Molecular and Functional Characterization of MobK Protein—A Novel-Type Relaxase Involved in Mobilization for Conjugational Transfer of Klebsiella pneumoniae Plasmid pIGRK

Conjugation, besides transformation and transduction, is one of the main mechanisms of horizontal transmission of genetic information among bacteria. Conjugational transfer, due to its essential role in shaping bacterial genomes and spreading of antibiotics resistance genes, has been widely studied for more than 70 years. However, new and intriguing facts concerning the molecular basis of this process are still being revealed. Most recently, a novel family of conjugative relaxases (Mob proteins) was distinguished. The characteristic feature of these proteins is that they are not related to any of Mobs described so far. Instead of this, they share significant similarity to tyrosine recombinases. In this study MobK—a tyrosine recombinase-like Mob protein, encoded by pIGRK cryptic plasmid from the Klebsiella pneumoniae clinical strain, was characterized. This study revealed that MobK is a site-specific nuclease and its relaxase activity is dependent on both a conserved catalytic tyrosine residue (Y179) that is characteristic of tyrosine recombinases and the presence of Mg2+ divalent cations. The pIGRK minimal origin of transfer sequence (oriT) was also characterized. This is one of the first reports presenting tyrosine recombinase-like conjugative relaxase protein. It also demonstrates that MobK is a convenient model for studying this new protein family

Architektura Miasto Piękno tom 2

"Wydział Architektury Politechniki Krakowskiej zawsze był miejscem pełnym osobowości, indywidualności, talentów. Gdy profesorowie i mistrzowie odchodzili z Wydziału, mieliśmy świadomość, że tu nigdy już nie będzie tak samo. Wydział jednak trwał i trwa jak piękno miasta, które jest przecież dziełem zbiorowym, o nakładających się warstwach, wzmacniających i tworzących jego tożsamość. Tak też kolejne pokolenia kontynuują i budują historię Wydziału. Wydział pełen jest wspomnień i anegdot. W fotografiach, księgach, obrazach, meblach kryją się opowieści."(...

Potential impact of HercepTest™ mAb PharmDx (Dako Omnis) (ge001) in breast cancer diagnosis

The HER2 gene is a biomarker for breast cancer prognosis and treatment. Overexpression of HER2 protein determined by immunohistochemistry (IHC) or amplification of the HER2 gene determined by fluorescence in situ hybridization (FISH) is a condition for qualifying patients for anti-HER2 therapy. Due to the high toxicity of anti-HER2 treatment, proper patient selection is essential. In our study we compared 40 cases with IHC staining of HER2 antibody determined by Ventana PATHWAY anti-HER2/neu antibody (4B5) as HER2 2+ with the new antibody (HercepTest™ mAb PharmDx [Dako Omnis] [GE001]). Then using a double-blind study we compared the (IHC) evaluation with FISH results. In 65% of cases (26/40) the IHC 2+ score remained unchanged, in 32.5% of cases (13/40) expression of HER2 protein after IHC with new antibody was indicated as 3+ score, and in one case we observed a decrease of HER2 protein expression to 1+. In all cases but one, in which we found IHC HER 3+ with new antibody, there was FISH amplification. We have reason to believe that the new antibody will reduce the diagnostic time and avoid unnecessary costs. Due to the small study group, further investigation is needed

Functional and morphological changes in the visual pathway in patients with Graves' orbitopathy

Background: The aim of the study was to perform a functional and structural evaluation of the anterior visual pathway in patients with Graves’ Orbitopathy (GO) using electrophysiological tests and OCT, as well as to identify potential parameters that could be useful in detecting early optic nerve damage. Methods: 47 GO patients were enrolled in the study and divided into three groups, depending on their disease severity: Group 1 with mild GO, Group 2 with moderate-to-severe GO, and Group 3 with dysthyroid optic neuropathy (DON). Pattern visual evoked potential (PVEP), flash visual evoked potential (fVEP), pattern electroretinogram (pERG), and optical coherence tomography (OCT) findings were compared between the groups. Results: In the DON Group (Group 3), N75, P100, and P2 latencies were significantly extended, whereas P100, P50, and N95 amplitudes were significantly reduced as compared to the non-DON group (Groups 1 and 2). Group 3 also had significantly thinner peripapillary retinal nerve fiber layer (RNFL) and macular ganglion cell complex (GCC). In Group 2, as compared to Group 1, P100 amplitudes were significantly reduced for all check sizes, while P100 latency was elongated for the check size of 0.9°. Group 2 also had a significantly thinner average GCC and GCC in the superior quadrant. Conclusions: Electrophysiological examinations may be of use in diagnosis of DON. OCT findings and electrophysiological responses vary in patients with different GO severity. Including regular electrophysiological evaluation and OCT in the examination of patients with GO could be of benefit. However, more research is needed to establish the true significance of pVEP, fVEP, pERG, and OCT in monitoring patients with GO

