Late rectal and bladder toxicity following radiation therapy for prostate cancer: Predictive factors and treatment results

Aim

This study aimed at investigating factors associated to late rectal and bladder toxicity following radiation therapy and the effectiveness of Hyperbaric Oxygen Therapy (HBOT) when toxicity is grade ≥2.

Background

Radiation is frequently used for prostate cancer, but a 5–20% incidence of late radiation proctitis and cystitis exists. Some clinical and dosimetric factors have been defined without a full agreement. For patients diagnosed of late chronic proctitis and/or cystitis grade ≥2 treatment is not well defined. Hyperbaric Oxygen Therapy (HBOT) has been used, but its effectiveness is not well known.

Materials and methods

257 patients were treated with radiation therapy for prostate cancer. Clinical, pharmacological and dosimetric parameters were collected. Patients having a grade ≥2 toxicity were treated with HBOT. Results of the intervention were measured by monitoring toxicity by Common Toxicity Criteria v3 (CTCv3).

Results

Late rectal toxicity was related to the volume irradiated, i.e. V50 > 53.64 (p = 0.013); V60 > 38.59% (p = 0.005); V65 > 31.09% (p = 0.002) and V70 > 22.81% (p = 0.012). We could not correlate the volume for bladder. A total of 24 (9.3%) patients experienced a grade ≥2. Only the use of dicumarinic treatment was significant for late rectal toxicity (p = 0.014). A total of 14 patients needed HBOT. Final percentage of patients with a persistent toxicity grade ≥2 was 4.5%.

Conclusion

Rectal volume irradiated and dicumarinic treatment were associated to late rectal/bladder toxicity. When toxicity grade ≥2 is diagnosed, HBOT significantly ameliorate symptoms.     

The impact of red and blue light-emitting diode illumination on radish physiological indices

The objective was to evaluate the effect of different combinations of red (638 nm) and blue (455 nm) light produced by solid-state light-emitting diodes (LEDs) on physiological indices (net assimilation rate, hypocotyl-to-leaf ratio, leaf area, leaf dry weight, hypocotyl length and diameter, plant length, developed leaves), variation of photosynthetic pigments and non-structural carbohydrates in radish (Raphanus sativus L., var. ‘Faraon’). Lighting experiments were performed under controlled conditions (total PPFD - 200 μmol m⁻² s⁻¹; 16 h photoperiod; 14/18°C night/day temperature). The LED conditions: 638 nm; 638 + 5% 455 nm; 638 + 10% 455 nm; 638 + 10% 455 + 731 nm; 638 + 10% 455 + 731 + 669 nm. Our results showed that radishes grown under red (638 nm) alone were elongated, and the formation of hypocotyl was weak. The net assimilation rate, hypocotyl-to-leaf ratio, and leaf dry weight also were low due to the low accumulation of photosynthetic pigments and non-structural carbohydrates in leaves. The supplemented blue (455 nm) light was necessary for the non-structural carbohydrates distribution between radish storage organs and leaves which resulted in hypocotyl thickening. Red alone (638 nm) or in combination with far-red (731 nm), or red669 for radish generative development was required.     

A Novel Role of the N Terminus of Budding Yeast Histone H3 Variant Cse4 in Ubiquitin-Mediated Proteolysis

Understanding the molecular basis of common traits is a primary challenge of modern genetics. One model holds that rare mutations in many genetic backgrounds may often phenocopy one another, together explaining the prevalence of the resulting trait in the population. For the vast majority of phenotypes, the role of rare variants and the evolutionary forces that underlie them are unknown. In this work, we use a population of Saccharomyces paradoxus yeast as a model system for the study of common trait variation. We observed an unusual, flocculation and invasive-growth phenotype in one-third of S. paradoxus strains, which were otherwise unrelated. In crosses with each strain in turn, these morphologies segregated as a recessive Mendelian phenotype, mapping either to IRA1 or to IRA2, yeast homologs of the hypermutable human neurofibromatosis gene NF1. The causal IRA1 and IRA2 haplotypes were of distinct evolutionary origin and, in addition to their morphological effects, associated with hundreds of stress-resistance and growth traits, both beneficial and disadvantageous, across S. paradoxus. Single-gene molecular genetic analyses confirmed variant IRA1 and IRA2 haplotypes as causal for these growth characteristics, many of which were independent of morphology. Our data make clear that common growth and morphology traits in yeast result from a suite of variants in master regulators, which function as a mutation-driven switch between phenotypic states.     

