Search results

1

Expression and polymorphism of defensins in farm animals

100%

Bagnicka E., Strzałkowska N., Jóźwik A., Krzyżewski J., Horbańczuk J., Zwierzchowski L.

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2010

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vol. 57

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issue 4

487-497

EN

Due to their activity against bacteria, viruses, and fungi, antimicrobial peptides are important factors in the innate resistance system of humans and animals. They are called "new generation antibiotics" for their potential use in preventive and therapeutic medicine. The most numerous group of antimicrobial peptides is a family of cationic peptides which include defensins and cathelicidins. Among them the most common are peptides with a beta-sheet structure containing three intra-molecular disulphide bonds, called defensins, comprising three classes: alpha, beta, and theta. The class of beta-defensins is the largest one. Their transcripts have been found in many tissues of humans and animals. The aim of this paper is to present the current knowledge about antimicrobial peptides from the defensin family in farm animals, their expression, polymorphism, as well as the potential of their use as genetic markers of health and production traits.

2

Inhibition of CYP17 expression by adrenal androgens and transforming growth factor β in adrenocortical cells.

88%

Biernacka-Łukanty J. M., Lehmann T. P., Trzeciak W. H.

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vol. 51

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issue 4

907-917

EN

Cytochrome P450c17, encoded by the CYP17 gene, is a component of the 17a-hydroxylase/17,20-lyase enzyme complex essential for production of adrenal glucocorticoids and androgens as well as gonadal androgens. The expression of CYP17 in adrenocortical cells is stimulated by corticotropin (ACTH) via the signal transduction pathway involving cAMP and protein kinase A (PKA). Thus, in addition to glucocorticoids, ACTH stimulates formation of adrenal androgens, which are known to induce transforming growth factor β (TGF-β) secretion. TGF-β in turn inhibits steroid hormone output by attenuating both basal and ACTH-dependent expression of CYP17. The present study revealed that treatment of bovine and human H295R adrenocortical cells with androgens resulted in a decrease in the basal level of CYP17 transcript and cortisol secretion, without affecting forskolin-stimulated levels. We also demonstrated that in H295R cells TGF-β inhibited both basal and forskolin-stimulated accumulation of CYP17 mRNA. Determination of promoter activity, directing luciferase reporter gene expression in H295R cells transfected with deletion fragments of bovine CYP17 promoter, indicated that the -483 to -433 bp fragment of the promoter was necessary for the inhibitory action of TGF-β on CYP17 expression. It is concluded that in bovine and human adrenocortical cells, androgens inhibit basal CYP17 expression probably at the transcriptional level and independently of the effect of TGF-β.

3

Regulatory mechanisms for the expression and activity of platelet-derived growth factor receptor.

88%

Funa K., Uramoto H.

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2003

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vol. 50

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issue 3

647-658

EN

PDGF is one of the most potent serum mitogens, and the signalling mechanism by way of its receptor tyrosine-kinase has been extensively studied since its first purification in 1979. The identification of homology between the simian sarcoma virus oncogene, v-sis, and the B-chain of PDGF, as well as the frequent over-expression of both the ligands and receptors in various tumours and stroma led to the proposal of the PDGF-mediated autocrine and paracrine hypothesis. Consistent with the important roles of PDGF in the growth and survival of cells, the expression and activity of PDGF receptors are tightly controlled by both positive and negative feedback mechanisms at different levels. The deregulation of the control system can result in serious pathological conditions such as chronic inflammation and tumours. Understanding the molecular mechanisms for the regulatory system and the signalling pathway of PDGF is essential in order to find effective therapies in the diseases where PDGF is involved.

4

Characterisation and molecular dynamic simulations of J15 asparaginase from Photobacterium sp. strain J15

88%

Yaacob M., Hasan W., Ali M., Rahman R., Salleh A., Basri M., Leow T.

