Diversity of partial geocomplexes types (morphotops) in the southern part of the Kościeliska Valley in the Western Tatra Mts

Poznanie i zrozumienie funkcjonowania zróżnicowanego systemu środowiska przyrodniczego obszarów wysokogórskich wymaga jego uporządkowania. Właściwa jest zatem analiza oparta o jedną cechę lub element środowiska. Badania na potrzeby niniejszego opracowania przeprowadzone zostały przez autorkę w południowej części Doliny Kościeliskiej w Tatrach Zachodnich. Rzeźba terenu jest jednym z najważniejszych elementów kształtujących środowisko przyrodnicze Tatr, dlatego też stanowi doskonałe „tło” dla zilustrowania zmienności pozostałych jego cech. Podstawą poniższego opracowana było wyznaczenie morfotopów (jednostek charakteryzujących się jednorodnością cech morfologicznych oraz zachodzących współcześnie procesów morfogenetycznych), na bazie których możliwe było przedstawienie zróżnicowania pozostałych cech środowiska – budowy geologicznej, klimatu, gleb i szaty roślinnej. Na podstawie przeprowadzonych prac terenowych i analiz, wyznaczone zostały 3 główne grupy morfotopów oraz opisane poszczególne ich rodzaje.The paper presents a way to classify the organization of the natural environment in high-mountain areas – a method of partial geocomplexes delimitation. The main concern was put on the terrain relief and morphogenetic processes being the most important factors influencing the landscape structure. In the southern part of the Kościeliska Valley in the Tatra Mts. 10 types of morphotops (partial geocomplexes with homogeneous terrain relief and recent morphogenetic processes) were identified 3 groups distinquished: morphotops of valley floors, those of slopes and those of ridge tops. For each type of morphotop basic characteristics of lithology, climate, soils, plant cover and recent morphogenetic processes were presented in the text

Zróżnicowanie typów geokompleksów częściowych (morfotopów) w południowej części Doliny Kościeliskiej w Tatrach Zachodnich

Poznanie i zrozumienie funkcjonowania zróżnicowanego systemu środowiska przyrodniczego obszarów wysokogórskich wymaga jego uporządkowania. Właściwa jest zatem analiza oparta o jedną cechę lub element środowiska. Badania na potrzeby niniejszego opracowania przeprowadzone zostały przez autorkę w południowej części Doliny Kościeliskiej w Tatrach Zachodnich. Rzeźba terenu jest jednym z najważniejszych elementów kształtujących środowisko przyrodnicze Tatr, dlatego też stanowi doskonałe „tło” dla zilustrowania zmienności pozostałych jego cech. Podstawą poniższego opracowana było wyznaczenie morfotopów (jednostek charakteryzujących się jednorodnością cech morfologicznych oraz zachodzących współcześnie procesów morfogenetycznych), na bazie których możliwe było przedstawienie zróżnicowania pozostałych cech środowiska – budowy geologicznej, klimatu, gleb i szaty roślinnej. Na podstawie przeprowadzonych prac terenowych i analiz, wyznaczone zostały 3 główne grupy morfotopów oraz opisane poszczególne ich rodzaje

Sposób prowadzenia badań archeologiczno-architektonicznych, a możliwości wykorzystania ich wyników w procesie projektowym na przykładzie byłego Szpitala im. Babińskiego we Wrocławiu.

