Concentrations Of Bleomycin Research Articles

Downregulation of PTEN mediates bleomycin-induced premature senescence in lung cancer cells by suppressing autophagy.

ObjectiveBleomycin is an important chemotherapeutic drug that activates premature senescence to decrease the tumorigenic process. We aimed to investigate the role of phosphatase and tensin homolog deleted on chromosome ten (PTEN) in bleomycin-induced premature senescence in lung cancer cells.MethodsHuman lung cancer A549 cells were incubated in the presence of different concentrations of bleomycin for 5 days. A lentivirus vector was used to silence the PTEN gene, followed by stimulation with bleomycin (1 µg/mL). Changes were evaluated by senescence-associated β-galactosidase staining, reverse transcription-polymerase chain reaction, and western blot.ResultsTreatment with bleomycin induced premature senescence. PTEN expression was decreased and key downstream molecules in the phosphoinositide 3-kinase (PI3K)/Akt/mammalian target of rapamycin (mTOR) pathway were gradually activated following bleomycin treatment. Silencing PTEN reduced autophagy and accelerated senescence of A549 cells. Autophagy levels were also increased and senescence markers were reduced after inhibiting mTOR.ConclusionsDownregulation of PTEN mediates bleomycin-induced premature senescence in lung cancer cells by suppressing autophagy via the PI3K/Akt/mTOR pathway. These findings provide new insights into the potential role of PTEN as a molecular target for cancer chemotherapy.

Prostatic Artery Embolization for Benign Prostatic Hyperplasia: Bleomycin-Eluting versus Bland Microspheres in a Canine Model

PurposeTo prospectively assess safety and efficacy of prostatic artery embolization (PAE) with bleomycin-eluting microspheres for benign prostatic hyperplasia (BPH) in a canine model. Materials and MethodsTwelve adult male beagles (mean age, 1.6 y ± 0.2; range, 1.2–2.0 y) were randomly assigned to group A (n = 6; PAE with bleomycin-eluting 30–60-μm HepaSphere microspheres) and group B (n = 6; PAE with bland 30–60-μm HepaSphere microspheres) between April 2017 and November 2018. Plasma bleomycin concentration in group A was measured within 7 days. Prostate volume (PV) and ischemic volume after PAE were measured by magnetic resonance imaging. Prostates and adjacent organs were harvested after the last magnetic resonance study and histopathologically examined. ResultsPlasma bleomycin concentration peaked at 10 minutes at 2,055.0 ng/mL ± 606.1 and lasted for 1,440 min at low levels after PAE. PV reduction percentage was greater in group A than in group B at 1 month (74.1% ± 4.3 vs 63.7% ± 3.5; P = .006) and 3 months (61.5% ± 6.7 vs 46.1% ± 3.8; P = .001) after PAE. Proportion of prostate ischemic volume was greater in group A than in group B (75.3% ± 3.0 vs 62.0% ± 7.1; P = .006) at 1 month after PAE. Proportion of prostate ischemic volume at 1 month positively correlated with PV percentage reduction at 3 months in group A (r = 0.840, P = .036) and group B (r = 0.844, P = .035). There were no complications or nontarget embolization to surrounding organs after the procedures. ConclusionsIn a canine model, PAE with bleomycin-eluting microspheres was feasible and well tolerated and caused ischemic necrosis and reduction in PV.

Bleomycin Polidocanol Foam (BPF) Stability – In Vitro Evidence for the Effectiveness of a Novel Sclerosant for Venous Malformations

This study investigated the invitro stability of a novel sclerosant, bleomycin polidocanol foam (BPF), for venous malformation (VM) sclerotherapy. The study was designed with control groups treated with polidocanol (0.5%, 1%, and 3%) only. Theexperimental groups included 21 BPFs, which was made by dissolving bleomycin at seven differentconcentrations (0.1%-1.5%) in polidocanol (0.5%, 1%, and 3%). The Tessari method was used to prepare sclerosant foam with a liquid:gas ratio of 1:4at room temperature invitro. The foam stability was measured for each group. The decay process, one component of foam stability, was recorded with acamera. Foam decay process experiments were performed 10 times per group. The stability indices included drainage rate, drainage time, half life, and microscopic measurement of the foams (mean bubble diameter, minimum and maximum bubble diameters, wall thickness, and bubble diameter distribution). Compared with the control groups, the half lives of BPFs mainly increased significantly with the addition of bleomycin (p<.001). BPF with 3% polidocanol and 0.1% bleomycin recorded the highest half life (246±1.6sec), and this group also achieved the smallest bubble diameter and wall thickness (69.9μm and 5.80μm) among the experimental groups. For the same polidocanol concentration, the bubble diameter and wall thickness increased when bleomycin was added. Bleomycin concentrations account for different BPF stability. BPF stability mainly increased significantly with the addition of a small amount of bleomycin but this advantage was no longer apparent with increasing bleomycin dose.