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2014

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vol. 61

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issue 4

745-752

EN

Genome mining revealed a 1011 nucleotide-long fragment encoding a type I l-asparaginase (J15 asparaginase) from the halo-tolerant Photobacterium sp. strain J15. The gene was overexpressed in pET-32b (+) vector in E. coli strain Rosetta-gami B (DE3) pLysS and purified using two-step chromatographic methods: Ni2+-Sepharose affinity chromatography and Q-Sepharose anion exchange chromatography. The final specific activity and yield of the enzyme achieved from these steps were 20 U/mg and 49.2%, respectively. The functional dimeric form of J15-asparaginase was characterised with a molecular weight of ~70 kDa. The optimum temperature and pH were 25°C and pH 7.0, respectively. This protein was stable in the presence of 1 mM Ni2+ and Mg2+, but it was inhibited by Mn2+, Fe3+ and Zn2+ at the same concentration. J15 asparaginase actively hydrolysed its native substrate, l-asparagine, but had low activity towards l-glutamine. The melting temperature of J15 asparaginase was ~51°C, which was determined using denatured protein analysis of CD spectra. The Km, Kcat, Kcat/Km of J15 asparaginase were 0.76 mM, 3.2 s-1, and 4.21 s-1 mM-1, respectively. Conformational changes of the J15 asparaginase 3D structure at different temperatures (25°C, 45°C, and 65°C) were analysed using Molecular Dynamic simulations. From the analysis, residues Tyr24, His22, Gly23, Val25 and Pro26 may be directly involved in the 'open' and 'closed' lid-loop conformation, facilitating the conversion of substrates during enzymatic reactions. The properties of J15 asparaginase, which can work at physiological pH and has low glutaminase activity, suggest that this could be a good candidate for reducing toxic effects during cancer treatment.

5

Heterologous expression and initial characterization of recombinant RbcX protein from Thermosynechococcus elongatus BP-1 and the role of RbcX in RuBisCO assembly

88%

Tarnawski M., Gubernator B., Kolesinski P., Szczepaniak A.

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2008

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vol. 55

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issue 4

777-785

EN

In the cyanobacterial RuBisCO operon from Thermosynechococcus elongatus the rbcX gene is juxtaposed and cotranscribed with the rbcL and rbcS genes which encode large and small RuBisCO subunits, respectively. It has been suggested that the rbcX position is not random and that the RbcX protein could be a chaperone for RuBisCO. In this study, the RbcX protein from T. elongatus was overexpressed, purified and preliminary functional studies were conducted. The recombinant protein purified from Escherichia coli extracts was predominantly present in a soluble fraction in a dimeric form. Coexpression experiments have demonstrated that RbcX can mediate RbcL dimer (L2) formation, and that it is essential for the L8 core complex assembly. This is the first characterization of the RbcX protein from a thermophilic organism.

6

Identification and properties of the Deinococcus grandis and Deinococcus proteolyticus single-stranded DNA binding proteins (SSB)

88%

Filipkowski P., Kur J.

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2007

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vol. 54

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issue 1

79-87

EN

To study the biochemical properties of SSB's from Deinococcus grandis (DgrSSB) and Deinococcus proteolyticus (DprSSB), we have cloned the ssb genes obtained by PCR and have developed Escherichia coli overexpression systems. The genes consist of an open reading frame of 891 (DgrSSB) and 876 (DprSSB) nucleotides encoding proteins of 296 and 291 amino acids with a calculated molecular mass of 32.29 and 31.33 kDa, respectively. The amino-acid sequence of DgrSSB exhibits 45%, 44% and 82% identity and the amino-acid sequence of DprSSB reveals 43%, 43% and 69% identity with Thermus aquaticus (TaqSSB), Thermus thermophilus (TthSSB) and Deinococcus radiodurans SSBs, respectively. We show that DgrSSB and DprSSB are similar to Thermus/Deinococcus SSBs in their biochemical properties. They are functional as homodimers, with each monomer encoding two single-stranded DNA binding domains (OB-folds). In fluorescence titrations with poly(dT), both proteins bind single-stranded DNA with a binding site size of about 33 nt per homodimer. In a complementation assay in E. coli, DgrSSB and DprSSB took over the in vivo function of EcoSSB. Thermostability with half-lives of about 1 min at 65-67.5°C make DgrSSB and DprSSB similar to the known SSB of Deinococcus radiodurans (DraSSB).

7

Molecular cloning, recombinant expression, and purification of osteocalcin in sika deer (Cervus nippon) antler

75%

Li X., Liu M., Bai X., Li Y., Zhao Y., Wang S., Wang J., Li X., Liu M., Bai X., Li Y., Zhao Y., Wang S., Wang J.

