In vivo skin dose measurement using MOSkin detectors in tangential breast radiotherapy

The purpose of this study is to measure patient skin dose in tangential breast radiotherapy. Treatment planning dose calculation algorithm such as Pencil Beam Convolution (PBC) and in vivo dosimetry techniques such as radiochromic film can be used to accurately monitor radiation doses at tissue depths, but they are inaccurate for skin dose measurement. A MOSFET-based (MOSkin) detector was used to measure skin dose in this study. Tangential breast radiotherapies (“bolus” and “no bolus”) were simulated on an anthropomorphic phantom and the skin doses were measured. Skin doses were also measured in 13 patients undergoing each of the techniques. In the patient study, the EBT2 measurements and PBC calculation tended to over-estimate the skin dose compared with the MOSkin detector (p < 0.05) in the “no bolus radiotherapy”. No significant differences were observed in the “bolus radiotherapy” (p > 0.05). The results from patients were similar to that of the phantom study. This shows that the EBT2 measurement and PBC calculation, while able to predict accurate doses at tissue depths, are inaccurate in predicting doses at build-up regions. The clinical application of the MOSkin detectors showed that the average total skin doses received by patients were 1662 ± 129 cGy (medial) and 1893 ± 199 cGy (lateral) during “no bolus radiotherapy”. The average total skin doses were 4030 ± 72 cGy (medial) and 4004 ± 91 cGy (lateral) for “bolus radiotherapy”. In some cases, patient skin doses were shown to exceed the dose toxicity level for skin erythema. Hence, a suitable device for in vivo dosimetry is necessary to accurately determine skin dose.     

Evaluation of cytotoxicity and immune modulatory activities of soyasaponin Ab: An in vitro and in vivo study

To improve the immune efficacy of protein subunit vaccines, novel adjuvants are needed to elicit a suitable protective immune response and to promote long term immunologic memory. In this work, soyasaponin Ab, a major constituent among group A soyasaponins in soybeans was purified and prepared from soy hypocotyls. The immunomodulatory effects of soyasaponin Ab both in vitro and in vivo were investigated, and its pro-immunomodulatory molecular mechanism was also studied. For in vitro assays, with mouse macrophage cell line RAW264.7 as the studying model, both cytotoxicity and immune stimulatory activity were investigated to evaluate the potential of soyasaponin Ab as the vaccine adjuvant. The results indicated that soyasaponin Ab could be significantly safer than Quillaja saponins (QS). Soyasaponin Ab showed no toxicities over the tested concentration ranges compared to QS. Soyasaponin Ab was proved able to promote releases of inflammatory cytokines like TNFα and IL-1β in a dose-dependent manner. Furthermore, NF-κB signalling was also activated by soyasaponin Ab effectively. In addition, with TLR4 gene expression of RAW264.7 cell inhibited by RNA interference, immune stimulatory effects by soyasaponin Ab dropped down significantly. On the other hand, the in vivo experiment results showed that anti-ovalbumin (OVA) IgG, IgG1, IgG2a, IgG2b were significantly enhanced by the soyasaponin Ab and QS groups (p < 0.05 or p < 0.01). The results suggested that compared to QS, soyasaponin Ab may represent a viable candidate for effective vaccine adjuvant. TLR4 receptor dependent pathway may be involved in immune stimulatory effects of soyasaponin Ab.     

Greater impact of dietary fat manipulation than apolipoprotein E genotype on ex vivo cytokine production – Insights from the SATgenε study

