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Introduction: Alpha-1-antitrypsin deficiency (AATD) is a rare hereditary condition, which is characterized by low alpha-1 antitrypsin (AAT) serum levels. While the association between AATD and COPD/emphysema is indisputed, the association to bronchiectasis and asthma is a matter of debate. Aims and Objectives: We compared the distribution of AAT genotypes between patients with COPD/emphysema, bronchiectasis and asthma. Methods: Between 2003 and 2019, 27,223 testing kits (AlphaKit©) were analyzed in the German AAT laboratory, University of Marburg. The diagnosis of AATD was made through a measurement of AAT serum levels, followed by genotyping, phenotyping or whole gene sequencing depending on availability and/or the need for more detailed interpretation of the AAT results. Respiratory symptoms were additionally recorded on the kit. Results: The distribution of the wild type (Pi*MM) was roughly comparable between the three groups. A higher percentage of COPD/emphysema and bronchiectasis patients exhibited the Pi*ZZ genotype as compared to asthma patients. In patients with bronchiectasis only, the distribution of alleles was as follows: M/M (59.4%), M/S (4%), M/Z (19.5%), S/S (0.8%), S/Z (2.8%), Z/Z (7.2%). Figure 1: Distribution of AAT alleles among patients with COPD/emphysema, bronchiectasis and asthma Conclusions: Not only patients with COPD/emphysema, but also patients with bronchiectasis (and asthma) should be screened for AATD.

Background Coronary Aneurysms are a focal dilatation of an artery segment >1.5-fold the normal size of adjacent segments. Although some series have suggested a prevalence of 0.3–12%, data are lacking. In addition, they are not mentioned in practice guidelines. Our aim was investigate its prevalence, management and long-term outcomes. Methods and results The coronary artery aneurysm registry (CAAR) involved 32 hospitals across 9 countries in America and Europe. We reviewed 436,467 consecutive angiograms performed over the period 2004–2016. Finally, 1565 patients were recruited. Aneurysm global prevalence was 0.35%. Most patients were male (78.5%) with a mean age of 65 years and frequent cardiovascular risk factors. The main indication for angiogram was an acute coronary syndrome, 966 cases. The number of aneurisms was ≤2 per patient in 95.8% of the cases, mostly saccular, most frequently found in the left anterior descending and with numbers proportional with coronary stenosis. Aortopathies were related with more aneurysms too. Most patients received any revascularization procedure (69%), commonly percutaneous (53%). After a median follow-up of 37.2 months, 485 suffered a combined event (MACE) and 240 died. Without major differences comparing CABG vs PCI, MACE and death were more frequent in patients who received bare metal stents. Conclusions Coronary artery aneurysms are not uncommon. Usually, they are associated with coronary stenosis and high cardiovascular risk. Antiplatelet therapy seems reasonable and a percutaneous approach is safe and effective.

Ein 67-jahriger Patient stellte sich mit seit funf Jahren bestehendem Auswurf, Belastungsdyspnoe und haufigem Rauspern auf einer pneumologischen Normalstation vor. Im Vorfeld angefertigte Rontgenbilder und eine Computertomografie (CT) des Thorax mit Verdacht auf eine chronische Bronchitis zeigten multiple pulmonale Zysten. Was steckt dahinter?

The cell cycle regulator p27 Kip1 (p27) is controlled by 26S proteasome-mediated proteolysis by two different pathways. From the S till the G2 phase of the cell cycle, degradation of p27 takes place in the nucleus and is initiated by CDK2-dependent phosphorylation of threonine 187 with subsequent ubiquitination by the SCF Skp2 ubiquitin ligase. During the Gl cell cycle phase (G1), p27 breakdown is cytosolic and is initiated by nuclear export with subsequent ubiquitination by a RING finger ligase called kipl ubiquitination complex. Here we show that the COP9 signalosome (CSN) is a regulator of p27 proteolysis during Gl. The CSN interacts with p27 and the CSN-associated kinase CK2 phosphorylates p27 at two regions. One is central to the protein (amino acids 101-113), and the other was mapped near to the C-terminus (amino acids 170-189). Elimination of the putative C-terminal phosphorylation sites stabilizes ectopic p27 towards proteasomal degradation and abolishes CSN-p27 binding. Inhibition of CSN-associated kinase activity by curcumin attenuates loss of p27 upon cell cycle re-entry. Similar but not additive effects of the phosphoinositol-3-kinase blocker LY 290042 may point to a common pathway of CSN-associated CK2 and protein kinase B/Akt (Akt) in regulating p27 abundance. Akt is found in Flag pulldowns of lysates obtained from cells permanently expressing Flag-tagged CSN2, indicating that Akt is a novel kinase associated with the CSN. Thus, the CSN seems to regulate p27 proteolysis at Gl downstream of Ras-mediated signal pathways.

