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Worldwide harvest of wild marine fisheries for fish oil cannot increase. However, the demand for fish oil is increasing due to a rapidly expanding aquaculture industry and is further increased by nutraceutical/biomedical and agricultural companies. Aquaculture uses fish oil as a source for essential fatty acids in particular omega-3 long chainpolyunsaturated fatty acids (˜ìv¢3 LC-PUFA) and for energy. Other novel sources of renewable, environmentally sustainable oil that provide these nutritional requirements for Atlantic salmon (Salmo salar L.) are needed. This research looked at alternate sources of oil containing the ˜ìv¢3 LC-PUFA that are associated with the many health benefits of eating Atlantic salmon. This thesis also contributed to the development of three techniques for use in aquaculture lipid nutrition research: 1) advanced chromatography and mass spectroscopy to examine intact molecular membrane lipids; 2) nuclear magnetic resonance (13C NMR) to assess the regiospecific distribution of ˜ìv¢3 LC-PUFA in oil, and 3) molecular RT-PCR to investigate endogenous ˜ìv¢3 LC-PUFA production.

© The Author 2015. study question: Are significant abnormalities of CatSper function present in IVF patients with normal sperm concentration and motility and if so what is their functional significance for fertilization success? summaryanswer: Sperm with a near absence of CatSper current failed to respond to activation of CatSper by progesterone and there was fertilization failure at IVF. what is known already: In human spermatozoa,Ca2+influx induced by progesterone is mediated by CatSper, a sperm-specificCa2+ channel.Asuboptimal Ca2+ influx is significantly associated with, and more prevalent in, men with abnormal semen parameters, and is associated with reduced fertilizing capacity. However, abnormalities in CatSper current can only be assessed directly using electrophysiology. There is only one report of a CatSper-deficient man who showed no progesterone potentiated CatSper current. ACatSper 2 genetic abnormality was present but there was no information on the [Ca2+]i response to CatSper activation by progesterone. Additionally, the semen samples had indicating significant abnormalities (oligoasthenoteratozoospermia) multiple suboptimal functional responses in the spermatozoon. As such it cannot be concluded that impaired CatSper function alone causes infertility or that CatSper blockade is a potential safe target for contraception. study design, size, duration: Spermatozoa were obtained from donors and subfertile IVF patients attenDing a hospital assisted reproductive techniques clinic between January 2013 and December 2014. In total 134 IVF patients, 28 normozoospermic donors and 10 patients recalled due to a history of failed/low fertilization at IVF took part in the study. participants/materials, setting, methods: Samples were primarily screened using the Ca2+ influx induced by progesterone and, if cell number was sufficient, samples were also assessed by hyperactivation and penetration into viscous media. A defective Ca2+ response to progesterone was defined using the 99% confidence interval fromthe distribution of response amplitudes in normozoospermic donors. Samples showing a defective Ca2+ response were further examined in order to characterize the potential CatSper abnormalities. In men where there was a consistent and robust failure of calcium signalling, a direct assessment of CatSper function was performed using electrophysiology (patch clamping), and a blood sample was obtained for genetic analysis. main results and the role of chance: Atotal of 101/102 (99%) IVF patients and 22/23 (96%) donors exhibited a normal Ca2+ response. The mean (+SD) normalized peak response did not differ between donors and IVF patients (2.57+0.68 [n 34 ejaculates from 23 different donors] versus 2.66+0.68 [n 102 IVF patients], P 0.63). In recall patients, 9/10 (90%) showed a normal Ca2+ response. Three men were initially identified with a defective Ca2+ influx. However, only one (Patient 1) had a defective response in repeat semen samples. Electrophysiology experiments on sperm from Patient 1 showed a near absence of CatSper current and exon screening demonstrated no mutations in the coDing regions of the CatSper complex. There was no increase in penetration of viscous media when the spermatozoa were stimulated with progesterone and importantly there was failed fertilization at IVF. limitations, reasons for caution: A key limitation relates to working with a specific functional parameter (Ca2+ influx induced by progesterone) in fresh sperm samples from donors and patients that have limited viability. Therefore, for practical, technical and logistical reasons, some men (22% of IVF patients) could not be screened. As such the incidence of significant Ca2+ abnormalities induced by progesterone may be higher than the 1% observed here. Additionally, we used a strict definition of a defective Ca2+ influx such that only substantial abnormalities were selected for further study. Furthermore, electrophysiology was only performed on one patient with a robust and repeatable defective calcium response. This man had negligible CatSper current but more subtle abnormalities (e.g. currents present but significantly smaller) may have been present in men with either normal or below normal Ca2+ influx. wider implications of the findings: These data add significantly to the understanDing of the role of CatSper in human sperm function and its impact on male fertility.Remarkably, these findings provide the first direct evidence that CatSper is a suitable and specific target for human male contraception. study funding/competing interest(s): Initial funDing was fromNHSTayside, Infertility Research Trust,TENOVUS, Chief Scientist Office NRS Fellowship, the Wellcome Trust, University of Abertay. The majority of the data were obtained using funDing from a MRC project grant (# 4190). The authors declare that there is no conflict of interest. trial registration number: Not applicable.