Apolipoprotein E (APOE) genotype is believed to play an important role in cardiovascular risk. APOE4 carriers have been associated with higher blood lipid levels and a more pro-inflammatory state compared with APOE3/E3 individuals. Although dietary fat composition has been considered to modulate the inflammatory state in humans, very little is known about how APOE genotype can impact on this response. In a follow-up to the main SATgenε study, we aimed to explore the effects of APOE genotype, as well as, dietary fat manipulation on ex vivo cytokine production. Blood samples were collected from a subset of SATgenε participants (n = 52/88), prospectively recruited according to APOE genotype (n = 26 E3/E3 and n = 26 E3/E4) after low-fat (LF), high saturated fat (HSF) and HSF with 3.45 g docosahexaenoic acid (DHA) dietary periods (each diet eight weeks in duration assigned in the same order) for the measurement of ex vivo cytokine production using whole blood culture (WBC). Concentrations of IL-1beta, IL-6, IL-8, IL-10 and TNF-alpha were measured in WBC supernatant samples after stimulation for 24 h with either 0.05 or 1 μg/ml of bacterial lipopolysaccharide (LPS). Cytokine levels were not influenced by genotype, whereas, dietary fat manipulation had a significant impact on TNF-α and IL-10 production; TNF-α concentration was higher after consumption of the HSF diet compared with baseline and the LF diet (P < 0.05), whereas, IL-10 concentration was higher after the LF diet compared with baseline (P < 0.05). In conclusion, our study has revealed the amount and type of dietary fat can significantly modulate the production of TNF-α and IL-10 by ex vivo LPS-stimulated WBC samples obtained from normolipidaemic subjects.     

Growth inhibition and pro-apoptotic activity of violacein in Ehrlich ascites tumor

The continuing threat to biodiversity lends urgency to the need of identification of sustainable source of natural products. This is not so much trouble if there is a microbial source of the compound. Herein, violacein, a natural indolic pigment extracted from Chromobacterium violaceum, was evaluated for its antitumoral potential against the Ehrlich ascites tumor (EAT) in vivo and in vitro. Evaluation of violacein cytotoxicity using different endpoints indicated that EAT cells were twofold (IC₅₀ = 5.0 μM) more sensitive to the compound than normal human peripheral blood lymphocytes. In vitro studies indicated that violacein cytotoxicity to EAT cells is mediated by a rapid (8–12 h) production of reactive oxygen species (ROS) and a decrease in intracellular GSH levels, probably due to oxidative stress. Additionally, apoptosis was primarily induced, as demonstrated by an increase in Annexin-V positive cells, concurrently with increased levels of DNA fragmentation and increased caspase-2, caspase-9 and caspase-3 activities up to 4.5-, 6.0- and 5.5-fold, respectively, after 72 h of treatment. Moreover, doses of 0.1 and 1.0 μg kg^?1 violacein, administered intraperitoneally (i.p.) to EAT-bearing mice throughout the lifespan of the animals significantly inhibited tumor growth and increased survival of mice. In view of these results, a 35-day toxicity study was conducted in vivo. Complete hematology, biochemistry (ALT, AST and creatinine levels) and histopathological analysis of liver and kidney indicated that daily doses of violacein up to 1000 μg kg^?1 for 35 days are well tolerated and did not cause hematotoxicity nor renal or hepatotoxicity when administered i.p. to mice. Altogether, these results indicate that violacein causes oxidative stress and an imbalance in the antioxidant defense machinery of cells culminating in apoptotic cell death. Furthermore, this is the first report of its antitumor activity in vivo, which occurs in the absence of toxicity to major organs.     

Growth-promoting effects of a bacterium on raphidophytes and other phytoplankton

On 29 April 2003, a Heterosigma akashiwo bloom (9.5 × 10⁴ cells mL⁻¹) associated with a fish kill (>10⁴ dead fishes estimated from aerial surveys) was observed offshore of Bulls Bay, McLellanville, South Carolina, USA. To assess a potential cause of this bloom event, we investigated the bacterial diversity and algal/bacterial interactions in the bloom microbial community. Thirty-five bacterial strains were isolated and screened for algicidal or algal growth-promoting activities. One strain (BBB25) had significant growth-promoting effects on all 7 algal species tested: three raphidophytes (Heterosigma akashiwo, Chattonella subsalsa, Fibrocapsa japonica), two diatoms (Chaetoceros neogracile, Nitzschia sp.), a cryptophyte (Cryptomonas sp.), and a chlorophyte, Ankistrodesmus sp. This strain (BBB25) is a Gram-positive, rod-shaped spore-forming bacterium. Partial 16S rDNA gene sequence and morphological characters indicated that BBB25 is related closely to the genus Bacillus. The general nature of the algal response indicates that the growth-promoting effects of BBB25 are not specific to H. akashiwo, and suggests potentially widespread effects. Since the presence or relative abundance of the other algal species was not assessed during the bloom initiation period, the selective stimulatory effect on H. akashiwo bloom formation in Bulls Bay is unknown. These results demonstrate, however, the potential for bacterial species to play a regulatory role in bloom formation.     