Mutant-enriched PCR and reverse dot blot hybridization in microplates were applied for examining K-ras status in stools and tissue samples from patients with pancreatic tumors and chronic pancreatitis. In tissue samples, K-ras mutations were found in 32 of 35 cases of ductal adenocarcinoma, in 5 of 7 periampullary cancers, in 1 cystadenocarcinoma, and in 3 of 5 patients with chronic pancreatitis. In stools, mutated K-ras was seen in 10 of 25 cases of ductal adenocarcinoma, in 1 case of cystadenocarcinoma, and in 2 of 6 cases of chronic pancreatitis. These data indicate that the K-ras status of stool samples may help identify pancreatic carcinoma and persons at risk for cancer development; however, it does not allow discrimination of malignant from nonmalignant diseases.

The diagnostic specificity of the detection of disseminated prostatic cells by reverse-transcriptase polymerase chain reaction (RT-PCR) of PSA mRNA was investigated. A sensitive nested PCR was developed. In blood samples from 10 healthy female and 10 healthy male persons examined by RT-PCR, mRNA of PSA was detected 3 times in each group. In the groups of patients suffering from benign prostate hyperplasia and prostate cancer, 6 of 11 and 5 of 12, respectively, gave positive RT-PCR results. With increasing analytical sensitivity of the RT-PCR of PSA mRNA, the diagnostic specificity of the assay is decreased. Further development of this diagnostic method requires the introduction of the quantitative PCR which may make possible discrimination between prostatic and non-prostatic source of PSA mRNA by quantification.

The isopeptide bonds formed by ubiquitin or its relatives are cleaved by hydrolases with active site cysteines. Recent studies have revealed that similar metalloprotease motifs – JAMMs – in the Rpn11 subunit of the 26S proteasome lid and in the Csn5 subunit of the COP9 signalosome are involved in deubiquitination and deneddylation, respectively.

We report the case of a 44-year-old man who underwent a partial medial meniscectomy for a meniscal tear whose postoperative course was complicated by the development of heterotopic ossification (HO) within the medial arthroscopic portal. Following a routine initial procedure, the patient presented with ongoing pain and a palpable, painful lump around the previous medial arthroscopy portal. Plain radiographs and MRI were suggestive of a bony structure within the soft tissues. Histopathological examination at repeat arthropscopy confirmed osseous tissue consistent with HO. Recovery after the second procedure was rapid and resulted in normal knee function and complete pain relief. HO within an arthroscopy portal is a rare complication following arthroscopic partial meniscectomy in the knee and has not previously been described in the literature.

