Synthesis and Characterization of γ‑Fe₂O₃ for H₂S Removal at Low Temperature

The performance of γ-Fe₂O₃ as sorbent for H₂S removal at low temperatures (20–80 °C) was investigated. First, γ-Fe₂O₃/SiO₂ sorbents with a three-dimensionally ordered macropores (3DOM) structure were successfully prepared by a colloidal crystal templating method. Then, the performance of the γ-Fe₂O₃-based material, e.g., reference γ-Fe₂O₃ and 3DOM γ-Fe₂O₃/SiO₂ sorbents, for H₂S capture was compared with that of α-Fe₂O₃ and the commercial sorbent HXT-1 (amorphous hydrated iron oxide). The results show that γ-Fe₂O₃ has an enhanced activity compared to that of HXT-1 for H₂S capture at temperatures over 60 °C, whereas α-Fe₂O₃ has little activity. Because of the large surface area, high porosity, and nanosized active particles, 3DOM γ-Fe₂O₃/SiO₂ sorbent shows the best performance in terms of sulfur capacity and utilization. Moreover, it was found that moist conditions favor H₂S removal. Furthermore, it was found that the conventional regeneration method with air at high temperature was not ideal for the composite regeneration because of the transmission of some amount of γ-Fe₂O₃ to α-Fe₂O₃. However, simultaneous regeneration by adding oxygen in the feed stream allowed the breakthrough sulfur capacity of FS-8 to increase up to 79.1%, which was two times the value when there was no O₂ in the feed stream

FISH analysis of DT40 clones carrying the corrected modified human chromosomes.

<p>For the hybrid DT40 clone SLK2, the presence of both hChr2 and hChr22 was detected by human COT-1 DNA as the probe; for DT40 clones carrying the translocated human chromosomes, a two-color FISH assays were conducted: the hChr2 painting probe was labeled with Rhodamine and the hChr22 painting probe was labeled with Fluorescein.</p

Activating hK-ras expression in LSL-hK-ras^G12D-IRES-HSV1-tk GFF cells by Adenovirus -Cre-GFP.

<p>A. Morphology of GFP positive cells 48-Cre-GFP. Phase contrast (Left panel) and florescent micrographs (Right panel) were shown at 100× magnification. B. Efficient excision of the LSL cassette in the cell culture. PCR analysis of genomic DNA prepared from the different GFF cells. The wild type allele was devoted the 279 bp fragment. The LSL-hKras^G12D-IRES-HSV1-TK allele was devoted 2.5 kb fragment. The Lox- hKras^G12D-IRES-HSV1-TK allele was devoted the 320 bp fragment. 1: Lox- hK-ras^G12D-IRES-HSV1-tk GFF cells, 2: LSL-hK-ras^G12D-IRES-HSV1-tk GFF cells, 3: wild-type GFF cells. C. mRNA of HSV1-tk in GFF cells. Marker: 100 bp ladder, 1: LSL-hK-ras^G12D-IRES-HSV1-tk GFF was infected with Adenovirus-Cre-GFP 48 h. 2: LSL-hK-ras^G12D-IRES-HSV1-tk GFF cells. 3: Lox- hK-ras^G12D-IRES-HSV1-tk GFF cells, 4: wild-type GFF cells, N: negative control (H₂O as template). D. Dependence of ganciclovir concentrations against GFF cell viability using MTT assays. The results were expressed as the percentage of living cells in treated conditions at various concentrations of ganciclovir with respect to ganciclovir-free cultures. E. Specific reduction of mutant hK-ras transcripts in GFF cells. BstNI digestion cuts the WT K-ras allele to produce a 156-bp DNA fragment, whereas the mutant hK-ras allele remains uncut to produce a 186-bp DNA fragment. 1: wild-type GFF cells, 2: LSL-hK-ras^G12D-IRES-HSV1-tk GFF cells, 3: Lox- hK-ras^G12D-IRES-HSV1-tk GFF cells, 4: LSL-hK-ras^G12D-IRES-HSV1-tk 48 h post infection.</p

Bovinization of hIgM CH2-TM Domain.

