A manually curated compendium of expression profiles for the microbial cell factory Corynebacterium glutamicum.

Corynebacterium glutamicum is the major host for the industrial production of amino acids and has become one of the best studied model organisms in microbial biotechnology. Rational strain construction has led to an improvement of producer strains and to a variety of novel producer strains with a broad substrate and product spectrum. A key factor for the success of these approaches is detailed knowledge of transcriptional regulation in C. glutamicum. Here, we present a large compendium of 927 manually curated microarray-based transcriptional profiles for wild-type and engineered strains detecting genome-wide expression changes of the 3,047 annotated genes in response to various environmental conditions or in response to genetic modifications. The replicates within the 927 experiments were combined to 304 microarray sets ordered into six categories that were used for differential gene expression analysis. Hierarchical clustering confirmed that no outliers were present in the sets. The compendium provides a valuable resource for future fundamental and applied research with C. glutamicum and contributes to a systemic understanding of this microbial cell factory. Measurement(s) Gene Expression Analysis Technology Type(s) Two Color Microarray Factor Type(s) WT condition A vs. WT condition B • Plasmid-based gene overexpression in parental strain vs. parental strain with empty vector control • Deletion mutant vs. parental strain Sample Characteristic - Organism Corynebacterium glutamicum Sample Characteristic - Environment laboratory environment Sample Characteristic - Location Germany.

Melanocyte antigen triggers autoimmunity in human psoriasis.

Psoriasis vulgaris is a common T cell-mediated inflammatory skin disease with a suspected autoimmune pathogenesis. The human leukocyte antigen (HLA) class I allele, HLA-C*06:02, is the main psoriasis risk gene. Epidermal CD8(+) T cells are essential for psoriasis development. Functional implications of HLA-C*06:02 and mechanisms of lesional T cell activation in psoriasis, however, remained elusive. Here we identify melanocytes as skin-specific target cells of an HLA-C*06:02-restricted psoriatic T cell response. We found that a Vα3S1/Vß13S1 T cell receptor (TCR), which we had reconstituted from an epidermal CD8(+) T cell clone of an HLA-C*06:02-positive psoriasis patient specifically recognizes HLA-C*06:02-positive melanocytes. Through peptide library screening, we identified ADAMTS-like protein 5 (ADAMTSL5) as an HLA-C*06:02-presented melanocytic autoantigen of the Vα3S1/Vß13S1 TCR. Consistent with the Vα3S1/Vß13S1-TCR reactivity, we observed numerous CD8(+) T cells in psoriasis lesions attacking melanocytes, the only epidermal cells expressing ADAMTSL5. Furthermore, ADAMTSL5 stimulation induced the psoriasis signature cytokine, IL-17A, in CD8(+) T cells from psoriasis patients only, supporting a role as psoriatic autoantigen. This unbiased analysis of a TCR obtained directly from tissue-infiltrating CD8(+) T cells reveals that in psoriasis HLA-C*06:02 directs an autoimmune response against melanocytes through autoantigen presentation. We propose that HLA-C*06:02 may predispose to psoriasis via this newly identified autoimmune pathway.

Analysis of the paired TCR α- and ß-chains of single human T cells.

Analysis of the paired i.e. matching TCR α- and ß-chain rearrangements of single human T cells is required for a precise investigation of clonal diversity, tissue distribution and specificity of protective and pathologic T-cell mediated immune responses. Here we describe a multiplex RT-PCR based technology, which for the first time allows for an unbiased analysis of the complete sequences of both α- and ß-chains of TCR from single T cells. We validated our technology by the analysis of the pathologic T-cell infiltrates from tissue lesions of two T-cell mediated autoimmune diseases, psoriasis vulgaris (PV) and multiple sclerosis (MS). In both disorders we could detect various T cell clones as defined by multiple T cells with identical α- and ß-chain rearrangements distributed across the tissue lesions. In PV, single cell TCR analysis of lesional T cells identified clonal CD8(+) T cell expansions that predominated in the epidermis of psoriatic plaques. An MS brain lesion contained two dominant CD8(+) T-cell clones that extended over the white and grey matter and meninges. In both diseases several clonally expanded T cells carried dual TCRs composed of one Vß and two different Vα-chain rearrangements. These results show that our technology is an efficient instrument to analyse αß-T cell responses with single cell resolution in man. It should facilitate essential new insights into the mechanisms of protective and pathologic immunity in many human T-cell mediated conditions and allow for resurrecting functional TCRs from any αß-T cell of choice that can be used for investigating their specificity.

