Bacteriophage-encoded virion-associated enzymes to overcome the carbohydrate barriers during the infection process

Bacteriophages are bacterial viruses that infect the host after successful receptor recognition and adsorption to the cell surface. The irreversible adherence followed by genome material ejection into host cell cytoplasm must be preceded by the passage of diverse carbohydrate barriers such as capsule polysaccharides (CPSs), O-polysaccharide chains of lipopolysaccharide (LPS) molecules, extracellular polysaccharides (EPSs) forming biofilm matrix, and peptidoglycan (PG) layers. For that purpose, bacteriophages are equipped with various virion-associated carbohydrate active enzymes, termed polysaccharide depolymerases and lysins, that recognize, bind, and degrade the polysaccharide compounds. We discuss the existing diversity in structural locations, variable architectures, enzymatic specificities, and evolutionary aspects of polysaccharide depolymerases and virion-associated lysins (VALs) and illustrate how these aspects can correlate with the host spectrum. In addition, we present methods that can be used for activity determination and the application potential of these enzymes as antibacterials, antivirulence agents, and diagnostic tools

Trudności insulinoterapii u chorej na cukrzycę typu 2 ze współistniejącym rozlanym zapaleniem powięzi z eozynofilią — opis przypadku

We describe problems of insulin therapy in 61-years old woman with type 2 diabetes treated for last 3 years with pre-mixed insulin given twice a day, in whom essential hyperglycemia occurred in the course of glucosteroid therapy (prednisone 35 mg per day). Corticosteroids were given last month by rheumatologists due to a rare connective tissue disease — diffuse fasciitis with eosinophilia, characterized by symmetrical and significant swelling, induration and thickening of the skin and soft tissues of the limbs, trunk, neck. Insulin was given in basal bolus schema and increased from 0.63 to 1.3 units per kg. Improvement was obtained by using NPH insulin given in the morning at much higher doses than in the evening (22 U v. 7 U) and with the highest doses of insulin aspart for the lunch time (30 U), than for the breakfast and dinner (16 U and 15 U). However, high glucose levels were still observed after lunch, in the afternoon and at dinner time. Injections of higher doses of insulin, especially needed for lunch, were impossible due to extreme induration of the skin, as backward flow of insulin was observed. Satisfactory glucose levels were obtained after introduction of a co-treatment with long acting sulphonylurea in the morning and acarbose for a lunch.W pracy przedstawiono trudności w prowadzeniu insulinoterapii u 61-letniej chorej z cukrzycą typu 2, leczonej od trzech lat mieszankami insulinowymi w dwóch wstrzyknięciach, u której dekompensacja cukrzycy nastąpiła w przebiegu steroidoterapii (prednizon 35 mg), wdrożonej w ostatnim miesiącu z powodu rzadkiej choroby tkanki łącznej — zapalenia powięzi z eozynofilią, charakteryzującej się znacznym obrzękiem, stwardnieniem i zgrubieniem skóry oraz tkanek miękkich, które objęło całe kończyny, tułów, szyję. Insulinę stosowano w schemacie baza–bolus, przy czym dobową dawkę insuliny zwiększono z 0,63 j. do 1,3 j. na kilogram masy ciała. Istotną poprawę uzyskano, stosując insulinę NPH rano w dawce 3-krotnie większej niż wieczorem (22 j. vs. 7 j.) oraz stosując większe dawki insuliny aspart do obiadu (30 j.) niż do śniadania i kolacji (16 j. i 15 j.). Problemem pozostawały hiperglikemie poposiłkowe — zwłaszcza po obiedzie, w godzinach popołudniowych i w porze kolacji. Istotną trudność w leczeniu stanowiła bardzo duża twardość skóry kończyn i tułowia spowodowana wspomnianym zapaleniem powięzi, ponieważ przy próbach zwiększenia dawek insuliny obserwowano wsteczny wypływ insuliny (szczególnie przy większych dawkach podawanych do obiadu). Dołączenie długodziałającej pochodnej sulfonylomocznika w godzinach rannych i akarbozy w porze obiadu pozwoliło na zadowalające wyrównanie glikemii

Can better hygiene generate an increase of type 1 diabetes frequency?