The paper discusses the issue of the mode and method of conducting archaeological and architectural research with particular emphasis on legal possibilities and analysis of potential effects (both: positive or negative) of the choice of individual forms on the way of carrying out the design works and the opportunity of displaying exposed relics. These issues will be discussed mainly on the example of excavations carried out since 2015 in the area of the former J. Babiński Regional Hospital at Jana Pawła II Square in Wrocław by a research team led by dr ing. arch. Piotr Kmiecik and dr Robert Szwed, with the participation of dr Czesław Lasota. The area of the former hospital was definitely one of the most interesting in terms of archeology and architecture in Wroclaw, but also one of the most complicated. The degree of complexity was influenced not so much by the accumulation of cultural layers and expected discoveries, but by the location and stratification of continuous historical transformations of the area, resulting in dense development, as well as in accumulation of active network buses of transit importance, which resulted in forming communication linkage within a vast area and the existence of urban greenery covered by legal protection. The simultaneous occurrence of many factors resulted in the inability to estimate scope and size of conducted works before their commencement, forced flexibility and the need of arranging excavations taking into account permanent participation of construction team during that time. The level of complexity also forced some modifications in the mode of research, which shows the need of verifying the approach towards restrictive division between different forms of architectural and archeological research and shows numbers of opportunities offered by closer cooperation between architect, archeologist, the office of monument conservation and the construction site. Keywords:Niniejszy artykuł stanowi omówienie zagadnienia trybu i sposobu prowadzenia badań archeologiczno-architektonicznych ze szczególnym uwzględnieniem możliwości prawnych oraz analizę skutków (pozytywnych i negatywnych) wyboru poszczególnych form na sposób, w jaki prowadzone są prace projektowe oraz możliwości ekspozycji odsłoniętych reliktów. Zagadnienia te zostaną omówione głównie na przykładzie prac prowadzonych od 2015 roku na obszarze byłego szpitala im. Babińskiego przy placu Jana Pawła II we Wrocławiu przez zespół badawczy pod kierunkiem dr inż. arch. Piotra Kmiecika oraz dr Roberta Szweda, przy współudziale dr Czesława Lasoty. Teren byłego Szpitala był zdecydowanie jednym z najciekawszych pod względem archeologiczno – architektonicznym we Wrocławiu, jednocześnie jednak również jednym z najbardziej skomplikowanych. Na stopień skomplikowania miało wpływ nie tyle nagromadzenie warstw kulturowych i spodziewanych odkryć, a samo położenie oraz nawarstwienie ciągłych przekształceń obszaru, skutkujące gęstą zabudową, a także nagromadzeniem czynnych magistrali sieciowych o znaczeniu tranzytowym, ukształtowaniem powiązań komunikacyjnych w obrębie rozległego obszaru i ukształtowaniem zieleni objętej ochroną prawną. Jednoczesne wystąpienie tylu czynników spowodowało niemożliwość oszacowania zakresu i wielkości prowadzonych prac przed ich rozpoczęciem, wymusiło elastyczność oraz konieczność zaaranżowania badań z uwzględnieniem udziału ekip budowlanych w ich trakcie. Poziom komplikacji wymuszał także pewne modyfikacje w trybie prowadzonych prac, co ukazuje konieczność weryfikacji podejścia do restrykcyjnego podziału, oraz ukazuje szereg możliwości, jakie daje ściślejsza współpraca na linii architekt-archeolog-urząd konserwatora zabytków-budowa

Combining NK cells and mAb9.2.27 to combat NG2-dependent and anti-inflammatory signals in glioblastoma

Glioblastoma is a deadly brain cancer with limited treatment options. Targeting chondroitin sulfate proteoglycan 4 (CSPG4, best known as NG2) with the monoclonal antibody mAb9.2.27 and activated natural killer (NK) cells abrogated the tumor growth and prolonged the survival of glioblastoma-bearing animals by favoring the establishment of a pro-inflammatory microenvironment. The combination of NK cells and mAb9.2.27 recruited ED1+CCR2low macrophages that stimulated ED1+ED2lowMHCIIhigh microglial cells to exert robust cytotoxicity. Our findings demonstrate the therapeutic potential of targeting salient tumor associated-antigens.publishedVersio

NK cells with KIR2DS2 immunogenotype have a functional activation advantage to efficiently kill glioblastoma and prolong animal survival

Glioblastomas (GBMs) are lethal brain cancers that are resistant to current therapies. We investigated the cytotoxicity of human allogeneic NK cells against patient-derived GBM in vitro and in vivo, as well as mechanisms mediating their efficacy. We demonstrate that KIR2DS2 immunogenotype NK cells were more potent killers, notwithstanding the absence of inhibitory killer Ig–like receptor (KIR)-HLA ligand mismatch. FACS-sorted and enriched KIR2DS2+ NK cell subpopulations retained significantly high levels of CD69 and CD16 when in contact with GBM cells at a 1:1 ratio and highly expressed CD107a and secreted more soluble CD137 and granzyme A. In contrast, KIR2DS2− immunogenotype donor NK cells were less cytotoxic against GBM and K562, and, similar to FACS-sorted or gated KIR2DS2− NK cells, significantly diminished CD16, CD107a, granzyme A, and CD69 when in contact with GBM cells. Furthermore, NK cell–mediated GBM killing in vitro depended upon the expression of ligands for the activating receptor NKG2D and was partially abrogated by Ab blockade. Treatment of GBM xenografts in NOD/SCID mice with NK cells from a KIR2DS2+ donor lacking inhibitory KIR-HLA ligand mismatch significantly prolonged the median survival to 163 d compared with vehicle controls (log-rank test, p = 0.0001), in contrast to 117.5 d (log-rank test, p = 0.0005) for NK cells with several inhibitory KIR-HLA ligand mismatches but lacking KIR2DS2 genotype. Significantly more CD56+CD16+ NK cells from a KIR2DS2+ donor survived in nontumor-bearing brains 3 wk after infusion compared with KIR2DS2− NK cells, independent of their proliferative capacity. In conclusion, KIR2DS2 identifies potent alloreactive NK cells against GBM that are mediated by commensurate, but dominant, activating signals.publishedVersio

Running performance at high running velocities is impaired but V'O_{2max} and peripheral endothelial function are preserved in IL-6^{−/−} mice