Intralesional injection of bleomycin in the treatment of xanthelasma palpebrarum: A clinical study.

Xanthelasma palpebrarum (XP) is the most common type of cutaneous xanthoma and has been treated with intralesional injection of pingyangmycin effectively. However, bleomycin, which has the same effect in antitumor activity as pingyangmycin, has not been applied in the treatment of XP. To explore and assess the treatment of xanthelasma by intralesional injection of bleomycin, which has been widely used as an antitumor antibiotic, for the replacement of pingyangmycin. Intralesional injection of different concentrations of bleomycin was administered to 44 xanthelasma lesions of 24 patients who have never been treated before, divided into two groups according to age. Photographs were taken and analyzed to assess the therapeutic efficiency. Patients were then followed up for 6-24months. All the lesions resolved after 1month of treatment with the intralesional injection of different concentrations of bleomycin. There was no significant difference observed between the two groups. No severe complications had occurred. The treatment of XP with intralesional injection of bleomycin is minimally invasive, safe, and effective. Consequently, it also has good cosmetic outcome with no adverse effects.

Dual-mode visible light-induced aptasensing platforms for bleomycin detection based on CdS–In2S3 heterojunction

CdS–In2S3 heterojunction with enhanced photoelectrochemical (PEC) performance was synthesized to construct dual-mode visible light-induced biosensors for highly sensitive and selective detection of bleomycin (BLM). Due to improved absorption in the visible region and suppressed recombination of electron-hole pairs in the heterojunction, CdS–In2S3 composite exhibited enhanced photocurrent response under visible light illumination. Using CdS–In2S3 as photoactive materials and BLM-binding aptamer as recognition element, a PEC aptasensor displaying a declined photocurrent response to BLM was facilely constructed, which was linear to BLM concentration in the range of 5.0–250 nM. On the other hand, the CdS–In2S3 photoanode was employed to construct a photofuel cell (PFC). In such a PFC, the oxidation of water on CdS–In2S3 photoanode under visible light illumination and the reduction of oxygen on Pt cathode led to the generation of electricity. When BLM-binding aptamer was immobilized on CdS–In2S3 photoanode, the output power of the PFC was inversely proportional to the logarithm of BLM concentration from 10 to 250 nM, offering a visible light-induced self-powered sensing platform for BLM detection. Both of the proposed sensors showed high selectivity, good reproducibility and high stability. They were successfully applied to the determination of BLM in human serum samples.

A Novel Method for Controlled Gene Expression via Combined Bleomycin and Plasmid DNA Electrotransfer.

Electrochemotherapy is an efficient method for the local treatment of cutaneous and subcutaneous metastases, but its efficacy as a systemic treatment remains low. The application of gene electrotransfer (GET) to transfer DNA coding for immune system modulating molecules could allow for a systemic effect, but its applications are limited because of possible side effects, e.g., immune system overactivation and autoimmune response. In this paper, we present the simultaneous electrotransfer of bleomycin and plasmid DNA as a method to increase the systemic effect of bleomycin-based electrochemotherapy. With appropriately selected concentrations of bleomycin and plasmid DNA, it is possible to achieve efficient cell transfection while killing cells via the cytotoxic effect of bleomycin at later time points. We also show the dynamics of both cell electrotransfection and cell death after the simultaneous electrotransfer of bleomycin and plasmid DNA. Therefore, this method could have applications in achieving the transient, cell death-controlled expression of immune system activating genes while retaining efficient bleomycin mediated cell killing.

Plasma Bleomycin Concentrations during Electrochemotherapeutic Treatment of Fibropapillomas in Green Turtles Chelonia mydas.