Efficacy of biogenic selenium nanoparticles against Leishmania major: In vitro and in vivo studies

Project: This study investigated the in vitro and in vivo effectiveness of biogenic selenium nanoparticles (Se NPs), biosynthesized by Bacillus sp. MSh-1, against Leishmania major (MRHO/IR/75/ER). Procedure: The 3-(4,5-dimethylthiozol-2-yl)-2,5-diphenyl-tetrazolium bromide (MTT) assay was used to evaluate the cytotoxicity effects of the biogenic Se NPs against both promastigote and amastigote forms of L. major. In a separate in vivo experiment, we also determined the preventive and therapeutic effects of biogenic Se NPs in BALB/c mice following subcutaneous infected with L. major. Results: The MTT assays showed that the highest toxicity occurred after 72 h against both promastigote and amastigote forms of L. major. The cytotoxicity of Se NPs was higher at all incubation times (24, 48, and 72 h) against the promastigote than the amastigote form (p < 0.05). The 50% inhibitory concentrations (IC₅₀) of the Se NPs were 1.62 ± 0.6 and 4.4 ± 0.6 μg ml^?1 against the promastigote and amastigote forms, respectively, after a 72-h incubation period. Apoptosis assays showed DNA fragmentation in promastigotes treated with Se NPs. In an animal challenge, prophylactic doses of biogenic Se NPs delayed the development of localized cutaneous lesions. Moreover, daily administration of Se NPs (5 or 10 mg kg^?1 day^?1) in similarly infected BALB/c mice that had not received prophylactic doses of Se NPs also abolished the localized lesions after 14 days. Conclusion: Based on these in vitro and in vivo studies, biogenic Se NPs can be considered as a novel therapeutic agent for treatment of the localized lesions typical of cutaneous leishmaniasis.     

Influence of ethanol on the release of growth factors in human blood-derived platelet gels

Platelet gels (PG) are new topical single-donor blood products which are attracting great interest in regenerative medicine. They are obtained by mixing a platelet-rich plasma fraction with thrombin to generate a fibrin gel enriched in platelet growth factors (GF). The type of thrombin preparation may affect PG reproducibility. We have determined the impact of 14.6% (v/v) ethanol-stabilized thrombin (EHT) on the release of GF by platelets. Various ratios of EHT and platelet concentrates were mixed to obtain from 2.43 to 7.96% ethanol concentration. Platelet-derived growth factor-AB (PDGF-AB), transforming growth factor-ß1 (TGF-ß1), vascular endothelium growth factor (VEGF), epidermal growth factor (EGF), and insulin-like growth factor-1 (IGF-1) were assessed at 5, 120, and 300 min after PG formation. Protein profiles of thrombin and PG releasates were analyzed by SDS-PAGE. The amount of PDGF-AB, TGF-ß1, and VEGF released per platelet decreased significantly (p < 0.05) with increasing ethanol concentrations but, however, not that of EGF. IGF-1 content was stable, consistent with its presence mostly in plasma. SDS-PAGE indicated that ethanol did not affect fibrin formation. In conclusion, ethanol has a significant impact on the amount of GF released by platelets and should be strictly controlled to standardize PG and optimize clinical benefits.     

