Evaluation of the relationship between two different methods for enumeration fecal indicator bacteria: Colony-forming unit and most probable number

Most probable number （MPN） and colony-forming unit （CFU） estimates of fecal indicator bacteria （FIB） concentration are common measures of water quality in aquatic environments. Thus, FIB intensively monitored in Yeongsan Watershed in an attempt to compare two different methods and to develop a statistical model to convert from CFU to MPN estimates or vice versa. As a result, the significant difference was found in the MPN and CFU estimates. The enumerated Escherichia coli concentrations in MPN are greater than those in CFU, except for the measurement in winter. Especially in fall, E. coli concentrations in MPN are one order of magnitude greater than that in CFU. Contrarily, enterococci bacteria in MPN are lower than those in CFU. However, in general, a strongly positive relationship are found between MPN and CFU estimates. Therefore, the statistical models were developed, and showed the reasonable converting FIB concentrations from CFU estimates to MPN estimates. We expect this study will provide preliminary information towards future research on whether different analysis methods may result in different water quality standard violation frequencies for the same water sample.

Protective Effect of phytolacca Acinosa Polysaccharides Ⅰ(PAP-Ⅰ)on Hematopoiesis in Cyclophosphamide-treated and 60￣Co-irradiated Mice

The protective effect of a kind of purified polysaccharides extracted from Radix of Phytolacca acinosa Roxb,with a molecular weight of 10 KDa,on hematopoiesis was investigated.Average survival time of mice treated with cyclophosphamide (CY) 300 mg/kg once alone was 13.3 ± 7.2d(n=7) however,average survival time of mice treated with CY 300 mg/kg in com-bination with PAP-1 10 mg/kg,3 times/wk was 36.7± 16.4d(n=7,P<0.01).PAP-1,ip had benefi-cial effect on the recovery of the CY induced decrease of peripheral leukocyte number,and the nu-cleated bone marrow cell(BMC)number and[3￣H]TdR uptaken by BMC induced by rmGM-CSF in S180 bearing mice treated with CY,In mice,after the first ip treatment with CY 100 mg/kg on d7,the peripheral leukocyte number decreased on d9 and recovered to normal level about d13 to d15. Such recovery was accelerated by administrating PAP-1,10mg/kg, 3 times/wk.A significant in-crease of the activity to form colony in spleen(colony-forming unit in spleen, CFU-S_8, CUF-S12) in mice irradiated with 550 rad 6O￣Co γ-rays and an enhancement of proliferative response of BMC to rmGM-CSF treated with PAP-1,10mg/kg,3 times/wk, ip were observed.After PAP-1,10 mg/kg,ip once,a significant increase in the number of peripheral blood leukocytes and a rise in the serum of colony stimulating factor(CSF) were also confirmed.The types of CSF in serum were M-CSF and other hematopoietic growth factors,which were confirmed by using McAb of IL-3, GM-CSF and PcAb of M-CSF. These beneficial effects of PAP-1 on hematopoiesis may be related to its activityinducing CSFs and other hematopoietic growth factors and warrant further evaluation of its use-fulness.

How old is too old? In vivo engraftment of human peripheral blood stem cells cryopreserved for up to 18 years-implications for clinical transplantation and stability programs

BACKGROUND Peripheral blood stem cells(PBSC)are commonly cryopreserved awaiting clinical use for hematopoietic stem cell transplant.Long term cryopreservation is commonly defined as five years or longer,and limited data exists regarding how long PBSC can be cryopreserved and retain the ability to successfully engraft.Clinical programs,stem cell banks,and regulatory and accrediting agencies interested in product stability would benefit from such data.Thus,we assessed recovery and colony forming ability of PBSC following long-term cryopreservation as well as their ability to engraft in NOD/SCID/IL-2 Rγnull(NSG)mice.AIM To investigate the in vivo engraftment potential of long-term cryopreserved PBSC units.METHODS PBSC units which were collected and frozen using validated clinical protocols were obtained for research use from the Cellular Therapy Laboratory at Indiana University Health.These units were thawed in the Cellular Therapy Laboratory using clinical standards of practice,and the pre-freeze and post-thaw characteristics of the units were compared.Progenitor function was assessed using standard colony-forming assays.CD34-selected cells were transplanted into immunodeficient mice to assess stem cell function.RESULTS Ten PBSC units with mean of 17 years in cryopreservation(range 13.6-18.3 years)demonstrated a mean total cell recovery of 88%±12%(range 68%-110%)and post-thaw viability of 69%±17%(range 34%-86%).BFU-E growth was shown in 9 of 10 units and CFU-GM growth in 7 of 10 units post-thaw.Immunodeficient mice were transplanted with CD34-selected cells from four randomly chosen PBSC units.All mice demonstrated long-term engraftment at 12 wk with mean34%±24%human CD45+cells,and differentiation with presence of human CD19+,CD3+and CD33+cells.Harvested bone marrow from all mice demonstrated growth of erythroid and myeloid colonies.CONCLUSION We demonstrated engraftment of clinically-collected and thawed PBSC following cryopreservation up to 18 years in NSG mice,signifying likely successful clinical transplantation of PBSC following long-term cryopreservation.