Fibropapillomatosis of sea turtles is traditionally treated with surgical debulking techniques that are often associated with prolonged healing and tumor recurrence. Electrochemotherapy was recently described for green turtles Chelonia mydas and can be an alternative to surgery and even general anesthesia. The objectives of this study were to replicate an electrochemotherapy protocol from a previous report and add plasma bleomycin analysis to the treatment. After bleomycin injection into similarly sized tumors of two green turtles and immediate electroporation at two time points, plasma bleomycin reached detectable concentrations that were considerably lower than those found in human studies. At 3months posttherapy, no healing complications or recurrences were encountered and only scar tissue remained. This study adds further support that electrochemotherapy with bleomycin has the potential to be used as an effective alternative treatment for this complex disease.

NMDA receptor activation inhibits the protective effect of BM‑MSCs on bleomycin‑induced lung epithelial cell damage by inhibiting ERK signaling and the paracrine factor HGF.

Endoplasmic reticulum (ER) stress in alveolar epithelial cells (AECs) is associated with the pathogenesis of pulmonary fibrosis. Bone marrow-derived mesenchymal stromal cells (BM-MSCs) can exert protective effects on ER-stressed AECs via paracrine signaling. In the present study, mouse lung epithelial (MLE)-12 cells were directly stimulated with various concentrations of bleomycin (BLM). MLE-12 cell apoptosis was detected by flow cytometry, and Ki67 expression was detected by immunofluorescence to reflect cell proliferation. The results revealed that BLM increased the protein expression levels of X-box binding protein 1 and immunoglobulin heavy chain-binding protein, thus inducing ER stress, and caused cell dysfunction by inhibiting proliferation and promoting apoptosis. In addition, MSC-derived conditioned medium (MSC-CM) protected MLE-12 cells from BLM-induced injury, by reducing ER stress, promoting cell proliferation and inhibiting cell apoptosis. Our previous studies reported that N-methyl-D-aspartate (NMDA) receptor activation partially inhibits the antifibrotic effect of BM-MSCs on BLM-induced pulmonary fibrosis through downregulating the paracrine factor hepatocyte growth factor (HGF). In the present study, the synthesis and secretion of HGF were detected by western blotting and ELISA, respectively. Results further demonstrated that NMDA inhibited the synthesis and secretion of HGF in BM-MSCs, and NMDA-preconditioned MSC-CM had no protective effects on BLM-induced injury in MLE-12 cells. In addition, activation of the NMDA receptor decreased the phosphorylation levels of extracellular signal-regulated kinase (ERK)1/2 in BM-MSCs. Using Honokiol and FR180204, the activator and inhibitor of ERK1/2, respectively, it was then revealed that Honokiol partially eliminated the decrease in HGF expression, whereas FR180204 further promoted the reduction in HGF caused by NMDA. Collectively, these findings suggested that NMDA receptor activation may downregulate HGF by inhibiting ERK signaling in BM-MSCs, thus weakening their protective effects on BLM-induced lung epithelial cell damage.

The effects of low irradiance long duration photochemical internalization on glioma spheroids.

Photodynamic therapy (PDT), if given over extended time periods (i.e. hours or days) and at very low irradiance in the μW/cm2 range, has been shown to be more effective than acute PDT (aPDT) administered over minutes. This has led to the concept of metronomic PDT (mPDT), which consists of ultra-low irradiance light illumination for extended periods of time along with either continuous or repetitive delivery of photosensitizer. Since the drug activating technology photochemical internalization (PCI) is based on PDT it seemed reasonable to expect that ultra-low irradiance, if administered over an extended period of time, could nevertheless result in effective metronomic PCI (mPCI) comparable to or more effective than that obtained with relatively high and short irradiance i.e. acute PCI (aPCI). Tumor spheroids consisting of F98 cells were used as in-vitro tumor models. The amphiphilic photosensitizer Al phthalocyanine disulfonate (AlPcS2a) was used for all PCI experiments. Light treatment was administered from a diode laser at λ=670 nm at various irradiance exposures of 2 mW/cm2 for aPCI and 0.05 - 0.2 mW/cm2 for mPCI with durations ranging from 3 to 12 min for aPCI and 120 min for mPCI. AlPcS2a fluorescence was seen throughout the cytosol following short or long light treatment, corresponding to aPCI and mPCI respectively. Spheroid growth was significantly inhibited or completely suppressed at a mPCI radiance of 0.05 or 0.72 J/cm2 respectively, with all bleomycin (BLM) concentrations used, compared to either BLM alone or aPCI at radiant exposure at these levels. The effects of BLM-aPCI and mPCI were comparable at radiance levels of 0.96 and 1.44 J/cm2. Results show that mPCI could effectively cause significant spheroid growth inhibition with the delivery of extremely low light irradiance rates delivered over an extended period of time. These findings suggest that effective implementation of mPCI can deliver adequate drug efficacy at depths necessary to reach infiltrating glioma cells in the surgical resection cavity wall.