Liczne badania epidemiologiczne wskazują na stały wzrost częstości zachorowań na cukrzycę typu 1, zwłaszcza w krajach rozwiniętych, obserwowany równolegle ze zmniejszeniem zachorowań na choroby infekcyjne. Wyraźny gradient północ–południe dotyczący zachorowań na cukrzycę typu 1 i inne schorzenia immunologiczne występuje w Stanach Zjednoczonych i Europie, dodatkowo w Europie obecny jest gradient zachód–wschód. W krajach rozwiniętych najmniejszą częstość zachorowań odnotowuje się na obszarach o najgorszych warunkach socjoekonomicznych i higienicznych. Istnieje kilka hipotez, które próbują tłumaczyć związek między większą częstością zachorowań na choroby alergiczne i autoimmunologiczne (w tym cukrzycę typu 1) a mniejszą ekspozycją na czynniki infekcyjne. Obejmują one między innymi koncepcję współzawodnictwa o antygen, zjawisko „bystander suppresion” czy hipotezę „polio”. Zgodnie z hipotezą higieny nadmierne przestrzeganie czystości może być przyczyną zaburzonej równowagi systemu immunologicznego, co z kolei może prowadzić do procesu autoimmunologicznego skierowanego przeciwko komórkom beta wysp trzustki i bezwzględnego niedoboru insuliny endogennej.Variety of epidemiological studies indicate a steady increase in the incidence of type 1 diabetes mellitus, especially in industrialized countries, contrasted by a gradual decrease in the incidence of infectious diseases. There is an overall North–South gradient for type 1 diabetes and other immune disorders in North America and Europe, and West–East gradient in Europe. In developed countries lowest type 1 diabetes frequency is reported in the areas with the poorest socio-economic and hygiene conditions. There are several hypotheses that attempt to explain the relationship between lower exposure to transmissible agents, and a higher incidence of cases of allergic or autoimmune diseases (including type 1 diabetes mellitus). These are: the concept of the antigen competition, the phenomenon of bystander suppresion or “polio” hypothesis. According to hygiene hypothesis increased hygiene may contribute to imbalance of the autoimmune system, facilitating autoimmune reactions leading to destruction of islet beta cells and lack of endogenous insulin

Intelligent organization in a local administrative unit : from theoretical design to reality

Purpose: The theoretical purpose of this paper is determining the essence of an intelligent organization (IO) in a local administrative unit (LAU) including identification of benefits and risks for local development connected with the development of IO in LAU. The practical purpose is presenting a concept of methods to identify the development level of IO in LAU. Design/Methodology/Approach: The work method is literature studies. In addition, two methods of evaluating the level of development of IO in LAU are proposed. The article makes use of statistical methods based on the vector of standardized sums. Findings: It is found that literature lacks a generally accepted definition of IO in LAU and many features characteristic of the development of IO in LAU are identified. The development of IO in LAU is a source of many benefits to local development, but it is also connected with the occurrence of some risks. The study proposes a set of features describing IO including their scoring. The paper also propose an indicator of the level of development of IO in LAU2 in Poland (municipality) in 2018 based on a set of variables (IO LAU2). Research has shown a positive correlation between the level of development of IO in LAU and the level of urbanization, investment attractiveness and economic growth. Practical Implications: The results of studies can be used by LAU for designing the strategy of developing an intelligent organization. They can provide valuable practical guidance about the type of activities to be implemented to ensure that LAU can evolve into IO. Originality/Value: The work proposes a definition of IO in LAU. Two methods of evaluating the level of development of IO in LAU are put forward. The first method allows pre-selection of LAU in terms the level of development of IO based on data available in public statistics. The next stage of the study can use the criteria of summary evaluation of the level of development of IO in the LAU, proposed in this work, according to a percentage scale. The presented methodological concept is universal – it can be used to evaluate the level of development of IO in taxonomic units at different levels and in different countries.peer-reviewe

The role of intelligent organisations in creating favourable conditions for the development of entrepreneurship

Purpose: The theoretical purpose of this study was to determine the essence of smart specialisation in local administrative units, together with the identification of its impact on the level of entrepreneurship and socio-economic development of spatial units. The empirical purpose was to indicate the relationship between the level of development of intelligent organisations (IO) in counties and the degree of entrepreneurship intensity. Design/Methodology/Approach: The paper uses the method of literature studies, the standardised sums method, the weighted correlation method and cartographic analysis. Findings: Intelligent specialisation in a local administrative unit (LAU) can contribute to the development of entrepreneurship and accelerate the socio-economic development of the area. Investment attractiveness is more important for stimulating the development of entrepreneurship than intelligent organisation in local administrative units at the county level in Poland. Practical Implications: Results of the research can be used by local administrative units to shape the development strategy of an intelligent organization as well as of a smart specialisation. They can provide valuable practical guidance and synergies. Originality/Value: The paper identifies key features of the smart specialization strategy. The paper defines the intelligent organizations development level index in local administrative units of the county level in Poland in 2018 based on a set of variables. A method of constructing typologies of local administrative units is based on indices determining the level of intelligent organization, investment attractiveness and entrepreneurship. The methodological concept can be used to assess the level of development of IOs in taxonomic units of different levels in different countries.peer-reviewe