Effect of genotype and medium composition on linseed (<Emphasis Type="Italic">Linum usitatissimum</Emphasis>) ovary culture

Breeding linseed (Linum usitatissimum L.) using haploid techniques allows breeders to develop new cultivars in a shorter time period. Many research groups successfully created new linseed genotypes through anther culture; however ovary culture has been the subject of only a few earlier studies. In the present study, the effect of genotype and growth regulators combination on callus induction and shoots regeneration in ovary culture of nine commercially important linseed cultivars was investigated. Ovaries were cultured on modified MS medium supplemented with three different combinations of plant growth regulators. Variable callogenic responses were expressed by all of the genotypes tested on different induction media. The results suggested that specific combination of growth regulators for callus induction must be designed for each genotype. Shoot regeneration from ovary derived callus is a critical phase of the whole gynogenetic process. Differences in adventitious shoot formation frequency among genotypes were demonstrated and four responsive genotypes have been selected. Ovary derived callus from cultivar ‘Mikael’ manifested the highest adventitious shoot formation frequency with a high number of shoots per explant. The optimum ratio of growth regulators for shoot regeneration was shown to depend on the genotype. Cultivars ‘Linola’, ‘Mikael’ and ‘Szaphir’ showed the highest shoot regeneration frequency when callus had originated on induction medium supplemented with 2 mg L⁻¹ BAP and 2 mg L⁻¹ NAA, while combination of 1 mg L⁻¹ BAP and 2 mg L⁻¹ IAA promoted shoot formation in ovary-derived callus of ‘Barbara’. The highest rate of shoots per explant has been obtained in second subculture.     

Cell Biology of the Endoplasmic Reticulum and the Golgi Apparatus through Proteomics