It has been reported that IL-6 knockout mice (IL-6^{−/−}) possess lower endurance capacity than wild type mice (WT), however the underlying mechanism is poorly understood. The aim of the present work was to examine whether reduced endurance running capacity in IL-6^{−/−} mice is linked to impaired maximal oxygen uptake (V′O_{2max}), decreased glucose tolerance, endothelial dysfunction or other mechanisms. Maximal running velocity during incremental running to exhaustion was significantly lower in IL-6−/− mice than in WT mice (13.00±0.97 m.min^{-1} vs. 16.89±1.15 m.min^{-1}, P<0.02, respectively). Moreover, the time to exhaustion during running at 12 m.min^{-1} in IL-6^{−/−} mice was significantly shorter (P<0.05) than in WT mice. V′O_{2max} in IL-6^{−/−} (n = 20) amounting to 108.3±2.8 ml.kg^{-1}.min^{-1} was similar as in WT mice (n = 22) amounting to 113.0±1.8 ml.kg^{-1}.min^{-1}, (P = 0.16). No difference in maximal COX activity between the IL-6^{−/−} and WT mice in m. soleus and m. gastrocnemius was found. Moreover, no impairment of peripheral endothelial function or glucose tolerance was found in IL-6^{−/−} mice. Surprisingly, plasma lactate concentration during running at 8 m.min−1 as well at maximal running velocity in IL-6^{−/−} mice was significantly lower (P<0.01) than in WT mice. Interestingly, IL-6^{−/−} mice displayed important adaptive mechanisms including significantly lower oxygen cost of running at a given speed accompanied by lower expression of sarcoplasmic reticulum Ca^{2+}-ATPase and lower plasma lactate concentrations during running at submaximal and maximal running velocities. In conclusion, impaired endurance running capacity in IL-6^{−/−} mice could not be explained by reduced V′O_{2max}, endothelial dysfunction or impaired muscle oxidative capacity. Therefore, our results indicate that IL-6 cannot be regarded as a major regulator of exercise capacity but rather as a modulator of endurance performance. Furthermore, we identified important compensatory mechanism limiting reduced exercise performance in IL-6^{−/−} mice

Targeting the NG2/CSPG4 Proteoglycan Retards Tumour Growth and Angiogenesis in Preclinical Models of GBM and Melanoma

Aberrant expression of the progenitor marker Neuron-glia 2 (NG2/CSPG4) or melanoma proteoglycan on cancer cells and angiogenic vasculature is associated with an aggressive disease course in several malignancies including glioblastoma multiforme (GBM) and melanoma. Thus, we investigated the mechanism of NG2 mediated malignant progression and its potential as a therapeutic target in clinically relevant GBM and melanoma animal models. Xenografting NG2 overexpressing GBM cell lines resulted in increased growth rate, angiogenesis and vascular permeability compared to control, NG2 negative tumours. The effect of abrogating NG2 function was investigated after intracerebral delivery of lentivirally encoded shRNAs targeting NG2 in patient GBM xenografts as well as in established subcutaneous A375 melanoma tumours. NG2 knockdown reduced melanoma proliferation and increased apoptosis and necrosis. Targeting NG2 in two heterogeneous GBM xenografts significantly reduced tumour growth and oedema levels, angiogenesis and normalised vascular function. Vascular normalisation resulted in increased tumour invasion and decreased apoptosis and necrosis. We conclude that NG2 promotes tumour progression by multiple mechanisms and represents an amenable target for cancer molecular therapy

Expression of the progenitor marker NG2/CSPG4 predicts poor survival and resistance to ionising radiation in glioblastoma

Glioblastoma (GBM) is a highly aggressive brain tumour, where patients respond poorly to radiotherapy and exhibit dismal survival outcomes. The mechanisms of radioresistance are not completely understood. However, cancer cells with an immature stem-like phenotype are hypothesised to play a role in radioresistance. Since the progenitor marker neuron-glial-2 (NG2) has been shown to regulate several aspects of GBM progression in experimental systems, we hypothesised that its expression would influence the survival of GBM patients. Quantification of NG2 expression in 74 GBM biopsies from newly diagnosed and untreated patients revealed that 50% express high NG2 levels on tumour cells and associated vessels, being associated with significantly shorter survival. This effect was independent of age at diagnosis, treatment received and hypermethylation of the O6-methylguanine methyltransferase (MGMT) DNA repair gene promoter. NG2 was frequently co-expressed with nestin and vimentin but rarely with CD133 and the NG2 positive tumour cells harboured genetic aberrations typical for GBM. 2D proteomics of 11 randomly selected biopsies revealed upregulation of an antioxidant, peroxiredoxin-1 (PRDX-1), in the shortest surviving patients. Expression of PRDX-1 was associated with significantly reduced products of oxidative stress. Furthermore, NG2 expressing GBM cells showed resistance to ionising radiation (IR), rapidly recognised DNA damage and effectuated cell cycle checkpoint signalling. PRDX-1 knockdown transiently slowed tumour growth rates and sensitised them to IR in vivo. Our data establish NG2 as an important prognostic factor for GBM patient survival, by mediating resistance to radiotherapy through induction of ROS scavenging enzymes and preferential DNA damage signalling

