavone glycoconjugates, designed as less biodegradable congeners of natural glycosides, is presented on example of daidzein linkage to a C-glycosidic synthon derived from l-rhamnose. 1,3-Dipolar cycloaddition was employed for chemical ligation of daidzein 7-O-propargyl ester and alkyl azide containing 2,3-unsaturated pyranoside moiety. The obtained constructs with opposite anomeric configuration both exhibited a considerable increase in cytotoxic activity against the HTC 116 cell line, in comparison to the parent isoflavone.

Enhanced Gene Expression Following Vaccination in Chronic Fatigue Syndrome/Myalgic Encephalomyelitis

Vaccines have been shown to cause differential expression of genes and increase antibody titers against antigens. Influenza vaccines may have an effect on unexplained disorders such as Chronic Fatigue Syndrome/Myalgic Encephalomyelitis (CFS/ME). Immunological changes have been identified following immunization with trivalent influenza vaccine (TIV). The objective of this pilot study was to examine the consequences of TIV on cytokine and cytotoxic genes in CFS/ME. Peripheral blood mononuclear cells were preferentially isolated from whole blood of 7 CFS/ME patients and 8 controls. Following total RNA extraction and synthesis of cDNA, reverse transcriptase-quantitative polymerase chain reaction (RT-qPCR) was used to determine the expression levels of mRNAs for cytotoxic genes (perforin (PRF1), granzyme A (GZMA), granzyme B (GZMB) and cytokine genes. GZMB was significantly increased overall in the CFS/ME patients compared to the controls. GZMA was significantly increased 28 days after vaccination while PRF1 was reduced pre-vaccination but increased 14 days post-vaccination in the CFS/ME patients. There were no significant changes in cytokine genes pre or post vaccination. Administration of TIV may increase the expression of lytic genes in CFS/ME and this may contribute to the increase in cytotoxic activity we observed in these patients post vaccination.

T and NK cells of B cell NHL patients exert cytotoxicity against lymphoma cells following binding of bispecific tetravalent antibody CD19×CD3 or CD19×CD16.

Bispecific tetravalent antibodies (TandAb) directed against the B cell surface marker CD19 and activating receptors on T or NK cells (CD19×CD3 or CD19×CD16) have shown promising effects in vitro and in preclinical studies. Here, we examine the cytotoxic efficacy of T and NK cells from patients with B cell Non-Hodgkin's Lymphoma (NHL) against B-lymphoma cells following the binding of the matching TandAb. The addition of CD19×CD16 TandAb led to a threefold increase in NK cell activation in the presence of B-lymphoma cells. Similarly, T cells displayed a sevenfold increase in cytotoxic activity after the addition of CD19×CD3 TandAb. Comparison of T and NK cell effector function of patients and healthy controls showed comparable levels of cytotoxic activity in response to lymphoma cells and no reduction in functional activity due to age, disease stage or the type and amount of previous therapy. Thus, T and NK cells of patients with B cell NHL are fully capable of being activated by therapeutic crosslinking antibodies. These results provide a rationale for the use of TandAbs for patients with B cell NHL, particularly in cases where remission with minimal residual disease could be achieved by cytotoxic chemotherapy.

Synthesis and antitumour activities of a novel class of dehydroabietylamine derivatives

Structural modification is still a popular and important route in the forest chemical field for finding novel tricyclic diterpenes with more potential bioactivities and broad bioactive spectra. In this study, a series of dehydroabietylamine derivatives containing tricyclic diterpene structures were synthesised through oxidation in the 7th position of ring B and nitrification in the 12th position of ring C using dehydroabietylamine as the starting material. Structures of the synthesised compounds were confirmed by IR, 1H-NMR, 13C-NMR, MS and HRMS. The cytotoxicities of these compounds against PC-3 (human prostate carcinoma cell line) and Hey-1B (human ovarian carcinoma cell line) cells by the MTT assay were investigated. The results showed that the presence of a nitro group at 12th position and a carbonyl group at 7th position resulted in an increase of cytotoxic activity. Our findings present more evidence, showing the relationship between the chemical structure and biological function.