PubMedCrossRef 9 McVay CS, Velasquez M, Fralick JA: Phage therap

PubMedCrossRef 9. McVay CS, Velasquez M, Fralick JA: Phage therapy of Pseudomonas aeruginosa infection in a mouse burn wound model.

MK 1775 Antimicrob Agents Chemother 2007, ACP-196 price 51:1934–1938.PubMedCrossRef 10. BergogneBerezin E, Towner KJ: Acinetobacter spp, as nosocomial pathogens: Microbiological, clinical, and epidemiological features. Clin Microbiol Rev 1996, 9:148–165. 11. Tsakris A, Pantazi A, Pournaras S, Maniatis A, Polyzou A, Sofianou D: Pseudo-outbreak of imipenem-resistant Acinetobacter baumannii resulting from false susceptibility testing by a rapid automated system. Clin Microbiol 2000, 38:3505–3507. 12. Peleg AY, Seifert H, Paterson DL: Acinetobacter baumannii: Emergence of a successful pathogen. Clin Microbiol Rev SB203580 cost 2008, 21:538–582.PubMedCrossRef 13. Perez F, Hujer AM, Hujer KM, Decker BK, Rather PN, Bonomo RA: Global challenge of multidrug-resistant Acinetobacter baumannii. Antimicrob Agents Chemother 2007, 51:3471–3484.PubMedCrossRef

14. Dijkshoorn L, Nemec A, Seifert H: An increasing threat in hospitals: multidrug-resistant Acinetobacter baumannii. Nat Rev Microbiol 2007, 5:939–951.PubMedCrossRef 15. Navon-Venezia S, Ben-Ami R, Carmeli Y: Update on Pseudomonas aeruginosa and Acinetobacter baumannii infections in the healthcare setting. Curr Opin Infect Dis 2005, 18:306–313.PubMedCrossRef 16. Ackermann HW, Brochu G, Konjin HPE: Classification Of Acinetobacter Phages. Arch Virol 1994, 135:345–354.PubMedCrossRef 17. Klovins J, Overbeek GP, van den Worm SHE, Ackermann HW, van Duin J: Nucleotide sequence of a ssRNA phage

from Acinetobacter: kinship to coliphages. J Gen Virol 2002, 83:1523–1533.PubMed 18. Petrov VM, Ratnayaka S, Nolan JM, Miller ES, Karam JD: Genomes of the T4-related bacteriophages as windows on microbial genome evolution. Virol J 2010, 7:292.PubMedCrossRef 19. Alisky J, Iczkowski K, Rapoport A, Troitsky N: Bacteriophages show promise as antimicrobial agents. J Infect 1998, 36:5–15.PubMedCrossRef 20. Lin NT, Chiou PY, Chang KC, Chen LK, Lai MJ: Isolation and characterization of phi AB2: a novel bacteriophage of Acinetobacter baumannii. Res Microbiol 2010, 161:308–314.PubMedCrossRef about 21. Soothill JS: Treatment of experimental infections of mice with bacteriophages. J Med Microbiol 1992, 37:258–261.PubMedCrossRef 22. Chibani-Chennoufi S, Bruttin A, Dillmann ML, Brussow H: Phage-host interaction: an ecological perspective. J Bacteriol 2004, 186:3677–3686.PubMedCrossRef 23. Yang HJ, Liang L, Lin SX, Jia SR: Isolation and Characterization of a Virulent Bacteriophage AB1 of Acinetobacter baumannii. BMC Microbiol 2010, 10:10.CrossRef 24. Nakagawa T, Ishibashi JI, Maruyama A, Yamanaka T, Morimoto Y, Kimura H, Urabe T, Fukui M: Analysis of dissimilatory sulfite reductase and 16 S rRNA gene fragments from deep-sea hydrothermal sites of the Suiyo Seamount, Izu-Bonin Arc, Western Pacific. Appl Environ Microbiol 2004, 70:393–403.PubMedCrossRef 25. Adams MH: Bacteriophages. Interscience, New York; 1959. 26.

