In vivo dosimetry of the rectum in image-guided adaptive interstitial-intracavitary brachytherapy of cervix cancer – A feasibility study

Aim and backgroundTo investigate the feasibility of in vivo rectal dosimetry in image-guided adaptive brachytherapy of cervical cancer.Materials and methodsError of measurement of dose rate in a semiconductor diode probe was investigated depending on the distance and angle in water, and on temperature in a polymethyl methacrylate phantom using an Ir-192 source. Furthermore, the difference between the measured and calculated dose was analysed in the interstitial brachytherapy of 30 cervix cancer patients. The relationship between in vivo measured dose, calculated dose in the point of the diode, calculated maximal dose in the point of the diodes and calculated maximal dose of the rectum were examined.ResultsThe dosimeter measured with 85% accuracy at more than 5 cm from the source, but within a closer distance the accuracy decreased significantly. At 45–90° angle, the device measured with a 15% error. The error increased with the temperature, 22% at 35 °C. In 8 cases (26.7%) the maximal dose was measured in the correct diode. The device measured 73% of the calculated dose in the point of the diode. The maximum of the calculated doses of diodes was 60% of the calculated maximal dose. The in vivo measured dose was 35% of the calculated maximal dose.ConclusionsUnder treatment conditions, the semiconductor diode does not provide reliable measured data. The probe pushes the rectal wall closer to the high dose areas and underestimates the dose of it. Semiconductor probe is not recommended for in vivo dosimetry of the rectum in image-guided brachytherapy of cervical cancer.     

Botulinum Neurotoxins A and E Undergo Retrograde Axonal Transport in Primary Motor Neurons

The striking differences between the clinical symptoms of tetanus and botulism have been ascribed to the different fate of the parental neurotoxins once internalised in motor neurons. Tetanus toxin (TeNT) is known to undergo transcytosis into inhibitory interneurons and block the release of inhibitory neurotransmitters in the spinal cord, causing a spastic paralysis. In contrast, botulinum neurotoxins (BoNTs) block acetylcholine release at the neuromuscular junction, therefore inducing a flaccid paralysis. Whilst overt experimental evidence supports the sorting of TeNT to the axonal retrograde transport pathway, recent findings challenge the established view that BoNT trafficking is restricted to the neuromuscular junction by highlighting central effects caused by these neurotoxins. These results suggest a more complex scenario whereby BoNTs also engage long-range trafficking mechanisms. However, the intracellular pathways underlying this process remain unclear. We sought to fill this gap by using primary motor neurons either in mass culture or differentiated in microfluidic devices to directly monitor the endocytosis and axonal transport of full length BoNT/A and BoNT/E and their recombinant binding fragments. We show that BoNT/A and BoNT/E are internalised by spinal cord motor neurons and undergo fast axonal retrograde transport. BoNT/A and BoNT/E are internalised in non-acidic axonal carriers that partially overlap with those containing TeNT, following a process that is largely independent of stimulated synaptic vesicle endo-exocytosis. Following intramuscular injection in vivo, BoNT/A and TeNT displayed central effects with a similar time course. Central actions paralleled the peripheral spastic paralysis for TeNT, but lagged behind the onset of flaccid paralysis for BoNT/A. These results suggest that the fast axonal retrograde transport compartment is composed of multifunctional trafficking organelles orchestrating the simultaneous transfer of diverse cargoes from nerve terminals to the soma, and represents a general gateway for the delivery of virulence factors and pathogens to the central nervous system.     

