Data for evaluation of the onshore Cretaceous Zululand Basin in South Africa for geological CO2 storage.

The world has set the goal of reducing CO2 emissions from burning fossil fuels by using carbon capture and storage (CCS) as one of the major solutions. A sudden and complete switch from fossil fuels to renewable resources cannot be achieved immediately. Therefore, CCS remains an essential techniques to reduce CO2. In this work, the 180 - 65 Ma old onshore part of the Zululand Basin in KwaZulu-Natal in South Africa was investigated for geological CO2 sequestration. A total of 160 core samples of sandstone, conglomerate, tuff, rhyolite, breccia, and siltstone were taken from NZA, ZA, ZB, and ZC drill cores. The wells were drilled in the 1960s by the South African Petroleum and Gas Corporation Company for hydrocarbon exploration. In order to examine the basin suitability for CO2 storage, porosity and permeability, mineralogy, geochemistry, geomechanical properties, and H2O-CO2-rock interactions were investigated using geological core logging, spectral scanning, petrography, X-ray diffraction (XRD), X-ray fluorescence (XRF), inductively coupled plasma mass spectrometry, uniaxial compressive stress, and scanning electron microscopy. The basin comprises clastic sedimentary rocks, pyroclastic deposits and carbonates from the Makatini, Mzinene and St. Lucia formations. Aptian and Cenomanian sandstones are identified as CO2 reservoirs, and the siltstone above is considered capstone. The sandstone comprises on average 34.45 wt% quartz, 32.91 wt% clays, 29.53 wt% feldspars, 4.44 wt% carbonates, 3.10 wt% Fe-oxides, 2.40 wt% micas, and 2.00 wt% organic materials as per XRD data, also contains trace amounts of sulphides and sulphates. Geochemical XRF data for sandstone are 29.72 - 62.51 wt% SiO2, 6.95 - 13.44 wt% Al2O3, 3.06 - 48.81 wt%, 1.90 - 4.51 wt% MgO, 1.04 - 2.19 wt% K2O, 1.00 - 3.67 Na2O wt%. The content of TiO2, Cr2O3 and P2O5 is below 0.01 wt% each. Siltstone has similar mineralogy and geochemistry as sandstone, but high clay content, fine-grained, impervious, with porosity <5%. The sandstone and siltstone are geomechanically soft and recorded 15 MPa on the Enerpac P141 device. CO2-H2O-rock interaction experiments performed at 100 °C and 100 bar using autoclaves showed that sandstone and siltstone react with scCO2.

ELISA-Based Crossmatching Allowing the Detection of Emerging Donor-Specific Anti-HLA Antibodies through the Use of Stored Donors' Cell Lysates.

About forty years ago the complement-dependent crossmatch assay (CDC-CM) was developed as standard procedure in order to select recipients without donor-specific antibodies directed against human leukocyte antigens of their given donors since the negative outcome of pretransplant crossmatching represents one of the most important requirements for a successful kidney graft survival. However, as a functional assay the CDC-CM strongly depends on the availability of donors' isolated lymphocytes and in particular on their vitality highly limiting its applicability for recipients treated with special drugs and therapeutic antibodies or suffering from underlying autoimmune diseases. In the great majority of these cases ELISA-based crossmatching has been demonstrated to be an adequate alternative procedure nevertheless leading to valid results. With these case reports we show for the first time that ELISA-based crossmatching is suitable to demonstrate the upcoming donor-specific anti-HLA antibodies as a consequence of allografting using deep-frozen deceased donor's material such as blood or spleen detergent lysate. Thus, this ELISA-based procedure first provides the option to routinely perform crossmatching using stored material of deceased donors in order to substitute or at least to complement virtual crossmatching, that is, the comparison of the recipients' anti-HLA antibody specificities with the donors' historically identified HLA types.

Artificially Positive Crossmatches Not Leading to the Refusal of Kidney Donations due to the Usage of Adequate Diagnostic Tools.

Allografting patients with human leukocyte antigens (HLA) which are recognized by preformed antibodies constitutes the main cause for hyper-acute or acute rejections. In order to select recipients without these donor-specific antibodies, the complement-dependent cytotoxicity crossmatch (CDC-CM) assay was developed as a standard procedure about forty years ago. The negative outcome of pretransplant crossmatching represents the most important requirement for a successful kidney graft survival. The artificially positive outcomes of CDC-based crossmatches due to the underlying disease Systemic Lupus Erythematosus (SLE), however, may lead to the unjustified refusal of adequate kidney grafts. Two prospective female recipients destined for a living as well as for a cadaver kidney donation, respectively, exhibited positive CDC-based crossmatch outcomes although for both patients no historical immunizing events were known. Furthermore, solid phase-based screening or antibody differentiation analyses never led to positive results. Immediate reruns of the CDC-based crossmatch assays using the alternative antibody monitoring system (AMS-)crossmatch ELISA resulted in unequivocally negative outcomes. Consequently both transplantations were performed without any immunological complications for the hitherto follow-up time of 25 and 28 months, respectively. We here show two case reports demonstrating an alternative methodical approach to circumvent CDC-based artefacts and point to the urgent need to substitute the CDC-based crossmatch procedure at least for special groups of patients.

