Acid hydrolysis of wheat gluten induces formation of new epitopes but does not enhance sensitizing capacity by the oral route: a study in "gluten free" Brown Norway rats.

BACKGROUND: Acid hydrolyzed wheat proteins (HWPs) are used in the food and cosmetic industry as emulsifiers. Cases of severe food allergic reactions caused by HWPs have been reported. Recent data suggest that these reactions are caused by HWPs produced by acid hydrolysis. OBJECTIVES: To examine the sensitizing capacity of gluten proteins per se when altered by acid or enzymatic hydrolysis relative to unmodified gluten in rats naïve to gluten. METHODS: High IgE-responder Brown Norway (BN) rats bred on a gluten-free diet were sensitized without the use of adjuvant to three different gluten products (unmodified, acid hydrolyzed and enzymatic hydrolyzed). Rats were sensitized by intraperitoneal (i.p.) immunization three times with 200 µg gluten protein/rat or by oral dosing for 35 days with 0.2, 2 or 20 mg gluten protein/rat/day. Sera were analyzed for specific IgG and IgE and IgG-binding capacity by ELISA. IgE functionality was measured by rat basophilic leukemia (RBL) assay. RESULTS: Regardless of the route of dosing, all products had sensitizing capacity. When sensitized i.p., all three gluten products induced a strong IgG1 response in all animals. Acid hydrolyzed gluten induced the highest level of specific IgE but with a low functionality. Orally all three gluten products induced specific IgG1 and IgE but with different dose-response relations. Sensitizing rats i.p. or orally with unmodified or enzymatic hydrolyzed gluten induced specific IgG1 responses with similar binding capacity which was different from that of acid hydrolyzed gluten indicating that acid hydrolysis of gluten proteins induces formation of 'new' epitopes. CONCLUSIONS: In rats not tolerant to gluten acid hydrolysis of gluten enhances the sensitizing capacity by the i.p. but not by the oral route. In addition, acid hydrolysis induces formation of new epitopes. This is in contrast to the enzymatic hydrolyzed gluten having an epitope pattern similar to unmodified gluten.

Dignity as an empirical lifeworld construction-in the field of surgery in Denmark.

Patient dignity is a complex yet central phenomenon. Disrespect for dignity can mean retention of sick role, loss of self-care and control, decreased participation and therefore influence healing. At the same time, nurses have an obligation to respect dignity, and patients expect it. In clinical practice, with the focus on efficiency and economy, dignity can be compromised. The surgical patient may be particularly vulnerable to loss of dignity, when focus is solely on surgical procedure, efficiency, and productivity. The aim of the article is to describe the characteristics of the importance of dignity perceived by four surgical patients at a university hospital in Denmark. The hermeneutic phenomenological approach of Van Manen is used to analyse and interpret data collected from in-depth semi-structured interviews. The interviews explored the lived experience with two women and two men who had undergone a surgical intervention in a Danish vascular surgery department. The thematic analysis led to the basic theme: "To be an important person" illustrated by the themes: "Being a co-player," "Over exposure," and "To swallow the bitter pill." The findings provide a better understanding of patient's perspective of dignity, which is characterized by a complex interaction of several factors. Nurses should be concerned with balancing expectations, values, and opinions to maintain dignity in nursing and create a common platform for collaboration. This collaboration makes it possible for patients to be involved and have a voice in relation to nursing, treatment, and administering of time even though it could be at the expense of the terms of the system.

Transfer of a two-tiered keratinocyte assay: IL-18 production by NCTC2544 to determine the skin sensitizing capacity and epidermal equivalent assay to determine sensitizer potency.