Scavenger receptors are associated with cellular interactions of S100A12 in vitro and in vivo

Increased plasma levels of S100 proteins and interaction of S100 proteins with receptor for advanced glycation end products (RAGE) have been associated with a number of disease states, including chronic inflammatory processes and atherosclerosis. However, data concerning the role of circulating S100 proteins in these pathologies in vivo are scarce and, furthermore, it is currently not known whether RAGE is the sole receptor for extracellular S100 proteins in vivo. We report a novel methodology using recombinant human S100 proteins radiolabelled with fluorine-18, particularly, ¹⁸F-S100A12, in receptor binding studies and cellular association studies in vitro, and in dynamic small animal positron emission tomography (PET) studies in rats in vivo. Association to both human aortic endothelial cells and macrophages revealed specific binding of ¹⁸F-S100A12 to RAGE, but, furthermore, provides evidence for interaction of ¹⁸F-S100A12 to various scavenger receptors (SR). PET data showed temporary association of ¹⁸F-S100A12 with tissues overexpressing RAGE (e.g., lung), and, moreover, accumulation of ¹⁸F-S100A12 in tissues enriched in cells overexpressing SR (e.g., liver and spleen). Blockade of overall SR interaction by maleylated BSA (malBSA) clearly shows diminished in vivo association of ¹⁸F-S100A12 to these tissues as well as a significant increment of the mean plasma residence time of ¹⁸F-S100A12 (4.8 ± 0.4 h vs. 2.3 ± 0.3 h). The present approach first demonstrates that besides RAGE also scavenger receptors contribute to distribution, tissue association and elimination of circulating proinflammatory S100A12.     

Leishmanicidal activity of Himatanthus sucuuba latex against Leishmania amazonensis

Himatanthus sucuuba (HsL) latex exhibited a potent leishmanicidal activity against intracellular amastigotes of Leishmania amazonensis, a causative agent of cutaneous leishmaniasis. HsL inhibited intracellular amastigote growth in a dose-dependent manner (IC₅₀ = 15.7 µg/mL). Moreover, HsL increased nitric oxide (NO) and Tumor Nuclear Factor-α (TNF-α) and decreased Transforming Growth Factor-β (TGF-β) production in macrophages. As assessed by plasma membrane integrity and mitochondrial activity, HsL showed low toxicity for host macrophages. HsL in vivo was active by the oral route, reducing the parasite load in established footpad lesions after only five doses. In summary, these findings support HsL as an interesting candidate for further evaluations regarding its potential application as a therapeutical agent against Leishmania.     

Anti-rheumatoid arthritic activity of flavonoids from Daphne genkwa

The aim of the study was to investigate the anti-rheumatoid arthritic activity of four flavonoids from Daphne genkwa (FFD) in vivo and in vitro. Flavonoids of D. genkwa were extracted by refluxing with ethanol and purified by polyamide resin. An in vivo carrageenan-induced paw edema model, tampon-granuloma model and Freund's complete adjuvant (FCA)-induced arthritis mouse model were used to evaluate the anti-rheumatoid arthritic activities of FFD. Moreover, nitric oxide (NO) release and neutral red uptake (NRU) in lipopolysaccharide (LPS)-induced murine macrophage RAW264.7 cells were used to evaluate the anti-inflammatory effect in vitro. In addition, antioxidant effect of FFD was determined using the 2,2-diphenyl-1-picrylhydrazyl (DPPH) method. A high dose of FFD significantly reduced the degree of acute inflammatory paw edema in mice as a response to carrageenan administration (p < 0.01). FFD displayed a dose-dependent inhibition of granuloma formation in mice (p < 0.05). FFD also inhibited chronic inflammation in adjuvant-induced arthritis rats when administered orally at the dose of 50 mg/kg/day (p < 0.001). In addition, FFD suppressed the production of NO and exhibited immunoregulatory function in LPS-activated RAW264.7 cells in a dose-related manner. Simultaneously, FFD revealed conspicuous antioxidant activity with IC₅₀ values of 18.20 μg/ml. FFD possesses significant anti-inflammatory and antioxidant activity, which could be a potential therapeutic agent for chronic inflammatory disorders such as rheumatoid arthritis.     