Background The COP9 signalosome (CSN) is a conserved protein complex in eukaryotic cells consisting of eight subunits (CSN1 to CSN8). Recent data demonstrate that the CSN is a regulator of the ubiquitin (Ub) proteasome system (UPS). It controls substrate ubiquitination by cullin-RING Ub ligases (CRLs), a process that determines substrate specificity of the UPS. The intrinsic deneddylating activity localized to CSN5 as well as the associated kinases and deubiquitinating activity are involved in the regulatory function of CSN. The exact mechanisms are unclear. In this study we knocked down CSN1 (siCSN1), CSN3 (siCSN3) and CSN5 (siCSN5) by specific siRNA oligos permanently expressed in HeLa cells. The analysis and comparison of siRNA cells revealed differential impact of individual subunits on CSN structure and function. Results Permanent knockdowns of CSN1 and CSN3 led to a reduction of the subunits to approximately 40%, which is accompanied by a proportional decrease of the CSN holocomplex. In contrast, downregulation of CSN5 in HeLa cells reduced the CSN5 protein below 20% without significant effects on the remaining complex. The CRL component Rbx1 was characterized by accelerated proteolysis in siCSN1 and siCSN3 and also in siCSN5 cells, however, with lesser extent. Immunoprecipitated CSN complex from siCSN5 cells was less effective in phosphorylating c-Jun and p27. Accelerated degradation of c-Jun in siCSN5 cells was rescued by overexpression of CSN5 as well as of the deneddylation mutant CSN5D151N. Overexpression of CSN5 cannot rescue c-Jun destabilization in siCSN1. Conclusion There exists a coordinated downregulation of CSN subunits in the CSN1 and CSN3 knockdowns. The underlying regulatory mechanisms are obscure. CSN5 seems to possess a specific status in HeLa cells. Its reduction is not connected with coordinated downregulation of other subunits. CSN knockdowns confirm that the stabilization of the CRL component Rbx1 is a major CSN function. In addition, downregulation of CSN subunits influences the stability of important cellular regulators such as c-Jun and p27.

HIV RNA was quantified in blood plasma from 209 patients and in control specimen comparing the NucliSens HIV-1 QT test (Organon Teknika), which is based on the nucleic acid sequence amplification procedure, and the Quantiplex 3.0 test (Bayer), which uses hybridization signal enhancement by branched DNA (bDNA) probes. A highly significant correlation (P=0.01) was found between the two methods with 88% of the samples showing similar results. In cases of discrepant findings, higher virus load was observed with either test (14xNASBA>bDNA; 12xbNDA>NASBA). Differences could neither be related to clinical features nor to divergent virus subtypes. Standard preparations containing 35000 and 222000 copies were quantified with intra-assay coefficients of variation of

Long-term changes of learning behavior and of the striatal dopaminergic system were observed in a rat model of early postnatal hypoxia. Striatal dopamine (DA) concentration, K+-stimulated DA release from slices, and DA uptake into crude synaptosomal preparations (S1 fractions) were used as markers of the striatal DAergic system. Active avoidance learning was tested as behavioral criterion. Cyclodextrin and flunarizine were found to produce long-term effects on the DAergic system in control animals. While cyclodextrin normalized hypoxia-induced effects in DA release, flunarizine prevented those in DA uptake and improved aviodance learning.

The dopamine (DA) uptake over a concentration range from 0.03 to 100 microM was studied in S1 fractions of the rat striatum prepared from control rats and those exposed for 14 h to hypobaric hypoxia. The uptake exhibited non-Michaelis-Menten kinetics, which were evaluated by applying an equation assuming two transport sites. The high-affinity uptake site was characterized by an apparent Michaelis-Menten constant of 0.47 microM and an apparent maximal transport rate of 113 pmol/mg protein/30 s. The respective constants of the low-affinity uptake site were 52.8 microM and 1490 pmol/mg protein/30 s. One hour after hypoxia kinetic constants of the high-affinity uptake were unchanged but the maximal transport rate of the low-affinity uptake was increased by 50%. The elevated low-affinity uptake capacity may represent a means of adaptation to hypoxia allowing a faster removal of high extracellular concentrations of DA.

Hypobaric hypoxia (10 h daily, pO2 10 kPa) and saline administration (2.5 microliters/g body wt) from the 2nd till the 11th day of life both induced a long-lasting increase of the low-affinity dopamine (DA) uptake capacity in S1-fractions of the rat striatum. Additionally, the potassium-stimulated DA release was enhanced in adult control rats postnatally injected with saline. The administration of a mixture of bovine brain gangliosides (30 micrograms/g body wt) was found to prevent these effects. However, the kinetic constants of the DA uptake of hypoxic rats treated with gangliosides were reduced in comparison to untreated controls. Thus, the effects of gangliosides appear to differ between hypoxic and control conditions. The modification of the dopaminergic activity during brain development is discussed as a possible mechanism of the preventive effects of gangliosides against long-term cerebral dysfunctions following hypoxia or stress.