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Ultrasonic Homogenizer Published Papers

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Article No. Title
UH1061 Singlet oxygen production in Chlamydomonas reinhardtii under heat stress
  Ankush Prasad, Ursula Ferretti, Michaela Sedlářová & Pavel Pospíšil
PDF icon In the current study, singlet oxygen formation by lipid peroxidation induced by heat stress (40 °C) was studied in vivo in unicellular green alga Chlamydomonas reinhardtii. Primary and secondary oxidation products of lipid peroxidation, hydroperoxide and malondialdehyde, were generated under heat stress as detected using swallow-tailed perylene derivative fluorescence monitored by confocal laser scanning microscopy and high performance liquid chromatography, respectively. Lipid peroxidation was initiated by enzymatic reaction as inhibition of lipoxygenase by catechol and caffeic acid prevented hydroperoxide formation. Ultra-weak photon emission showed formation of electronically excited species such as triplet excited carbonyl, which, upon transfer of excitation energy, leads to the formation of either singlet excited chlorophyll or singlet oxygen. Alternatively, singlet oxygen is formed by direct decomposition of hydroperoxide via Russell mechanisms. Formation of singlet oxygen was evidenced by the nitroxyl radical 2,2,6,6-tetramethylpiperidine-1-oxyl detected by electron paramagnetic resonance spin-trapping spectroscopy and the imaging of green fluorescence of singlet oxygen sensor green detected by confocal laser scanning microscopy. Suppression of singlet oxygen formation by lipoxygenase inhibitors indicates that singlet oxygen may be formed via enzymatic lipid peroxidation initiated by lipoxygenase.
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UH1062 Transient receptor potential melastatin 3 is a phosphoinositide-dependent ion channel
  Doreen Badheka, Istvan Borbiro, and Tibor Rohacs
PDF icon Phosphoinositides are emerging as general regulators of the functionally diverse transient receptor potential (TRP) ion channel family. Phosphatidylinositol 4,5-bisphosphate (PI(4,5)P2) has been reported to positively regu- late many TRP channels, but in several cases phosphoinositide regulation is controversial. TRP melastatin 3 (TRPM3) is a heat-activated ion channel that is also stimulated by chemical agonists, such as pregnenolone sulfate. Here, we used a wide array of approaches to determine the effects of phosphoinositides on TRPM3. We found that channel activity in excised inside-out patches decreased over time (rundown), an attribute of PI(4,5)P2-dependent ion channels. Channel activity could be restored by application of either synthetic dioctanoyl (diC8) or natural ara- chidonyl stearyl (AASt) PI(4,5)P2. The PI(4,5)P2 precursor phosphatidylinositol 4-phosphate (PI(4)P) was less ef- fective at restoring channel activity. TRPM3 currents were also restored by MgATP, an effect which was inhibited by two different phosphatidylinositol 4-kinase inhibitors, or by pretreatment with a phosphatidylinositol-specific phospholipase C (PI-PLC) enzyme, indicating that MgATP acted by generating phosphoinositides. In intact cells, reduction of PI(4,5)P2 levels by chemically inducible phosphoinositide phosphatases or a voltage-sensitive 5-phosphatase inhibited channel activity. Activation of PLC via muscarinic receptors also inhibited TRPM3 channel activity. Overall, our data indicate that TRPM3 is a phosphoinositide-dependent ion channel and that decreasing PI(4,5)P2 abundance limits its activity. As all other members of the TRPM family have also been shown to require PI(4,5)P2 for activity, our data establish PI(4,5)P2 as a general positive cofactor of this ion channel subfamily.
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UH1063 Phenotypic plasticity in Bacillus cereus strains isolated from various Antarctic habitats
  Rupesh Kumar Sinha & Kottekkatu Padinchati Krishnan
PDF icon We studied five strains of psychrotolerant Bacillus cereus (B. cereus) isolated from Antarctic snow (BCsn), ice (BCic), lake water (BCwr), sediment (BCsd), and soil (BCsl) samples in terms of their growth, biochemical properties, and heat shock re- sponses. Analyses of growth kinetics at 4°C showed that BCsn had the fastest generation time (16.1 h), whereas BCwr had the slowest (30.8 h). Strain BCsd formed the largest zone of lipid hydrolysis (18 mm) whereas BCsn formed the smallest zone (3 mm).Only BCsd produced gelatinase. These physiological differences illustrate adaptations of B. cereus isolates to different niches. Strains BCsl and BCwr were resistant to all 12 of the antibiotics tested. Strains BCsn, BCic, and BCsd were resistant to cell wall synthesis inhibitors (penicillin and ampicillin) and susceptible to protein synthesis inhibitors (tetracycline and streptomycin). A carbon-substrate utilization assay revealed that BCsn, BCic, and BCwr could specifically utilize D-glucose-6-phosphate, salicin, and 2’-deoxyadenosine, respectively, indicating a degree of metabolic diversity among these Antarctic B. cereus strains. An analy- sis of heat shock proteins (HSPs) produced in response to a 60°C heat treatment revealed significant variations in the amounts of HSP33 (p = 0.01, df = 4), HSP44 (p = 0.003, df = 4), and HSP60 (p = 0.04, df = 4) among the strains. This emphasizes the impor- tance of HSPs in bacterial taxonomy. These results show that there are considerable adaptive variations among B. cereus strains from extremophilic environments. This could be significant in evaluating the taxonomy and evolution of this species.
