High superoxide dismutase and low glutathione peroxidase activities in red blood cells predict susceptibility of lung cancer patients to radiation pneumonitis

Radiation pneumonitis is an unpredictable complication of radiotherapy for lung cancer and a condition which can cause significant morbidity. The ability to identify patients at a high risk of developing pneumonitis is critical, since it will enable the individualization of the treatment plan. Because the cytotoxic effect of radiation is propagated through reactive oxygen species (ROS) and ROS-driven oxidative stress, the role of antioxidant defense systems in radiation pneumonitis was investigated. Using the pneumonitis-sensitive C3H/HeN mice as a model, we demonstrated that the antioxidant response of the lung correlated well with that of red blood cells (RBC). We then proceeded to test whether differences of RBC antioxidant response would predict the pneumonitis development in patients. Superoxide dismutase (SOD), glutathione peroxidase (GPX), and catalase (CAT) activities and glutathione in RBC were measured at baseline and then weekly for 6 weeks of treatment in 15 eligible patients receiving concurrent chemo-radiotherapy for unresectable stage III NSCLC. Striking differences were found in the antioxidant activities of RBC with respect to the pneumonitis development. Those who developed pneumonitis showed higher SOD and lower GPX activities at baseline compared to those who did not (3.7 vs 6.8 units/mg for median SOD, 16.5 vs 10.7 nmol/min/mg for median GPX). The functional imbalance of SOD and GPX was displayed consistently throughout the treatment period. The sensitivity and specificity of pneumonitis prediction were further increased when the GPX/SOD ratio was analyzed (pretreatment P = 0.0046). Our results provide a strong rationale to monitor SOD and GPX activities of RBC to identify patients who are at risk of developing pneumonitis, and to implement a strategy of increasing the GPX/SOD ratio in order to lower the risk.     

Assessment and evaluation of MV image guidance system performance in radiotherapy

Background and aim

The clinical use of imaging system in image guided radiotherapy (IGRT) necessitates performing periodic quality assurance of the system to be confident in applying corrections for patient set-up errors. We aim to develop and implement a quality assurance (QA) programme for megavoltage (MV) based image guidance system and assess its long term performance for a period of 3 years.

Materials and methods

Periodic QA tests were performed for the MV planar and cone beam computed tomography (CBCT) imaging system to assess the system safety, mechanical and geometrical accuracy, image quality and dose. The tests were performed using the equipment supplied by the manufacturer along with the image guidance system and using simple methods developed in-house. The test results were compared with expected or baseline values established during commissioning.

Results

The safety system was found to be functional. The results of mechanical and geometrical tests were in good agreement with the expected results. The system mechanical positioning was stable and reproducible within ±2 mm accuracy. The image quality and the imaging dose of the planar and CBCT imaging were found to agree with the baseline values and the manufacturer specifications.

Discussion

Throughout the three-year period, all the QA tests were within the specification. The mechanical and geometrical tests are most crucial as they directly affect the patient positioning accuracy.

Conclusion

We conclude that the MV image guidance system is efficient to perform IGRT and insist to perform periodic QA tests and calibration for the system.     

Effect of hydrogen bond strength on the redox properties of phylloquinones: a two-dimensional hyperfine sublevel correlation spectroscopy study of photosystem I

The phylloquinones of photosystem I (PS I), A(1A) and A(1B), exist in near-equivalent protein environments but possess distinct thermodynamic and kinetic properties. Although the determinants responsible for the different properties of the phylloquinones are not completely understood, the strength and geometry of hydrogen bond interactions are significant factors in tuning and control of function. This study focuses on characterizing the hydrogen-bonding interactions of the phylloquinone acceptor, A(1A), by (1)H and (14)N HYSCORE spectroscopy. Photoaccumulation of PS I complexes at pH 8.0 results in the trapping of the phyllosemiquinone anion, A(1A)(-), on the A-branch of cofactors. The experiments described here indicate that A(1A)(-) forms a single H-bond. Using a simple point dipole approximation, we estimate its length to be 1.6 ± 0.1 ?. The value of the (1)H isotropic hyperfine coupling constant suggests that the H-bond has significant out-of-plane character. The (14)N HYSCORE spectroscopy experiments support the assignment of a H-bond wherein, the (14)N quadrupolar coupling constant is consistent with a backbone amide nitrogen as the hydrogen bond donor.     

Poly <Emphasis Type="Italic">N</Emphasis>-acetyllactosamine substitutions on <Emphasis Type="Italic">N</Emphasis>- and not <Emphasis Type="Italic">O</Emphasis>-oligosaccharides or Thomsen–Friedenreich antigen facilitate lung specific metastasis of melanoma cells via galectin-3

Galectin-3 on vascular endothelium has been shown to facilitate lung specific metastasis. Metastatic variants of B16 melanoma were chosen to identify specific ligands that mediate lung colonization via galectin-3. Flow cytometry showed that, galectin-3 binding to cells correlates with surface expression of poly N-acetyllactosamine (polylacNAc) but not with other reported ligands, e.g. Thomsen-Friedenreich (T/Tn) antigen. Immobilized galectin-3 promoted adhesion of melanoma cells in a metastasis dependent manner. Moreover, adhesion and galectin-3 binding to cells were specifically inhibited with lactose. These properties together with lung metastasis were inhibited with N-glycosylation inhibitor Swainsonine (SW), whereas, O-glycosylation inhibitor Benzyl-α-N-acetylgalactosamine (BG) had no effect. BG treatment significantly increased expression of T/Tn antigen on low metastatic cells; however, had no effect on their metastatic potential. The studies very comprehensively demonstrate the importance of polylacNAc substitutions on N-oligosaccharides in galectin-3 mediated lung metastasis. Electronic supplementary material  The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