The influence of habitat conditions on the performance of two invasive,annuals — Impatiens glandulifera and Bidens frondosa

The investigations of habitat conditions on the variability of selected population features in the two invasive, annuals with different life-history traits were conducted in the years 2008–2010, in the Polish part of the Carpathian Mountains in communities characterized by the gradual decrease of light availability. The individuals of Impatiens glandulifera were surveyed along roadsides, in willow thickets, as well as inside and along the edges of the riparian forest, whereas the individuals of Bidens frondosa were observed in riverside gravels characterized by a different species composition. Each year, the number and density of individuals (stems) occurring in the particular sites were examined, as well as the height and the fruit production in 30 randomly chosen stems were surveyed. Moreover, the number of seeds per fruit, the diaspore dimensions and the seedling recruitment in laboratory conditions were examined during each season. As the values of height of individuals, number of fruits per stem, number of seeds per fruit, as well as the seedling abundance in some groups were not consistent with the normal distribution and the variances were not homogeneous, the non-parametric Kruskal-Wallis test was used in statistical analyses. Much greater number and density of individuals of Impatiens glandulifera were found in riparian forest, than in willow thickets and along the roadsides. In all sites the number and density of individuals increased steadily in consecutive seasons. The great number and density of Bidens frondosa individuals observed in shady and partly shady sites during the first year of studies raised in the second year and subsequently dramatically decreased in the third season. The lowest number and density of individuals noted in the first season in unshaded site raised substantially in subsequent years. The height of stems, as well as seed and fruit production of both taxa diminished with a decrease of height of neighboring plants. The seed dimensions presented the spatial and temporal variability, whereas the number of seedlings among consecutive years and in successive sites did not differ. The considerable height of the individuals of Impatiens glandulifera, high production of large fruits and seeds in open and dry roadside areas can contribute to more effective ballistic dissemination, while substantial seedling recruitment enables the colonization of new, perhaps more advantageous sites. On the other hand, lower individual height, as well as fruit and seed production and considerable seedling emergence allow the population to last and to gradually extend the area in forest communities. The considerable abundance of the high-statured Bidens frondosa individuals, substantial production of large capitula and achenes contribute to long persistence of populations in open and sun-lit sites. The gradually decrease in the height of the stems, achene number and size observed in partly-shaded and shaded places might allow to long-distance dispersal of seeds by animals, while substantial seedling recruitment might contribute to establishment in new areas.     

Physiology and methane productivity of Methanobacterium thermaggregans

Applied Microbiology and Biotechnology - Accumulation of carbon dioxide (CO2), associated with global temperature rise, and drastically decreasing fossil fuels necessitate the development...     

Nicotinic acid suppresses sebaceous lipogenesis of human sebocytes via activating hydroxycarboxylic acid receptor 2 (HCA2)

Nicotinic acid (NA) activates hydroxycarboxylic acid receptor 2 (HCA₂), and it is widely used in treating dyslipidaemias. Since its side effects include skin dryness, whereas its deficiency can be accompanied by dyssebacia, characterized by sebaceous gland enlargement, we asked if HCA₂ is expressed on human sebocytes, and if NA influences sebocyte functions. By using human immortalized SZ95 sebocytes, we found that non‐cytotoxic (≤100 μmol/L; MTT‐assay) concentrations of NA had no effect on the homeostatic sebaceous lipogenesis (SLG; Nile Red), but normalized excessive, acne‐mimicking SLG induced by several lipogenic agents (arachidonic acid, anandamide, linoleic acid + testosterone; Nile Red; 48‐hr treatments). Moreover, it exerted significant anti‐proliferative actions (CyQUANT‐assay), and increased [Ca²⁺]_IC (Fluo‐4 AM‐based Ca²⁺‐measurement). Although NA did not prevent the lipopolysaccharide‐induced pro‐inflammatory response (up‐regulation [Q‐PCR] and release [ELISA] of several pro‐inflammatory cytokines) of the sebocytes, collectively, these data support the concept that NA may be effective in suppressing sebum production in vivo. While exploring the mechanism of the sebostatic actions, we found that sebocytes express HCA₂ (Q‐PCR, immunofluorescent labelling), siRNA‐mediated silencing of which prevented the NA‐induced Ca²⁺‐signal and the lipostatic action. Collectively, our data introduce NA, and HCA₂ activators in general, as novel, potent and most likely safe sebostatic agents, with possible anti‐acne potential.     