Indeed, overall complications are lowered, so as ileus and need f

Indeed, overall complications are lowered, so as ileus and need for analgesics. Hospital stay, in-hospital costs, and return to work are subject to personal differences and are biased by unblinded randomization. The better cosmetics and patients’ perceived quality

of life tend to converge with OA in a long term follow-up, similarly to other BLZ945 concentration disease treatments (i.e. colectomies) [6]. One thing is for sure: wound infections in LA are significantly and constantly less than in OA, even if OA is always less time-consuming [7]. As for the former, superficial wound infections are minor complications according to Clavien’s classification, but they indeed heighten costs, outpatients’ accesses and worsen quality of life in the first two-three weeks after the procedure [8]. Laparoscopic operative time is approximately 10 minutes

longer (confidence interval 6-15 min) than the open operation, and this difference cannot influence significantly the outcome nor the economics [9]. A potential but unstudied further advantage could regard the rate of post-operative adhesions and that of incisional hernias. Some low grade evidence suggests that in certain find more age groups (younger and females) laparoscopy could lower the occurrence of small bowel obstruction and infertility in patients who undergo appendectomy [10]. These are key points in planning a comparative study between single port and three-port appendectomy. Factors involving operative time, length of hospital stay, analgesic requirement, improvement in cosmetics and port-site hernias have to be related to a substantial equivalence or lessening on morbidity and costs. Different devices have been approved for single access-multiport surgery.

The oldest is the side-view 10 mm camera with a 3 mm operative channel used by gynaecologists. This system requires a 10 mm access, the very same as the usual umbilical optical access used in three port surgery; this modality did not gain popularity between general surgeons, due to the its absolute lack of triangulation for it generally requires a suspension for the appendix (trans-parietal stitches or supplemental miniport). The quality of view RVX-208 and the limited operability makes complicated appendicitis difficult to complete [11]. Anyway the so-called “”video-assisted appendectomy”", consisting in a mobilization and extraction of the organ via the single umbilical Selleck EPZ-6438 trocar, and subsequent open appendectomy, gained some popularity [12, 13]. The first releases from the industry, beginning in the second half of the last decade, regarded multichannel ports, requiring a 1.5 to 2 cm incision of the fascia. They are disposable, have three-channels (usually two 5 mm and one 10/12 mm), recently broadened to 4-6 (due to the need for application to more complex operations), and generally require a longer 5 mm angulated camera.

Mean values and standard errors (95% confidence) were calculated

Mean values and standard errors (95% confidence) were calculated from three independent experiments. Considering all the results described here, we propose the

following working hypothesis which is illustrated in Figure 5: Tep1 participates in the efflux of small compounds such as chloramphenicol and aminosugars which are core Nod factor precursors. Although these compounds have different structures, secondary multidrug (Mdr) transporters of the Major Facilitator Superfamily are known to be promiscuous in substrate recognition and transport [22]. In the tep1 mutant, chloramphenicol and Nod factor precursors accumulate inside the bacteria to concentrations which either hamper growth (chloramphenicol accumulation) or affect maximal nod gene expression (aminosugar accumulation). At the same time, the EPZ015666 mw diminished efflux of aminosugars in the transport mutant leads to improved nodulation efficiency. Elafibranor Figure 5 Working model showing possible roles for Tep1 and their substrates. Cm, chloramphenicol;

IM, inner membrane; OM, outer membrane. Conclusion The results obtained in this work suggest that the tep1 gene encodes a transport protein belonging to the MFS family of permeases able to confer chloramphenicol resistance in S. meliloti by expelling the antibiotic outside the cell. A tep1-linked gene in S. meliloti, fadD, plays a role in swarming motility and in nodule formation efficiency on alfalfa Ivacaftor datasheet plants. We have demonstrated that tep1 is not involved in swarming motility but like fadD affects the establishment of the S. meliloti-alfalfa symbiosis. A tep1 loss-of-function mutation leads to increased nodule formation efficiency but reduced nod gene expression suggesting that Tep1 transports compounds which influence different steps of the nodule formation process. Whether these effects are caused by the same Loperamide or different compounds putatively transported by Tep1, still needs to be investigated. Curiously, nod gene expression is reduced in a S. meliloti nodC mutant with the same intensity as in the tep1 mutant. This has implications