Evaluation of efficacy and safety of intralesional bleomycin in the treatment of common warts: Results of a pilot study.

This study evaluated the efficacy and safety of intralesional bleomycin in the treatment of common warts in 50 (32 men, 18 women) patients aged between 14 and 80 (mean ± SD, 28.5 ± 13.27) years. The warts were present over dorsal hands, feet, palms, soles and periungual skin for 1 month to 10 years. They were infiltrated with bleomycin (1 mg/ml) till blanching. The total cumulative dose did not exceed 2 mg in one session. The treatment was repeated after paring of eschar at 2 weeks in case there was no or partial response. The patients were reviewed at 4, 12 and 24 weeks for cure, adverse effects or recurrences and outcome satisfaction levels. Complete cure without recurrence occurred in 40 (80%) patients and partial response occurred in 7 (14%) patients at the end of the 24-week study period. Three patients did not complete follow-up. No major systemic or local adverse effects other than injection site pain for 2-3 days were noted. All cured patients were very satisfied (Likert scale 5). Intralesional bleomycin appears to be an effective and safe treatment for common warts including palmoplantar and periungual warts. It carries the advantage of low dose, no significant adverse effects and high patient satisfaction. Small number of patients, lack of a control group, comparing different bleomycin concentrations and a short follow-up are a few limitations of this study. Better designed studies are warranted for this useful treatment modality.

Bleomycin-induced genotoxicity in vitro in human peripheral blood lymphocytes evidenced as complex chromosome- and chromatid-type aberrations

Bleomycin is a chemotherapeutic and a radiomimetic drug which induces single and double-strand breaks in DNA by forming free radicals. We demonstrate in this study the capacity of bleomycin in inducing complex chromosome- and chromatid-type aberrations. Human peripheral blood was exposed to different concentrations of bleomycin (0, 10, 20, 30 and 40 μg/mL) and the aberrations induced were studied. The chromosomal-type aberrations studied were dicentrics, tricentrics, tetracentrics, centric rings and acentric fragments. The chromatid-type aberrations studied were double minutes, terminal lesions and terminal deletions. Though the overall trends that we obtained in the dose-dependent mitotic index and the chromosome- and chromatid-type aberrations conform to the reported literature, we could observe enhanced numbers and the types of such damages in this study. We could notice that chromosome-type aberrations were more than the chromatid-type aberrations. The enhanced numbers and the types of aberrations induced pave way for enhancing the sensitivity of genotoxic assays. Also, with more numbers and type of aberrations available, it would be useful to study the mechanisms of genotoxicity of drugs and in understanding phenomena such as “tolerance induction” to chronic exposure to such mutagens.

Core-Shell Heterostructured CuFe@FeFe Prussian Blue Analogue Coupling with Silver Nanoclusters via a One-Step Bioinspired Approach: Efficiently Nonlabeled Aptasensor for Detection of Bleomycin in Various Aqueous Environments.

We synthesized novel core-shell heterostructured Prussian blue analogue (PBA) nanospheres coupled with silver nanoclusters (AgNCs) via a one-step bioinspired approach and further exploited these as aptasensors for the detection of a trace antibiotic, bleomycin (BLM). Using FeFe Prussian blue (FeFe PB) as the core, a bimetallic CuFe@FeFe PBA layer was prepared by coupling with AgNCs synthesized by taking the BLM-targeted aptamer as a template (denoted by AgNCs/Apt@CuFe@FeFe). The coupling of AgNCs/Apt via a one-step bioinspired approach not only can improve the sensing performance of CuFe@FeFe-based aptasensors but also can shorten the aptasensor fabrication procedure. Due to the strong coordination interaction between abundant Fe(II) ions contained in CuFe@FeFe PBA nanospheres and BLM (represented by Fe(II)·BLM), the Fe(II)·BLM complex formed enables aptamer strands to undergo an irreversible cleavage event that can result in a significant change in electrochemical activity. Electrochemical results displayed that both CuFe@FeFe- and AgNCs/Apt@CuFe@FeFe-based aptasensors exhibited high sensitivity and selectivity, good stability and reproducibility, and acceptable applicability toward BLM. In comparison with the pristine CuFe@FeFe-based aptasensor (limit of detection (LOD) = 0.49 fg mL-1 within the BLM concentration from 1.0 to 2.0 ng mL-1), the as-prepared AgNCs/Apt@CuFe@FeFe-based aptasensor gave a extremely lower LOD value of 0.0082 fg mL-1 within a relatively narrow BLM concentration range (0.01 fg mL-1 to 0.1 pg mL-1). The proposed method can broaden the application of PBA nanomaterials in food safety and biosensing fields and provides a potential determination method for rapidly detecting BLM in various aqueous environments.

