Evaluation of combining bony anatomy and soft tissue position correction strategies for IMRT prostate cancer patients

Background

Radiotherapy treatment requires delivering high homogenous dose to target volume while sparing organs at risk. That is why accurate patient positioning is one of the most important steps during the treatment process. It reduces set-up errors which have a strong influence on the doses given to the target and surrounding tissues.

Aim

The aim of this study was to investigate the efficiency of combining bony anatomy and soft tissue imaging position correction strategies for patients with prostate cancer.

Materials and methods

The study based on pre-treatment position verification results determined for 10 patients using kV images and CBCT match. At the same patients’ position, two orthogonal kV images and set of CT scans were acquired. Both verification methods gave the information about patients’ position changes in vertical, longitudinal and lateral directions.

Results

For 93 verifications, the mean values of kV shifts in vertical, longitudinal and lateral directions equaled: −0.11 ± 0.54 cm, 0.26 ± 0.38 cm and −0.06 ± 0.47 cm, respectively. The same values achieved for CBCT matching equaled: 0.07 ± 0.62 cm, 0.22 ± 0.36 cm and −0.02 ± 0.45 cm. Statistically significant changes between the values of shifts received during the first week of treatment and the rest time of the irradiation process were found for 2 patients in the lateral direction and 2 patients in vertical direction among kV results and for 3 patients in the longitudinal direction among CBCT results. A significant difference between kV and CBCT match results was found in the vertical direction.

Conclusions

In clinical practice, CBCT combined with kV or even portal imaging improves precision and effectiveness of prostate cancer treatment accuracy.     

Decreased hypertrophic differentiation accompanies enhanced matrix formation in co-cultures of outer meniscus cells with bone marrow mesenchymal stromal cells

Introduction

The main objective of this study was to determine whether meniscus cells from the outer (MCO) and inner (MCI) regions of the meniscus interact similarly to or differently with mesenchymal stromal stem cells (MSCs). Previous study had shown that co-culture of meniscus cells with bone marrow-derived MSCs result in enhanced matrix formation relative to mono-cultures of meniscus cells and MSCs. However, the study did not examine if cells from the different regions of the meniscus interacted similarly to or differently with MSCs.

Methods

Human menisci were harvested from four patients undergoing total knee replacements. Tissue from the outer and inner regions represented pieces taken from one third and two thirds of the radial distance of the meniscus, respectively. Meniscus cells were released from the menisci after collagenase treatment. Bone marrow MSCs were obtained from the iliac crest of two patients after plastic adherence and in vitro culture until passage 2. Primary meniscus cells from the outer (MCO) or inner (MCI) regions of the meniscus were co-cultured with MSCs in three-dimensional (3D) pellet cultures at 1:3 ratio, respectively, for 3 weeks in the presence of serum-free chondrogenic medium containing TGF-β1. Mono-cultures of MCO, MCI and MSCs served as experimental control groups. The tissue formed after 3 weeks was assessed biochemically, histochemically and by quantitative RT-PCR.

Results

Co-culture of inner (MCI) or outer (MCO) meniscus cells with MSCs resulted in neo-tissue with increased (up to 2.2-fold) proteoglycan (GAG) matrix content relative to tissues formed from mono-cultures of MSCs, MCI and MCO. Co-cultures of MCI or MCO with MSCs produced the same amount of matrix in the tissue formed. However, the expression level of aggrecan was highest in mono-cultures of MSCs but similar in the other four groups. The DNA content of the tissues from co-cultured cells was not statistically different from tissues formed from mono-cultures of MSCs, MCI and MCO. The expression of collagen I (COL1A2) mRNA increased in co-cultured cells relative to mono-cultures of MCO and MCI but not compared to MSC mono-cultures. Collagen II (COL2A1) mRNA expression increased significantly in co-cultures of both MCO and MCI with MSCs compared to their own controls (mono-cultures of MCO and MCI respectively) but only the co-cultures of MCO:MSCs were significantly increased compared to MSC control mono-cultures. Increased collagen II protein expression was visible by collagen II immuno-histochemistry. The mRNA expression level of Sox9 was similar in all pellet cultures. The expression of collagen × (COL10A1) mRNA was 2-fold higher in co-cultures of MCI:MSCs relative to co-cultures of MCO:MSCs. Additionally, other hypertrophic genes, MMP-13 and Indian Hedgehog (IHh), were highly expressed by 4-fold and 18-fold, respectively, in co-cultures of MCI:MSCs relative to co-cultures of MCO:MSCs.

