Endometrial carcinoma: Preliminary assessment of postoperational tolerance in pulsatile brachytherapy

W grupie 110 chorych operowanych z powodu raka trzonu macicy oceniono tolerancję uzupełniającej pulsacyjnej brachyterapii dopochwowej. Wczesny odczyn popromienny o lekkim przebiegu w pochwie i w odbytnicy wystapił łąpznie u 9 chorych (8.2%), późny u 6 chorych (5.5%). U jednej chorej wystapił odczyn popromienny w odbytnicy w stopniu czwartym. W okresie obserwacji nie stwierdzono wznowy w pochwie.Pulsacyjna brachyterapia dopochwpwa jest bezpieczną i skuteczną metodą uzupełniającego leczenia chorych operowanych z powodu raka trzonu macicy.In a group of 110 patients operated on for endometrial carcinoma the tolerance of a supplementary pulsatile intravaginal brachytherapy was assessed. Early light post-radiation reaction in the vagina and in the anus was found in a total of 9 (8.2%) patients, whereas late reaction occurred in 6 (5.5%) patients. One patient showed the fourth degree post-radiation reaction in the anus. During the follow-up no remission was observed in the vagina.Pulsatile brachytherapy is a safe and effective supplementary method of treatment in patients operated on for endometrial carcinoma

Usefulness of RAPD, RFLP and SCAR molecular markers and AGPaseB gene methylation level in the screening of resistance to the golden cyst nematode (Globodera rostochiensis) pathotype Ro1 in different Polish potato genotypes

Abstract The aim of this study was to assess the application of various DNA markers as well as the ADP-glucose pyrophosphorylase small subunit (AGPaseB) gene methylation for the screening of potato cultivars and breeding lines with different resistance to Globodera rostochiensis. Tetraploid genotypes were included. The 2 kb and 0.7 kb random amplified polymorphic (RAPD) fragments as well as a PCR-amplified 1.6 kb fragment of AGPaseB gene positioned close to the Gro1 locus were tested. Two novel sequence characterised amplified region (SCAR) fragments were generated from 2 kb RAPD product of susceptible and resistant genotypes. Significant correlation (r=0.32) was detected only between the presence of SCARI and SCARII fragments markers in all the investigated genotypes. Three different allelic forms of the AGPaseB gene were detected in tetraploid potato genotypes. One of these allelic forms may be closely linked with Gro1. A strong methylation occurred at promoter region and the 5' part of AGPaseB gene which was generally absent at its 3' UTR region. The methylation pattern of AGPaseB investigated by restriction with MspI and HpaII varied between some G. rostochiensis resistant and sensitive genotypes. We concluded that methylation profiling of AGPaseB gene should be promising for the development of practical assays for resistance to nematodes in potato

Exogenously induced expression of ethylene biosynthesis, ethylene perception, phospholipase D, and Rboh-oxidase genes in broccoli seedlings

In higher plants, copper ions, hydrogen peroxide, and cycloheximide have been recognized as very effective inducers of the transcriptional activity of genes encoding the enzymes of the ethylene biosynthesis pathway. In this report, the transcriptional patterns of genes encoding the 1-aminocyclopropane-1-carboxylate synthases (ACSs), 1-aminocyclopropane-1-carboxylate oxidases (ACOs), ETR1, ETR2, and ERS1 ethylene receptors, phospholipase D (PLD)-α1, -α2, -γ1, and -δ, and respiratory burst oxidase homologue (Rboh)-NADPH oxidase-D and -F in response to these inducers in Brassica oleracea etiolated seedlings are shown. ACS1, ACO1, ETR2, PLD-γ1, and RbohD represent genes whose expression was considerably affected by all of the inducers used. The investigations were performed on the seedlings with (i) ethylene insensitivity and (ii) a reduced level of the PLD-derived phosphatidic acid (PA). The general conclusion is that the expression of ACS1, -3, -4, -5, -7, and -11, ACO1, ETR1, ERS1, and ETR2, PLD-γ 1, and RbohD and F genes is undoubtedly under the reciprocal cross-talk of the ethylene and PAPLD signalling routes; both signals affect it in concerted or opposite ways depending on the gene or the type of stimuli. The results of these studies on broccoli seedlings are in agreement with the hypothesis that PA may directly affect the ethylene signal transduction pathway via an inhibitory effect on CTR1 (constitutive triple response 1) activity

Structure, catalytic activity and evolutionary relationships of 1-aminocyclopropane-1-carboxylate synthase, the key enzyme of ethylene synthesis in higher plants.