Celiac disease in adult patients with type 1 diabetes mellitus

Cukrzycy typu 1 mogą towarzyszyć inne schorzenia autoimmunologiczne, w tym celiakia (choroba trzewna, choroba glutenowa). U około 15% dorosłych chorych na cukrzycę typu 1 może być obecna celiakia. Zwykle przebiega skrycie, może jednak prowadzić do licznych epizodów hipoglikemii lub kwasicy ketonowej. Współistnienie celiakii zwiększa ryzyko wystąpienia retinopatii, nefropatii i osteoporozy. W pracy omówiono aktualnie obowiązujące schematy diagnostyczne celiakii przeznaczone dla osób z grup dużego ryzyka, do których zaliczani są również chorzy na cukrzycę typu 1. Zwrócono szczególną uwagę na zalecenia dotyczące badań serologicznych i ich interpretację oraz badania genetyczne, które są niezwykle przydatne w celu wykluczenia tego schorzenia.Type 1 diabetes may be accompanied by other autoimmune diseases, including celiac disease (sprue, gluten disease). Celiac disease may be present inapproximately 15% of adult patients with type 1 diabetes. It is usually asymptomatic, however it may lead to a number of episodes of hypoglycemia or ketoacidosis. The coexistence of celiac disease increases the risk of retinopathy, nephropathy and osteoporosis. The paper discusses current diagnostic procedures of celiac disease in the high-risk population, including type 1 diabetes patients. Particular attention was paid to the recommended serological tests, their interpretation, and genetic studies, which are extremely useful to exclude this disease

Phage-borne depolymerases decrease Klebsiella pneumoniae resistance to innate defense mechanisms

Klebsiella pneumoniae produces capsular polysaccharides that are a crucial virulence factor protecting bacteria against innate response mechanisms of the infected host. Simultaneously, those capsules are targeted by specific bacteriophages equipped with virion-associated depolymerases able to recognize and degrade these polysaccharides. We show that Klebsiella phage KP32 produces two capsule depolymerases, KP32gp37 and KP32gp38, with a high specificity for the capsular serotypes K3 and K21, respectively. Together, they determine the host spectrum of bacteriophage KP32, which is limited to strains with serotype K3 and K21. Both depolymerases form a trimeric beta-structure, display moderate thermostability and function optimally under neutral to alkaline conditions. We show that both depolymerases strongly affect the virulence of K. pneumoniae with the corresponding K3 and K21 capsular serotypes. Capsule degradation renders the otherwise serum-resistant cells more prone to complement-mediated killing with up to four log reduction in serum upon exposure to KP32gp37. Decapsulated strains are also sensitized for phagocytosis with a twofold increased uptake. In addition, the intracellular survival of phagocytized cells in macrophages was significantly reduced when bacteria were previously exposed to the capsule depolymerases. Finally, depolymerase application considerably increases the lifespan of Galleria mellonella larvae infected with K. pneumoniae in a time- and strain-dependent manner. In sum, capsule depolymerases are promising antivirulence compounds that act by defeating a major resistance mechanism of K. pneumoniae against the innate immunity

Czy nieprawidłowe rozpoznanie zespołu Wolframa jako cukrzycy typu 1 i jej powikłań może być przyczyną rzadkiego rozpoznawania tego zespołu?