Conclusions

Co-culture of primary MCI or MCO with MSCs resulted in enhanced matrix formation. MCI and MCO increased matrix formation similarly after co-culture with MSCs. However, MCO was more potent than MCI in suppressing hypertrophic differentiation of MSCs. These findings suggest that meniscus cells from the outer-vascular regions of the meniscus can be supplemented with MSCs in order to engineer functional grafts to reconstruct inner-avascular meniscus.     

Synthesis of immunoreagents for detection of deoxypyrrololine, a cross-link of bone collagen

Two immunogens (5,6) and two probes (fluorescent 7 and chemiluminescent 8) were prepared from benzyl ester (-)-10. These immunoreagents (5,6 and 7,8) are useful for detection of collagen cross-link (+)-deoxypyrrololine (Dpl, 4), and for development of assays for osteoporosis.     

Minicollagen-15, a novel minicollagen isolated from Hydra, forms tubule structures in nematocysts

Minicollagens constitute a family of unusually short collagen molecules isolated from cnidarians. They are restricted to the nematocyst, a cylindrical explosive organelle serving in defense and capture of prey. The nematocyst capsule contains a long tubule inside of its matrix, which is expelled and everted during an ultrafast discharge process. Here, we report the cloning and characterization of a novel minicollagen in Hydra, designated minicollagen-15 (NCol-15). NCol-15, like NCol-3 and NCol-4, shows deviations from the canonical cysteine pattern in its terminal cysteine-rich domains (CRDs). Minicollagens share common domain architectures with a central collagen sequence flanked by polyproline stretches and short N- and C-terminal CRDs. The CRDs are involved in the formation of a highly resistant cysteine network, which constitutes the basic structure of the nematocyst capsule. Unlike NCol-1, which is part of the capsule wall, NCol-15 is localized to tubules, arguing for a functional differentiation of minicollagens within the nematocyst architecture. NMR analysis of the altered C-terminal CRD of NCol-15 showed a novel disulfide-linked structure within the cysteine-containing region exhibiting similar folding kinetics and stability as the canonical CRDs. Our data provide evidence for evolutionary diversification among minicollagens, which probably facilitated alterations in the morphology of the nematocyst wall and tubule.     

Microbial production host selection for converting second-generation feedstocks into bioproducts

Although many secondary metabolites with diverse biological activities have been isolated from myxobacteria, most strains of these biotechnologically important gliding prokaryotes remain difficult to handle genetically. In this study we describe the new fast growing myxobacterial thermophilic isolate GT-2 as a heterologous host for the expression of natural product biosynthetic pathways isolated from other myxobacteria. According to the results of sequence analysis of the 16S rDNA, this moderately thermophilic isolate is closely related to Corallococcus macrosporus and was therefore named C. macrosporus GT-2. Fast growth of moderately thermophilic strains results in shorter fermentation and generation times, aspects which are of significant interest for molecular biological work as well as production of secondary metabolites. Development of a genetic manipulation system allowed the introduction of the complete myxochromide biosynthetic gene cluster, located on a transposable fragment, into the chromosome of GT-2. Genetic engineering of the biosynthetic gene cluster by promoter exchange leads to much higher production of myxochromides in the heterologous host C. macrosporus GT-2 in comparison to the original producer Stigmatella aurantiaca and to the previously described heterologous host Pseudomonas putida (600 mg/L versus 8 mg/L and 40 mg/L, respectively).     