At present, the identification of potentially sensitizing chemicals is carried out using animal models. However, it is very important from ethical, safety and economic point of view to have biological markers to discriminate allergy and irritation events, and to be able to classify sensitizers according to their potency, without the use of animals. Within the Sens-it-iv EU Frame Programme 6 funded Integrated Project (LSHB-CT-2005-018681), a number of in vitro, human cell based assays were developed which, when optimized and used in an integrated testing strategy, may be able to distinguish sensitizers from non-sensitizers. This study describes two of these assays, which when used in a tiered strategy, may be able to identify contact sensitizers and also to quantify sensitizer potency. Tier 1 is the human keratinocyte NCTC2544 IL-18 assay and tier 2 is the Epidermal Equivalent potency assay. The aim of this study is to show the transferability of the two-tiered approach with training chemicals: 3 sensitizers (DNCB, resorcinol, pPD) and 1 non sensitizer (lactic acid) in tier 1 and 2 sensitizers with different potency in tier 2 (DNCB; extreme and resorcinol; moderate). The chemicals were tested in a non-coded fashion. Here we describe the transferability to naïve laboratories, the establishment of the standard operating procedure, critical points, acceptance criteria and project management. Both assays were successfully transferred to laboratories that had not performed the assays previously. The two tiered approach may offer an unique opportunity to provide an alternative method to the Local Lymph Node Assay (LLNA). These assays are both based on the use of human keratinocytes, which have been shown over the last two decades, to play a key role in all phases of skin sensitization.

Chiral pool synthesis of calystegine A3 from 2-deoxyglucose via a Brown allylation.

Calystegine A(3) is a naturally occurring nortropane iminosugar of which there previously have been three total syntheses. Inspired by our previous work we here report on a fourth approach using 17 steps from 2-deoxy-d-glucose applying a diastereoselective allylation protocol.

Synthesis of uronic-noeurostegine--a potent bacterial ß-glucuronidase inhibitor.

Inhibition of ß-glucuronidases has recently been shown to be useful in alleviating drug toxicity for common colon cancer chemotherapeutic CPT-11 (also called Irinotecan). We have prepared a new compound of the nortropane-type, uronic-Noeurostegine, and demonstrated that this is a competitive and potent E. coli ß-glucuronidase inhibitor, while inhibition of the mammalian ß-glucuronidase from bovine liver was found to be less significant. Although not intended, two other compounds having N-ethyl and N-(4-hydroxybutyl) substituents were also prepared in this study due to the sluggish debenzylation in the final step. The N-substituents are believed to come from reaction with the solvents used being ethanol and THF, respectively. These compounds also inhibited the two ß-glucuronidases albeit to a lesser extent compared to the parent compound. Noeurostegine and the three uronic-noeurostegines were additionally evaluated as inhibitors against a wide panel of glycosidases with the former showing potent inhibition of rat intestinal lactase and trehalase, whereas the latter was found to be inactive.

Synthesis of N-alkylated noeurostegines and evaluation of their potential as treatment for Gaucher's disease.

The potent and selective inhibitor of ß-glucosidases, noeurostegine, was evaluated as an inhibitor of glucocerebrosidase (GCase) to give an IC(50) value of 0.4 µM, being 250- and 150-fold better than N-butyl and N-nonyl noeurostegine, respectively. The parent noeurostegine and its N-butyl and N-nonyl alkylated congeners were also tested as pharmacological chaperones against a N370S GCase mutant. Of these, only noeurostegine, was found to increase enzyme activity, which in potency was comparable to that previously reported for isofagomine.

Effects of exposure to the xenoestrogen octylphenol and subsequent transfer to clean water on liver and gonad ultrastructure during early development of Zoarces viviparus embryos.

Female eelpouts (Zoarces viviparus L.) are exposed during early pregnancy to nominal concentrations of 100 microg/L of 4-tert-octylphenol (OP) or 0.5 microg/L of 17beta-estradiol (E2). Effects on maternal metabolism and on liver and gonad development in embryos were examined and compared with controls (C) during exposure and after transfer to clean water (depuration). In the mother fish, significantly higher concentrations of plasma vitellogenin (vtg) and calcium were found in the two exposed groups, when compared with the C group after exposure and depuration. When compared, however, with the respective values after exposure, vtg had decreased significantly after depuration. The hepatosomatic index was normalized after depuration. In both exposed groups, the hepatocytes were rounded and not distinctly polygonal as in the controls. The amount of glycogen was considerably less while the number of mitochondria increased, and the rER significantly proliferated after exposure as well as after depuration. The gonads of nine of more than 28 embryos in the group treated with OP exhibited a number of abnormalities as compared with the normal gonad development in both sexes. Feminization of the male gonads in the exposed specimens and a number of histopathological features were observed in all the abnormal gonads, whereas reliable male features, such as formation of seminiferous tubules or spermioduct, were not observed. This study showed that 4t-tert-OP and 17beta-estradiol exert estrogenic effects during very early development of the embryos and that depuration had a positive effect on the motherfish and her embryos.