for nod gene regulation in S. meliloti as it rules out the existence of a feedback regulation as described for B. japonicum. On the other hand, it could indicate that Tep1 is involved in the transport of Nod factors or its precursors. Indeed, increased concentrations of the core Nod factor precursor N-acetyl glucosamine reduced nod gene expression. Moreover, both glucosamine and N-acetyl glucosamine inhibit nodulation at high concentrations. Therefore, this constitutes the first work which attributes a role for core Nod factor precursors as regulators for nodulation of the host plant by S. meliloti. Furthermore, the results suggest that the activity of Tep1 can modulate the nodule formation efficiency of the bacteria by controlling the transport of core Nod factor precursors.

Toluidine blue-stained longitudinal sections of S nodorum SN15 p

Toluidine blue-stained longitudinal sections of S. nodorum SN15 pycnidia identified the presence of a number of enlarged cells that form the opening of the ostiole from which the cirrhus of spores are released from the pycnidial cavity (Figure 8). Analysis of the pycnidia formed by each of the G-protein mutants by cold induction revealed the structures to be comparable to those produced by the wildtype PF-01367338 manufacturer strain under the same conditions. It was observed that the ostiole failed to differentiate on the mutant pycnidia (Figure 9). This observation was consistent with the requirement that the pycnidiospores within these mutant pycnidia could

NCT-501 manufacturer not be released by water (as typically observed in wildtype pycnidia) and required manual disruption. The pycnidia of gba1-6 and gga1-25 were also nearly always observed as multiple structures

fused FRAX597 purchase together and were almost never seen individually (Figure 9). Although the pycnidia of SN15 and gna1-35 often developed fused, it was uncommon for the pycnidia to form indistinct from one another. The pycnidia of gga1-25 and gba1-6 were also comparatively misshapen and less mature in appearance than those of SN15 and gna1-35. Figure 8 A longitudinal section of a wax embedded excision from an asexually sporulating culture of S. nodorum SN15 -stained with toluidine blue. Pictured are pycnidiospores being released from a mature pycnidium. Arrows point to the masses of enlarged cells producing the ostiole, formed in the development of the mature pycnidium, from which the pycnidiospores are released from the pycnidial cavity as a cirrus. Cv, pycnidial cavity; W, pycnidial wall; Ch, cirrus. Figure 9 Longitudinal sections of

a wax embedded excision from asexually sporulating cultures of S. nodorum -stained with toluidine blue. Pictured are conidiogenous cells and pycnidiospores contained within the mature pycnidia of wild-type strain SN15, and the (potentially less) ‘matured’ pycnidia of mutant strains gna1-35, gba1-6 and gga1-25. The pycnidia of SN15 and gna1-35 often develop fused, but the pycnidial cavities remain visually distinct, by comparison to those of gba1-6 and gga1-25 tuclazepam which often form a single body of pycnidia. Cv, pycnidial cavity; W, pycnidial wall. Discussion The deactivation of the Gα subunit Gna1 from S. nodorum has proven fruitful to further understanding the pathogenesis of this fungal pathogen [9]. The lack of sporulation and reduced pathogenicity of the resulting gna1 strain sparked further investigation into the molecular and phenotypic attributes of this mutant strain largely because a determination of the molecular processes underpinning the phenotype could lead to more targeted control of the pathogen. Subsequent analysis of the gna1 strain to identify downstream-regulated targets and processes has uncovered many interesting aspects of the disease including mycotoxin production.

Cancer Res 2005, 65:8366–8371 PubMedCrossRef 17 Pan Q, Bao LW, T

Cancer Res 2005, 65:8366–8371.PubMedCrossRef 17. Pan Q, Bao LW, Teknos TN, Merajver SD: Targeted disruption of protein kinase C epsilon reduces cell invasion and motility through inactivation of RhoA and RhoC GTPases in head and neck squamous cell carcinoma. Cancer Res 2006, 66:9379–9384.PubMedCrossRef 18. Bae KM, Wang H, Jiang G, Chen MG, Lu L, Xiao L: Protein kinase C epsilon is overexpressed in primary human non-small cell lung cancers and functionally required for proliferation of non-small cell lung cancer cells in a p21/Cip1-dependent manner. Cancer Res 2007, 67:6053–6063.PubMedCrossRef 19. Brenner W, Benzing F, Gudejko-Thiel