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UH1064 Rheological Properties of Alginate–Essential Oil Nanodispersions
  Martina Cofelice, Francesca Cuomo, and Francesco Lopez
PDF icon Due to its favorable structural properties and biocompatibility, alginate is recognized as a suitable versatile biopolymer for use in a broad range of applications ranging from drug delivery, wound healing, tissue engineering, and food formulations such as nanodispersions. Rheological analysis plays a crucial role in the design of suitable nanoemulsion based coatings. Different essential oil and alginate nanodispersion compositions stabilized by Tween 80 were analyzed for rheological and conductometric properties. The results confirmed that the nanoformulations shared a pseudoplastic non-Newtonian behavior that was more evident with higher alginate concentrations (2%). Nanodispersions made of alginate and essential oil exhibited a slight thixotropic behavior, demonstrating the aptitude to instantaneously recover from the applied stress or strain. Oscillatory frequency sweep tests showed a similar fluid-like behavior for 1% and 2% alginate nanodispersions. Finally, it was demonstrated that advantages coming with the use of the essential oil are added to the positive aspects of alginate with no dramatic modification on the flow behavior.
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UH1065 Polymeric Nanoparticles for Pulmonary Protein and DNA Delivery
  Jyothi U. Menon, Priya Ravikumar, Amruta Pise, Dipendra Gyawali, Connie C.W. Hsia, and Kytai T. Nguyen
PDF icon Polymeric nanoparticles (NPs) are promising carriers of biological agents to lung due to advantages including biocompatibility, ease of surface modification, localized action and reduced systemic toxicity. However, there have been no studies extensively characterizing and comparing the behavior of polymeric NPs for pulmonary protein/DNA delivery both in vitro and in vivo. We screened six polymeric NPs: gelatin, chitosan, alginate, poly lactic-co-glycolic acid (PLGA), PLGA-chitosan, and PLGA-polyethylene glycol (PEG), for inhalational protein/ DNA delivery. All NPs except PLGA-PEG and alginate were <300 nm in size with bi-phasic core compound release profile. Gelatin, PLGA NPs and PLGA-PEG NPs remained stable in deionized water, serum, saline and simulated lung fluid (Gamble’s solution) over 5 days. PLGA-based NPs and natural polymer NPs exhibited highest cytocompatibility and dose-dependent in vitro uptake respectively by human alveolar type-1 epithelial cells. Based on these profiles, gelatin and PLGA NPs were used to encapsulate a) plasmid DNA encoding yellow fluorescent protein (YFP) or b) rhodamine-conjugated erythropoietin (EPO) for inhalational delivery to rats. Following a single inhalation, widespread pulmonary EPO distribution persisted for up to 10 days while increasing YFP expression was observed for at least 7 days for both NPs. The overall results support both PLGA and gelatin NPs as promising carriers for pulmonary protein/DNA delivery.
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UH1066 Enhancement of Mucus Production in Eukaryotic Cells and Quantification of Adherent Mucus by ELISA
  Christian Reuter and Tobias A. Oelschlaeger
PDF icon The mucosal surfaces of the gastrointestinal, respiratory, reproductive, and urinary tracts, and the surface of the eye harbor a resident microflora that lives in symbiosis with their host and forms a complex ecosystem. The protection of the vulnerable epithelium is primarily achieved by mucins that form a gel-like structure adherent to the apical cell surface. This mucus layer constitutes a physical and chemical barrier between the microbial flora and the underlying epithelium. Mucus is critical to the maintenance of a homeostatic relationship between the microbiota and its host. Subtle deviations from this dynamic interaction may result in major implications for health. The protocol in this article describes the procedures to grow low mucus-producing HT29 and high mucus-producing HT29-MTX-E12 cells, maintain cells and use them for mucus quantification by ELISA. Additionally, it is described how to assess the amount of secreted adherent mucus. This system can be used to study the protective effect of mucus, e.g., against bacterial toxins, to test the effect of different culture conditions on mucus production or to analyze diffusion of molecules through the mucus layer. Since the ELISA used in this protocol is available for different species and mucus proteins, also other cell types can be used.
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UH1067 CCR2-Dependent Dendritic Cell Accumulation in the Central Nervous System during Early Effector Experimental Autoimmune Encephalomyelitis Is Essential for Effector T Cell Restimulation In Situ and Disease Progression
  Benjamin D. Clarkson, Alec Walker, Melissa G. Harris, Aditya Rayasam, Matyas Sandor and Zsuzsanna Fabry