Generation of donor-specific anti-human leukocyte antigen antibodies after the transplantation of a fully matched kidney allograft and its impact on the selection of a subsequent renal regraft: a case report.

Detection of anti-human leukocyte antigen (HLA) antibodies and identification of their specificities represent important tasks for patients awaiting kidney allografts. Regarding patients immunized by pregnancies, transfusions, or previous transplantations of solid organs, the immunization status must be observed carefully because grafting them with HLA phenotypes recognized by their antibodies represents the main cause for hyper-acute or acute rejection episodes, often leading to transplant loss. A 10-year-old patient with end-stage renal insufficiency of HLA type A3, 25; B8, 18, (Bw6); Cw7,12; DR15,17; DR51,52; DQ2,6 received a deceased donor graft showing no HLA mismatch in 1998. It lost function after 8 years, resulting in the patient's re-entry onto the waiting list for kidney transplantation in 2006. Antibody screening detected anti-HLA-A25, A26, A34, and A66 (broad A10) antibodies using various techniques (DynaChip, Single Antigen enzyme-linked immunosorbent assay [ELISA]). Additionally, a kidney offer expressing the HLA-A25 phenotype was not acceptable for the patient due to a positive complement-dependent cytotoxicity assay (CDC)-based cross-match. The question arose whether this reactivity might be due to auto-reactive antibodies directed against the HLA-A25 phenotype. However, no auto-reactive antibodies were detectable using either the CDC-based or the antibody monitoring system-ELISA-based cross-match assays. Consequently the patient was re-examined at high resolution showing the rare HLA-A*25:14 genotype. This case showed that rare alleles may result in allele-specific antibodies directed against the common variants, thus leading to unexpected positive cross-match results against apparently matched allografts.

General insufficiency of the classical CDC-based crossmatch to detect donor-specific anti-HLA antibodies leading to invalid results under recipients' medical treatment or underlying diseases.

Antibodies directed against HLA antigens of a given donor represent the most prominent cause for hyper-acute and acute rejections. In order to select recipients without donor-specific antibodies the complement-dependent cytotoxicity (CDC-) crossmatch as the standard procedure was established. As a functional assay it strongly depends on the availability of isolated donor lymphocytes and in particular on their vitality. However, due to several diseases or pharmacological treatment of a given recipient unexpected "false-positive" results of the CDC-crossmatch may arise. We here present three groups of patients which demonstrate the limits of the conventional crossmatch. 1) Kidney recipients before living donations exhibited positive CDC-reactions due to their conditioning using the therapeutical anti-CD20 mAb Rituximab (n=7), routinely used to deplete B-cells, or the anti-CD25 mAb basiliximab (n=2) to inhibit the proliferation of activated T-cells. 2) Recipients suffering from various leukaemias (n=5) exhibited "positive" CDC-crossmatches using PBL of the donors, although formerly these patients had never shown anti-HLA antibodies. Instead of donor-specific allo-antibodies, cytostatic agents such as 6-mercaptopurine led to an unspecific cell death. 3) Patients projected for post mortem or living kidney donations (n=44) exhibited "positive" CDC-crossmatch results which were not in accordance with their former antibody status and, partially, with high degrees of HLA-matching. These implausible results were due to underlying auto-immune diseases, mainly of the systemic immune complex type III such as lupus erythematosus, mainly leading to false-positive B-cell crossmatches by immune complexes binding to Fcγ-receptors. In all these 58 cases the alternatively performed ELISA-based "Antibody Monitoring System" (AMS-) crossmatch assay was not artifically affected, suggesting that this assay may be comprehensively established at least for the cases described.

The del/del genotype of the nuclear factor-kappaB -94ATTG polymorphism and its relation to aggressive periodontitis.