Enriched endoplasmic reticulum (ER) and Golgi membranes subjected to mass spectrometry have uncovered over a thousand different proteins assigned to the ER and Golgi apparatus of rat liver. This, in turn, led to the uncovering of several hundred proteins of poorly understood function and, through hierarchical clustering, showed that proteins distributed in patterns suggestive of microdomains in cognate organelles. This has led to new insights with respect to their intracellular localization and function. Another outcome has been the critical testing of the cisternal maturation hypothesis showing overwhelming support for a predominant role of COPI vesicles in the transport of resident proteins of the ER and Golgi apparatus (as opposed to biosynthetic cargo). Here we will discuss new insights gained and also highlight new avenues undertaken to further explore the cell biology of the ER and the Golgi apparatus through tandem mass spectrometry.Most biosynthetic proteins destined for the plasma membrane and secretion, as well as resident proteins of endosomes, lysosomes, the endoplasmic reticulum (ER), and the Golgi apparatus originate with cotranslational translocation into the ER. This is followed by carefully controlled folding and quality control. The ER is also a major site for sensing cellular stress and for cholesterol and phospholipid biosynthesis and constitutes a vast continuous endomembrane system that often pervades the entire cytoplasm. The ER commences as the nuclear envelope extends to rough (ribosome-studded) membranes, and ends with tripartite-like structures of tubular smooth (ribosome-free) ER.The ratio of rough and smooth ER is cell type specific. Stem cells have little rough ER and smooth ER (Murphy et al. 1971; Cheng and Leblond 1974), whereas cells highly specialized in protein secretion, such as in the exocrine pancreas, have extensive rough ER with little to no smooth ER (Palade and Siekevitz 1956; Jamieson and Palade 1967). By contrast, liver parenchymal hepatocytes have a near equal abundance of rough and smooth ER with the latter associated with glycogen storage and elimination of exogenous steroids through a multitude of P450-driven oxidation pathways (Bruni and Porter 1965; Loud 1968; Estabrook et al. 1971; Blouin et al. 1977; Reed and Backes 2012). At the other extreme, Leydig cells of the testis are specialized in cholesterol metabolism to form testosterone and are enriched with smooth ER (Mori and Christensen 1980). Emanating from the ER of all cell types in mammals are discrete export sites that are distributed throughout the cell, sometimes numbering in the hundreds. ER export is controlled by the COPII coat machinery, which has been characterized through proteomics analyses in yeast (Otte et al. 2001) and is coupled to microtubule- and dynein/dynactin-dependent transport directed toward the central juxta-nuclear Golgi apparatus (COPII vesicles are discussed in Lord et al. [2013]). This occurs in vesicular/tubular clusters (VTCs) that seemingly undergo a maturation/distillation process (Saraste and Kuismanen 1984) such that when arriving at the cis-face Golgi stacks, biosynthetic cargo appears more concentrated. This is achieved, at least in part, through a continuous removal of ER export machinery components (e.g., p58, p24, and SNARE proteins) that are then returned to the ER via vesicular transport intermediates controlled by the COPI coat machinery. This apparent “distillation” process continues throughout the secretory pathway from as far as the trans-part of the Golgi apparatus (Miesenbock and Rothman 1995). Whether or not such distillation occurs through intra-Golgi cisternal transport or through direct delivery to the ER is part of ongoing investigation, including the extent of recycling (see below).The Golgi apparatus is morphologically distinct from the ER because of its juxtanuclear position, and at the ultrastructural level, appears as a ribbon-like structure of laterally interconnected stacks of flattened cisternae, each having a network of extensive fenestrated membranes associated both at their cis- and trans-face. Biosynthetic cargo here, undergo extensive posttranslational modifications including the maturation of N-linked oligosaccharides, and the addition of O-linked oligosaccharides. At the trans-face, cargo is sorted and packaged for transport either to the plasma membrane or to the endosomal endomembrane system. Some specialized cargo is also packaged and concentrated into dedicated membrane structures for regulated secretion.The functional and morphological demarcation between the ER and the Golgi apparatus is usually assumed to be complete, giving rise to the notion of two independent organelles—each with their own distinct functions. Because of the extensive recycling that takes place, however, the two organelles appear functionally intertwined. Indeed, inhibition of the Golgi-located ARF1 guanine nucleotide exchange factor GBF1 results in a rapid collapse of the Golgi apparatus into the ER (Misumi et al. 1986; Oda et al. 1987; Fujiwara et al. 1988; Lippincott-Schwartz et al. 1989; Claude et al. 1999). As ARF1^GDP to ARF1^GTP conversion is obligatory to COPI recruitment and vesicle formation and subsequent recycling, the effect of BFA seems at first counter intuitive, but can be explained through the additional role of COPI as a protective coat on cisternal membranes, i.e., BFA-induced dissociation of COPI coat from Golgi membranes enables uncontrolled formation of Golgi-derived tubules that within minutes fuse with the ER, collapsing most of the Golgi into the ER. Strikingly, removal of BFA equally rapidly leads to the reformation of individual functional Golgi stacks positioned at the various ER exit sites and after microtubule-dependent transport, coalesce into the central and juxtanuclear Golgi apparatus. Thus, the two organelles are temporally as well as spatially linked to enable productive communication and trafficking for the process of protein secretion, as well as lipid biosynthesis. This link gives rise to some unexpected functions (see below).     

LED irradiance level affects growth and nutritional quality of Brassica microgreens

This study examines the effect of irradiance level produced by solid-state light-emitting diodes (LEDs) on the growth, nutritional quality and antioxidant properties of Brassicaceae family microgreens. Kohlrabi (Brassica oleracea var. gongylodes, ‘Delicacy Purple’) mustard (Brassica juncea L., ‘Red Lion’), red pak choi (Brassica rapa var. chinensis, ‘Rubi F₁’) and tatsoi (Brassica rapa var. rosularis) were grown using peat substrate in controlled-environment chambers until harvest time (10 days, 21/17°C, 16 h). A system of five lighting modules with 455, 638, 665 and 731 nm LEDs at a total photosynthetic photon flux densities (PPFD) of 545, 440, 330, 220 and 110 µmol m^?2s^?1 respectively were used. Insufficient levels of photosynthetically active photon flux (110 µmol m^?2 s^?1) suppressed normal growth and diminished the nutritional value of the Brassica microgreens studied. In general, the most suitable conditions for growth and nutritional quality of the microgreens was 330–440 µmol m^?2 s^?1 irradiation, which resulted in a larger leaf surface area, lower content of nitrates and higher total anthocyanins, total phenols and 2,2-diphenyl-1-picrylhydrazyl (DPPH) free-radical scavenging capacity. High light levels (545 µmol m^?2 s^?1), which was expected to induce mild photostress, had no significant positive impact for most of investigated parameters.     