“Fuzzy oil drop” model applied to individual small proteins built of 70 amino acids

The proteins composed of short polypeptides (about 70 amino acid residues) representing the following functional groups (according to PDB notation): growth hormones, serine protease inhibitors, antifreeze proteins, chaperones and proteins of unknown function, were selected for structural and functional analysis. Classification based on the distribution of hydrophobicity in terms of deficiency/excess as the measure of structural and functional specificity is presented. The experimentally observed distribution of hydrophobicity in the protein body is compared to the idealized one expressed by a three-dimensional Gauss function. The differences between these two distributions reveal the specificity of structural/functional characteristics of the protein. The residues of hydrophobicity deficiency versus the idealized distribution are assumed to indicate cavities with the potential to bind ligands, while the residues of hydrophobicity excess are interpreted as potentially participating in protein-protein complexation. The distribution of hydrophobicity irregularity seems to be specific for particular structures and functions of proteins. A comparative analysis of such profiles is carried out to identify the potential biological activity of proteins of unknown function.     

Pennogenyl Saponins from Paris quadrifolia L. Induce Extrinsic and Intrinsic Pathway of Apoptosis in Human Cervical Cancer HeLa Cells

Pennogenyl saponins are the active compounds of large number of plant species and consequently many polyherbal formulations. Hence, great interest has been shown in their characterization and in the investigation of their pharmacological and biological properties, especially anticancer. This present study reports on the evaluation of cytotoxic effects and explanation of the molecular mechanisms of action of the two pennogenyl saponins (PS 1 and PS 2) isolated from Paris quadrifolia L. rhizomes on human cervical adenocarcinoma cell line HeLa. To determine the viability of the cells treated with the compounds we used real-time cell proliferation analysis and found that the pennogenyl saponins PS 1 and PS 2 strongly inhibited the tumor cells growth with IC50 values of 1.11 ± 0.04 μg/ml and 0.87 ± 0.05 μg/ml, respectively. The flow cytometry analysis indicated that the two compounds induced apoptosis in a dose-dependent manner and decreased mitochondrial membrane potential in HeLa cells in the early stage of apoptosis. Quantitative PCR and Western Blot analysis showed that the two saponins significantly increased mRNA expression of FADD and BID as well as induced caspase-8 via increased of procaspase-8 processing in the treated cells. The results of this study suggest that both the extrinsic death receptor and intrinsic mitochondrial pathways are involved in the programmed cell death.     

Shugoshin is a Mad1/Cdc20-like interactor of Mad2

Mammalian centromeric cohesin is protected from phosphorylation-dependent displacement in mitotic prophase by shugoshin-1 (Sgo1), while shugoshin-2 (Sgo2) protects cohesin from separase-dependent cleavage in meiosis I. In higher eukaryotes, progression and faithful execution of both mitosis and meiosis are controlled by the spindle assembly checkpoint, which delays anaphase onset until chromosomes have achieved proper attachment to microtubules. According to the so-called template model, Mad1-Mad2 complexes at unattached kinetochores instruct conformational change of soluble Mad2, thus catalysing Mad2 binding to its target Cdc20. Here, we show that human Sgo2, but not Sgo1, specifically interacts with Mad2 in a manner that strongly resembles the interactions of Mad2 with Mad1 or Cdc20. Sgo2 contains a Mad1/Cdc20-like Mad2-interaction motif and competes with Mad1 and Cdc20 for binding to Mad2. NMR and biochemical analyses show that shugoshin binding induces similar conformational changes in Mad2 as do Mad1 or Cdc20. Mad2 binding regulates fine-tuning of Sgo2's sub-centromeric localization. Mad2 binding is conserved in the only known Xenopus laevis shugoshin homologue and, compatible with a putative meiotic function, the interaction occurs in oocytes.