The TetR-type transcriptional regulator FasR of Corynebacterium glutamicum controls genes of lipid synthesis during growth on acetate.

The addition of fatty acids to either Escherichia coli or Bacillus subtilis elicits an elaborate cellular response of the lipid metabolism. We found that in Corynebacterium glutamicum the expression of accD1 encoding the ß-subunit of the essential acetyl-CoA carboxylase is repressed in acetate-grown cells without the addition of fatty acids. The TetR-type transcriptional regulator NCgl2404, termed FasR, was identified and deleted. During growth on acetate, but not on glucose, 17 genes are differentially expressed in the deletion mutant, among them accD1, and fasA and fasB both encoding fatty acid synthases, which were upregulated. Determination of the 5' ends of accD1, fasA, fasB and accBC together with the use of isolated FasR protein identified the FasR binding site, fasO, which is located within the accD1 and fasA transcript initiation site thus blocking transcription by RNA polymerase binding directly. The identified fasO motif is present in C. efficiens or C. diphtheriae, too, and it is actually similarly positioned in these bacteria within the 5' ends of the accD1 and fasA transcripts, and a fasR orthologue is also present. The identification of the FasR-fasO system in Corynebacteriaceae might indicate a conserved transcriptional control of the unique lipid synthesis in these mycolic acid-containing bacteria.

The 3' ends of mature transcripts are generated by a processosome complex in fission yeast mitochondria.

In this article, we report on the genetic analysis of the Schizosaccharomyces pombe open reading frames SPCC1322.01 and SPAC637.11, respectively, which encode proteins that are similar to the exoribonuclease Dss1p and the RNA helicase Suv3p, respectively, forming the mitochondrial degradosome of Saccharomyces cerevisiae. While the helicase Suv3p is exchangeable between S. cerevisiae and S. pombe, the functions of Dss1p and the putative fission yeast RNase protein are specific for each species. Unlike S. cerevisiae mutants lacking a functional degradosome, the major defect of fission yeast knock-out strains is their inability to perform downstream processing of transcripts. In addition, the lack of pah1 results in instability of mitochondrial RNA ends. Overexpression of par1 and pah1 has no significant effect on the steady-state levels of mitochondrial RNAs. The Pet127p-stimulated RNA degradation activity is independent of Par1p/Pah1p in fission yeast mitochondria. The results presented herein indicate that both fission yeast proteins play only a minor role (if at all) in mitochondrial RNA degradation. We assume that the RNA-degrading function was taken over by other enzymes in fission yeast mitochondria, while the former degradosome proteins were recruited to new cellular pathways, for example, RNA processing in fission yeast (as discussed in this article) or mitochondrial DNA replication, apoptosis, or chromatin maintenance in eukaryotes, during evolution.

[Differential diagnosis of tailgut cyst in the case of a rectal carcinoma with presacral mass]. / Differentialdiagnose Tailgut-Zyste bei einem Rektumkarzinom mit präsakraler Raumforderung.

Cystic masses of various different origins and degrees of malignancy occur in the presacral space. Apart from benign, malignant and acute inflammatory masses, the benign lesions include a group of cystic structures to which the dysgenetic cysts belong. The tailgut cyst is a relatively rare type of gastrointestinal origin from this group. We present the case of a stenosing rectal carcinoma in which a presacral mass was additionally found, which did not get smaller compared to the rectal tumour during combined preoperative radiochemotherapy, with a lymphoma being suspected. After extirpation of the tumour, histopathological processing revealed a stenosing rectal carcinoma with an adjacent dysgenetic lesion--a tailgut cyst. Although dysgenetic lesions are rare as presacral masses, this must also be taken into account as a differential diagnosis in the various imaging procedures such as endosonography, CT and MRT when a rectal carcinoma is present.

[Selective embolisation of posttraumatic intractable epistaxis]. / Selektive Embolisation bei posttraumatisch unstillbarer Epistaxis.