Immunological mechanisms of tumour progression. The rationale for natural killer cell based immunotherapy for glioblastoma

Glioblastoma (GBM) is the most frequent and malignant brain tumour, where the patients’ median survival after diagnosis is only 14.6 months. Therefore, concerted research is required to develop novel treatments that will improve patients’ outcome. Immunotherapy is one of the promising strategies for novel treatment. However, GBM develops multiple mechanisms of immune suppression and escape from immune surveillance. Moreover, steroids, chemotherapy and radiotherapy render GBM patients immunocompromised. These aspects need to be taken into consideration prior to development of immunotherapy tailored to GBM patients. Natural killer (NK) cells are large, granular lymphocytes that are able to recognise and kill tumour cells and virus – infected cells without prior sensitization and costimulation. Therefore, they might be one of the most potent effectors for use in an immunotherapy. However, there is currently a great paucity of studies investigating their potential as therapeutic agents for GBM. This study confirmed the prognostic significance of elevated T cell infiltration into the tumour in GBM patients. However, NK cells represented a minor population in the tumour microenvironment. Several mechanisms of the tumour’s escape from immune attack were identified, including the induction of a novel population of CD8+CD28- Foxp3+ regulatory T cells and expression of CD73 and CD39 ectonucleotidases. Tumour cells expressed classical HLA class I molecules that are ligands for inhibitory killer immunoglobulin-like receptors (KIRs) mediating NK cells tolerance towards self cells. Tumour infiltrating macrophages/microglia displayed phenotypic features that were indicative of their tolerisation by the CD8+CD28-Foxp3+ regulatory T cells. These features include: (1) down-regulated expression of CD40, CD80 and CD86 costimulatory molecules and (2) up-regulated expression of immunoglobulin-like transcripts 2, 3, 4 (ILT2,3,4). In the systemic circulation, decreased T helper (Th) cells, up-regulated expression of the inhibitory receptor CTLA-4 on the Th cells and increased plasma concentration of IL-10 were identified in GBM patients’ blood compared to healthy controls. Despite these integrated mechanisms of tolerance and immunological escape, a proportion of the tumour cells might be susceptible to immune cell – mediated cytotoxicity due to their expression of Fas ligand, and MICA stress – induced ligand for NK cell NKG2D activating receptor. Thus, taking into consideration the GBM patients’ local and systemic immune suppression and escape, we hypothesised that the use of allogeneic NK cells in a KIR receptor – HLA ligand mismatch setting might be an amenable strategy against GBM cells. We demonstrated that allogeneic, ex vivo cultured NK cells efficiently lysed GBM cells in vitro and in vivo. However, the NK cell efficacy was donor – dependent. The presence of KIR2DS2 and KIR2DS4 genes in donors’ NK cells correlated with increased cytotoxicity in vitro and this effect was partially independent of the inhibitory KIR genes repertoire. Intracranial injection of a single dose of 106 allogeneic ex vivo cultured NK cells improved the survival of GBM – bearing mice compared to controls. NK cells obtained from donor possessing KIR2DS2 and KIR2DS4 genes were more effective than NK cells missing these genes and manifested in prolonged animals’ median survival, increased tumour cell apoptosis, decreased proliferation and diminished angiogenesis. In contrast, treatment with double doses of 106 NK cells from each donor did not result in improved survival. We observed increased recruitment of macrophages into the brain parenchyma of all NK cell treated animals compared to controls. However, the animals receiving single dose of NK cells exhibited higher proportions of F4/80+ macrophages that expressed the IL-7 receptor (CD127), while higher proportions of microglia expressed CD40 co-stimulatory molecules compared to control group. The tumour cells up-regulated the expression of nestin and HLA-ABC after single dose NK cell treatment as a response to inflammation. In summary, this work provides the rationale for using allogeneic NK cells against GBM and indicates possible targets for adjuvant immunotherapy, such as the CTLA-4 inhibitory receptor and the targets of CD8+CD28-Foxp3+ regulatory T cells – induced immunotolerisation. Moreover, the results of the effect of KIR2DS2 and KIR2DS4 in NK cell – mediated cytotoxicity against GBM provide a novel insight into tumour immunology. However, further research is required to evaluate the safety and efficacy of allogeneic NK cell – based therapy and to confirm the role of activating KIR receptors in determining the potency of NK cells against solid tumours