Ingestible insecticides for spotted wing Drosophila control: a polyol,Erythritol, and an insect growth regulator,Lufenuron

Bioassays tested insecticidal activity of Erythritol from the nutritive sweetener, Truvia, and an insect growth regulator, Lufenuron, against life stages (eggs, larvae, pupae, adults) of Drosophila melanogaster (Meigen) and Drosophila suzukii (Matsumura), the spotted wing Drosophila (SWD). These compounds were chosen for their demonstrated acute toxicity to adult and larval Drosophila and potential use on organic fruit farms. D. melanogaster fed on standard Drosophila diet media moistened with water containing known concentrations of Erythritol. Likewise, SWD consumed standard diet media as well as thawed host fruit (blackberries and blueberries) treated with solutions of Erythritol, Lufenuron or both. During the first bioassay, Erythritol at lower concentrations between 0 and 500 mm (~61 000 ppm) in water and mixed with instant diet media increased adult survival from ~80% to 97% for D. melanogaster and SWD. However, from aqueous concentrations ranging from 1750 (~414 000 ppm) to 2000 mm (~244 000 ppm), Erythritol killed 100% of adult Drosophila in culture vials. One hundred per cent mortality for SWD and D. melanogaster occurred at ≥0.5 m (~61 000 ppm) Erythritol added to diet media or topically applied to host fruit. In a second bioassay, 0.013–1.000 ppm of aqueous Lufenuron, a chitin synthase inhibitor, when added to dry diet media prevented 90–99% of SWD from reaching the pupal stage. In another assay, ~67% of SWD eggs or neonates (early first instars) died inside blackberries pre‐treated with (dipped in) a soapy solution of 10 ppm Lufenuron. Pre‐treating blackberry fruit with an Erythritol–Lufenuron mixture reduced SWD brood survival by 99%. Likewise, during our last fruit‐based bioassay, 98% of eggs and neonates died inside blueberries similarly pre‐treated. During the last experiment, Lufenuron in diet media also rendered adult females sterile. Sterility, however, dissipated over 7 days once females began feeding on a Lufenuron‐free diet media.     

Splinting or surgery for carpal tunnel syndrome? Design of a randomized controlled trial [ISRCTN18853827]

Background  

Carpal tunnel syndrome is a common disorder, which can be treated with surgery or conservative options. However, there is insufficient evidence and no consensus among physicians with regard to the preferred treatment for carpal tunnel syndrome. Therefore, a randomized controlled trial is conducted to compare the short- and long-term efficacy of surgery and splinting in patients with carpal tunnel syndrome. An attempt is also made to avoid the (methodological) limitations encountered in earlier trials on the efficacy of various treatment options for carpal tunnel syndrome.     

O-(Fluoresceinylmethyl)hydroxylamine (OFMHA): A fluorescent reagent for detection of damaged nucleic acids

5- and 6-O-(Fluoresceinylmethyl)hydroxylamine (OFMHA, 5a, b) were prepared from the corresponding bis-pivaloyl-protected hydroxymethylfluoresceins (1a, b) in 50–70% yield. The hydroxylamine derivatives reacted smoothly with the abasic sites present in acid/heat stressed calf thymus DNA.     

The genus Pyrus L. (Rosaceae) in south-west Europe and North Afkica

A multivariate morphometric study of the genus Pyrus in south-west Europe and North Africa shows that five species may be recognized in the area: P. bourgaeana Decne., P. communis L., P. cordata Dew., P. spinosa Forssk, and P. nivalis Jacq. Some valuable characters for identification of these species are proposed. In particular the width of fruit peduncle, petal size, leaf width and petiole length served to discriminate the taxa. Several names such as P. gharbiona Trab., P. cossonii Rehder (|M= P. longipes Balansa ex Coss. & Durieu) and P. boisseriana Buhse, are regarded as synonyms of P. cordata , while P. marnormis Trab. of P. bourgaeana. Consequently a check-list and a key to these species are provided.