Fourier transform infrared (FT-IR) spectroscopy and improved principal component regression (PCR) for quantification of solid analytes in microalgae and bacteria

In bioanalytical chemistry, a detailed chemical understanding of biomaterials is often difficult to obtain due to the sheer number of analytes contained in a sample along with the samples' generally low reproducibility. This study presents a Fourier transform infrared (FT-IR) spectroscopic technique in conjunction with innovations in sample preparation and chemometric data preprocessing to overcome these limitations. These methodologies were applied to quantitative analyses of 31 representative compounds commonly found in biomaterial, which have been incorporated into a spectroscopic calibration database, that is, albumin (protein); D-alanine, glycine, histidine, valine, arginine, cysteine, phenylalanine, tyrosine, methionine, L-glutamine, and glutamic acid, (amino acids); glucose, fructose, galactose, mannose, sucrose, lactose, glycogen, agarose, and starch (carbohydrates); DNA (salmon sperm), sulphonoquinovosyl diglyceride ( sulpho-lipid ), and 1,2-diacyl-sn-glycero-3-phospho-L-serine ( phospho-lipid ); succinic acid and malic acid ( carboxylic acids ); glycolic acid (a -hydroxy acid), sodium pyruvate, b -carotene, frustules (microalgae silica-shells), and ammonium formate. Two proof-of-principle applications were based on calibration models incorporating these solids, i.e., characterization of E. coli and microalgae. The former aims for detection of bacterial contamination and the latter to enable investigations of changes in chemical composition of microalgae cells in response to shifting environmental conditions. Chemometric preprocessing steps have been developed for handling sample-to-sample fluctuations of absorption path lengths and baselines; the former incorporated mass normalization while the latter utilized a novel baseline correction method that requires no a priori information. Data preprocessing, chemometric calibration, and evaluation algorithms have been combined, together with an extensive spectral database of the aforementioned compounds (～1500 samples), for quantitative calibration purposes through the remotely accessible Virtual Chemometrics Lab , which can be utilized for a multitude of applications through a graphical user interface.     

Resistance to nitric oxide-mediated apoptosis in HL-60 variant cells is associated with increased activities of Cu,Zn-superoxide dismutase and catalase

Nitric oxide (NO) released from (Z)-1-[N-(2-aminoethyl)-N-(2-ammonioethyl)amino]diazen-1- ium-1,2-diolate (DETA/NO or NOC-18) induces apoptosis in human leukemia HL-60 cells. In this study, we isolated a HL-60 variant cell line, HL-NR6, that is resistant to DETA/NO toxicity as assessed by DNA fragmentation, morphology, and colony forming ability. The variant cells also showed resistance to reactive oxygen species (ROS) such as superoxide and hydrogen peroxide as well as NO donors, but not to anti-tumor drugs. We found that HL-NR6 cells when compared with HL-60 cells possessed twice the activities of Cu,Zn-superoxide dismutase (Cu,Zn-SOD) and catalase, but no change in Mn-SOD nor in glutathione peroxidase. Immunoblotting confirmed the high levels of both enzymes in the variant cell. We also observed that ROS generation following DETA/NO exposure was substantially higher in HL-60 cells than in HL-NR6 cells, using the 2',7'-dichlorofluorescein fluorometric method. Moreover, the SOD mimetic Mn(III) tetrakis(1-methyl-4-pyridyl) porphyrin and exogenous catalase effectively attenuated DETA/NO-elicited DNA fragmentation in HL-60 cells. Taken together, these data suggested that the NO resistance in HL-NR6 cells is associated with the increased Cu,Zn-SOD/catalase and that NO-mediated apoptosis in HL-60 cells is correlated with the generation of ROS and derived molecules like peroxynitrite.     

Photoperiodic effects on gonadotropin-releasing hormone (GnRH) content and the GnRH-immunoreactive neuronal system of male Siberian hamsters

Despite profound photoperiodic differences in circulating gonadotropin levels, consistent differences in the GnRH system have not been observed in Siberian hamsters (Phodopus sungorus) housed chronically in short or long days. During the transition from short to long days, however, male hamsters exhibit a transient increase in the number of cells expressing prepro-GnRH mRNA on the morning of the second long day. Here, we present two experiments designed to examine whether or not this change in mRNA level is associated with changes in GnRH protein synthesis. In the first experiment, we used RIA to measure GnRH content in preoptic area-mediobasal hypothalamic homogenates. We observed a significant increase in GnRH protein levels on the morning of the second long day relative to short- and long-day controls. The latter two groups did not differ from one another. In the second experiment, we used immunocytochemistry to quantify GnRH cell number in the various treatment groups. GnRH-immunoreactive (-ir) cell number did not increase significantly after long-day transfer relative to that in short-day controls; however, both of these groups had significantly more GnRH-ir neurons than long-day controls. We hypothesize that during the transition from short to long days, male Siberian hamsters experience a transient increase in GnRH production in a stable population of neurons. When GnRH secretion subsequently increases on long days, peptide content within neuronal cell bodies declines, leading to a decrease in the number of immunoreactive neurons detected. The rapid response of the hypothalamo-pituitary-gonadal axis in Siberian hamsters to a change in day length defines a narrow temporal window in which to identify the physiological, cellular, and molecular mechanisms mediating the photoperiodic regulation of reproduction.     