A pilot study on the use of ultra‐deformable liposomes containing bleomycin in the treatment of equine sarcoid

SummaryThe efficacy of a novel topical liposome‐encapsulated preparation of bleomycin (Bleosome) was studied in 118 clinical cases of equine sarcoid and efficacy was compared with two other standard conventional treatments, tazarotene and 5‐fluorouracil (5‐FU) as well as with the Bleosome in combination with each of these two conventional treatments. Treatments were arbitrarily assigned. Fifty‐two of the 118 sarcoids (44%) were resolved after 12 months but there were significant differences between treatment groups with the combinations of either 5‐FUand bleomycin, or tazarotene and bleomycin resulting in significantly superior resolution of 77 and 78% of the lesions, respectively. The preliminary results suggest that bleomycin in a liposomal carrier may be a useful treatment modality for superficial, diffuse and verrucose sarcoids. The treatment process was simple, and outcomes were functionally and cosmetically excellent. Liposome‐encapsulated bleomycin is economic compared to radiation and other treatment options and can be applied effectively by the owner under guidance. Further studies with different administration protocols and higher concentrations of bleomycin are warranted.

S100A9 aggravates bleomycin-induced dermal fibrosis in mice via activation of ERK1/2 MAPK and NF-κB pathways.

Objective(s):This study aims to investigate the pathogenicity and possible mechanisms of S100A9 function in mice models of scleroderma.Materials and Methods:The content of S100A9 in the skin tissues of mice with scleroderma was determined. Different concentrations of bleomycin (BLM) and S100A9 were subcutaneously injected into the backs of mice simultaneously, and then pathological changes in the skin of these mice were monitored. Specifically, the levels of inflammatory cytokines and alpha smooth muscle actin (α-SMA), the activation of extracellular regulated kinase 1/2 (ERK1/2), mitogen-activated protein kinase (MAPK) and nuclear factor-kappa B (NF-κB) pathways, and the expression of the receptor for advanced glycation end-product (RAGE) in the skin were determined.Results:The content of S100A9 in the skin tissues of mice with scleroderma was determined. Different concentrations of BLM and S100A9 were subcutaneously injected into the backs of mice simultaneously, and then pathological changes in the skin of these mice were monitored. Specifically, the levels of inflammatory cytokines and alpha smooth muscle actin (α-SMA), the activation of extracellular regulated kinase 1/2 (ERK1/2) mitogen-activated protein kinase (MAPK) and nuclear factor-kappa B (NF-κB) pathways, and the expression of the receptor for advanced glycation end-product (RAGE) in the skin were determined.Conclusion:S100A9 aggravates dermal fibrosis in BLM-induced scleroderma (BIS) mice, and its mechanisms might be mediated by RAGE, ERK1/2, and NF-κB pathway.

Electrochemotherapy with bleomycin and cisplatin enhances cytotoxicity in primary and metastatic uveal melanoma cell lines in vitro.

