Molecular Markers of Yolk Sac Fry Development in Nine Families of Lake Trout

Salmonids in certain areas of North America and northern Europe suffer from reproductive disturbances manifested through the death of yolk sac fry. These disturbances are referred to as early mortality syndrome (EMS) in the Great Lakes region and M74 in the Baltic Sea. Both of these syndromes have been associated with reduced concentrations of thiamine in affected females and their eggs. However, large variations in signs and mortality, both within and between the individual syndromes, have been reported. Yolk sac fry mortality (M74) in Atlantic salmon Salmo salar has been shown to be associated with reduced DNA binding of the hypoxia-inducible transcription factor 1 (HIF-1), reduced production of vascular endothelial growth factor (VEGF) protein, decreased capillary density, and down-regulation of adult-type globin gene transcription (which is responsible for the protein part of adult hemoglobin). One of the main effects of all of these changes is reduced oxygen transport to the tissues of affected fry. In this study, the developmental patterns of HIF-1 DNA binding, VEGF protein expression, and adult-type globin gene transcription were analyzed in nine family groups of Lake Michigan lake trout Salvelinus namaycush. The results indicate that HIF-1 DNA binding and globin gene transcription increase from hatch to the end of yolk sac stage. Interindividual and between-family biological variations were detected, especially in VEGF protein expression and globin gene transcription. Our results demonstrate the possibility of using these molecular markers in investigating the etiology of EMS and making comparisons between the mechanisms of different salmonid yolk sac fry mortalities

Copepod reproductive effort and oxidative status as responses to warming in the marine environment

The marine ecosystems are under severe climate change-induced stress globally. The Baltic Sea is especially vulnerable to ongoing changes, such as warming. The aim of this study was to measure eco-physiological responses of a key copepod species to elevated temperature in an experiment, and by collecting field samples in the western Gulf of Finland. The potential trade-off between reproductive output and oxidative balance in copepods during thermal stress was studied by incubating female Acartia sp. for reproduction rate and oxidative stress measurements in ambient and elevated temperatures. Our field observations show that the glutathione cycle had a clear response in increasing stress and possibly had an important role in preventing oxidative damage: Lipid peroxidation and ratio of reduced and oxidized glutathione were negatively correlated throughout the study. Moreover, glutathione-s-transferase activated in late July when the sea water temperature was exceptionally high and Acartia sp. experienced high oxidative stress. The combined effect of a heatwave, increased cyanobacteria, and decreased dinoflagellate abundance may have caused larger variability in reproductive output in the field. An increase of 7 degrees C had a negative effect on egg production rate in the experiment. However, the effect on reproduction was relatively small, implying that Acartia sp. can tolerate warming at least within the temperature range of 9-16 degrees C. However, our data from the experiment suggest a link between reproductive success and oxidative stress during warming, shown as a significant combined effect of temperature and catalase on egg production rate.Peer reviewe

Identification of modifiable pre- and postnatal dietary and environmental exposures associated with owner-reported canine atopic dermatitis in Finland using a web-based questionnaire

A cross-sectional hypothesis generating study was performed to investigate modifiable exposures such as whether feeding pattern (a non-processed meat based diet, NPMD, or an ultra-processed carbohydrate based diet, UPCD), certain environmental factors and their timing of exposure might be associated with the development of canine atopic dermatitis (CAD). Also, genetic and demographic factors were tested for associations with CAD. The data was collected from the validated internet-based DogRisk food frequency questionnaire in Finland. A total of 2236 dogs were eligible for the study (the owners reported 406 cases and 1830 controls). Our main interest was to analyze modifiable early risk factors of CAD, focusing on nutritional and environmental factors. We tested four early life periods; prenatal, neonatal, early postnatal and late postnatal periods. Twenty-two variables were tested for associations with CAD using logistic regression analysis. From the final models we identified novel dietary associations with CAD: the NPMD during the prenatal and early postnatal periods had a significant negative association with the incidence of CAD in adult dogs (age above 1 year). Oppositely, UPCD was associated with a significantly higher risk for CAD incidence. Other variables that were associated with a significantly lower risk for CAD were maternal deworming during pregnancy, sunlight exposure during early postnatal period, normal body condition score during the early postnatal period, the puppy being born within the same family that it would stay in, and spending time on a dirt or grass surface from 2 to 6 months. Also, the genetic factors regarding maternal history of CAD, allergy-prone breeds and more than 50% white-colored coat all showed a significant positive association with CAD incidence in agreement with previous findings. Although no causality can be established, feeding NPMD early in life seemed to be protective against CAD, while UPCD could be considered a risk factor. Prospective intervention studies are needed to establish the causal effects of the protective role of NPMD on prevalence of CAD during the fetal and early postnatal life.Peer reviewe

Diet and dog characteristics affect major and trace elements in hair and blood of healthy dogs