<p><b>The upper line shows the genomic configuration of the CH 1-TM2 domain of hIgM, the middle line shows the gene targeting vector pCH2CAGzeoDT, and the bottom line depicts the modified CH2-TM2 domain of hIgM in a DT40 cell clone, CH2D4</b>. </p

Schematic representation of the conditional hK-Ras^G12D construct.

<p>A, Targeting strategy of knock-in of the hK-ras^G12D to goat K-ras exon 1 locus and its activation by Cre; B, Schematic outline of the cloning strategy for the construction of targeting vector: pKO2.1-LSL-hK-ras^G12D-IRES-HSV1-tk.</p

Analysis of constructed isKcHACΔ vector.

<p>(<i>A</i>) Extensive genomic PCR for genotyping of the isKcHACΔ vector. Location of each genomic PCR primer pair is depicted in relation to the isKcHACΔ vector structure. (<i>B</i>) CGH analysis among three different CHO clones containing the isKcHACΔ vector. DNA from isKCDC15-8 was used as a reference. There was no apparent structural difference in the isKcHACΔ among the three cell lines.</p

Modification of hChr14.

<p>(A) A lox511 sequence along with the promoter-less <i>puro</i> cassette was integrated at the AL512355 locus with gene targeting vector p14CEN(FR)hygpuro^lox511DT, and a lox511 sequence along with a CAG promoter and a hygromycin (hyg) selection cassette was integrated at locus AL512355 with gene targeting vector pSC355CAG^lox511hisDDT. Following Cre expression, the ~85 Mb genomic sequence was removed rendering <i>puro</i> expression. A loxP sequence and a GFP reporter cassette was then integrated at the <i>RNR2</i> locus to generate 14D. (B) FISH analysis of a DT40 clone, 14D, containing the correctly modified hChr14.</p

Trans-class switched bovine IgG levels in a series of Tc cattle.

<p>(A) The ratio of t-bIgG to total IgG (human IgG and t-bIgG) in percentage. The number of Tc <b>cattle</b> evaluated at each time points was noted on top of each bar in brackets. (B) Serum t-bIgG concentration in Tc bovines. The serum t-bIgG concentrations of Tc bovines with TKO background were evaluated at ages of 5–6 months and at 11–12 months. The number of Tc bovines evaluated at each time points was noted on top of the each bar in brackets.</p

Human IgG production profile in cKSL-HACΔ/DKO Tc cattle. (A) Comparison of average total hIgG (left panel) and fully hIgG (hIgG/hIgκ) concentrations in the sera of κHAC/DKO and cKSL-HACΔ/DKO calves at 5-6 months of age. (B) IgG subclass distribution in the plasma of cKSL-HACΔ/DKO calves. (C) Mean fluorescence intensity (MFI) of tumor cells stained with the sera from immunized cattle. Left panel shows the MFI with anti-hIgG antibodies (measuring total hIgG); right panel shows the MFI with anti-hIgκ antibodies (measuring fully hIgG/hIgκ).

<p>Human IgG production profile in cKSL-HACΔ/DKO Tc cattle. (A) Comparison of average total hIgG (left panel) and fully hIgG (hIgG/hIgκ) concentrations in the sera of κHAC/DKO and cKSL-HACΔ/DKO calves at 5-6 months of age. (B) IgG subclass distribution in the plasma of cKSL-HACΔ/DKO calves. (C) Mean fluorescence intensity (MFI) of tumor cells stained with the sera from immunized cattle. Left panel shows the MFI with anti-hIgG antibodies (measuring total hIgG); right panel shows the MFI with anti-hIgκ antibodies (measuring fully hIgG/hIgκ).</p

Construction of the istHAC vector.

<p>A flow of the istHAC vector construction is illustrated. The <i>attP</i> sequence is integrated at 5’ side of the hI_γ1 exon 1 and 3’ side of the h<i>IGHG1</i> TM2 by the targeting vectors phI_γ1FRTCAGattPhisDDT and ph_γ1TMNeoattPDT, respectively. Then, the replacement vector pBAC-istHAC is introduced with the ΦC31 recombinase to bring about the <i>attP</i>/<i>attB</i> recombination to replace the flanked region. The successful replacement causes the CAG promoter-driven DsRed gene to be reconstituted to provide red fluorescence for sorting. Finally, the DsRed cassette is removed by the FLP expression.</p