Design,synthesis and biological screening of new 4-thiazolidinone derivatives with promising COX-2 selectivity,anti-inflammatory activity and gastric safety profile

Two series of new thiazolidin-4-one derivatives 4a–c and 8a–e were designed and prepared. All the synthesized compounds were evaluated for their in vitro COX-2 selectivity and anti-inflammatory activity in vivo. Compounds 8c and 8d showed the best overall in vitro COX-2 selectivity (selectivity indexes of 4.56 and 5.68 respectively) and in vivo activities (edema inhibition % = 61.8 and 67 after 3 h, respectively) in comparison with the reference drug celecoxib (S.I. = 7.29, edema inhibition % = 60 after 3 h). In addition, 8c and 8d were evaluated for their mean effective anti-inflammatory doses (ED₅₀ = 27.7 and 18.1 μmol/kg respectively, celecoxib ED₅₀ = 28.2 μmol/kg) and ulcerogenic liability (reduction in ulcerogenic potential versus celecoxib = 85%, 92% respectively. Molecular docking studies were performed and the results were in agreement with that obtained from the in vitro COX inhibition assays.     

Strong activation of cyclooxygenase I and II catalytic activity by dietary bioflavonoids

Cyclooxygenases (COXs) catalyze the conversion of arachidonic acid to prostaglandins (PGs), thromboxanes, and hydroxyeicosatetraenoic acids. In the present study, we investigated several dietary bioflavonoids for their ability to modulate the catalytic activity of COX I and II in vitro and also in cultured cells. We found that some of them are the most powerful direct stimulators of the catalytic activity of COX I and II known to date, increasing the formation of prostaglandin products in vitro by up to 11-fold over the controls. This stimulatory effect of bioflavonoids is enzyme specific because none of them stimulates the catalytic activity of a number of lipooxygenases tested. Compared with phenol, a prototypical COX stimulator commonly used in vitro, the naturally occurring bioflavonoids are up to 29 times more efficacious in stimulating the COX activity. Additional studies using intact cells in culture showed that some of the dietary compounds that were active in the biochemical assays also activated the formation of PGE(2) (a representative PG) when they were present at 0.01 to 1 muM concentrations. The stimulatory effect of dietary compounds on COX-mediated PG formation is far more potent in intact cells than in the in vitro assays. Mechanistically, bioflavonoids mainly acted to slow down the suicidal inactivation of the COX enzymes, but they did not appear to reactivate the inactivated enzymes. The finding of this study suggests that some of the bioflavonoids likely will serve as the naturally occurring cofactors for the COX enzymes in humans.     

Nutritional value of Chlorella vulgaris: Effects of ultrasonication and electroporation on digestibility in rats

Three processed products derived from the green algae C. vulgaris were investigated: (1) spray-dried only (S-DA); (2) spray-dried and electroporated (ES-DA); (3) spray-dried; ultrasonicated treated (US-DA). A nitrogen-balance study was performed. Male growing Wistar rats, housed separately in metabolism cages, were fed the three algal products as the sole protein source at 150 mg N per 100 g of body weight. A control group of rats was fed with casein at a level to give the same protein nitrogen intake. The coefficients of total intestinal tract apparent crude protein digestibility for the different C. vulgaris products were: S-DA = 0.47 ± 0.127% (mean ± S.D.), ES-DA = 0.44 ± 0.075%, US-DA = 0.57 ± 0.137%. Protein efficiency ratio was 1.4 ± 0.3, 1.0 ± 0.5 and 2.1 ± 0.3, respectively. N-balance was 41.86 ± 32.8 mg, 31.3 ± 17.3 mg and 66.7 ± 30.1 mg, respectively. The biological value was 93 ± 9.5%, 93.6 ± 10%, and 101 ± 5%, respectively. The coefficient of total intestinal tract apparent crude protein digestibility and biological value of C. vulgaris was enhanced by ultrasonic treatment and reduced by electroporating, thus ultrasonication may be a helpful technological process in practical processing of green algae in food industry.     