Immunocytochemical identification of crustacean hyperglycemic hormone-producing cells in the brain of a freshwater fairy shrimp,Streptocephalus dichotomus Baird (Crustacea: Anostraca)

Localization of crustacean hyperglycemic hormone (CHH) activity in the brain ganglia of Streptocephalus dichotomus was demonstrated by immunocytochemical method. For this, two different rabbit antisera, one raised against a crayfish Orconectus limosus, and the other with a crab Carcinus maenas, were used. Positive immunoreactivity was recorded with the antibody raised against Orconectus limosus CHH. However, the antibody raised against the CHH of the crab, Carcinus maenas failed to show any cross-reactivity. The biological specificity of these peptides was further confirmed by immunoblotting and immunodiffusion studies performed with the partially purified CHH. The results obtained with the immunodot-blotting and immunodiffusion studies with CHH, further confirmed the immunocytochemical studies. Bioassay experiments performed with a freshwater prawn, Macrobrachium rosenbergii indicate the interspecific biological activity of this neuropeptide. The present study indicates that besides the sinus gland, there are other sites of CHH synthesis and release in the brain ganglia of S. dichotomus.     

Expression of epithelial calcium transport system in rat cochlea and vestibular labyrinth

Background  

The low luminal Ca²⁺ concentration of mammalian endolymph in the inner ear is required for normal hearing and balance. We recently reported the expression of mRNA for a Ca²⁺-absorptive transport system in primary cultures of semicircular canal duct (SCCD) epithelium.     

Biochemical characterisation of grain mould resistant and susceptible genotypes and PGPR-induced resistance in the host to Curvularia lunata and Fusarium proliferatum

Resistance to biotic stresses in plants is either due to the presence of preformed biochemical compounds or induced in response to external stimulus. In this study, 13 grain mould resistant and seven susceptible lines of sorghum were analysed for biochemical defence mechanism. The levels of total phenols and phenylalanine ammonia lyase were almost same in the resistant and susceptible genotypes. However, two additional isoforms of peroxidase were found in the three of the 13 resistant genotypes. The isoform peroxidase corresponding to the R _f value of 0.25 was found in the resistant genotypes IS 13969, ICSB 377 and IS 8219-1, and two genotypes IS 13969 and ICSB 377 had an additional isoform corresponding to the R _f value of 0.32. The results indicated the genotype specific association of peroxidases with grain mould resistance in sorghum. Nine bacterial strains (Bacillus pumilus SB 21, Bacillus megaterium HiB 9, Bacillus subtilis BCB 19, Pseudomonas plecoglossicida SRI 156, Brevibacterium antiquum SRI 158, B. pumilus INR 7, P. fluorescens UOM SAR 80, P. fluorescens UOM SAR 14, B. pumilus SE 34) were tested to induce systemic resistance in sorghum cultivars 296B and Bulk Y against the highly pathogenic grain mould pathogens Curvularia lunata and Fusarium proliferatum, respectively. The bacterial isolates were effective in inducing resistance in sorghum. Among the strains tested, SRI 158 was found highly effective in reducing grain mould severity in both the genotypes.     

Screening of endophytic bacteria isolated from leaves of Sambung Nyawa [Gynura procumbens (Lour.) Merr.] for cytokinin-like compounds

Endophytic bacteria are harmless in most plant species; and known to boost the growth and development of the host plants probably by secreting growth hormones. The isolation, identification and screening of endophytic bacteria for the plant growth regulators like cytokinin are needed to get the leads for their applications in agriculture sector. We describe the isolation and identification of the bacterial endophytes from the leaves of Sambung Nyawa [Gynura procumbens (Lour.) Merr.] and their screening for cytokinin-like compounds. We isolated three endophytic bacteria from the leaves of G. procumbens collected from the forest research institute of Malaysia (FRIM). They were further identified using amplified 16S rRNA gene sequence based method of bacterial identification. The ethyl acetate extracts of the isolates-broth were analyzed using cucumber cotyledon greening bioassay (CCGB) to determine the presence of cytokinin-like compounds. Consequently, the bacterial putative endophytes were identified as Psuedomonas resinovorans, Paenibacillus polymaxa, and Acenitobacter calcoaceticus. Broth-extracts from two (Psuedomonas resinovorans and Paenibacillus polymaxa) of the three putative bacterial endophytes show the positive results in their screening for cytokinin-like compounds using CCGB. Thus, we hypothesize that the bacterial putative endophytes of G. procumbens that produce cytokinin-like compounds might have a role in the growth and development of G. procumbens. ABBREVIATIONS: CCGB - Cucumber cotyledon greening bioassay, rDNA - Ribosomal DNA, K12, BAP - 6-Benzylaminopurine, Db1, MSA - Multiple sequence alignment. 8081.