A Novel Lentinula edodes Laccase and Its Comparative Enzymology Suggest Guaiacol-Based Laccase Engineering for Bioremediation

Laccases are versatile biocatalysts for the bioremediation of various xenobiotics, including dyes and polyaromatic hydrocarbons. However, current sources of new enzymes, simple heterologous expression hosts and enzymatic information (such as the appropriateness of common screening substrates on laccase engineering) remain scarce to support efficient engineering of laccase for better “green” applications. To address the issue, this study began with cloning the laccase family of Lentinula edodes. Three laccases perfectio sensu stricto (Lcc4A, Lcc5, and Lcc7) were then expressed from Pichia pastoris, characterized and compared with the previously reported Lcc1A and Lcc1B in terms of kinetics, stability, and degradation of dyes and polyaromatic hydrocarbons. Lcc7 represented a novel laccase, and it exhibited both the highest catalytic efficiency (assayed with 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) [ABTS]) and thermostability. However, its performance on “green” applications surprisingly did not match the activity on the common screening substrates, namely, ABTS and 2,6-dimethoxyphenol. On the other hand, correlation analyses revealed that guaiacol is much better associated with the decolorization of multiple structurally different dyes than are the two common screening substrates. Comparison of the oxidation chemistry of guaiacol and phenolic dyes, such as azo dyes, further showed that they both involve generation of phenoxyl radicals in laccase-catalyzed oxidation. In summary, this study concluded a robust expression platform of L. edodes laccases, novel laccases, and an indicative screening substrate, guaiacol, which are all essential fundamentals for appropriately driving the engineering of laccases towards more efficient “green” applications.     

Clinical Significance of Frizzled Homolog 3 Protein in Colorectal Cancer Patients

Frizzled homolog 3 receptor was up-regulated in several gastrointestinal cancers such as esophageal and gastric cancers. Moreover, frizzled homolog 3 has recently reported to be expressed in colorectal adenoma specimens. In the present study, we investigated the clinical significance of frizzled homolog 3 protein in colorectal cancer patients. Using immunocytochemical staining, frizzled homolog 3 expression was examined in 186 colorectal cancer specimens, 79 colorectal adenoma specimens, 133 colorectal polyp specimens, 127 colorectal cancer specimens with lymph node and/or distant metastasis, 310 specimens of various non-colorectal cancer metastatic carcinomas and 40 specimens with simultaneous occurrence of colorectal cancer, colorectal adenoma and colorectal polyp. Statistical analysis was used to correlate frizzled homolog 3 protein expression to the clinicohistopathological factors, recurrence/metastasis and survival after follow-up for 42 months in colorectal cancer patients. Frizzled homolog 3 protein was expressed in 100% colorectal cancer specimens, 89% colorectal adenoma specimens, 75% colorectal polyp specimens and 69% normal colorectal epithelial tissues. Moreover, frizzled homolog 3 immunocytochemical scores were highly correlated with colorectal cancer progression. Furthermore, frizzled homolog 3 was expressed in a comparatively lower percentage of metastatic hepatocellular carcinoma and metastatic renal clear cell carcinoma with focal and very weak staining than other metastatic tumor types. On the other hand, the frizzled homolog 3 immunocytochemical scores of colorectal adenomas with synchronous colorectal carcinomas were significantly higher than those of pure colorectal adenomas. Statistical analysis showed that frizzled homolog 3 immunocytochemical scores were associated with Dukes stage and lymph node status. Finally, stratified groups of colorectal cancer patients had significant differences in their recurrence/metastasis and survival. In conclusion, the present large-scale study has clearly showed that frizzled homolog 3 protein can generate clinically important information for colorectal cancer patients.