Polish Journal of Veterinary Sciences

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2019

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issue 1

8

Cloning, sequence, expression and characterization of human β-mannosidase

75%

Samra Z. Q., Athar M. A.

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2008

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vol. 55

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issue 3

479-490

EN

β-mannosidase (EC 3.2.1.25, MANB) dissects the non-reducing end of N-linked mannose moieties of glycoproteins in eukaryotic cells. The human β-mannosidase gene was amplified by RT-PCR, cloned and sequenced. The DNA sequence was compared with reported human β-mannosidase DNA sequence and sixteen nucleotide differences were found. The deduced amino-acid sequence showed that seven codons coded the same amino acids and nine codons coded different amino acids with reference to nucleotide substitution positions but did not affect recombinant MANB enzyme activity. No splice mutation was observed after comparison with reported MANB DNA sequences. A 75% homology of deduced amino-acid sequence was observed with mouse, goat and bovine β-mannosidase amino-acid sequences. The cloned β-mannosidase gene was subcloned into pET22b+ and pET28a+ expression vectors to transform the BL21-codon plus cells for expression of recombinant MAN22 and MAN28 enzymes, respectively. The optimized conditions for overexpression of recombinant β-mannosidase enzyme were induction with 1 mM IPTG for 12 h at 37°C. The expressed β-mannosidase enzyme was purified to homogeneity by a combination of DEAE-ion exchange and size exclusion chromatography. The molecular mass of MAN22 and MAN28 enzymes is 97 kDa by SDS/PAGE and is confirmed by western blot analysis. The recombinant enzymes are active at 37°C and at pH 5.0 and showed activity with p-nitrophenyl-β-d-mannopyranoside and not with p-nitrophenyl-α-d-mannopyranoside. The Km value of enzymes was 2.53 mM. The enzyme activity was inhibited by Zn2+, Co2+, Cu2+, Pb2+, Ag1+, iodoacetate, SDS, DMF, DMSO and ethanol. Fe3+, Ca2+ Mg2+, Mn2+, Triton X-100 and PMSF did not inhibit the enzyme activity. Northern blot analysis showed a transcript of about 3.7 kb in all cells and tissues studied. This is the first report on the expression and characterization of recombinant human MANB enzyme.

9

Changes in kinesin expression in the CNS of mice with dynein heavy chain 1 mutation

75%

Kuźma-Kozakiewicz M., Kaźmierczak B., Usarek E., Barańczyk-Kuźma A.

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2013

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vol. 60

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issue 1

51-55

EN

Dysfunction of fast axonal transport, vital for motor neurons, may lead to neurodegeneration. Anterograde transport is mediated by N-kinesins (KIFs), while retrograde transport by dynein 1 and, to a minor extent, by C-kinesins. In our earlier studies we observed changes in expression of N- and C-kinesins (KIF5A, 5C, C2) in G93ASOD1-linked mouse model of motor neuron degeneration. In the present work we analyze the profile of expression of the same kinesins in mice with a dynein 1 heavy chain mutation (Dync1h1, called Cra1), presenting similar clinical symptoms, and in Cra1/SOD1 mice with milder disease progression than SOD1 transgenics. We found significantly higher levels of mRNA for KIF5A and KIF5C but not the KIFC2 in the frontal cortex of symptomatic Cra1/+ mice (aged 365 days) compared to the wild-type controls. No changes in kinesin expression were found in the spinal cord of any age group and only mild changes in the hippocampus. The expression of kinesins in the cerebellum of the presymptomatic and symptomatic mice (aged 140 and 365 days, respectively) was much lower than in age-matched controls. In Cra1/SOD1 mice the changes in KIFs expression were similar or more severe than in the Cra1/+ groups, and they also appeared in the spinal cord. Thus, in mice with the Dync1h1 mutation, which impairs dynein 1-dependent retrograde transport, expression of kinesin mRNA is affected in various structures of the CNS and the changes are similar or milder than in mice with double Dync1h1/hSOD1G93A mutations.

10

Tissue- and stage-specific expression of a fatty acid binding protein-like gene from amphioxus Branchiostoma belcheri

75%

Wang Y., Zhang Y., Zhang S., Tian J., Jiang S.