J, Fischer R, Färber G, Hengstler JG, Seliger B, Thüroff Entospletinib in vitro JW: Regulation of beta1 integrin expression by PKCepsilon in renal cancer cells. Int Evofosfamide J Oncol 2004, 25:1157–1163.PubMed 20. Engers R, Mrzyk S, Springer E, Fabbro D, Weissgerber G, Gernharz CD, Gabbert HE: Protein kinase C in human renal cell carcinomas: role in invasion and differential isoenzyme expression. Br J Cancer 2000, 82:1063–1069.PubMedCrossRef 21. Green FL, Page DL, Fleming ID, et al.: AJCC Cancer Staging Manual. 6th edition. Springer: New York; 2002. 22. Fuhrman SA, Lasky LC, Limas C: Prognostic significance of morphologic parameters

in renal cell carcinoma. Am J Surg Pathol 1982, 6:655–663.PubMedCrossRef 23. Yamada S, Yanamoto S, Kawasaki G, Rokutanda S, Yonezawa H, Kawakita A, Nemoto TK: Overexpression of CRKII increases selleck migration and invasive potential in oral squamous cell carcinoma. Cancer Letters 2011,

303:84–91.PubMedCrossRef 24. Fu L, Qin YR, Xie D, Chow HY, Ngai SM, Kwong DL, Li Y, Guan XY: Identification of alpha-actinin 4 and 67 kDa laminin receptor as stage-specific markers in esophageal cancer via proteomic approaches. Chloroambucil Cancer 2007, 110:2672–2681.PubMedCrossRef 25. Guo S, Mao X, Chen J, Huang B, Jin C, Xu Z, Qiu S: Overexpression of Pim-1 in bladder cancer. J Exp Clin Cancer Res 2010, 29:161.PubMedCrossRef 26. Pedram A, Razandi M, Wallace DC, Levin ER: Functional estrogen receptors in the mitochondria of breast cancer cells. Mol Biol Cell 2006, 17:2125–37.PubMedCrossRef 27. Lu D, Huang J, Basu A: Protein kinase C epsilon activates protein kinase B/Akt via DNA-PK to protect against tumor necrosis factor-alpha-induced cell death. J Biol Chem 2006, 281:22799–22807.PubMedCrossRef 28. Hu B, Shen B, Su Y, Geard CR, Balajee AS: Protein kinase C ε is involved in ionizing radiation induced bystander response in human cells. Int J Biochem Cell Biol 2009, 41:2413–2421.PubMedCrossRef 29. Wei X, Juan ZX, Min FX, Nan C, Hua ZX, Qing FZ, Zheng L: Recombinant immunotoxin anti-c-Met/PE38KDEL inhibits proliferation and promotes apoptosis of gastric cancer cells. J Exp Clin Cancer Res 2011, 30:67.PubMedCrossRef 30.

However, flocculation in response to FeSO4 was less pronounced at

However, flocculation in response to FeSO4 was less pronounced at that iron concentration compared to 30 μM FeCl3 as quantified by measuring sedimentation rates (Figure 1B) as previously described [33]. Figure 1 Iron induced concentration dependent flocculation of C. albicans cells. (A) Microscopic AG-120 price analysis. C. albicans SC5314 (WT) was incubated with different FeCl3 concentrations (indicated at the top left hand of each sub panel) or with 30 μM FeSO4 in RPMI at 30°C for 2 h. (B) Relative sedimentation rates of WT cells. Flocculation of cells was triggered

by 30 μM FeCl3 or 30 μM FeSO4 in RPMI and sedimentation rates were determined after incubation at 30°C for 2 h. Means and standard Mocetinostat datasheet deviations of three independent samples are shown (n = 3). ** denotes P < 0.01 (student’s t-test). (C) Relative sedimentation rates of WT cells pre-cultured in the sufficient iron (YPD) or restricted iron medium (RIM) at 30°C for 3 h. Flocculation of cells was triggered by 30 μM FeCl3 in RPMI and sedimentation rates were determined after incubation at 30°C for 2 h.