Factors affecting the course of Graves' Orbitopathy and poor response to glucocorticoid treatment followed by orbital radiotherapy

Graves’ orbitopathy is a rare autoimmune disorder characterized by the inflammation of orbital tissues. The course of disease can be described in terms of its activity and severity. Aim: The aim of our study was to determine the factors affecting the activity and severity of Graves’ orbitopathy, as well as to identify the predictive factors of poor response to glucocorticoid treatment followed by orbital irradiation. Methods: We performed a prospective observational study of 214 patients with Graves’ orbitopathy who were divided into two groups depending on the treatment they had previously obtained for their Graves’ disease. They received i.v. methylprednisolone pulses followed by orbital radiotherapy. They were examined and had their TSH, TRAb and FT4 levels evaluated prior to treatment and after 1, 6 and 12 months. Results: A pre-treatment TRAb concentration higher by one unit (U/L) implied a mean increase in the relative risk of active orbitopathy by 4.7% (p = 0.0362). A TRAb concentration higher by one U/L 1 month after treatment implied a mean increase in the relative risk of moderate-to-severe and severe GO by 8.7% (p = 0.0167) 6 months after treatment. As regards poor response to treatment, patients with moderate-to-severe and severe Graves’ orbitopathy on admission carried a higher risk of being non-responders. Each point scored on the NOSPECS scale prior to treatment increased the relative risk of the patient being a non-responder by 30%. Conclusions: Patients with higher TRAb levels have a higher risk of active Graves’ orbitopathy and moderate-to-severe and severe Graves’ orbitopathy. Monitoring TRAb serum concentration in those patients is of great importance. Patients with more severe Graves’ orbitopathy carry a higher risk of being poor responders to immunosuppressive treatment. Therefore, careful monitoring of patients with Graves’ orbitopathy and their early referral to specialized centers is essential

Wcześniejsze leczenie choroby Gravesa lekami tyreostatycznymi lub radiojodem nie wpływa na skuteczność leczenia immunosupresyjnego orbitopatii tarczycowej

  Introduction: We studied the efficacy of immunosuppressive treatment of GO in a group of patients who had been treated with antithyroid drugs (the ATD group) and in another group that had undergone radioiodine therapy (the 131-I group). Material and methods: A total of 214 patients with exacerbation of GO were studied; the ATD group consisting of 168 patients, and the 131-I group consisting of 46 patients. All patients were treated with methylprednisolone IV pulses (total dose 8.0 g) followed by orbital irradiation (20 Gy in 10 fractions). CAS and IO indices, TSH, fT4, and TRAb levels were evaluated prior to, and 1, 6, and 12 months after treatment. Results: One month after treatment the CAS index decreased significantly in both groups, against values before treatment, p < 0.05. In the ATD group the median level of TRAb-0 before treatment was 5.6 IU/L (min = 0.1; max = 114.0), and 12 months later (TRAb-12) it was 1.4 IU/L (min = 0.1; max = 75.3) (p < 0.05). In the 131-I group the median level of TRAb-0 was 14.3 IU/L (min = 0.6; max = 90.0) vs. TRAb-12 of 3.65 IU/L (min = 0.1; max = 41.0) (p < 0.05). In the ATD group the median value of IO-0 before treatment was 5.0 (min = 1.0; max = 12.0) vs. IO-12 of 2.0 (min = 0.0; max = 8.0) (p < 0.05). In the 131-I group the median value of IO-0 was 5.0 (min = 2.0; max = 9.0) vs. IO-12 of 2.0 (min = 0.0; max = 6.0) (p < 0.05). Conclusions: The severity of GO in the ATD and 131-I groups did not differ significantly over the course of observation despite differences noted in their TRAb levels. The efficacy of GO treatment did not differ between these groups. (Endokrynol Pol 2016; 67 (6): 554–561)    Wstęp: Porównano skuteczność leczenia immunosupresyjnego orbitopatii tarczycowej (GO) u pacjentów leczonych wcześniej doustnymi lekami tyreostatycznymi (grupa ATD) oraz u pacjentów po leczeniu radiojodem (grupa 131-I). Materiał i metody: Przebadano 214 pacjentów z zaostrzeniem GO. Grupa ATD składała się ze 168 pacjentów leczonych lekami tyreostatycznymi. Grupa 131-I składała się z 46 pacjentów leczonych radiojodem. Wszyscy pacjenci byli leczeni pulsami dożylnymi methylprednisolonu (łączna dawka 8,0 g), a następnie poddawani radioterapii oczodołów (20 Gy w 10 frakcjach). Indeksy CAS i IO, stężenia TSH, fT4 oraz TRAb oceniano przed leczeniem, a następnie 1, 6 i 12 miesięcy po leczeniu. Wyniki: W obu grupach miesiąc po leczeniu indeks CAS istotnie zmniejszył się w porównaniu z wartościami wyjściowymi, p < 0,05. W grupie ATD mediana stężenia TRAb-0 wynosiła 5,6 U/l (min = 0,1; max = 114,0) przed leczeniem i 1,4 U/l (min = 0,1; max = 75,3) (p < 0,05) 12 miesięcy po leczeniu (TRAb-12). W grupie 131-I mediany stężenia TRAb-0 i TRAb-12 wynosiły odpowiednio 14,3 IU/L (min = 0,6; max = 90,0) i 3,65 IU/l (min = 0,1; max = 41,0) (p < 0,05). Mediana wartości indeksu IO przed leczeniem (IO-0) w grupie ATD wynosiła 5,0 (min = 1,0; max = 12,0), a po leczeniu (IO-12) 2,0 (min = 0,0; max = 8,0) (p < 0,05). W grupie 131-I mediany wartości IO-0 i IO-12 wynosiły odpowiednio 5,0 (min = 2,0; max = 9,0) i 2,0 (min = 0,0; max = 6,0) (p < 0,05). Wnioski: Stopień zaawansowania GO u pacjentów w grupie ATD i 131-I nie różnił się znamiennie statystycznie przez cały okres obserwacji. Mimo że stężenia TRAb w grupie 131-I zawsze przekraczały górny zakres wartości prawidłowych i były wyższe niż w grupie ATD, skuteczność leczenia GO w obu grupach była taka sama. (Endokrynol Pol 2016; 67 (6): 554–561)