Electrochemotherapy (ECT) enhances responsiveness to cytotoxic drugs in numerous cell lines in vitro. Clinically ECT is widely applied for skin tumor ablation and has shown efficacy in treating non-resectable colorectal liver metastases. There is limited experience of ECT for ocular tumor therapy. We investigated the cytotoxic effect of bleomycin and cisplatin in combination with electroporation on chemoresistant human uveal melanoma (UM) cell lines in vitro. Four UM cell lines (Mel 270, 92-1, OMM-1, OMM-2.5) were treated with electroporation (pulse amplitude 300-1000 V/cm, 8-80 pulses, 100 μs, 5 Hz) and increasing concentrations of bleomycin and cisplatin (0-7.5 μg/ml). Cell survival was analyzed by MTT viability assay after 36 hours. UM cell lines were resistant to both bleomycin and cisplatin. In combination with electro- poration, the effects of bleomycin and cisplatin were increased 8-70 fold and 3-15 fold, respectively, in all UM cell lines. At the lowest concentration of bleomycin tested (1 μg/ml), viability was maximally reduced in all UM cell lines by ≥69% with electroporation conditions of 750 V/cm and 20 pulses. All UM cell lines were more resistant to cisplatin; however, electro- poration of 1000 V/cm and 8 pulses resulted in similar reductions in cell viability of 92-1, Mel270 with 2.5 μg/ml cisplatin, OMM2-5 cells with 5 μg/ml cisplatin and OMM1 cells with 1 μg/ml cisplatin. In vitro ECT with bleomycin or cisplatin is more effective than the highest concentration of the antineoplastic drug or electroporation alone, opening new perspectives in primary and metastatic UM treatment.

HRP-Mimicking DNAzyme-Catalyzed in Situ Generation of Polyaniline To Assist Signal Amplification for Ultrasensitive Surface Plasmon Resonance Biosensing

It is well-known that the horseradish peroxidase- (HRP-) mimicking DNAzyme, namely, hemin/G-quadruplex, can effectively catalyze the polymerization of aniline to form DNA-guided polyaniline. Meanwhile, polyaniline exhibits extraordinary electrical, electrochemical, and redox properties, as well as excellent SPR signal-enhancing ability. Herein, we report a novel ultrasensitive surface plasmon resonance (SPR) biosensor based on HRP-mimicking DNAzyme-catalyzed in situ formation of polyaniline for signal amplification, using bleomycin (BLM) as the proof-of-concept analyte. The recognition and the subsequent cleavage of DNA probe P1 by BLM switches off the hybridization between P1 and the G-rich DNA probe P2, resulting in less hemin/G-quadruplex complexes and reduced DNA-guided polyaniline deposition on the SPR Au disk surface. As compared to the case when BLM is absent, a significant shift in SPR angle is observed, which is dependent on the BLM concentration. Therefore, ultrasensitive SPR detection of the target BLM is realized, with a detection limit down to 0.35 pM, much lower than those reported in the literature. Moreover, the proposed SPR biosensor has been successfully applied for the detection of BLM spiked in human serum samples. The HRP-mimicking DNAzyme-catalyzed in situ polyaniline deposition and polyaniline-assisted signal amplification not only significantly improves the specificity and the sensitivity of the BLM assay but also allows the ultrasensitive detection of other biomolecules by simply changing the specific target recognition DNA sequences, thus providing a versatile SPR biosensing platform for the ultrasensitive detection of a variety of analytes and showing great potential for application in the fields of bioanalysis and clinical biomedicine.

Bleomycin increases neutrophil adhesion to human vascular endothelial cells independently of upregulation of ICAM-1 and E-selectin

ABSTRACTAim of the Study: Bleomycin-induced lung disease is a serious complication of therapy characterized by alveolar injury, cytokine release, inflammatory cell recruitment, and eventually pulmonary fibrosis. The mechanisms underlying bleomycin-induced pulmonary fibrosis may be relevant to other progressive scarring diseases of the lungs. Pulmonary vascular endothelial cells are critically involved in immune cell extravasation at sites of injury through adhesion molecule expression and cytokine release. We sought to determine the effects of bleomycin on adhesion molecule expression and cytokine release by pulmonary vascular endothelial cells, and their functional relevance to inflammatory cell recruitment. Materials and Methods: The effects of pharmacologically relevant concentrations of bleomycin on adhesion molecule expression and cytokine release by human vascular endothelial cells in vitro were studied by flow cytometry, quantitative polymerase chain reaction, and enzyme-linked immunosorbent assay. A flow chamber model was used to assess the functional consequences on adhesion of flowing human neutrophils to endothelial cell monolayers. Results: Bleomycin increased intercellular adhesion molecule 1 (ICAM-1; CD54), vascular cell adhesion molecule (VCAM-1; CD106), and E-selectin (CD62E) expression, and increased monocyte chemoattractant protein (MCP-1) and interleukin (IL-8) release by endothelial cells. Increases in protein expression were accompanied by increased mRNA transcription. In contrast, there was no direct effect of bleomycin on the profibrotic cytokines transforming growth factor-beta (TGF-β), platelet-derived growth factor-BB (PDGF-BB), or endothelin-1. Under flow conditions, endothelial cells exposed to bleomycin supported increased neutrophil adhesion which was independent of ICAM-1 or E-selectin. Conclusion: Our findings demonstrate that bleomycin promotes endothelial-mediated inflammation and neutrophil adhesion. These mechanisms may contribute to the development of pulmonary fibrosis by supporting immune cell recruitment in the lungs.