Means and standard deviations of three independent samples are shown (n = 3). *** denotes P < 0.001 (student’s t-test). (D) Microscopic analysis of cycloheximide (CHX) or MeOH pre-treated cells. C. albicans SC5314 was pre-treated either with 500 μg ml-1 CHX or MeOH in RPMI at 30°C for 15 min. Iron or water were subsequently added and cells Savolitinib manufacturer were incubated at 30°C for 2 h. Flocculation was also induced in yeast nitrogen base (YNB) medium containing 30 μM FeCl3 compared to 1.2 μM basal Fe3+ concentration (information given by the manufacturer), thus showing that the induction of flocculation was independent from the medium used (see Additional file 1). Cells may possess internal iron stores from pre-cultivation in an iron sufficient medium. Thus, Idoxuridine we investigated whether the iron content of the medium used during pre-cultivations influenced

the dependence of the flocculent phenotype on the iron concentration in RPMI. C. albicans was either pre-cultivated in a medium with sufficient iron, i.e. the rich yeast extract-peptone-dextrose (YPD) medium, or starved for iron by pre-cultivation in a medium with restricted iron availability (restricted iron medium: RIM). RIM resulted from addition of the iron chelator bathophenanthroline disulfonate (BPS) to YPD medium. As shown in Figure 1C, flocculation due to exposure to 30 μM Fe3+ was independent on the pre-cultivation medium: WT cells starved for iron by pre-cultivation in RIM flocculated upon exposure to 30 μM Fe3+ with a similar sedimentation rate as cells pre-cultivated in YPD. During all later experiments, we pre-cultivated C. albicans in YPD and added 30 μM FeCl3 as iron source to the respective medium of the working culture unless it is mentioned otherwise.

And right now, no one lives in it, it’s a no person’s


And right now, no one lives in it, it’s a no person’s

land” (PU3). The main role of these translators was seen by some participants as condensing information to deliver accessible, check details clear and robust messages. In addition, translators could go further and help MCC950 supplier scientists understand better the complex and fuzzy policy making context, and open the complexities of biodiversity and ecosystem services issues to policy makers (Cash and Moser 2000). This could be done for instance by arranging sessions to familiarize policy makers with models and concepts developed by scientists (Haas 2004), and familiarising scientists with the needs and constraints of policy-makers (an example is that of the problems of communicating uncertainty). One such individual therefore described his role as “actually understanding what the question is and what the person wants to try to do…the point the person is trying to make, you need to be able to hear that and translate that, and then to be able to read the facts and translate those and try and marry the

two together” (U4). They have a key role therefore in overcoming the language boundaries on both sides and linking communities—leading one participant to note the potential of having science translators talking to policy translators. Within research organisations such individuals EPZ5676 mw may be knowledge exchange specialists, or within policy departments these may be specialist scientific advisors. The challenge could be training or recruiting scientists who have

high profiles within their own disciplines crotamiton and who are able to efficiently communicate with counterparts from other disciplines, as well as with the media, policy makers, and popular audiences (Haas 2004). ‘Translation’ roles are, however, at present not always formally recognised or rewarded. The organisational support of these staff would be partly aided by the development of organisations’ communication strategies, which would outline their objectives and their timescales for various information needs. These strategies will of course vary according to the organisation’s outputs and strengths, and will need to reflect different priorities over time. However, the existence of translators (also called mediators or linkers) should not (and could not) absolve individuals in science and policy from having some role to play in seeking out translation, dialogue, learning and sharing opportunities. Otherwise, a risk is that dialogue can become overly vulnerable to the continuity of key personnel. The challenge will be to promote translators, but also train and incentivise scientists and policy makers wanting to engage themselves in translation roles in addition to their scientific and policy roles.