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2008

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vol. 55

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issue 1

27-34

EN

A cDNA clone encoding an amphioxus fatty acid binding protein-like (AmphiFABPL) protein was isolated from a gut cDNA library of Branchiostoma belcheri. It contained a 423 bp open reading frame corresponding to a deduced protein of 140 amino acids with a predicted molecular mass of approximately 15.9 kDa. Phylogenetic analysis showed that AmphiFABPL fell outside the vertebrate clade of fatty acid binding proteins (FABPs), being positioned at the base of the chordate lineage, and was almost equally homologous to various vertebrate FABPs, suggesting that it may be the archetype of vertebrate FABPs. Both northern blotting and in situ hybridization analyses demonstrated that AmphiFABPL was expressed in the hepatic caecum and hind-gut, and although at a much lower level, it was also present in the endostyle, ovary and testis. In addition, whole-mount in situ hybridization revealed that AmphiFABPL was initially expressed in the posterior two thirds of the primitive gut, including the mid-gut where the hepatic caecum will form later, in 2-day larvae. The expression pattern is closely similar to that of the L-FABP and I-FABP genes in vertebrates, supporting the hypothesis that the hepatic caecum in the amphioxus is homologous to the vertebrate liver.

11

An Accent-Based Approach to Automatic Rendering of Piano Performance: Preliminary Auditory Evaluation

75%

Bisesi E., Parncutt R.

Archives of Acoustics

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2011

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vol. 36

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issue 2

12

Podłoże molekularne rdzeniaka wieku dziecięcego

63%

Zakrzewska M., Liberski P. P.

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EN

Medulloblastoma is the most common type of embryonal tumour in paediatric population. This entity represents up to 4% of all intracranial neoplasms in the whole population and 20-25% of all brain tumours in children. The majority of tumours is located within vermis and fourth ventricle. Due to its frequency, histological aggressiveness, and unfavourable outcome, the treatment of children with medulloblastoma is a great clinical problem. According to the newest achievements of molecular biology, the significant heterogeneity of this tumour was confirmed. The most frequent types of molecular abnormalities detected in medulloblastoma were SHH and WNT pathways activation, MYC amplification and isochromosome i17q presence. Recently, on the basis of modern molecular analyses comprising gene expression profiling, several molecular subtypes of that tumour were described. Among them there were two subgroups connected with SHH and WNT pathways activation. SHH type medulloblastomas showed frequent CTNNB1 gene mutations and monosomy of chromosome 6. The PTCH and SUFU mutations accompanied by loss of chromosome 9 were identified in WNT type.Remaining subgroups distinguished on the basis of transcriptome analyses were not so clearly characterized and their number varied in particular molecular studies. This paper is a review of the latest data describing molecular background of medulloblastoma.

PL

Rdzeniak jest najczęstszym nowotworem ośrodkowego układu nerwowego pochodzenia zarodkowego występującym w wieku dziecięcym. W populacji ogólnej guz ten stanowi około 4%, natomiast u dzieci – 20-25% wszystkich nowotworów ośrodkowego układu nerwowego. Rdzeniak rozwija się tylko w obrębie tylnego dołu czaszki (podnamiotowo). Najczęściej umiejscowiony jest w robaku móżdżku i komorze IV mózgu. Ze względu na częstość występowania, wysoki stopień złośliwości histologicznej i związany z tym niekorzystny przebieg choroby rdzeniak stanowi poważny problem kliniczny. Ostatnio dzięki zaawansowanej diagnostyce molekularnej wykazano znaczną heterogenność tych nowotworów. Wśród najczęściej obserwowanych zaburzeń molekularnych wymienia się aktywację szlaków przekazywania sygnałów komórkowych SHH i WNT, amplifikację genu MYC i obecność izochromosomu i17q. W oparciu o wykorzystanie nowoczesnych metod biologii molekularnej, m.in. profilowania genomowego, wyodrębniono kilka podtypów molekularnych tego nowotworu. Wśród nich wyróżniają się dwie spójne podgrupy związane z aktywacją szlaków WNT i SHH. Nowotwory uwarunkowane aktywacją pierwszego ze szlaków cechują częste mutacje genu CTNNB1 i zmiany pod postacią monosomii chromosomu 6. Rdzeniaki, których profil genowy wskazuje na aktywację drugiego ze szlaków, prezentują częste mutacje genów PTCH i SUFU oraz delecję ramienia długiego chromosomu 9. Pozostałe podtypy molekularne określone na podstawie analizy transkryptomów nie są już tak dobrze scharakteryzowane i w zależności od opracowania różnią się liczbą wyróżnionych podgrup. Praca jest aktualnym przeglądem piśmiennictwa opisującego podłoże molekularne rdzeniaka, które w niedalekiej przyszłości może zostać wykorzystane w praktyce klinicznej do indywidualizacji terapii u dzieci z tym nowotworem.