Both ethylene and the enzymes of ethylene synthesis are subjects of intensive scientific investigation. The present review discusses structure, catalytic activity and evolutionary relationships of 1-aminocyclopropane-1-carboxylate synthase, identified for the first time in ripening tomato in 1979. This enzyme is responsible for the conversion of S-adenosyl-L-methionine to 1-aminocyclopropane-1-carboxylic acid, which is the key step of ethylene synthesis in higher plants. The role of this enzyme (especially in the fruit ripening) was demonstrated in 1991 in transgenic tomato plants, expressing 1-aminocyclopropane-1-carboxylate synthase antisense RNA. On the basis of mutagenesis and crystallization of the enzyme, new data were provided on the three-dimensional structure and amino-acid residues which are critical for catalysis. The control of ethylene production is of great interest for plant biotechnology because it can delay senescence and overmaturation. These processes are responsible for large loss of vegetables and fruit on storage. Detailed structural and biochemical data are necessary to help design 1-aminocyclopropane-1-carboxylate synthase inhibitors, whose application is expected to have immense agricultural effects

Operacja wtórnego wszczepienia soczewki transskleralnej metodą bezszwową – obserwacje własne

Aim of the study: Presentation of the results of Carlevale™ intraocular lens implantation with a transscleral sutureless fixation method, based on our own experience. Material and methods: The sutureless transscleral implantation of a Carlevale™ lens was performed in 39 people (40 eyes) with postoperative aphakia after dislocation of lens, including 10 post-traumatic ones, and in one patient with a dislocated transscleral intraocular lens fixated with non-absorbable sutures. Patients were followed for 6 to 18 months. Results: 80% of patients after surgery achieved visual acuity equal to or better than before surgery (20% achieved Snellen acuity 1.0 without correction). Intraoperative and postoperative complications were observed in slightly more than half of the patients. The most common were keratopathy, slight bleeding into the vitreous chamber, conjunctival wound dehiscence, and intraocular pressure fluctuations. Most of these complications did not have any lasting consequences. Conclusions: Sutureless transscleral implantation of an intraocular lens seems to be an effective and relatively safe method of treating aphakia. However, the assessment of the durability of sutureless fixation requires further research.Cel pracy: Przedstawienie wyników operacji wszczepiania soczewki wewnątrzgałkowej typu Carlevale™ o fiksacji transskleralnej metodą bezszwową na podstawie własnych doświadczeń. Materiał i metody: Operacji wszczepienia metodą bezszwową soczewki transskleralnej typu Carlevale™ poddano 39 osób (40 oczu) z afakią po operacji usunięcia podwichniętej soczewki własnej lub sztucznej, w tym 10 pourazowych i jednego pacjenta z podwichniętą sztuczną soczewką o fiksacji transskleralnej z użyciem szwów niewchłanialnych. Pacjenci pozostawali pod obserwacją w okresie od 6 do 18 miesięcy. Wyniki: U 80% pacjentów po operacji uzyskano ostrość wzroku taką jak przed zabiegiem lub lepszą (20% osiągnęło 1,0 w skali dziesiętnej bez korekcji). U nieco ponad połowy obserwowano powikłania śródoperacyjne i pooperacyjne. Do najczęstszych należały: keratopatia, niewielkie krwawienie do komory ciała szklistego, rozejście się rany spojówki oraz zaburzenia ciśnienia wewnątrzgałkowego. W większości nie pozostawiły one trwałych następstw. Wnioski: Wszczepienie sztucznej soczewki o fiksacji transskleralnej bez użycia szwów wydaje się efektywną i stosunkowo bezpieczną metodą leczenia afakii. Jednakże ocena trwałości fiksacji bezszwowej wymaga dalszych badań

The effect of vanadate on Pichia pastoris growth, protein kinase A activity and ribosomal protein phosphorylation.