Obtaining correct amounts of essential elements, and avoiding toxic metals are key factors in dog health. Through analyzing major and trace elements in hair and blood of 50 healthy companion dogs using ICP-MS, we study their associations with dog characteristics and diet, hypothesizing that eating the same diet long-term results in strong correlations between hair and blood element concentrations, and that dog characteristics and diet affect element status. The correlation between hair and blood was significant for Hg (R = 0.601, p = 0.000) and Pb (R = 0.384, p = 0.010). The following associations were significant (p < 0.05): Dark hair had higher Ca and Mg compared to light hair. Females had higher hair Zn, blood Mn, and blood As compared to males. Blood Mn and Se increased, while blood Pb decreased with age. Raw diet fed dogs had higher hair Zn and Se compared to dry or mixed diet fed dogs, and lower blood Mn compared to dry diet fed dogs. Dry and mixed diet fed dogs had higher blood Cd compared to raw diet fed dogs. Mixed diet fed dogs had higher hair Ca and Mg compared to raw or dry diet fed dogs, and higher hair Pb compared to dry diet fed dogs. Wild game consumption was associated with higher blood Pb, and rice consumption with higher blood As. In conclusion, hair provides an alternative for assessing Hg and Pb exposure, and major and trace elements status is affected by hair color, sex, age, and diet.Peer reviewe

Copepod reproductive effort and oxidative status as responses to warming in the marine environment

The marine ecosystems are under severe climate change-induced stress globally. The Baltic Sea is especially vulnerable to ongoing changes, such as warming. The aim of this study was to measure eco-physiological responses of a key copepod species to elevated temperature in an experiment, and by collecting field samples in the western Gulf of Finland. The potential trade-off between reproductive output and oxidative balance in copepods during thermal stress was studied by incubating female Acartia sp. for reproduction rate and oxidative stress measurements in ambient and elevated temperatures. Our field observations show that the glutathione cycle had a clear response in increasing stress and possibly had an important role in preventing oxidative damage: Lipid peroxidation and ratio of reduced and oxidized glutathione were negatively correlated throughout the study. Moreover, glutathione-s-transferase activated in late July when the sea water temperature was exceptionally high and Acartia sp. experienced high oxidative stress. The combined effect of a heatwave, increased cyanobacteria, and decreased dinoflagellate abundance may have caused larger variability in reproductive output in the field. An increase of 7 degrees C had a negative effect on egg production rate in the experiment. However, the effect on reproduction was relatively small, implying that Acartia sp. can tolerate warming at least within the temperature range of 9-16 degrees C. However, our data from the experiment suggest a link between reproductive success and oxidative stress during warming, shown as a significant combined effect of temperature and catalase on egg production rate

Whole blood trace element and toxic metal concentration in dogs with idiopathic epilepsy and healthy dogs : A case-control study

Peer reviewe

Assessing the association between supplemented puppyhood dietary fat sources and owner-reported epilepsy in adulthood, among Finnish companion dogs

IntroductionEpilepsy is a serious and common neurological condition in dogs, despite the wide number of antiepileptic drugs available, in approximately one third of the patients, epilepsy remains unsatisfactorily controlled. We aim to analyze whether feeding dietary fat sources during puppyhood was associated with canine epilepsy in adulthood.MethodsA nested case–control study was compiled from the validated DogRisk food frequency questionnaire (DogRisk FFQ). DogRisk FFQ collected feeding, disease, and background data about the dog. The study sample consisted of 108 owner-reported epileptic cases and 397 non-epileptic controls. Each case was matched with up to four controls for the key confounding factors of sex, breed, and age. We analyzed associations between feeding as a puppy and owner-reported epilepsy as an adult dog using Cox regression. We tested 55 different food variables.ResultsWe found that feeding fish fat from dietary sources at least once a week during puppyhood was inversely associated with epilepsy in later life in the unadjusted analysis [OR 0.46 (95% CI 0.25–0.83), p=0.01], while when adjusting for keeping conditions and dog characteristics the association was [OR 0.45 (95% CI 0.23–0.88), p=0.02]. When adjusted for keeping conditions, dog characteristics, and other feeding factors, the association was of similar magnitude but not significance [OR 0.56 (95% CI 0.27–1.15), p=0.12].DiscussionThe study indicates possible protective associations of feeding the dog with dietary sources of fish fat against epilepsy, although the result could be confounded by other feeding factors. Findings are compatible with current knowledge regarding the role of omega-3 fatty acids and ketogenic diet, a low carbohydrate, high fat diet as supportive treatments of epilepsy. As our findings are based on observations, we suggest the possibility of causality but do not prove it. Dietary intervention studies should now be conducted to confirm our findings

Early life programming by diet can play a role in risk reduction of otitis in dogs