Wolfram syndrome (WS), also known as DIDMOAD (Diabetes Insipidus, Diabetes Mellitus, Optic Atrophy and Deafness), is a rare autosomal recessive syndrome (1/770,000 in the United Kingdom), characterised by juvenile onset of diabetes mellitus, optic nerve atrophy, diabetes insipidus, sensorineural deafness, renal tract and neurological abnormalities, and primary gonadal atrophy. WS is caused mainly by biallelic mutations in the WFS1 gene, which encodes wolframin. Wide tissue distribution of wolframin and many mutations in the wolframin gene resulting in Wolfram syndrome may contribute to different phenotypes and the unusual combinations of clinical features. We describe a female patient with Wolfram syndrome diagnosed at the age of 25, with a previous false diagnosis of type 1 diabetes mellitus and misdiagnosed diabetic complications. The patient was found to be a compound heterozygote for two novel mutations in exon 8 of WFS1 gene: a 2-bp deletion AT at nt 1539 leading to a frameshift (Y513fs) and a single-base substitution 1174C > T resulting in a stop codon (Q392X). A detailed analysis of the patient’s medical history and a review of the literature suggest that many cases of Wolfram syndrome may remain undiagnosed due to misdiagnosis as type 1 diabetes mellitus and incorrect interpretation of clinical symptoms of neurodegenerative abnormalities, especially in their early stages. (Endokrynol Pol 2014; 65 (5): 398–400)Zespół Wolframa (WS), znany również jako DIDMOAD (moczówka prosta, cukrzyca, atrofia nerwu wzrokowego i głuchota), jest rzadkim zespołem dziedziczonym autosomalnie recesywnie (1/770000 w Wielkiej Brytanii), charakteryzującym się wystąpieniem cukrzycy w wieku młodzieńczym, zanikiem nerwu wzrokowego, moczówką prostą, głuchotą, niewydolnością oddechową i zaburzeniami neurologicznymi oraz pierwotną atrofią gonad. WS jest spowodowany głównie mutacją w genie WFS1, który koduje wolframinę. Obecność wolframiny w wielu tkankach organizmu oraz wiele różnych mutacji w genie wolframiny, których skutkiem jest wystąpienie zespołu Wolframa, może stanowić przyczynę różnych fenotypów tego zespołu oraz różnych kombinacji cech klinicznych. W poniższej publikacji opisano przypadek pacjentki z zespołem Wolframa, której choroba początkowo była błędnie zdiagnozowana jako cukrzyca typu 1 i jej powikłania. W badaniach genetycznych wykazano, że pacjentka była heterozygotą w zakresie dwóch nowych mutacji w egzonie 8 genu WFS1: 2-bp delecji AT w regionie nt 1539, prowadzącej do mutacji zmiany ramki odczytu (Y513fs) oraz substytucji pojedynczej zasady 1174 C > T, czego skutkiem był stop kodon (Q392X). Ze szczegółowej analizy historii medycznej pacjentki oraz przeglądu piśmiennictwa wynika, że duża liczba przypadków zespołu Wolframa może zostać niewłaściwie rozpoznana jako cukrzyca typu 1 lub zaburzenia neurodegeneracyjne, zwłaszcza w początkowej fazie ich rozwoju. (Endokrynol Pol 2014; 65 (5): 398–400

Production of medical Sc radioisotopes with an alpha particle beam

The internal α-particle beam of the Warsaw Heavy Ion Cyclotron was used to produce research quantities of the medically interesting Sc radioisotopes from natural Ca and K and isotopically enriched 42Ca targets. The targets were made of metallic calcium, calcium carbonate and potassium chloride. New data on the production yields and impurities generated during the target irradiations are presented for the positron emitters 43Sc, 44 gSc and 44 mSc. The different paths for the production of the long lived 44 mSc/44 gSc in vivo generator, proposed by the ARRONAX team, using proton and deuteron beams as well as alpha-particle beams are discussed. Due to the larger angular momentum transfer in the formation of the compound nucleus in the case of the alpha particle induced reactions, the isomeric ratio of 44 mSc/44 gSc at a bombarding energy of 29 MeV is five times larger than previously determined for a deuteron beam and twenty times larger than for proton induced reactions on enriched CaCO3 targets. Therefore, formation of this generator via the alpha-particle route seems a very attractive way to form these isotopes. The experimental data presented here are compared with theoretical predictions made using the EMPIRE evaporation code. Reasonable agreement is generally observed

Cyclotron production of 43Sc for PET imaging

Recently, significant interest in 44Sc as a tracer for positron emission tomography (PET) imaging has been observed. Unfortunately, the co-emission by 44Sc of high-energy γ rays (Eγ = 1157, 1499 keV) causes a dangerous increase of the radiation dose to the patients and clinical staff. However, it is possible to produce another radionuclide of scandium—43Sc—having properties similar to 44Sc but is characterized by much lower energy of the concurrent gamma emissions. This work presents the production route of 43Sc by α irradiation of natural calcium, its separation and purification processes, and the labeling of [DOTA,Tyr3] octreotate (DOTATATE) bioconjugate. Methods: Natural CaCO3 and enriched [40Ca]CaCO3 were irradiated with alpha particles for 1 h in an energy range of 14.8–30 MeV at a beam current of 0.5 or 0.25 μA. In order to find the optimum method for the separation of 43Sc from irradiated calcium targets, three processes previously developed for 44Sc were tested. Radiolabeling experiments were performed with DOTATATE radiobioconjugate, and the stability of the obtained 43Sc-DOTATATE was tested in human serum. Results: Studies of natCaCO3 target irradiation by alpha particles show that the optimum alpha particle energies are in the range of 24–27 MeV, giving 102 MBq/μA/h of 43Sc radioactivity which creates the opportunity to produce several GBq of 43Sc. The separation experiments performed indicate that, as with 44Sc, due to the simplicity of the operations and because of the chemical purity of the 43Sc obtained, the best separation process is when UTEVA resin is used. The DOTATATE conjugate was labeled by the obtained 43Sc with a yield >98 % at elevated temperature. Conclusions: Tens of GBq activities of 43Sc of high radionuclidic purity can be obtainable for clinical applications by irradiation of natural calcium with an alpha beam