PDF icon Dendritic cells (DCs)—although absent from the healthy CNS parenchyma—rapidly accumulate within brain and spinal cord tissue during neuroinflammation associated with experimental autoimmune encephalomyelitis (EAE; a mouse model of multiple sclerosis). Yet, although DCs have been appreciated for their role in initiating adaptive immune responses in peripheral lymphoid organ tissues, how DCs infiltrate the CNS and contribute to ongoing neuroinflammation in situ is poorly understood. In this study, we report the following: 1) CD11c+ bone marrow–derived DCs and CNS-infiltrating DCs express chemokine receptor CCR2; 2) compared with CCR2+/+ cells, adoptively transferred CCR22/2 bone marrow–derived DCs or DC precursors do not accumulate in the CNS during EAE, despite abundance in blood; 3) CCR22/2 DCs show less accumulation in the inflamed CNS in mixed bone marrow chimeras, when compared with CCR2+/+ DCs; and 4) ablation of CCR2+/+ DCs during EAE clinical onset delays progression and attenuates cytokine production by infiltrating T cells. Whereas the role of CCR2 in monocyte migration into the CNS has been implicated previously, the role of CCR2 in DC infiltration into the CNS has never been directly addressed. Our data suggest that CCR2-dependent DC recruitment to the CNS during ongoing neuroinflammation plays a crucial role in effector T cell cytokine production and disease progression, and signify that CNS-DCs and circulating DC precursors might be key therapeutic targets for suppressing ongoing neuroinflammation in CNS autoimmune diseases. The Journal of Immunology, 2015, 194: 000–000.
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UH1068 Nanofluids as Novel Alternative Smart Fluids for Reservoir Wettability Alteration
  Lezorgia Nekabari Nwidee, Ahmed Barifcani, Mohammad Sarmadivaleh and Stefan Iglauer
PDF icon This chapter presents an account of two metal oxide nanoparticles (zirconium and nickel oxide) on basis of their structure, morphology, crystallinity phases, and their wetting effect on solid-liquid interface. As a preliminary step to sound understanding of process mechanisms; wettability, nanoparticles, and their relations thereof were scrutinized. To investigate the nanofluids wetting inclinations, complex mixtures of the nanoparticles and NaCl brine (ZrO2/NaCl; NiO/NaCl) were formulated and their technical feasibility as wetting agents tested via contact angle measurement. The result shows that the nanopar- ticles exhibit different structural and morphological features and capable of addressing reservoir wettability challenges owing to favorable adsorption behavior on the surface of the calcite which facilitated the wetting changes quantified by contact angle. We believe this study will significantly impact the understanding of wetting at solid-liquid interface which is crucial for recovery process optimization.
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UH1069 CCR7 deficient inflammatory Dendritic Cells are retained in the Central Nervous System
  Benjamin D. Clarkson, Alec Walker, Melissa G. Harris, Aditya Rayasam, Martin Hsu, Matyas Sandor & Zsuzsanna Fabry
PDF icon Dendritic cells (DC) accumulate in the CNS during neuroinflammation, yet, how these cells contribute to CNS antigen drainage is still unknown. We have previously shown that after intracerebral injection, antigen-loaded bone marrow DC migrate to deep cervical lymph nodes where they prime antigen- specific T cells and exacerbate experimental autoimmune encephalomyelitis (EAE) in mice. Here, we report that DC migration from brain parenchyma is dependent upon the chemokine receptor CCR7. During EAE, both wild type and CCR7−/− CD11c-eYFP cells infiltrated into the CNS but cells that lacked CCR7 were retained in brain and spinal cord while wild type DC migrated to cervical lymph nodes. Retention of CCR7-deficient CD11c-eYFP cells in the CNS exacerbated EAE. These data are the first to show that CD11chigh DC use CCR7 for migration out of the CNS, and in the absence of this receptor they remain in the CNS in situ and exacerbate EAE.
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UH1070 Synthesis of 3-Aryl Coumarin Derivatives Using Ultrasound
  Shubashini Krishnan Sripathi, Kandasamy Logeeswari
PDF icon Coumarins occupy an important place in the realm of natural products and synthetic organic chemistry. A fast and highly efficient green method for synthesizing 3-aryl coumarin derivatives from salicylaldehyde and phenyl acetyl chloride in the presence of tetrahydrofuran and K2CO3 using ultrasound irradiation is reported. Ultrasound assisted re- actions have resulted in better yields and faster reaction time of the desired products than when prepared under conven- tional conditions. The resulting coumarin derivatives were characterized by IR spectrum.
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Ultrasonic Homogenizers
Sometimes referred to as, Sonicator, Cell Disruptor or Cell Disrupter, Probe Sonicator, Ulrasonicator, Sonifier®, Sonic Dismembrator, and Ultrasonic Liquid Processor.

Sonifier is a registered trademark of Branson Ultrasonics Corporation

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Our Ultrasonic Homogenizer product line is being expanded to include thermo-conductive tube pods. KoldPods are available for 1.5ml Micro Tubes, 15ml and 50ml Conical tubes, and will decrease thermal variability with conventional temperature sources.

Ultrasonic Homogenizers are also referred to as Sonicators, Cell Disruptors, Probe Sonicators, Sonifiers®, and Sonic Dismembrators

Sonifier is a registered trademark of Branson Ultrasonics Corporation

 
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