13

The use of basophil activation tests in clinical-laboratory investigations in patients with beta-lactam allergy (pilot study)

51%

Havrylyuk A., Zubchenko S., Lomikovska M., Kril I., Lishchuk-Yakymovych K., Pukalyak R., Chopyak V.

Alergia Astma Immunologia - przegląd kliniczny

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2023

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vol. 28

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issue 1

8-16

PL

WPROWADZENIE: Antybiotyki beta(β)-laktamowe (BLA) są terapią pierwszego rzutu w pozaszpitalnych i szpitalnych zakażeniach bakteryjnych i są najczęściej zgłaszane jako wywołujące reakcje alergiczne. Około 50% wszystkich alergików w Europie i USA cierpi na alergie na leki, w tym na alergie na BLA. Celem badania była ocena reakcji krzyżowych między cefalosporyną II i III generacji u pacjentów z klinicznym wywiadem reakcji na BLA i ryzyka wystąpienia niepożądanych reakcji BLA na podstawie wyników testu aktywacji bazofilów. MATERIAŁ I METODY: przebadaliśmy 48 kobiet i 8 mężczyzn (razem 56) z pierwotnymi reakcjami BLA w wieku od 26 do 61 lat oraz 24 zdrowych ochotników (grupa kontrolna). Pacjentów 19 (34%) bylo leczonych amoksycyliną, 18 (32,1%) – amoksycylina + kwasem klawulanowym, 6 (10,7%) – cefuroksymem i 13 (23,2%) – ceftriaksonem. W celu oceny markera degranulacji bazofili CD 63 po stymulacji antygenem w pełnej krwi wykorzystano oznaczenie Flow CAST (FK-CCR) (Bühlmann Laboratories AG, Szwajcaria). Na podstawie wyników ВАТ pacjentów podzielono na dwie podgrupy: w pierwszej grupie było 33 pacjentów z dodatnim niższym wynikiem indeksu stymulacji (<10%), a w drugiej grupie było 15 pacjentów z istotnie wyższym poziomem CD63 ekspresja CD63 (>10%). WYNIK: Nasze wyniki wykazały, że pacjenci z drugiej podgrupy mieli najwyższe wyniki ekspresji CD63 i wskaźnika stymulacji dla amoksycyliny, następnie dla ceftriaksonu, a ostatni dla cefuroksymu. Byli leczeni amoksycyliną i odpowiadali na nią, jak wykazały wysokie wartości ekspresji CD63 w BAT, ci pacjenci mieli również wysoką ekspresję CD63 po stymulacji ceftriaksonem i cefuroksymem. W pierwszej podgrupie u 51,5% pacjentów pokrzywka i skurcz oskrzeli ustąpiły w ciągu 3 godzin od wystąpienia objawów, u 42,4% pacjentów z pokrzywką i obrzękiem naczynioruchowym objawy utrzymywały się przez 2-3 dni, a u 6,1% osutka plamkowo-grudkowa - przez ponad tydzień. Pacjenci z pierwszej podgrupy (z niską ekspresją CD63) wykazywali klinicznie słabe objawy reakcji. Po leczeniu antybiotykami pacjenci z drugiej podgrupy wykazywali silniejsze objawy: u 60,0% - anafilaksja; 6,7% - zespół Stevensa-Johnsona. Wykazaliśmy, że u pacjentów z reakcja nadwrażliwości na leczenie antybiotykami im wyższe wyjściowe wyniki testu po stymulacji in vitro, tym bardziej nasilone są ich objawy kliniczne. WNIOSEK: u pacjentów z klinicznymi objawami BLA w przypadku sprzecznych danych z wywiadu zaleca się ustalenie rzeczywistego uczulenia na antybiotyki i przewidywanie występowania reakcji krzyżowych między penicylinami i cefalosporynami nie tylko drugiej, ale i trzeciej generacji. Wyniki BAT z antybiotykami mogą być wykorzystane do opracowania zaleceń przyszłych terapii przeciwbakteryjnych.