Oxidation of cyanobenzocycloheptatrienes: Synthesis,photooxygenation reaction and carbonic anhydrase isoenzymes inhibition properties of some new benzotropone derivatives

The oxidation of some cyanocycloheptatrienes with CrO₃ and pyridine was investigated and a few new nitrile functionalised benzotropone derivatives were obtained. Photooxygenation reaction of these products was also studied. The structures of the formed products were determined on the basis of NMR spectroscopy and the formation mechanism of unusual products was discussed. Human carbonic anhydrase isoenzymes I, and II (hCA I and hCA II) inhibition properties of nitrile functionalized new benzotropone derivatives were also studied. Both CA isozymes were inhibited in the low micromolar range by these nitrile functionalized benzotropone analogues. The newly synthesized benzotropone derivatives showed inhibition constants in the sub-micromolar range (2.51–4.06 μM). The best hCA I inhibition was observed in 5H-benzocycloheptene-7-carbonitrile (K_i: 2.88 ± 0.86 μM). On the other hand, 5-oxo-5H-benzocycloheptatriene-7-carbonitrile showed the powerful inhibitory effect against hCA II (K_i: 2.51 ± 0.34 μM).     

Discovery of benzo[g]indol-3-carboxylates as potent inhibitors of microsomal prostaglandin E2 synthase-1

Selective inhibition of pro-inflammatory prostaglandin (PG)E₂ formation via microsomal PGE₂ synthase-1 (mPGES-1) might be superior over inhibition of all cyclooxygenase (COX)-derived products by non-steroidal anti-inflammatory drugs (NSAIDs) and coxibs. We recently showed that benzo[g]indol-3-carboxylates potently suppress leukotriene biosynthesis by inhibiting 5-lipoxygenase. Here, we describe the discovery of benzo[g]indol-3-carboxylates as a novel class of potent mPGES-1 inhibitors (IC₅₀ ? 0.1 μM). Ethyl 2-(3-chlorobenzyl)-5-hydroxy-1H-benzo[g]indole-3-carboxylate (compound 7a) inhibits human mPGES-1 in a cell-free assay (IC₅₀ = 0.6 μM) as well as in intact A549 cells (IC₅₀ = 2 μM), and suppressed PGE₂ pleural levels in rat carrageenan-induced pleurisy. Inhibition of cellular COX-1/2 activity was significantly less pronounced. Compound 7a significantly reduced inflammatory reactions in the carrageenan-induced mouse paw edema and rat pleurisy. Together, based on the select and potent inhibition of mPGES-1 and 5-lipoxygenase, benzo[g]indol-3-carboxylates possess potential as novel anti-inflammatory drugs with a valuable pharmacological profile.     

Analysis of hydroxylation and nitration products of d-phenylalanine for in vitro and in vivo radical determination using high-performance liquid chromatography and photodiode array detection

d-Phenylalanine is capable of trapping reactive oxygen species (ROS) and reactive nitrogen species (RNS) by forming three major hydroxylation (o-, m-, p-tyrosine) and two major nitration products (nitrophenylalanine, nitrotyrosine). Here, we show how a method for the analysis of these phenylalanine derivatives was established using isocratic HPLC (Nucleosil120, C18 column) coupled with photodiode array detection and validated for cell-free in vitro and in vivo determination of radical formation. An ideal separation was achieved using a mobile phase consisting of 5% acetonitrile, 50 mM KH₂PO₄, pH 3.0, a column temperature of 35 °C and a flow rate of 1.0 mL/min. Limits of detection were in the range of 5–100 nM. Linearity was given within 5 nM–100 μM (correlation coefficient >0.999). Retention times as well as peak heights exhibited a high precision (RSD: ≤0.1% and <1.5%, respectively). The feasibility of d-phenylalanine for ROS/RNS measurement was demonstrated in a cell-free in vitro assay using peroxynitrite and by analysis of brain samples of mice treated with the dopaminergic neurotoxin 6-hydroxydopamine.     