Formation of antibiotic,biodegradable/bioabsorbable polymers by processing with neomycin sulfate and its inclusion compound with β‐cyclodextrin

Samples of pure neomycin sulfate and its inclusion compound (IC) with β‐cyclodextrin were implanted into films of poly(L ‐lactic acid) (PLLA) and poly(ε‐caprolactone) (PCL). Both polymers have been widely used commercially to make sutures. The antibacterial activity of these films against Escherichia coli was tested. Films made by either solution casting or melt pressing were divided into the following three groups: (1) plain polymer films, (2) those embedded with pure neomycin sulfate, and (3) those embedded with neomycin sulfate‐β‐cyclodextrin IC. Filter paper treated with 1.5 μL of 10 mg/μL Kanamycin and neomycin were used as controls and resulted in 11‐ and 8‐mm zones of inhibition/or antibacterial activity, respectively. Small discs (ca. 2% of total area) cut from solution‐cast films of PLLA and PCL containing 50 wt % neomycin sulfate IC had 17‐ and 16‐mm zones of inhibition, and PLLA and PCL containing 50 wt % pure neomycin sulfate deterred bacterial growth, resulting in 19‐mm zones of inhibition. Melt‐pressed films containing 10 wt % pure neomycin sulfate or its IC, showed 17‐ and 11‐mm zones of inhibition for PLLA films, respectively, while PCL films showed 13‐ and 9‐mm zones of inhibition, respectively. For melt‐pressed films that contain 0.01 wt % pure neomycin sulfate or its IC, PLLA films showed 11‐ and 9.5‐mm zones of inhibition, respectively, while PCL films showed 11‐ and 10‐mm zones of inhibition, respectively. Since an antibiotic, bioabsorbable suture does not require surgical removal, implanting an inclusion compound in the suture might allow the slow release of antibiotic, thereby guarding against postsurgical infection and also protecting the antibiotic from degradation during the melt‐spinning process used to make the suture. © 1999 John Wiley & Sons, Inc. J Appl Polym Sci 74: 937–947, 1999     

A review of algorithms for molecular sequence comparison

Computers have recently become an essential component of research in molecular biology. Most computer analyses of nucleic acid and protein sequences depend on comparisons between sequences. These comparisons, depending on their purpose, may differ not only in the kinds of comparisons that are done, but also in the way the results of the comparison are used by molecular biologists or by other computer programs. This paper reviews algorithms currently in use to solve comparison problems in molecular biology. Each algorithm is explained in detail and discussed in terms of the molecular biology problems it is most suited to solve.     

ITER operational space for full plasma current H-mode operation

Sensitivity studies performed as part of the ITER IO design review highlighted a very stiff dependence of the maximum Q attainable on the machine parameters. In particular, in the considered range, the achievable Q scales with I_p^{^}4. As a consequence, the achievement of the ITER objective of Q = 10 requires the machine to be routinely operated at a nominal current I_p of 15 MA, and at full toroidal field BT of 5.3 T. This paper analyses the capabilities of the poloidal field (PF) system (including the central solenoid) of ITER against realistic full current plasma scenarios. An exploration of the ITER operational space for the 15 and 17 MA inductive scenario is carried out. An extensive analysis includes the evaluation of margins for the closed loop shape control action. The overall results of this analysis indicate that the control of a 15 MA plasma in ITER is likely to be adequate in the range of li 0.7–0.9 whereas, for a 17 MA plasma, control capabilities are strongly reduced. The ITER operational space, provided by the reference pre-2008 PF system, was rather limited if compared to the range of parameters normally observed in present experiment. Proposals for increasing the current and field limits on PF2, PF5 and PF6, adjustment on the number of turns in some of the PF coils, changes to the divertor dome geometry, to the conductor of PF6 to Nb3Sn, moving PF6 radially and/or vertically are described and evaluated in the paper. Some of them have been included in 2008 ITER revised configuration.     

Hydrogen Storage in Partially Exfoliated Magnesium Diboride Multilayers

Metal boride nanostructures have shown significant promise for hydrogen storage applications. However, the synthesis of nanoscale metal boride particles is challenging because of their high surface energy, strong inter- and intraplanar bonding, and difficult-to-control surface termination. Here, it is demonstrated that mechanochemical exfoliation of magnesium diboride in zirconia produces 3–4 nm ultrathin MgB₂ nanosheets (multilayers) in high yield. High-pressure hydrogenation of these multilayers at 70 MPa and 330 °C followed by dehydrogenation at 390 °C reveals a hydrogen capacity of 5.1 wt%, which is ≈50 times larger than the capacity of bulk MgB₂ under the same conditions. This enhancement is attributed to the creation of defective sites by ball-milling and incomplete Mg surface coverage in MgB₂ multilayers, which disrupts the stable boron–boron ring structure. The density functional theory calculations indicate that the balance of Mg on the MgB₂ nanosheet surface changes as the material hydrogenates, as it is energetically favorable to trade a small number of Mg vacancies in Mg(BH₄)₂ for greater Mg coverage on the MgB₂ surface. The exfoliation and creation of ultrathin layers is a promising new direction for 2D metal boride/borohydride research with the potential to achieve high-capacity reversible hydrogen storage at more moderate pressures and temperatures.