IntroductionOtitis in dogs is often chronic while local treatment primarily consists of flushing, antibiotics, and/or antifungals. We were interested in finding early life variables that associate with otitis later in life, preferably some that could be modified.MethodsA cross-sectional hypothesis-driven study with longitudinal data was performed to search for associations between pre- and postnatal exposures, and the incidence of owner-reported otitis in dogs at over 1 year of age. The multivariate logistic regression analysis study included data from 3,064 dogs and explored 26 different early life variables at four early life stages: prenatal, neonatal, postnatal, and puppyhood. We compared two feeding patterns, a non-processed meat-based diet (NPMD, raw) and an ultra-processed carbohydrate-based diet (UPCD, dry).ResultsWe report that eating a NPMD diet significantly decreased the risk of otitis later in life, while eating a UPCD diet significantly increased the risk. This was seen in different life stages of mother or puppy: The maternal diet during pregnancy (p=0.011) and the puppies’ diet from 2 to 6 months of age (p=0.019) were both significantly associated with otitis incidence later in life, whereas the puppies’ first solid diet, was associated in the same way, but did not reach significance (p=0.072). Also, analyzing food ratios showed that when puppies were consuming &gt;25% of their food as NPMD it significantly decreased their incidence of otitis later in life, while a ratio of &gt;75% UPCD in their diet significantly increased their risk of otitis. Also, if the dog was born in the current family, was exposed to sunlight for more than 1 hour daily, and was raised on a dirt floor during puppyhood, there was a lower risk of otitis development later in life.DiscussionThe findings only suggest causality, and further studies are required. However, we propose that veterinarians, breeders, and owners can impact otitis risk by modifying factors such as diet and environment

Novel HTS Strategy Identifies TRAIL-Sensitizing Compounds Acting Specifically Through the Caspase-8 Apoptotic Axis

Tumor Necrosis Factor-Related Apoptosis-Inducing Ligand (TRAIL) is potentially a very important therapeutic as it shows selectivity for inducing apoptosis in cancer cells whilst normal cells are refractory. TRAIL binding to its cognate receptors, Death Receptors-4 and -5, leads to recruitment of caspase-8 and classical activation of downstream effector caspases, leading to apoptosis. As with many drugs however, TRAIL's usefulness is limited by resistance, either innate or acquired. We describe here the development of a novel 384-well high-throughput screening (HTS) strategy for identifying potential TRAIL-sensitizing agents that act solely in a caspase-8 dependent manner. By utilizing a TRAIL resistant cell line lacking caspase-8 (NB7) compared to the same cells reconstituted with the wild-type protein, or with a catalytically inactive point mutant of caspase-8, we are able to identify compounds that act specifically through the caspase-8 axis, rather than through general toxicity. In addition, false positive hits can easily be “weeded out” in this assay due to their activity in cells lacking caspase-8-inducible activity. Screening of the library of pharmacologically active compounds (LOPAC) was performed as both proof-of-concept and to discover potential unknown TRAIL sensitizers whose mechanism is caspase-8 mediated. We identified known TRAIL sensitizers from the library and identified new compounds that appear to sensitize specifically through caspase-8. In sum, we demonstrate proof-of-concept and discovery of novel compounds with a screening strategy optimized for the detection of caspase-8 pathway-specific TRAIL sensitizers. This screen was performed in the 384-well format, but could easily be further miniaturized, allows easy identification of artifactual false positives, and is highly scalable to accommodate diverse libraries

Exosome-mediated MIR211 modulates tumor microenvironment via the DUSP6-ERK5 axis and contributes to BRAFV600E inhibitor resistance in melanoma

The microRNA MIR211 is an important regulator of melanoma tumor cell behavior. Previous studies suggested that in certain tumors, MIR211 acted as a tumor suppressor while in others it behaved as an oncogenic regulator. When MIR211 is expressed in BRAFV600E-mutant A375 melanoma cells in mouse xenografts, it promotes aggressive tumor growth accompanied by increased cellular proliferation and angiogenesis. We demonstrate that MIR211 is transferred to adjacent cells in the tumor micro-environment via exosomes. Cross-species genome-wide transcriptomic analysis showed that human tumor-derived MIR211 interacts with the mouse transcriptome in the tumor microenvironment, and activates ERK5 signaling in human tumor cells via the modulation of a feedback loop. Human miR211 directly inhibits human DUSP6 protein phosphatase at the post-transcriptional level. We provide support for the hypothesis that DUSP6 inhibition conferred resistance of the human tumor cells to the BRAF inhibitor vemurafenib and to the MEK inhibitor cobimetinib, with associated increases in ERK5 phosphorylation. These findings are consistent with a model in which MIR211 regulates melanoma tumor proliferation and BRAF inhibitor resistance by inducing ERK5 signaling within the complex tumor microenvironment. We propose that the MIR211-ERK5 axis represents an important and sensitive regulatory arm in melanoma with potential theranostic applications