Speciation of zinc in fish feed by size exclusion chromatography coupled to inductively coupled plasma mass spectrometry - using fractional factorial design for method optimisation and mild extraction conditions.

Zinc (Zn) is an element essential to all living organisms and it has an important role as a cofactor of several enzymes. In fish, Zn deficiency has been associated with impaired growth, cataracts, skeletal abnormalities and reduced activity of various Zn metalloenzymes. Fish meal and fish oil traditionally used in salmon feed preparation are being replaced by plant-based ingredients. Zinc additives are supplemented to salmon feed to ensure adequate Zn levels, promoting good health and welfare in Atlantic salmon (Salmo salar). The main objective of the present study was to evaluate Zn species found in an Atlantic salmon feed. This work describes a Zn extraction method that was optimized using a fractional factorial design (FFD), whereby the effect of six factors could be studied by performing only eight experiments. The effects of the type of extraction solution and its molar concentration, pH, presence of sodium dodecyl sulphate, temperature and extraction time on Zn extraction were investigated. Mild extraction conditions were chosen in order to keep the Zn species intact. Total Zn (soluble fractions and non-soluble fractions) was determined by inductively coupled plasma mass spectrometry (ICP-MS). The highest Zn recovery was obtained using 100 mM Tris-HCl, pH 8.5 at a temperature of 4 °C for 24 h where the total Zn in soluble fraction and non-soluble fraction was 9.9 ±â€¯0.2% and 98 ±â€¯6%, respectively. Zinc speciation analysis (on the soluble fractions) was further conducted by size exclusion inductively coupled plasma mass spectroscopy (SEC-ICP-MS). The SEC-ICP-MS method provided qualitative and semi-quantitative information regarding Zn species present in the soluble fractions of the feed. Four Zn-containing peaks were found, each with different molecular weights: Peak 1 (high molecular weight - ≥600 kDa), peak 2 and peak 3 (medium molecular weight - 32 to 17 kDa) were the least abundant (1-6%), while peak 4 (low molecular weight - 17 to 1.36 kDa) was the most abundant (84-95%).

Sensitivity and toxic mode of action of dietary organic and inorganic selenium in Atlantic salmon (Salmo salar).

Depending on its chemical form, selenium (Se) is a trace element with a narrow range between requirement and toxicity for most vertebrates. Traditional endpoints of Se toxicity include reduced growth, feed intake, and oxidative stress, while more recent finding describe disturbance in fatty acid synthesis as underlying toxic mechanism. To investigate overall metabolic mode of toxic action, with emphasis on lipid metabolism, a wide scope metabolomics pathway profiling was performed on Atlantic salmon (Salmo salar) (572±7g) that were fed organic and inorganic Se fortified diets. Atlantic salmon were fed a low natural background organic Se diet (0.35mg Se kg-1, wet weight (WW)) fortified with inorganic sodium selenite or organic selenomethionine-yeast (SeMet-yeast) at two levels (∼1-2 or 15mgkg-1, WW), in triplicate for 3 months. Apparent adverse effects were assessed by growth, feed intake, oxidative stress as production of thiobarbituric acid-reactive substances (TBARS) and levels of tocopherols, as well as an overall metabolomic pathway assessment. Fish fed 15mgkg-1 selenite, but not 15mgkg-1 SeMet-yeast, showed reduced feed intake, reduced growth, increased liver TBARS and reduced liver tocopherol. Main metabolic pathways significantly affected by 15mgkg-1 selenite, and to a lesser extent 15mgkg-1 SeMet-yeast, were lipid catabolism, endocannabinoids synthesis, and oxidant/glutathione metabolism. Disturbance in lipid metabolism was reflected by depressed levels of free fatty acids, monoacylglycerols and diacylglycerols as well as endocannabinoids. Specific for selenite was the significant reduction of metabolites in the S-Adenosylmethionine (SAM) pathway, indicating a use of methyl donors that could be allied with excess Se excretion. Dietary Se levels to respectively 1.1 and 2.1mgkg-1 selenite and SeMet-yeast did not affect any of the above mentioned parameters. Apparent toxic mechanisms at higher Se levels (15mgkg-1) included oxidative stress and altered lipid metabolism for both inorganic and organic Se, with higher toxicity for inorganic Se.