The sequence analysis of mgoC prompted us to search the superfami

The sequence analysis of mgoC prompted us to search the superfamily protein domains, revealing a similarity to the N-oxygenase domain. This domain was identified in the protein PrnD, which is derived from the pyrrolnitrin biosynthesis gene cluster of Pseudomonas fluorescens. MgoC is also similar to AurF from Streptomyces thioluteus, which produces the starter unit p-nitrobenzoic Proteasome inhibitor acid (PNBA) for the polyketide synthase of the aureothin biosynthesis pathway [25]. The gene mgoA, which is Selleck PLX4032 homologous to non-ribosomal peptide synthetases, is the largest gene in the mgo

operon, and its disruption produces a mutant that is defective in mangotoxin production. Its structure, participation in mangotoxin production and influence on the virulence of the wild-type bacterium has been discussed previously [15]. The final gene studied was mgoD; a domain localisation analysis indicated that mgoD could be a Polyketide_cyc2 belonging to the star-related lipid-transfer (START) domain superfamily. The START superfamily includes bacterial polyketide cyclase/aromatases and two families of previously uncharacterised proteins that are present only in plants and the cyanobacterium Prochlorococcus [26]. After analysing the elements that composed the putative mgo operon, we evaluated whether the four genes

were transcribed together in a single transcript. RT-PCR experiments using the wild-type RNA showed that the four genes were connected in the single transcript (Figure 2). Moreover, the transcript AZD1390 nmr size was analysed by hybridisation, which confirmed the presence of a single transcript with a sufficient size (about 6 kb) to contain the genes mgoBCAD; however, the exact size of the transcript could not be determined. Following the identification of the mgo operon, the promoter and transcription terminator were identified and studied. The in silico analysis of the sequence identified two putative promoters. Promoter activity was detected only in a minimal medium, the same culture

medium that is traditionally used for antimetabolite toxin assays [2, 13]. Promoter activity occurred in the wild-type strain at both temperatures and in the ORF2 insertion mutant at 22°C only. The other Pseudomonas spp. experimental strains, Thymidylate synthase which do not produce mangotoxin, did not exhibit any β-Gal activity. The promoter activity in the wild-type strain was more intense at 28°C than 22°C. When the promoter activity was assayed at 22°C, the activity of the mutant UMAF0158::ORF2 was statistically comparable with that of the wild-type strain. These results suggest a possible influence of ORF2 on the mgo operon during its regulation in response to temperature variations. The promoter inactivity in the other two strains of Pseudomonas spp. may be due to the absence of genes homologous to the mgo operon in P.

Robin got him to spend much of his time with plant material… Ret

Robin got him to spend much of his time with plant material…. Returning to the United States in 1956, Tom joined the faculty of the University of Rochester where he stayed for 7 years. His research efforts were focused

primarily in photosynthesis, but he also published a paper with his wife, Hope (one of the authors of this Tribute), in Nature, on a leukocyte growth factor isolated from red beans (Punnett and Punnett 1963; Punnett et al. 1962). Later, Punnett et al. (1980) did an analysis of hydrozoan sperm attractant. His understanding of biochemical Pifithrin-�� ic50 techniques including processes for the purification of proteins was exceptional. The primary focus of Tom’s research life remained an unquenchable interest in photosynthesis, stemming from the early experiments of Robert Emerson on photosynthetic processes in plants. Emerson and Lewis (1943) had found that the quantum yield of photosynthesis dropped precipitously when algae were illuminated beyond 685 nm (the so-called Red Drop). A major breakthrough came when Emerson et al. (1957) discovered a synergistic effect by illuminating algae with two beams together,

one in the red drop region and another on the short-wave side of the spectrum. This phenomenon, now known as the Emerson Enhancement Effect, implied that there were two photosystems involved in the photosynthetic GDC-0449 process. Emerson’s enhancement experiment was the seminal experiment for establishing the two light system hypothesis in plant photosynthesis (also see Govindjee and Rabinowitch 1960; Myers and French 1960). During this period, Punnett (1959) continued his experiments with broken chloroplasts along with their uncertainties, and this moved him toward techniques