BACKGROUND AND OBJECTIVE: Periodontitis is influenced by specific host-dependent immune responses. Periodontopathogens induce innate immune responses, amongst others, via toll-like receptor 2 (TLR2), resulting in activation of the nuclear transcription factor nuclear factor-kappaB (NF-kappaB). The aim of this case-control study was to evaluate links between genetic variants of these genes and chronic/aggressive periodontitis in a multivariate model. MATERIAL AND METHODS: A total of 141 patients with periodontitis (63 with chronic periodontitis and 78 with aggressive periodontitis) and 81 controls without periodontitis were included in the study. Polymorphisms in TLR2 (Arg677Trp, Arg753Gln) and in NF-kappaB (-94ins/delATTG) were determined by restriction fragment length polymorphism and fragment length analyses, respectively. Subgingival bacterial colonization was evaluated using a PCR/DNA probe test (micro-Ident). RESULTS: Although there was no association of the TLR2 polymorphism Arg753Gln with periodontitis, heterozygous carriers (Arg/Gln) were at a higher risk for colonization with bacteria of the 'red complex' (corrected p-value = 0.042). The del/del genotype of the NF-kappaB polymorphism was associated with aggressive periodontitis considering age, gender, smoking and approximal plaque index as potential confounders (odds ratio = 2.81, p = 0.035, 95% confidence interval: 1.08-7.33). del/del carriers had a higher risk for subgingival colonization with Aggregatibacter actinomycetemcomitans (odds ratio = 2.36, p = 0.030, 95% confidence interval: 1.09-5.1; adjusted for age, gender, smoking and pocket depth(bacteria)). CONCLUSIONS: The del/del genotype of NF-kappaB was shown to be associated with the occurrence of aggressive periodontitis.

Impact of genetic variants of CD14 and TLR4 on subgingival periodontopathogens.

CD14 and toll-like receptor 4 (TLR4) are involved in host's immune response to bacterial pathogens including periodontal bacteria. Functional important gene polymorphisms are described for both genes. The aim of this study was to evaluate links between genetic polymorphisms of CD14 and TLR4 and risk markers of periodontitis in a multivariate model. One hundred and thirty-three periodontitis patients (chronic: n = 60, aggressive: n = 73) and 80 healthy controls without periodontitis were included in the study. Polymorphisms in CD14 c.-159C>T and in TLR4 Asp299Gly, Thr399Ile were determined by restriction fragment length polymorphism analyses. The clinical investigation included smoking status, plaque and bleeding indexes, pocket depth and attachment loss. Subgingival bacterial colonization was analysed molecularbiologically using the micro-Ident test. Prevotella intermedia occurred less frequently in individuals positive for the TT genotype of CD14 in bivariate analysis (odds ratio = 0.36%, confidence interval: 0.14-0.91, P = 0.045). In binary logistic regression analyses, the occurrence of this bacterium was significantly decreased in TT carriers (odds ratio = 0.31%, confidence interval: 0.81-0.12, P = 0.017) considering age, smoking and maximum clinical attachment loss at microbial test site as confounding factors. However, no significant association with chronic and or aggressive periodontitis and polymorphisms in CD14 and TLR4 could be proven. Although the CD14 c.-159C>T polymorphism could be shown to be associated with subgingival colonization with P. intermedia, there is no evidence that CD14 and TLR4 polymorphisms investigated are independent risk factors for chronic or aggressive periodontitis in German periodontitis patients.

Expression and regulation of non-classical HLA-G in renal cell carcinoma.

Under physiological conditions, the non-classical major histocompatibility complex class Ib molecule human leukocyte antigen G (HLA-G) is selectively expressed in placental trophoblasts, thymus and cornea. In pathological situations, HLA-G expression was frequently found in tumour cells of distinct origin, thereby allowing these tumour cells to escape immune surveillance. Although HLA-G expression occurs at a relatively high frequency in renal cell carcinoma (RCC) of the clear cell subtype, the molecular mechanisms of its aberrant expression in RCC has not yet been determined. Therefore, the constitutive and cytokine-mediated HLA-G expression as well as its mode of regulation was investigated. In addition to HLA-G-specific mRNA expression, membrane-bound and soluble/shed HLA-G protein was determined. Eight of 14 RCC cell lines analysed (57%) exhibited HLA-G-specific transcripts, whereas only 6 of 14 RCC cell lines (43%) expressed HLA-G protein, suggesting a post-transcriptional control of HLA-G in some cases. Treatment of RCC cell lines with either interferon-gamma or interleukin-10, respectively, increased HLA-G-specific mRNA and protein in six of eight HLA-G(+) RCC lines (75%), but not in HLA-G(-) RCC cells. A 5'-aza-2-deoxycytidine (5-Aza-dC)-mediated demethylation of the HLA-G promoter DNA resulted in an enhanced HLA-G expression in four of six RCC cell lines, whereas a de novo induction of HLA-G was only observed in one HLA-G(-) RCC cell line on treatment with 5-Aza-dC. Thus, there exist multiple mechanisms controlling HLA-G expression in RCC, which might also have an impact on the development of RCC-specific immunotherapies.