Harnessing the potential of the environmental microbiome in forensic science

Konsorcjum naukowe pod przewodnictwem Centralnego Laboratorium Kryminalistycznego Policji podjęło się opracowania metody analizy DNA mikrobiomu gleby, która znajdzie zastosowanie w badaniach kryminalistycznych. Celem projektu o akronimie SMAFT (Soil Microbiome Analysis Forensic Tool, http://smaft.eu/), finansowanego przez Narodowe Centrum Badań i Rozwoju (DOB-BIO10/03/01/2019), jest stworzenie nowego narzędzia umożliwiającego powiązanie śladu w postaci próbki gleby z określoną lokalizacją geograficzną. W pierwszej części artykułu przybliżono pojęcie mikrobiomu oraz przedstawiono możliwości wykorzystania analiz DNA mikrobiomu w kryminalistyce. W jego drugiej części szczegółowo opisano etapy realizowanego projektu, począwszy od zbierania próbek gleby z różnych miejsc Polski w czterech porach roku i izolacji z nich DNA mikrobiomów, poprzez oparte na technologii MPS (ang. Massively Parallel Sequencing) sekwencjonowanie izolatów oraz opracowanie testu genetycznego zawierającego zestaw markerów metagenomicznych pozwalających na skuteczną indywidualizację próbek gleby, aż po stworzenie systemu informatycznego umożliwiającego analizę i interpretację otrzymanych wyników, który obejmuje bazę danych profili DNA mikrobiomów gleb pochodzących z różnych miejsc Polski.A scientific consortium led by the Central Forensic Laboratory of the Police has undertaken to develop a method for DNA analysis of the soil microbiome to be used in forensic investigations. The aim of the project entitled Soil Microbiome Analysis Forensic Tool – SMAFT (http://smaft.eu/), financed by the National Center for Research and Development (DOB-BIO10/03/01/2019), is to develop a new tool that enables the association of a trace in the form of a soil sample with a specific geographical location. The first part of the paper introduces the concept of the microbiome and presents the possibilities of using microbiome DNA analysis in forensic science. In the second part, the stages of the SMAFT project are described in detail, beginning from the collection of soil samples from different sites in Poland across all seasons and isolation of microbiome DNA through massively parallel sequencing (MPS) technology-based analysis of isolates and the development of a genetic test containing a set of metagenomic markers allowing for effective individualization of soil samples, up to the creation of an IT system enabling analysis and interpretation of the obtained results, which includes a database of soil microbiome DNA profiles from various locations in Poland