Idiopathic or trauma-related nose-bleeds can in rare cases be an intractable clinical picture in which normal conservative measures fail. Further therapeutic options that can be applied are surgical vascular ligature or selective endovascular occlusion. We report on a patient who developed hemodynamically relevant epistaxis due to splanchnocranial injuries after falling from his bicycle. Spiral-CT showed a fracture of the posterior nasal septum and of the inferior nasal concha. Despite anterior and posterior nasal tamponade, anemia occurred requiring transfusion and resulting in the interdisciplinary decision to perform transcatheter embolization. Via a trans femoral incision, a 5 F Vitek catheter was introduced into the maxillary artery, and a tracker system was then advanced through the catheter supraselectively and placed in the peripheral branches of the vascular system of the sphenopalatine artery. Selective DSA clearly showed the source of bleeding to be the area supplying the sphenopalatine artery. Following documentation of the localization of bleeding, a total of six (3/30 mm) complex heliacal mini-coils were inserted. After intravascular occlusion of the sphenopalatine artery, bleeding completely stopped within a few minutes. This shows that supraselective embolization is an easily conducted, effective and low-complication method for the definitive treatment of severe posttraumatic nosebleeds, even in emergency situations.

[Eagle syndrome: diagnostic imaging and therapy]. / Eagle Syndrom-- Bildgebende Diagnostik und Therapie.

In the case of clinical symptoms such as dysphagia, foreign-body sensation and chronic neck or facial pain close to the ear, an Eagle syndrome should be considered in the differential diagnosis. Rational diagnostics and therapy are elucidated on the basis of four case reports. Four patients presented in the outpatients clinic with chronic complaints on chewing and a foreign-body sensation in the tonsil region. Upon specific palpation below the mandibular angle, pain radiating into the ear region intensified. In all patients, local anaesthesia with lidocaine only led to a temporary remission of symptoms. Imaging diagnostics then performed initially included cranial survey radiograms according to Clementschitsch as well as in the lateral ray path and an OPTG. An axial spiral-CT was then performed using the thin-layer technique with subsequent 3-D reconstruction. Therapy consisted of elective resection with a lateral external incision from the retromandibular. From a symptomatic point of view, the cranial survey radiograms and the OPTG revealed hypertrophic styloid processes. The geometrically corrected addition of the axial CT images produced an absolute length of 51-58 mm. The 3-D reconstruction made it possible to visualise the exact spatial orientation of the styloid processes. An ossification of the stylohyoid ligament could definitely be ruled out on the basis of the imaging procedures. After resection of the megastyloid, the patients were completely free of symptoms. Spiral-CT with subsequent 3-D reconstruction is the method of choice for exact determination of the localisation and size of a megastyloid, while cranial survey radiograms according to Clementschitsch and in the lateral ray path or an OPTG can provide initial information. The therapy of choice is considered to be resection of the megastyloid, whereby an external lateral incision has proved effective.

[Diagnostic imaging and therapy of chondroblastoma]. / Bildgebung und Therapie eines Chondroblastoms.

The chondroblastoma is a rare lytic osseous lesion, which is typically to be found in the epiphyses of the long tubular bones. We present a case report with the differential diagnostic imaging and the surgical therapy of a chondroblastoma of the proximal tibial epiphysis. A 16-year-old male presented in the traumatological accident and emergency centre with knee pain of unclear origin that had been increasing for several months. He had no memory of any acute trauma. For further clarification, a conventional radiograph in two planes, a thin-layer CT, a multiplanar MRI before and after Gd-DTPA, and a bone scan were performed. The conventional radiological diagnostics show a smoothly circumscribed, osteolytic, eccentric lesion with marginal sclerosis, which not exceeds the epiphyseal seam of the proximal tibia. The thin-layer CT reveals an eccentric osteolysis, with a typical, narrow sclerotic seam and central calcifications. In the high-resolution MRI, the T2-weighted sequences show a locally limited, epiphyseal, lobulated lesion with a heterogeneous, in part raised signal. A further signal increase can be seen in the T1-weighted sequences after administration of Gd-DTPA. Perifocally, there is an epiphyseal edema and discrete (intraarticular fluid collection) fluid in the inner knee. In the skeletal scintigraphy, there is a strong focal and diffuse increased activity from the proximal, lateral tibial epiphysis. After the diagnosis had been rendered, curettage of the defect was performed and then, after histological confirmation of the diagnosis, completing spongiosaplasty was conducted. Taking into account the localisation and the patient's age, a virtually certain diagnosis can already be rendered using conventional imaging. A supplementary MRI or CT can be helpful for final clarification, while 3-phase skeletal scintigraphy is superfluous. The therapy of choice is curettage and subsequent filling of the defect with autologous spongiosa.