It was found that wild type yeast Pichia pastoris can tolerate vanadate concentration as high as 25 mM in the growth medium. Moreover, four vanadate-resistant P. pastoris strains designated JC100/1, JC100/3, JC100/9 and JC100/15 exhibiting tolerance up to 150 mM vanadate were selected. Growth of P. pastoris was correlated with vanadate to vanadyl reduction and its accumulation in the growth medium. In two selected strains, JC100/9 and JC100/15, protein kinase A activity was much higher in comparison to the wild type strain even without vanadate addition to the growth medium. Moreover, in the presence of vanadate, protein kinase A activity was significantly increased in the wild type and the vanadate-resistant JC100/1 and JC100/3 strains. It was also found that phosphorylation of a 40 kDa protein associated with ribosomes occured in all vanadate-resistant strains from the logarithmic, while in the wild type strain from the stationary growth phase. From the presented results it can be concluded that a protein kinase A signalling pathway(s) might be involved in the mechanism of P. pastoris vanadate resistance. The results also indicate a possible role of the 40 kDa protein in protection of P. pastoris against vanadate toxicity

The effect of cAMP and cGMP on the activity and substrate specificity of protein kinase A from methylotrophic yeast Pichia pastoris.

Cyclic AMP dependent protein kinase (PKA) from Pichia pastoris yeast cells was found to be activated by either cAMP or cGMP. Analogs of cAMP such as 8-chloro-cAMP and 8-bromo-cAMP were as potent as cAMP in PKA activation while N6,2'-O-dibutyryl-cAMP did not stimulate the enzyme activity. It was shown that protamine sulfate was almost equally phosphorylated in the presence of 1-2 × 10-6 M cAMP or cGMP while other substrates such as Kemptide, ribosomal protein S6, were phosphorylated to a lower extent in the presence of cGMP. It was demonstrated that pyruvate kinase is a substrate of PKA which co-purified with the P. pastoris enzyme. Moreover, pyruvate kinase was phosphorylated by PKA in the presence of cAMP and cGMP to comparable levels

The involvement of protein kinase A in the immune response of Galleria mellonella larvae to bacteria

The role of protein kinase A (PKA) in the humoral immune response of the greater wax moth Galleria mellonella larvae to live Gram-positive bacteria Micrococcus lysodeikticus and Gram-negative bacteria Escherichia coli was investigated. The immune challenge of larvae with both kinds of bacteria caused an increase in fat body PKA activity depending on the injected bacteria. Gram-positive M. lysodeikticus was a much better inducer of the enzyme activity than Gram-negative E. coli. The PKA activity was increased about 2.5-fold and 1.5-fold, after M. lysodeikticus and E. coli injection, respectively. The in vivo inhibition of the enzyme activity by a cell permeable selective PKA inhibitor, Rp-8-Br-cAMPS, was correlated with considerable changes of fat body lysozyme content and hemolymph antimicrobial activity in bacteria-challenged insects. The kinetics of changes were different and dependent on the bacteria used for the immune challenge of G. mellonella larvae

Protein kinases CKI and CKII are implicated in modification of ribosomal proteins of the yeast Trichosporon cutaneum

Phosphorylation of acidic ribosomal proteins P1/P2-P0 is a common phenomenon in eukaryotic organisms. It was found previously that in Trichosporon cutaneum, unlike in other yeast species, in addition to the two acidic ribosomal proteins, two other proteins of 15 kDa and 19 kDa of the small ribosomal subunit were phosphorylated. Here we describe two protein kinases: CKI and CKII, which are engaged in the modification of T. cutaneum ribosomal proteins. The acidic ribosomal proteins and the protein of 19 kDa were modified by CKII associated with ribosomes, while the protein of 15 kDa was modified by CKI. Protein kinase CKI was purified from cell-free extract (CKIC) and from ribosomal fraction (CKIR). The molecular mass of CKIC was established at 33 kDa while that of CKIR at 35-37 kDa. A protein of 40 kDa copurified with CKIR but not CKIC. Heparin significantly increased 40 kDa protein phosphorylation level by CKIR. Microsequencing analysis revealed the presence of CKI recognition motifs in the N-terminal fragment of the 40 kDa protein