Synthesis and glycosidase inhibitory activity of noeurostegine-a new and potent inhibitor of beta-glucoside hydrolases.

A new, stable hemi-aminal nor-tropane christened noeurostegine was synthesised in 22 steps from levoglucosan and tested for inhibitory activity against glycoside hydrolases. Sweet almond and Thermotoga maritimabeta-glucosidases, coffee bean alpha-galactosidase, and Asp. oryzaebeta-galactosidase were inhibited in the low micromolar region but significant tightening of binding to K(i) 50 nM for almond beta-glucosidase was found to occur after pre-incubation. Yeast alpha-glucosidase and E. colibeta-galactosidase were not inhibited at 1 mM.

Modeling neuro-vascular coupling in rat cerebellum: characterization of deviations from linearity.

We investigated the quantitative relation between neuronal activity and blood flow by means of a general parametric mathematical model which described the neuro-vascular system as being dynamic, linear, time-invariant, and subjected to additive noise. The model was constructed from measurements by means of system identification methods and validated across experiments. We sought to cover the system response to multiple stimulation frequencies and durations by a single model. We used the model to investigate the transport delay, the linear order, the deviations from linearity, and conditions for linearizability. We exercised the model on data from rat cerebellar cortex. In anesthetized rats, stimulation of the inferior olive caused climbing fiber activity and blood flow changes. Field potential amplitudes were used as an indicator of neuronal activity and blood flow was measured by laser-Doppler flowmetry. In one set of experiments, stimulation frequencies were in the range 2-20 Hz and the stimulation durations were 60 s and 600 s. The transport delay was estimated to be nearly zero, the linear order to be two. The deviations from linearity were consistently characterized as frequency saturation and dips in blood flow responses to stimulation for 60 s, and overgrowth of blood flow responses to stimulation for 600 s. In another set of experiments, stimulation frequencies were in the range 0.5-10 Hz and the stimulation duration was 15 s. The neuro-vascular system could be approximated by the linear model when the stimulation frequencies were restricted to the range 0.5-7 Hz. In conclusion, our model could predict blood flow responses to stimuli of low frequency and short duration.

Gonadal morphogenesis and sex differentiation in intraovarian embryos of the viviparous fish Zoarces viviparus (Teleostei, Perciformes, Zoarcidae): a histological and ultrastructural study.

It is essential to know the timing and process of normal gonadal differentiation and development in the specific species being investigated in order to evaluate the effect of exposure to endocrine-disrupting chemicals on these processes. In the present study gonadal sex differentiation and development were investigated in embryos of a viviparous species of marine fish, the eelpout, Zoarces viviparus, during their intraovarian development (early September to January) using light and electron microscopy. In both sexes of the embryos at the time of hatching (September 20) the initially undifferentiated paired bilobed gonad contains primordial germ cells. In the female embryos, ovarian differentiation, initiated 14 days posthatch (dph), is characterized by the initial formation of the endoovarian cavity of the single ovary as well as by the presence of some early meiotic oocytes in a chromatin-nucleolus stage. By 30 dph, the endoovarian cavity has formed. By 44 dph and onward, the ovary and the oocytes grow in size and at 134 dph, just prior to birth, the majority of the oocytes are at the perinucleolar stage of primary growth and definitive follicles have formed. In the presumptive bilobed testis of the male embryos, the germ cells (spermatogonia), in contrast to the germ cells of the ovary, remain quiescent and do not enter meiosis during intraovarian development. However, other structural (somatic) changes, such as the initial formation of the sperm duct (30 dph), the presence of blood vessels in the stromal areas of the testis (30 dph), and the appearance of developing testicular lobules (102 dph), indicate testicular differentiation. Ultrastructually, the features of the primordial germ cells, oogonia, and spermatogonia are similar, including nuage, mitochondria, endoplasmic reticulum, and Golgi complexes.

[Accelerated course after operation for ovarian cancer]. / Accelereret forløb efter operation for ovariecancer.