[Decisions concerning applications for termination of pregnancy during the period from 1 April 1970 to 31 March 1979]. / Samrådsafgørelser på landsbasis ved ansøgning om svangerskabsafbrydelse i perioden 1.IV.1970-31.III.1979.

PIP: An analysis is undertaken of abortions performed between 1970 and 1979 under paragraphs 3 and 4 of the Danish abortion law, i.e. abortions granted by a consultant committee. This period is divided into 18 half-year periods. The increase in abortions performed under paragraph 6 increased 45.9% between 1970 and 1973. This increase was 108% for pregnancies 12 weeks and 39.4% for pregnancies 12 weeks. During this period, the percentage of refusals of applications for abortion decreased from ca. 12% to ca. 4% before the 12th week of pregnancy and from 30-40% to ca. 20% after the 12th week of pregnancy. Of the cases after the 12th week of pregnancy, the greatest increase in the granting of abortions occurred in cases after the 16th week of pregnancy. The revision of the abortion law in 1973, in spite of identical guidelines concerning abortions in the second trimester of pregnancy, caused an increase in the number of abortion applications that were rejected after the 12th week of pregnancy, instead of a decrease.^ieng

[Peroral treatment with natural human estrogens in the climacteric]. / Peroral behandling med naturlige, humane østrogener i klimakteriet.

PIP: Various aspects of climacteric treatment with natural human estrogens are discussed. Estradiol, estradiol valerate, estron sulfate, or estriol are used separately or together in various preparations to treat the symptoms of approaching menopause. Estrogen treatment causes proliferation of the endometrium and causes a decrease in LHRH, FSH, and LH secretion. Treatment can take the form of continuous or cyclic treatment with estrogens alone, or sequential estrogne/gestagen preparations can be used. Ovarian function decreases as menopause approaches and results in the cessation of ovulation. Then the hypolutein phase begins, during which the secretion of progesterone is reduced and menstrual bleeding irregularities begin to occur. Eventually, estrogen production decreases so much that menstruation ceases completely, and symptoms such as heat flashes are experienced. Women who want treatment for climacteric symptoms but who want no regular menstrual bleeding can be administered low doses of pure estrogen. Regular abrasio control of endometrial development should be performed, however. Pure estrogen treatment can also be used in the case of hysterectomized women. Otherwise, a sequential treatment is generally indicated. Possible side effects of estrogen substitution therapy are changes in the genitalia, breasts, menstrual bleeding, blood pressure, and weight. There is also an indication that estrogen use can induce endometrial cancer. Besides the definite contraindication of endometrial cancer, relative contraindications of estrogen therapy include breast cancer, reduced liver function, thromboembolic disease, and serious hypertension. Estrogen therapy is to be used to solve acute climacteric symptoms; women should be well informed about possible side effects and that the therapy is no panacea for all menopausal problems.^ieng

[Contraception in general practice. A research project from general practice. II. Presentation of cases with special attention to the contraceptive ttern]. / Svangerskabsforebyggelse i almen laegepraksis. Et forskningsprojekt fra almen praksis. II. Praesentation af materialet specielt med henblik på antikonceptionsmønstret.

PIP: A study is presented of 3722 women in Denmark receiving contraception from 64 doctors in the course of 1 year. 76% of the women received oral contraceptives. IUDs were used more often by women over 25, while the diaphragm was used by the younger women. 78% of the women under 20 were given oral contraceptives. 20% of the women had not used contraceptives before. Of those who had used contraceptives, most had used the condom or the diaphragm previously. When compared with earlier studies of the same nature, there was a shift from demand for the diaphragm to demand for oral contraception with unmarried women. With married women, the shift was from demand for the IUD to demand for oral contraception. 30% of the women were unmarried. Oral contraceptives were asked for by the women with the greatest need for effective contraception. Women from lower social strata lived more often in urban areas and more often received oral contraception; the diaphragm increased in usage among higher social strata. As the number of pregnancies increased, the demand for the diaphragm decreased and the demand for the IUD tended to increase. The doctor took the initiative in giving contraceptive counseling in about 18% of the cases, more frequently for married women.^ieng

[Contraception in general practice. A research project from general practice. Presentation of the project and participating physicians]. / Svangerskabsforebyggelse i almen laegepraksis. Et forskningsprojekt fra almen praksis. I. Praesentation af projektet og dets laeger.