A novel HLA-B*57 (B*5714) allele in a healthy male Caucasian individual.

A novel human leucocyte antigen (HLA)-B57 (HLA-B*5714) allele has been identified in a male Caucasian individual from Middle Europe using single allele-specific sequencing strategy. This allele is identical to the HLA-B*570101 allele except for two point mutations in exon 3 at codon 138 (ACG-->ACC) with no amino acid change [persisting threonine (T)] and at codon 171 (TAC-->CAC), resulting in an amino acid change from tyrosine (Y) to histidine (H).

Soluble CD30 serum level--an adequate marker for allograft rejection of solid organs?

The CD30 molecule, a 120 kDa cell surface glycoprotein, is a member of the tumor necrosis factor receptor (TNF-R) superfamily and was originally identified on the surface of Reed-Sternberg cells and anaplastic large cell lymphomas in Hodgkin's disease patients. In addition to lymphoproliferative disorders the expression of CD30 was found in both activated CD8+ and CD4+ Th2 cells which lead to the activation of B-cells and consequently to the inhibition of the Th1-type cellular immunity. The membrane-bound CD30 molecule can be proteolytically cleaved, thereby generating a soluble form (sCD30) of about 85 kDa. Low serum levels of soluble CD30 were found in healthy humans, whereas increased sCD30 serum concentrations were detected under pathophysiological situations such as systemic lupus erythematosus, rheumatoid arthritis, certain viral infections and adult T cell leukaemia/lymphoma. In addition, it has recently been suggested that pre- or post-transplant levels of sCD30 represent a biomarker for graft rejection associated with an impaired outcome for transplanted patients. We here review (i) the current knowledge of the clinical significance of sCD30 serum levels for solid organ transplantations and (ii) our own novel data regarding inter- and intra-individual variations as well as time-dependent alterations of sCD30 levels in patients. (iii) Based on this information the implementation of sCD30 as predictive pre-transplant or post-transplant parameter for solid organ transplantation is critically discussed.

A novel HLA-A*33 (A*3309) allele in a Caucasian family that differs at protein position 66 from HLA-A*3308.

A novel human leucocyte antigen (HLA)-A33 (HLA-A*3309) allele has been identified in a Caucasian family from Middle Europe using single allele-specific sequencing strategy. This allele is identical to the HLA-B*3308 allele except for one point mutation in exon 2 at codon 66 (AAA-->AAT), resulting in an amino acid change from lysine (K) to asparagine (N).

A novel HLA-B*35 (B*3570) allele of a Caucasian family differing with one amino acid mismatch from HLA-B*3503 allele in exon 4.

A novel human leucocyte antigen (HLA)-B35 (HLA-B*3570) allele has been identified in a Caucasian family from Middle Europe using single allele-specific sequencing strategy. This allele is identical to the HLA-B*3503 allele except for one point mutation in exon 4 at codon 188 (CAC-->CGC), resulting in an amino acid change from histidine to arginine.

A novel HLA-A*0119 allele of a Caucasian family containing a mutation in a highly conserved region.

A novel human leucocyte antigen (HLA)-A (HLA-A*0119) allele has been identified in two individuals of a Caucasian family from Middle Europe using a single-allele-specific sequencing strategy. This allele is identical to the HLA-A*0101 allele except for one point mutation in the highly conserved codon 92 (TCT --> GCT) resulting in an amino acid change from serine to alanine.

Comparison of the established standard complement-dependent cytotoxicity and flow cytometric crossmatch assays with a novel ELISA-based HLA crossmatch procedure.

The detection of donor-specific anti-HLA antibodies by standard procedures such as complement-dependent cytotoxicity assay (CDC) or flow cytometric (FACS) analysis is limited by its low sensitivity and the quality of the donor cells. Therefore, an ELISA-based technique was employed using solid phase-immobilized monoclonal antibodies to capture HLA class I or class II molecules of the donor, respectively. In this HLA class I and class II antibody monitoring system (AMS) the donor-specific anti-HLA antibodies from the sera of recipients bind to the HLA molecules of the donor which have been immobilized by monoclonal antibodies (mAb) recognizing non-polymorphic epitopes. Upon binding of donor-specific anti-HLA antibodies they are recognized by secondary enzyme-conjugated anti-human immunoglobulin (Ig) antibodies. A newly established modification of the standard protocol allows the differentiation between bound antibodies of the IgG and IgM isotype. Furthermore, this assay was adapted for investigating small amounts of solid tissue of donors from whom no other cells (e.g. from blood) were available. We here provide an overview of the classical crossmatch methods with their advantages and limits. In addition, the design of the novel AMS-ELISA is described in terms of quality and sensitivity of the approach using exemplary cases of different application. The selected cases show that the AMS-ELISA represents a valuable tool for the post-transplantation monitoring of donor-specific anti-HLA antibodies during reaction crisis, after transfusion reactions and in particular cases of tissue transplantations lacking single cells.