Point-of-Care Immunoassays with Tunable Detection Range for Detecting Infection in Intensive Care Unit

One of the difficulties of a multiplexed analytical assay is matching the concentration range of different markers.Here,the authors develop an automatic,multiplexed point-of-care(POC)immunoassay that allows detecting C-reactive protein(CRP),procalcitonin(PCT),and interleukin 6(IL-6)in 50μL serum samples with limits of detection 1.87μg/mL,0.17 ng/mL,and 49.75 pg/mL,respectively.The authors use electrospun fibers and surface chemistry of gold nanoparticles(AuNPs)to adjust the detection range of different biomarkers.The authors have proposed some new diagnostic indicators(mScore and mScorePlus)that combine the results of CRP,PCT,IL-6,and complete blood counts(CBCs).The authors also tracked changes in CRP,PCT,and IL-6 of patients in an intensive care unit(ICU).The authors find that mScore and mScorePlus have advantages in improving the diagnostic accuracy and providing more analytical information.mScore and mScorePlus are effective tools to detect infections,differentiate bacterial and viral infections,and monitor disease.Integrating multiple markers into one straightforward parameter is an effective method for analytical applications.The authors believe that this method provides a general way for chemists to develop increasingly accurate detection methods and indicators.

Comparison of the Content,Uniformity of Dosage Units and Dissolution Rate of Triphasic Oral Contraceptives from Two Pharmaceutical Factories

Objective To compare the content, uniformity of dosage units and dissolution rate of triphasic oral contraceptives from two pharmaceutical factories A and B. Methods A High Performance Liquid Chromatography （HPLC） method for the simultaneous determination of levonorgestrel and ethinylestradiol was used. The content of levonorgestrel （LNG） was monitored by an UV detector at 247 nm, while ethinylestradiol （EE） was monitored by fluorescence detector with the excitation of 285 nm and emission wavelengths of 310 nm. The dissolution test was performed using the paddle method. Results The content of levonorgestrel （LNG） and ethinylestradiol （EE） in product A was within 100.5%-122.4% while product B within 120.6%-140.9%. The uniformity value of dosage units of tablets from two factories was more than 15. The dissolution rate of tables from two factories was more than 60% within 60 min. Conclusion Only the content of product A was in the ±25% range of label claim. The uniformity of two products was not up to standard. The dissolution rate of the tablets from two products met the requirement of ChP2005.

Adenovirus-mediated and tumor-specific transgene expression of the sodium-iodide symporter from the human telomerase reverse transcriptase promoter enhances killing of lung cancer cell line in vitro

Background The sodium-iodide symporter （NIS） protein can mediate the active radioiodine uptake.The human telomerase reverse transcriptase （hTERT） promoter is known to be selectively reactivated in majority of tumors and hence could be used for tumor targeting.We constructed a recombinant adenovirus containing the human sodium iodide symporter （hNIS） gene directed by the hTERT promoter, characterized the ability of infected cells in uptaking iodide, and explored the therapeutic efficacy of 131I in a lung cancer cell line in vitro.Methods The hTERT promoter was amplified by PCR from DNA isolated from log-phase HepG2 cells, subcloned into lineralized FL＊-hNIS/pcDNA3, and then the hTERT-hNIS sequence was subcloned into the shuttle plasmid pAdTrack.The recombinant adenovirus Ad-hTERT-hNIS was constructed by AdEasy system.A positive control adenovirusAd-CMV-hNIS and a negative control adenovirus Ad-CMV were created similarly.A549 cells were transduced with recombinant adenoviruses.125I uptake studies and sodium perchlorate suppression studies were used to confirm hNIS expression and function.Toxic effects of 131I on tumor cells were studied by in vitro clonogenic assay.Results We first successfully constructed an adenovirus mediated transgene expression system of the hNIS under the control of hTERT promoter.When infected with recombinant adenovirus constructs expressing hNIS directed by hTERTand CMV-promoters （Ad-hTERT-hNIS and Ad-CMV-hNIS, respectively）, the lung cancer cell line A549 had increased ability to uptake radioiodide up to 23- and 30- fold compared to the control parental cells, respectively.The radioiodide uptake ability of both the Ad-CMV-hNIS and Ad-hTERT-hNIS transduced cell lines were repressed 11-fold by sodium perchlorate （NaCIO4）.The subsequent in vitro clonogenic assay of the infected A549 cell line was further repressed to 23% （Ad-CMV-hNIS） and 30% （Ad-hTERT-hNIS） of the control group after receiving radioiodide for 7 hours （P 〈0.001）.Conclusion Our preliminary study indicates that an adenovirus mediated transgene expression system of the hNIS under the control of hTERT promoter has the potential to become an effective wide-spectrum yet highly specific anti-cancer strategy.