for proper isolation of chloroplasts. Tom moved to the Biology Department at Temple in 1963 (Fig. 4), serving twice as Acting Chair in his long tenure there. In the early 1960s, the department was becoming more engaged in research and the young, active plant physiologist was just the addition the department needed. During this period, Tom published the work he had done earlier on improved methods for studying the Hill reaction (Punnett 1957; Punnett et al. 1964) and on Liothyronine Sodium an enhancement of the Hill reaction and photophosphorylation by CO2 (Punnett and Iyer 1964; cf. Govindjee et al. 1964 for Emerson Enhancement in NADP Hill reaction by different wavelengths of light). The new effect of CO2 on photophosphorylation was called “Punnett Effect” by Govindjee and van Rensen (1978). Fig. 4 Tom Punnett in his office, with a photograph of Bob Emerson; on the book shelf are Volume 1, Volume 2 (Part 1) and Volume 2 (Part 2) of Rabinowitch’s classic monograph (1945–1956) on “Photosynthesis”; in the Preface of Volume 2 (Part 1, 1951), Rabinowitch thanked Tom Punnett for his “valuable aid in the reading of the proofs and the checking of the bibliography”.

Following the FDA approval of anti-CD20 mAb Rituximab for CD20+ N

Following the FDA approval of anti-CD20 mAb Rituximab for CD20+ NHL treatment, monoclonal antibody (mAb)-based targeting therapy has revolutionized the treatment of malignancies for the specific antitumor activity and low cytotoxicity against normal tissues [22, 23]. In the last decade, more and more studies have confirmed that the combination of mAb-based learn more active targeting and nanoparticle-based passive targeting can improve drug concentration in tumor tissues and tumor cells in a shorter time with greater accuracy [7, 24, 25]. In this study, we have developed an adriamycin (ADR)-loaded Selleck MS-275 liposome using the diacetylenic

phosphatidylcholine 1,2-bis(10,12-tricosadiynoyl)-sn-glycero-3-phosphocholine (DC8,9PC, hereafter referred to as PC), which can form intermolecular cross-linking through the diacetylenic group to produce a conjugated polymer within the hydrocarbon region of the bilayer by ultra-violet (UV) irradiation (Additional file 1: Figure S1) [26, 27]. For the sake of active targeting, the Fab fragments of rituximab were conjugated onto the liposomal surface. Our experimental results demonstrate that this well-modified liposome, which owns good serum stability and prolonged circulation time, can accumulate in

the tumor tissues and malignant cells with high specificity and sufficient amount, which can bring out exceptional excellent and durable therapeutic efficacy against CD20-positive lymphomas. Methods Cell lines and materials Two human B cell lymphoma cell lines, Raji and Daudi, were obtained from the American Type Culture Collection (ATCC). Cells were propagated and maintained in RPMI 1640 supplemented with 10% (v/v) fetal bovine serum (FBS, GIBCO, Invitrogen, Carlsbad, CA, USA) in a controlled atmosphere GNAT2 incubator at 37°C with 5% CO2. The DC8,9PC and 1,2-distearoyl-sn-glycero-3-phosphoethanolamine-N-[maleimide(polyethylene

glycol)-2000] (Mal-PEG) were purchased from Avanti Polar Lipids (Williamsport, PA, USA). The anti-CD20 antibodies rituximab was purchased from Roche (Basel, Switzerland). Fabrication of Fab fragment-conjugated liposome (Figure 1) Figure 1 Fabrication of rituximab Fab fragment-decorated liposomes. Formation of drug-loaded liposomes Total lipids mixtures of 2 mg DC8,9PC and 0.25 mg Mal-PEG were dissolved in 500 μL mixed solvent of chloroform and methyl alcohol with the volume ratio at 1:1. Then, the solvent was evaporated under vortex and flashed with nitrogen to obtain the lipid film, followed by washing-out with 2 mL of ADR (doxorubicin HCl, Melonepharma CO. LTD., Dalian, China) solution (0.5 mg/mL in PBS) to obtain ADR-loaded multilamellar vesicles [26]. The collected liposome solution was dialyzed against PBS using a membrane (molecular weight cutoff 3 kDa) at 4°C for 12 h to remove uncombined ADR resulting in the ADR-loaded liposome stocking solutions. Thiolation of mAbs The Fab fragment of rituximab was prepared as reported previously [25].