EN

BACKGROUND: Beta (β)-lactam antibiotics (BLAs) are the first-line therapy for non-nosocomial and nosocomial bacterial infections and are most commonly reported to cause allergic reactions. Approximately 50% of all allergic patients in Europe and the USA suffer from drug allergies and BLA allergies. The AIM of the study was to assess cross-reactivity reactions between 2nd and 3rd generation cephalosporins in patients with a medical history of BLA reactions and the risk of adverse reactions to BLAs based on the results of the basophil activation test. MATERIALS AND METHODS: we examined 48 females and 8 males (in all 56 patients) aged 26 to 61 with primary reactions to BLAs and 24 healthy volunteers (control group). 19 (34%) patients were treated with amoxicillin, 18 (32,1%) patients were receiving amoxicillin+clavulanic acid, 6 (10,7%) patients were treated with cefuroxime, and 13 (23,2%) patients with ceftriaxone. Quantitative determination of the CD63 marker of basophil degranulation upon antigen stimulation in whole blood was performed with the use of Flow CAST (FK-CCR) (Bühlmann Laboratories AG, Switzerland). Based on the obtained ВАТ results, the patients were divided into two subgroups: the first group included 33 patients with positive stimulation index but lower CD63 expression (<10%), and the second group included 15 patients with a significantly higher level of CD63 expression (>10 %). THE RESULT: We showed that patients from the second subgroup had the highest level of CD63 expression and stimulation index when amoxicillin, whereas the level of CD63 expression and stimulation index were lower whith ceftriaxone; at the same time, the level of CD63 expression and stimulation index were the lowest with cefuroxime. The patients who treated with and reacted to amoxicillin, as shown by high BAT, also had high CD63 expresiion after ceftriaxone and cefuroxime stimulation. In the first subgroup, urticarial and bronchospasm disappeared within 3 hours of the onset of symptoms in 51.5% of patients, the symptoms persisted for 2-3 days in 42.4% of patients with urticaria and angioedema, whereas maculopapular exanthema persisted for more than a week in 6.1% of the patients. Patients from the first subgroup (with low CD63 expression) had a weak reaction to the culprit antibiotic. Patients from the second subgroup had the strongest reaction to culprit antibiotics: anaphylaxis – 60.0%; Stevens-Johnson syndrome – 6.7%. We established that in patients with hypersensitivity to antibiotics the higher the baseline test scores after in vitro stimulation, the more severe clinical symptoms. CONCLUSION: for patients with clinical manifestations of BLA in case of conflicting anamnesis data, it is recommended to establish true sensitization to antibiotics and to predict the occurrence of cross-reactions between penicillins and cephalosporins not only of the 2nd but also of the 3rd generation. The results of BAT for antibiotics can be used to formulate future antibacterial treatments recommendation.

Refine search results

Expression and polymorphism of defensins in farm animals

Inhibition of CYP17 expression by adrenal androgens and transforming growth factor β in adrenocortical cells.

Regulatory mechanisms for the expression and activity of platelet-derived growth factor receptor.

Characterisation and molecular dynamic simulations of J15 asparaginase from Photobacterium sp. strain J15

Heterologous expression and initial characterization of recombinant RbcX protein from Thermosynechococcus elongatus BP-1 and the role of RbcX in RuBisCO assembly

Identification and properties of the Deinococcus grandis and Deinococcus proteolyticus single-stranded DNA binding proteins (SSB)

Molecular cloning, recombinant expression, and purification of osteocalcin in sika deer (Cervus nippon) antler

Cloning, sequence, expression and characterization of human β-mannosidase

Changes in kinesin expression in the CNS of mice with dynein heavy chain 1 mutation

Tissue- and stage-specific expression of a fatty acid binding protein-like gene from amphioxus Branchiostoma belcheri

An Accent-Based Approach to Automatic Rendering of Piano Performance: Preliminary Auditory Evaluation

Podłoże molekularne rdzeniaka wieku dziecięcego

The use of basophil activation tests in clinical-laboratory investigations in patients with beta-lactam allergy (pilot study)