Allogeneic compact bone-derived mesenchymal stem cell transplantation increases survival of mice exposed to lethal total body irradiation： a potential immunological mechanism

Background Radiation-induced injury after accidental or therapeutic total body exposure to ionizing radiation has serious pathophysiological consequences,and currently no effective therapy exists.This study was designed to investigate whether transplantation of allogeneic murine compact bone derived-mesenchymal stem cells （CB-MSCs） could improve the survival of mice exposed to lethal dosage total body irradiation （TBI）,and to explore the potential immunoprotective role of MSCs.Methods BALB/c mice were treated with 8 Gy TBI,and then some were administered CB-MSCs isolated from C57BL/6 mice.Survival rates and body weight were analyzed for 14 days post-irradiation.At three days post-irradiation,we evaluated IFN-Y and IL-4 concentrations; CD4＋CD25＋Foxp3＋ regulatory T cell （Treg） percentage; CXCR3,CCR5,and CCR7 expressions on CD3＋T cells; and splenocyte T-bet and GATA-3 mRNA levels.CB-MSC effects on bone marrow hemopoiesis were assessed via colony-forming unit granulocyte/macrophage （CFU-GM） assay.Results After lethal TBI,compared to non-transplanted mice,CB-MSC-transplanted mice exhibited significantly increased survival,body weight,and CFU-GM counts of bone marrow cells （P＜0.05）,as well as higher Treg percentages,reduced IFN-Y,CXCR3 and CCR5 down-regulation,and CCR7 up-regulation.CB-MSC transplantation suppressed Th1 immunity.Irradiated splenocytes directly suppressed CFU-GM formation from bone marrow cells,and CB-MSC co-culture reversed this inhibition.Conclusion Allogeneic CB-MSC transplantation attenuated radiation-induced hematopoietic toxicity,and provided immunoprotection by alleviating lymphocyte-mediated CFU-GM inhibition,expanding Tregs,regulating T cell chemokine receptor expressions,and skewing the Th1/Th2 balance toward anti-inflammatory Th2 polarization.

Reconstruction of hematopoietic function in irradiated mice promoted by polysaccharide peptide

Objective: To study the effect of polysaccharide peptide (PSP) on the reconstruction of the hematopoietic function in mice irradiated by lethal dose. Methods: The characteristics of proliferation of colony forming unit granulocyte macrophage (CFU GM) and colony forming unit spleen (CFU S) were measured after continuous injection of PSP in mice for seven days with different doses. Results: Injection of 25 mg/kg PSP in mice could promote and increase CFU GM proliferation and cell mitosis, markedly enhance survival ratio, survival time and CFU S. Conclusions: PSP has significant regulative effects on the reconstruction of the hematopoietic and the immunologic functions in mice irradiated by lethal dose.

Real-time 3D Microtubule Gliding Simulation Accelerated by GPU Computing

A microtubule gliding assay is a biological experiment observing the dynamics of microtubules driven by motor proteins fixed on a glass surface. When appropriate microtubule interactions are set up on gliding assay experiments, microtubules often organize and create higher-level dynamics such as ring and bundle structures. In order to reproduce such higher-level dynamics on computers, we have been focusing on making a real-time 3D microtubule simulation. This real-time 3D microtubule simulation enables us to gain more knowledge on microtubule dynamics and their swarm movements by means of adjusting simulation paranleters in a real-time fashion. One of the technical challenges when creating a real-time 3D simulation is balancing the 3D rendering and the computing performance. Graphics processor unit （GPU） programming plays an essential role in balancing the millions of tasks, and makes this real-time 3D simulation possible. By the use of general-purpose computing on graphics processing units （GPGPU） programming we are able to run the simulation in a massively parallel fashion, even when dealing with more complex interactions between microtubules such as overriding and snuggling. Due to performance being an important factor, a performance n, odel has also been constructed from the analysis of the microtubule simulation and it is consistent with the performance measurements on different GPGPU architectures with regards to the number of cores and clock cycles.