SERIES “ATS/ERS TASK FORCE: STANDARDISATION OF LUNG FUNCTION TESTING” Edited by V. Brusasco, R. Crapo and G. Viegi Number 1 in this Series ⇓In preparing the joint statements on lung function testing for the American Thoracic Society (ATS) and the European Respiratory Society (ERS), it was agreed by the working party that the format of the statements should be modified so that they were easier to use by both technical and clinical staff. This statement contains details about procedures that are common for many methods of lung function testing and, hence, are presented on their own. A list of abbreviations used in all the documents is also included as part of this statement. All terms and abbreviations used here are based on a report of the American College of Chest Physicians/ATS Joint Committee on Pulmonary Nomenclature 1. The metrology definitions agreed by the International Standards Organization (ISO) are recommended 2 and some important terms are defined as follows. Accuracy is the closeness of agreement between the result of a measurement and the conventional true value. Repeatability is the closeness of agreement between the results of successive measurements of the same item carried out, subject to all of the following conditions: same method, same observer, same instrument, same location, same condition of use, and repeated over a short space of time. In previous documents, the term reproducibility was used in this context, and this represents a change towards bringing this document in line with the ISO. Reproducibility is the closeness of agreement of the results of successive measurements of the same item where the individual measurements are carried out with changed conditions, such as: method of measurement, observer, instrument, location, conditions of use, and time. Thus, if a technician tests a subject several times, this is looking at the …

[⇓][1] SERIES “ATS/ERS TASK FORCE: STANDARDISATION OF LUNG FUNCTION TESTING” Edited by V. Brusasco, R. Crapo and G. Viegi Number 2 in this Series [1]: #F13

Udgivelsesdato: Sep

Udgivelsesdato: Oct

The objective of the study was to examine the effect of farming exposure, respiratory symptoms and smoking on the shape of the MEFV-curve in 1,691 male farming students and 407 male controls and to relate the slope ratio with FEV(1) and FEV(1) /FVC. Each subject underwent a medical interview and the slope ratios from the MEFV-curve at 75 (SR75), 50 (SR50) and 25 (SR25) %FVC together with FEV1 and FVC were recorded. Histamine bronchial reactivity (Yan method) was measured and skin prick test with inhalant allergens was performed. In smokers, SR75 increased with increasing exposure to: general farming, swine and dairy cattle (por =0.020). SR50 increased with increasing exposure to farming (p=0.015). In non-smokers, SR25 increased with increasing exposure to swine and dairy cattle (p=0. 021) and increased SR25 was associated with sensitisation to house dust mite (p=0.017). Data revealed an interaction between smoking and exposure to farming. FEV(1) and FEV(1)/FVC was not associated with farming exposure or production animals. FEV1 and FEV(1)/FVC (por =0.003) were lower among subjects wi bronchial hyperresponsiveness and asthma (FEV(1) and asthma only in smokers). SR75 (p=0.037) and SR50 (p=0.024) were increased in subjects with asthma and SR75 was increased in subjects with bronchial hyperresponsiveness, but only in smokers (p=0.002). In conclusion, exposure to farming seems to influence the shape of the MEFV-curve and there are indications of interaction between exposure to organic dust and smoking. These changes are seen only in the slope ratios and not in FEV(1) and FEV(1)/FVC. However, FEV(1) and FEV(1)/FVC are superior to slope ratios in differentiating healthy subjects from those with respiratory symptoms.

Little is known about the linearity of portable peak flow meters, or about physical gas factors affecting peak expiratory flow (PEF) readings. We therefore tested five portable peak flow meters of three types in an altitude chamber (sea level to 5,500 m) and in a climate chamber at sea level (7-37 degrees C) to determine the influence of the physical conditions of the gas on the reading of the meters. The nonlinear response of the variable orifice meters was confirmed and, when this was corrected for, the readings of these meters were found to be significantly reduced by higher altitude and lower temperature. The readings from a turbine type of peak flow meter were not affected by altitude but were reduced at low temperature. A mathematical model for the variable orifice meters could correct for both their nonlinear behaviour and the effect of gas density (altitude, temperature and humidity). The model showed that correction is not necessary for the differences in gas conditions between calibration and taking of measurements under normal laboratory conditions. All the meters tested had impedances higher than recommended (0.05 kPa.l-1.s) and this may influence PEF at high flows. The mean uncorrected PEF of six healthy subjects when measured with a Mini Wright peak flow meter at sea level and at 3,000 m fell by 5%, but the mean corrected PEF increased by 12%. This increase in PEF was about 60% of that predicted for fully density-dependent flow and agreed with the findings of other similar studies.(ABSTRACT TRUNCATED AT 250 WORDS)

The involvement of DNA polymerases {alpha}, {beta}, and {delta} in DNA repair synthesis induced by N-methyl-N{prime}-nitro-N-nitrosoguanidine (MNNG) was investigated in human fibroblasts (HF). The effects of anti-(DNA polymerase {alpha}) monoclonal antibody, (p-n-butylphenyl)deoxyguanosine triphosphate (BuPdGTP), dideoxythymidine triphosphate (ddTTP), and aphidicolin on MNNG-induced DNA repair synthesis were investigated to dissect the roles of the different DNA polymerases. A subcellular system (permeable cells), in which DNA repair synthesis and DNA replication were differentiated by CsCl gradient centrifugation of BrdUMP density-labeled DNA, was used to examine the effects of the polymerase inhibitors. Another approach investigated the effects of several of these inhibitors of MNNG-induced DNA repair synthesis in intact cells by measuring the amount of ({sup 3}H)thymidine incorporated into repair DNA as determined by autoradiography and quantitation with an automated video image analysis system. In permeable cells, MNNG-induced DNA repair synthesis was inhibited 56% by 50 {mu}g of aphidicolin/mL, 6% by 10 {mu}M BuPdGTP, 13% by anti-(DNA polymerse {alpha}) monoclonal antibodies, and 29% by ddTTP. In intact cells, MNNG-induced DNA repair synthesis was inhibited 57% by 50 {mu}g of aphidicolin/mL and was not significantly inhibited by microinjecting anti-(DNA polymerase {alpha}) antibodies into HF nuclei. These results indicate that both DNA polymerase {delta} and {beta}more » are involved in repairing DNA damage caused by MNNG.« less