INTRODUCTION: Introduction of principles for postoperative multimodal rehabilitation (fast track surgery) has decreased hospital stay from about 8-10 days to 2-4 days after colonic resection. The aim of this study was to investigate the effect of a similar fast track regimen in patients operated for ovarian cancer. METHOD: 72 consecutive patients operated with a conventional perioperative treatment regimen (group 1) were compared with the initial 69 consecutive patients (group 2) with a multimodal rehabilitation regimen and the next 50 consecutive patients (group 3) where the fast track regimen was implemented as a routine. RESULTS: Patients demographics and surgical characteristics were comparable between groups. Median postoperative hospital stay was reduced from six days in group 1, to five days in group 2, and four days in group 3 (p < 0,05). Surgical complications were similar while medical complications were reduced from 12% to 1% (p < 0,05) and readmissions from 10% to 2% (p < 0,05) with the fast track regimen. CONCLUSION: Principles for postoperative multimodal rehabilitation from colonic surgery lead to faster rehabilitation, decreased risk of medical complications and hospital stay in patients operated for ovarian cancer.

The effect of accelerated rehabilitation on recovery after surgery for ovarian malignancy.

BACKGROUND: In patients undergoing colonic surgery the postoperative hospital stay has been reduced from 8-12 days to 2-4 days with multimodal rehabilitation programs. The aim of this study was to evaluate the postoperative outcome after surgery for ovarian malignancy with conventional care compared to fast-track multimodal rehabilitation. METHODS: Seventy-two consecutive patients receiving conventional care (group 1) were compared with 69 consecutive patients receiving multimodal, fast-track rehabilitation with a planned care program including continuous epidural analgesia, early oral feeding and mobilization (group 2) in the same department. Outcome was postoperative hospital stay and morbidity during the first postoperative month. RESULTS: Median age was 63 years (group 1) and 62 years (group 2). Median postoperative hospital stay was reduced from 6 days in group 1 (mean 7.3) to 5 days in group 2 (mean 5.4) (p < 0.05). There was no difference in the overall complication rate, although severe medical complications were reduced in group 2 (14% versus 2%; p < 0.01). Readmission rate was 10% in group 1 and 3% in group 2 (p > 0.05). CONCLUSIONS: The concept of fast-track multimodal rehabilitation appears to be beneficial in patients operated for ovarian malignancy, as hospital stay and medical morbidity are reduced.

Anti-estrogen prevents xenoestrogen-induced testicular pathology of eelpout (Zoarces viviparus).

Estrogenic alkylphenols have been shown to affect the reproductive system of male fish causing induction of vitellogenin synthesis and altered testis structure. However, it is still unknown whether the histopathological effects on the testes is mediated by the estrogen receptor or if it represent general toxicopathological effects. In the present study, the effects of different concentrations of the estrogenic chemical 4-tert-octylphenol on vitellogenin (Vtg) synthesis and testicular structure were investigated in the eelpout Zoarces viviparus during spermatogenesis. Adult male eelpout were exposed to 4-tOP (nominal concentrations: 10, 50 or 100 microg l(-1)) or 17beta-estradiol (E2; 0.5 microg l(-1)) in a continuous flow-through system for 3 weeks. A group of fish were exposed to 4-tOP (50 microg l(-1)) concomitantly with the anti-estrogen ZM 189,154 (20 microg g(-1) week(-1), i.p.). The Vtg concentration in plasma was measured by enzyme-linked immunosorbent assay. The testicular structure was examined by light microscopy and gamma-glutamyl transpeptidase (gamma-GTP) activity was measured in the testes. The testicular localization of gamma-GTP was analysed by enzyme histochemistry. A marked increase in the plasma Vtg concentration was observed after exposure to the actual concentration of 35 microg l(-1) 4-tOP (nominal concentration, 50 microg l(-1)), 63 microg l(-1) 4-tOP (nominal concentration, 100 microg l(-1)) or E2. Co-treatment with ZM 189,154 totally abolished the 4-tOP-dependent induction of Vtg synthesis. Exposure to 4-tOP or E2 caused a marked reduction in the testis mass and severely affected the testicular development and structure including the Sertoli cells (based on histology and gamma-GTP activity), resulting in impairment of spermatogenesis and degeneration of lobular structures. Other cellular abnormalities such as accumulations of yellowish-brown pigmented cells and increased interstitial fibrosis in the testes was also observed in the exposed fish. In the groups exposed to the nominal concentrations of 50 or 100 microg l(-1) all fish had severely affected testes, while both normal, moderately and severely affected testes were found in the group exposed to the nominal concentration of 10 microg l(-1). Co-treatment with ZM 189,154 abolished part of these 4-tOP-induced effects on the testicular growth and histological structure. The study demonstrates that an anti-estrogen can abolish effects on the testis caused by estrogenic chemicals, providing evidence that some of the effects are mediated by the estrogen receptor.