A bicyclo-hairpin probe mediated strand displacement amplification strategy for label-free and sensitive detection of bleomycin

Monitoring of bleomycin concentration is critical in clinical therapy. Here, a bicyclo-hairpin probe mediated strand displacement amplification strategy was developed for bleomycin detection. Firstly, bleomycin cleaved bicyclo-hairpin probe to release a trigger sequence. Then the trigger sequence initiated strand displacement amplification reaction to produce numerous G-quadruplex forming sequences. Finally, G-quadruplexes bound with N-methyl mesoporphyrin IX molecules and an enhanced fluorescent signal could be detected. In this strategy, a bicyclo-hairpin probe was ingeniously designed for bleomycin detection, which could silence the trigger sequence to reduce non-specific hybridization of detection method. In addition, based on bleomycin-induced DNA scission, a label-free fluorescent sensor for bleomycin detection was established. Our strategy could determine bleomycin sensitively with a detection limit of 0.34nM. Fluorescent response of detection system toward four different anti-tumor drugs proved that this strategy had admirable selectivity. Satisfactory recoveries in human serum samples revealed that our method had a great potential in real sample assay.

Bleomycin pharmacokinetics of bolus bleomycin dose in elderly cancer patients treated with electrochemotherapy.

With the aim to determine effective therapeutic window of electrochemotherapy, we analyzed bleomycin pharmacokinetic parameters in elderly patients. In prospective clinical study in the treatment of tumors with electrochemotherapy, blood samples of patients older than 65years were collected after the bolus intravenous injection of bleomycin (15,000IU/m(2)). In serum samples, quantitative analysis was performed with liquid chromatography coupled to high-resolution mass spectrometry. Based on the data, the pharmacokinetic parameters of bleomycin elimination were determined. Pharmacokinetic analysis of the data revealed a monophasic serum clearance curve, which demonstrates slow elimination of bleomycin, being less than 500ml/min and a half-time of 30min. Slow monophasic elimination of bleomycin from serum in elderly patients implies on the longer therapeutic window, from 8 to up to 40min or even longer post-bleomycin injection for electrochemotherapy. However, prolonged therapeutic bleomycin serum concentrations may also affect the possible adverse effects, such as lung fibrosis and extensive necrosis of tumors due to the uptake of toxic bleomycin concentrations into the tumors. This may imply on lowering of bleomycin dosage, in particular in the elderly patients.

Enhancement of Bleomycin Sensitivity in Human Lung Cancer Cell Line using &lt;i&gt;Centella asiatica&lt;/i&gt; Leaf Extract

Purpose: To demonstrate the effectiveness of Centella asiatica aqueous extract in augmenting the cytotoxic effect of bleomycin in the adenocarcinoma human alveolar basal epithelial A549 cell line. Methods: The inhibitory effect of bleomycin on A549 cells was determined by incubating the cells for 24 h in different concentrations of bleomycin. The effect of C. asiatica extract on A549 cells was determined after exposing cells to 5, 10, 15, 20, 25, and 30 % (v/v) bleomycin diluted from an initial 10% stock solution. Based on the 50 % inhibitory (IC 50 ) values of bleomycin, 80, 100, and 120 μg/mL concentrations were tested with 15, 20, and 25 % (v/v) of C. asiatica extract. Results: IC 50 value of bleomycin on A549 cells was 100 μg/mL, and was reduced to 80 μg/mL when cells were co-incubated with 20 % v/v of C. asiatica extract. The results show that C. asiatica extract reduces the concentration of bleomycin necessary for inhibition of A459 cell growth (IC 50 ). While the IC 50 value of bleomycin was 100 μg/ml, it fell to 80 μg/ml when used along with 20 % v/v CA extract (p < 5) Conclusions: This study demonstrates that compounds extracted from C. asiatica may have potential for use as adjuvants in bleomycin cancer chemotherapy. Keywords: Centella asiatica , A549 cells, Bleomycin, Lung cancer, Increased susceptibility, Cell viability