PIP: A study is presented of 64 Danish physicians who gave contraceptive counseling. 3 journals were kept for each patient: a journal containing biographic data and an anamnesis, a control journal, and a journal for those patients who discontinued contraceptive use. Most of the participating physicians were 40-49 years old, were male, and had private practice. Most of the doctors had practices consisting of city dwellers. 3726 patients were counseled in 1 year. Women doctors gave comparatively more counseling than men. The amount of contraceptive counseling given in this project is not representative of doctors for the entire country of Denmark. A 4-fold increase in contraceptive counseling, due to revisions of the pregnancy hygiene law in 1966, is evident.^ieng

[Differentiation of benign and malignant ovarian tumors by transluminal ultrasound scanning]. / Differentiering af benigne og maligne ovarietumorer med transabdominal ultralydskanning.

We intended to evaluate ultrasonographic criteria for differentiation of benign and malignant ovarian tumours, and to estimate the risk of malignancy in unilocular ovarian cysts. The files of 186 women aged 40 or above, who had undergone surgery in the department of gynaecology and obstetrics at Hillerød hospital between 01.01.1988-31.12.1990 and where pathological ovarian histology was found were reviewed. Preoperative ultrasonographic examination concerning size and morphology was compared to histological diagnosis. Tumours were classified according to the ultrasonographic internal structure into a) unilocular cysts, b) unilocular cysts with solid areas, c) multilocular cysts, d) multilocular cysts with solid areas and e) solid tumours. Criteria for suspicion of malignancy were unilocular cysts > 10 cm and complex and solid tumours, whereas smaller unilocular cysts were thought to be benign. Twenty-one tumours, diagnosed by ultrasound as unilocular cysts were all histologically benign, independently of size. For complex and solid tumors there was an increased risk of malignancy, and a statistically significant correlation between tumour size and risk of malignancy was found. Using the ultrasonographic criteria for malignancy a sensitivity of 97% and a specificity of 28% was found. The suggested criteria for ultrasonographic suspicion of malignancy were found usable in differentiating benign and malignant ovarian tumours. The risk of malignancy for unilocular cysts seems to be low, independently of size.

[The birth rate in 1982. Assessment of the prediction of birth rate in 1981 and a prognosis for 1982 on the basis of registered pregnancies in the county of Fredriksborg]. / Fødselstallet i 1982. Vurdering af prognosen for fødselstallet i 1981 samt prognose for 1982 på grundlag af registrerede graviditeter i Frederiksborg amtskommune.

PIP: Predictions of the birth rate in Denmark for 1981 based on a centralized register of pregnant women in Frederiksborg county are reviewed, and their accuracy is confirmed. Similar predictions for 1982 using the same source of data are presented with suggestions of a smaller decline in fertility than that which occurred in 1981. (summary in ENG)^ieng

[Birth rate in 1981. A prognosis based on registered pregnancies in the county of Fredriksborg]. / Fødselstallet i 1981. En prognose på grundlag af registrerede graviditeter i Frederiksborg amtskommune.

PIP: On the basis of a centralized registration of pregnant women early in pregnancy, it is possible with adequate certainty to predict the birth rate 1/2-1 year in advance. On this basis, it has been calculated that the birth rate in the county of Frederiksborg, Denmark (population as of January 1, 1980--329,141 persons) will fall by 16% during the first 1/2 of 1981 as compared with the first 1/2 of 1980. As far as alterations in the birth rate for the county are concerned, they follow, by and large, the alterations seen in Denmark as a whole; thus a fall in birth rate is to be anticipated in the country as a whole. (author's)^ieng