Estrogenic octylphenol affects seminal fluid production and its biochemical composition of eelpout (Zoarces viviparus).

Estrogenic chemicals such as alkylphenols (APs) have been shown to disrupt the reproductive system of male fish. In the present study, the effects of the estrogenic chemical octylphenol (100 microg g(-1)) and 17 beta-estradiol on the semen production and the biochemical composition of the seminal fluid of the viviparous eelpout (Zoarces viviparus) were investigated at the time of spawning. After 10 days of octylphenol or estrogen treatment, vitellogenin (Vtg) synthesis was induced as indicated by increased plasma vitellogenin concentration. In accordance with the increased vitellogenin concentration, hepatosomatic index (HSI), total protein concentration, and total calcium concentration were also increased, and free amino acids concentration was decreased in blood plasma. Octylphenol treatment caused a decrease in the gonadosomatic index (GSI) and the milt volume and an increase in the spermatocrit. The histological examination revealed that octylphenol affected the normal lobular structure, including the Sertoli cells. In the majority of the octylphenol-treated fish, trapped sperm cells were observed in parts of the seminiferous lobules and the sperm ducts. The biochemical composition of the seminal fluid was also affected by the octylphenol or estrogen. The seminal plasma concentrations of magnesium, calcium, and total protein were elevated, and the concentration of free amino acids was reduced in the treated fish. This study indicates that octylphenol inhibits the seminal fluid production and changes its biochemical composition in eelpouts.

Effects of waterborne exposure of octylphenol and oestrogen on pregnant viviparous eelpout (Zoarces viviparus) and her embryos in ovario.

Exposure to oestrogenic chemicals (xeno-oestrogens) may have severe effects on embryonic development. The present study investigates whether the oestrogenic endocrine disruptor 4-tert-octylphenol (4-tOP) or 17beta-oestradiol (E(2)) is accumulated in the viviparous fish the eelpout (Zoarces viviparus) and transferred to the embryos in ovario and subsequently affects embryonic development, including gonadal differentiation. Pregnant eelpouts were exposed to nominal concentrations of 25 micro gl(-1) or 100 micro gl(-1) 4-tOP (OP25 or OP100, respectively) or 0.5 micro gl(-1) E(2) in water. During 4-tOP exposure, the compound accumulated in both plasma and ovarian fluid in a concentration-dependent manner. In the mother fish, the oestrogenic biomarkers, vitellogenin (Vtg) in plasma, Vtg mRNA in liver and oestrogen-binding activity in liver, were all induced by 4-tOP (and by E(2)) at an actual concentration of 14 micro gl(-1). E(2) and 4-tOP were examined for their potency to disturb the maternal-foetal trophic relationship by disturbing the physiology of the ovary and by changing the distribution of essential nutrients normally transported to embryos during pregnancy. After exposure to E(2) or 4-tOP, calcium was depleted from the ovarian fluid and the level of free amino acids available in maternal plasma was decreased. A marked overall effect on ovarian components, including the ovarian sac, ovarian fluid and embryonic mass, was evident. Embryonic growth was significantly decreased, which might in part be attributed to disturbances of the maternal-foetal trophic relationship. Marked inductions of Vtg mRNA and Vtg protein, determined by RT-PCR and immunohistochemistry, respectively, were found in embryos from the OP100 group - the only group to show considerable accumulation of an oestrogenic compound in the ovarian fluid. A different pattern of gonadal development was found in embryos from the OP100 group compared with embryos from the control, OP25 or E(2) groups, in which approximately 50% had normal ovaries and 50% had normal presumptive male gonads. In the OP100 group, 46% had normal ovaries but, in contrast to controls, only 22% had normal presumptive male gonads, whereas the remaining 32% had abnormal male gonads with structures resembling the endo-ovarian cavity of a female gonad. As oestrogen receptor (ER) expression was detected by in situ hybridisation in early differentiating gonads, these effects could be mediated by direct interaction of the xeno-oestrogens with gonadal ER. In conclusion, this study indicates that the xeno-oestrogen 4-tOP can be transferred from the water via the mother fish to the ovarian fluid and can subsequently disturb the maternal-foetal trophic relationship and cause severe effects on embryonic development, including gonadal differentiation in ovario.