A simple but reliable method for measuring 3D Achilles tendon moment arm geometry from a single,static magnetic resonance scan

Current methods for measuring in vivo 3D muscle-tendon moment arms generally rely on the acquisition of magnetic resonance imaging (MRI) scans at multiple joint angles. However, for patients with musculoskeletal pathologies such as fixed contractures, moving a joint through its full range of motion is not always feasible. The purpose of this research was to develop a simple, but reliable in vivo 3D Achilles tendon moment arm (ATMA) technique from a single static MRI scan. To accomplish this, for nine healthy adults (5 males, 4 females), the geometry of a cylinder was fit to the 3D form of the talus dome, which was used to estimate the talocrural flexion/extension axis, and a fifth-order polynomial fit to the line of action of the Achilles tendon. The single static scan in vivo 3D ATMA estimates were compared to estimates obtained from the same subjects at the same ankle joint angles using a previously validated 3D dynamic MRI based in vivo ATMA measurement technique. The ATMA estimates from the single scan in vivo 3D method (52.5 mm ± 5.6) were in excellent agreement (ICC = 0.912) to the validated in vivo 3D method (51.5 mm ± 5.1). These data show reliable in vivo 3D ATMA can be obtained from a single MRI scan for healthy adult populations. The single scan, in vivo 3D ATMA technique provides researchers with a simple, but reliable method for obtaining subject-specific ATMAs for musculoskeletal modelling purposes.     

Anti-angiogenic activity of julibroside J8, a natural product isolated from Albizia julibrissin

PurposeThe purpose of this study was to investigate the anti-angiogenic properties of julibroside J₈, a triterpenoid saponin isolated from Albizia julibrissin.MethodsIn the presence of juliborside J_8, the growth of human microvascular endothelial cells (HMEC-1), four human tumor cell lines, and a normal cell line (MRC-5) was evaluated by MTT assay. The in vivo anti-angiogenic effect of julibroside J₈ was evaluated on a chorioallantoic membrane (CAM) and in transplanted colon carcinoma cells in a nude mice neovascularisation model.ResultsTreatment with 0.5–4 μg/ml julibroside J₈ resulted in dose-dependent inhibition of growth, migration, and tube formation in HMEC-1 cells; julibroside J₈ also inhibited the formation of microvessels on CAM at a concentration of 10–50 μg/egg and reduced vessel density within tumor at a concentration of 0.5–3 mg/kg.ConclusionsJulibroside J₈ may be a potent anti-angiogenetic and cytotoxic drug; further investigation is warranted.     

A phase II study of allogeneic natural killer cell therapy to treat patients with recurrent ovarian and breast cancer

BackgroundNatural killer (NK) cells derived from patients with cancer exhibit diminished cytotoxicity compared with NK cells from healthy individuals. We evaluated the tumor response and in vivo expansion of allogeneic NK cells in recurrent ovarian and breast cancerMethodsPatients underwent a lymphodepleting preparative regimen: fludarabine 25 mg/m² × 5 doses, cyclophosphamide 60 mg/kg × 2 doses, and, in seven patients, 200 cGy total body irradiation (TBI) to increase host immune suppression. An NK cell product, from a haplo-identical related donor, was incubated overnight in 1000 U/mL interleukin (IL)-2 prior to infusion. Subcutaneous IL-2 (10 MU) was given three times/week × 6 doses after NK cell infusion to promote expansion, defined as detection of ≥100 donor-derived NK cells/μL blood 14 days after infusion, based on molecular chimerism and flow cytometryResultsTwenty (14 ovarian, 6 breast) patients were enrolled. The median age was 52 (range 30–65) years. Mean NK cell dose was 2.16 × 10⁷cells/kg. Donor DNA was detected 7 days after NK cell infusion in 9/13 (69%) patients without TBI and 6/7 (85%) with TBI. T-regulatory cells (Treg) were elevated at day +14 compared with pre-chemotherapy (P = 0.03). Serum IL-15 levels increased after the preparative regimen (P = < 0.001). Patients receiving TBI had delayed hematologic recovery (P = 0.014). One patient who was not evaluable had successful in vivo NK cell expansionConclusionsAdoptive transfer of haplo-identical NK cells after lymphodepleting chemotherapy is associated with transient donor chimerism and may be limited by reconstituting recipient Treg cells. Strategies to augment in vivo NK cell persistence and expansion are needed.