Recombinant generation of two fragments of the rat complement inhibitory factor H [FH(SCR1-7) and FH(SCR1-4)] and their structural and functional characterization in comparison to FH isolated from rat serum.

Factor H (FH) is the predominant soluble inhibitor of the complement system. With a concentration of 200-800 microg/ml in human and rat plasma it acts as a cofactor for the soluble factor I (FI)-mediated cleavage of the component C3b to iC3b. Furthermore it competes with factor B for binding to C3b and C3(H2O) and promotes the dissociation of the C3bBb complex. FH is a monomer of about 155 kDa which comprises 20 short consensus repeats (SCR), each of which is composed of approximately 60 amino acid (aa) residues. Two functional fragments of FH comprising the SCR1-4 or SCR1-7 were generated using either the Baculovirus system or stably transfected human embryonal kidney cells, respectively. These fragments, as well as FH purified from rat serum, were first analyzed for their relative molecular weights (Mr) using non-reducing or reducing SDS-PAGE. The Mr of the FH variants differed by about 20% depending on the experimental conditions employed. Only the Mr of proteins separated under reducing conditions were in accordance with the MW calculated from the aa sequence. Analyses of the glycosylation patterns using PAS-staining showed a lack of staining of the recombinant variants (SCR1-4 and SCR1-7) in contrast to FH(SCR1-20) from serum. Using a complement hemolysis assay (CH50-assay) all three variants exhibited a molar complement inhibitory activity of FH(1-20)/FH(1-7)/FH(1-4) of about 3/1/1. These data support the postulated model of FH bearing three binding sites for its ligand C3b, from which one is located in the SCR1-4, whereas the other two are located in the SCR8-20.

[Heparin-induced type II thrombocytopenia within the scope of high dose chemotherapy with subsequent stem cell rescue]. / Heparin-induzierte Thrombozytopenie Typ II im Rahmen einer Hochdosis-Chemotherapie mit anschliessender Stammzellrescue.

We report on a 17-year-old young man with rhabdomyosarcoma in the right parotid area. Relapse therapy was performed with high dose chemotherapy and consecutive autologous stem cell rescue. During this therapy heparin-induced thrombocytopenia was diagnosed. Because of its proven antithrombotic activity Danaparoid-Sodium, a natural low molecular glycosaminoglycan preparation, was used for further antithrombotic prophylaxis. We discharged our patient from the laminar air flow unit six months ago. The alternative antithrombotic therapy was tolerated without any problems. No bleeding events occurred, thrombotic complications and veno-occlusive disease of the liver were avoided.

Microfossils from the Neoarchean Campbell Group, Griqualand West Sequence of the Transvaal Supergroup, and their paleoenvironmental and evolutionary implications.

The oldest filament- and colonial coccoid-containing microbial fossil assemblage now known is described here from drill core samples of stromatolitic cherty limestones of the Neoarchean, approximately 2600-Ma-old Campbell Group (Ghaap Plateau Dolomite, Lime Acres Member) obtained at Lime Acres, northern Cape Province, South Africa. The assemblage is biologically diverse, including entophysalidacean (Eoentophysalis sp.), probable chroococcacean (unnamed colonial coccoids), and oscillatoriacean cyanobacteria (Eomycetopsis cf. filiformis, and Siphonophycus transvaalensis), as well as filamentous fossil bacteria (Archaeotrichion sp.); filamentous possible microfossils (unnamed hematitic filaments) also occur. The Campbell Group microorganisms contributed to the formation of stratiform and domical to columnar stromatolitic reefs in shallow subtidal to intertidal environments of the Transvaal intracratonic sea. Although only moderately to poorly preserved, they provide new evidence regarding the paleoenvironmental setting of the Campbell Group sediments, extend the known time-range of entophysalidacean cyanobacteria by more than 400 million years, substantiate the antiquity and role in stromatolite formation of Archean oscillatoriacean cyanobacteria, and document the exceedingly slow (hypobradytelic) evolutionary rate characteristic of this early evolving prokaryotic lineage.