Health status of fathead minnow (Pimephales promelas) populations in a municipal wastewater effluent-dominated stream in the Canadian prairies, Wascana Creek, Saskatchewan.

Their unique hydrological and climatic conditions render surface water systems in the southern Canadian Prairies at an elevated risk from exposure to contaminants released from municipal wastewater effluents (MWWEs). The aim of this study was to characterize the potential health effects and their underlying molecular mechanisms in populations of fathead minnow (Pimephales promelas; FHM) in Wascana Creek, an effluent dominated stream in Southern Saskatchewan, Canada. Studies were conducted during the spawning season in 2014 and 2015 to assess responses in terms of overall health, reproductive functions, plasma sex steroid hormone levels, and expression of selected genes along the hypothalamus-pituitary-gonadal axis. FHM downstream of the effluent fallout had lower gonadosomatic indices and significantly greater hepatosomatic indices compared to upstream populations. In both male and female FHMs, significantly greater occurrence and severity of gonadal degradation and delayed maturation were observed in downstream fish compared to upstream fish. Downstream males also displayed lower scores of secondary sexual characteristics and a decreasing trend in plasma 11-ketotestosterone levels. Interestingly, no indications of exposure to estrogenic compounds, such as occurrence of testicular oocytes were observed, which was in accordance with the lack of presence of key biomarkers of estrogenic exposure, such as induction of vitellogenin. In general, expression of the majority of transcripts measured in FHMs downstream of the effluent fallout was significantly downregulated, which supports observations of the general deterioration of the health and reproductive status of these fish. Chemical analysis indicated that 10 pharmaceuticals and personal care products (PPCPs) were present at the downstream site, some at sufficiently great concentrations that may present a risk to aquatic organisms. With continuous exposure to a diverse number of stressors including high nutrient and ammonia levels, the presence of a variety of PPCPs and other contaminants, Wascana Creek should be considered as an ecosystem at risk.

Evaluation of Anatomical and Functional Hip Joint Center Methods: The Effects of Activity Type, Gender, and Proximal Reference Segment.

Accurate hip joint center (HJC) location is critical when studying hip joint biomechanics. The HJC is often determined from anatomical methods, but functional methods are becoming increasingly popular. Several studies have examined these methods using simulations and in vivo gait data, but none has studied high-range of motion activities, such a chair rise, nor has HJC prediction been compared between males and females. Furthermore, anterior superior iliac spine (ASIS) marker visibility during chair rise can be problematic, requiring a sacral cluster as an alternative proximal segment; but functional HJC has not been explored using this approach. For this study, the quality of HJC measurement was based on the joint gap error (JGE), which is the difference in global HJC between proximal and distal reference segments. The aims of the present study were to: (1) determine if JGE varies between pelvic and sacral referenced HJC for functional and anatomical methods, (2) investigate which functional calibration motion results in the lowest JGE and if the JGE varies depending on movement type (gait versus chair rise) and gender, and (3) assess whether the functional HJC calibration results in lower JGE than commonly used anatomical approaches and if it varies with movement type and gender. Data were collected on 39 healthy adults (19 males and 20 females) aged 14-50 yr old. Participants performed four hip "calibration" tests (arc, cross, star, and star-arc), as well as gait and chair rise (activities of daily living (ADL)). Two common anatomical methods were used to estimate HJC and were compared to HJC computed using a published functional method with the calibration motions above, when using pelvis or sacral cluster as the proximal reference. For ADL trials, functional methods resulted in lower JGE (12-19 mm) compared to anatomical methods (13-34 mm). It was also found that women had significantly higher JGE compared to men and JGE was significantly higher for chair rise compared to gait, across all methods. JGE for sacrum referenced HJC was consistently higher than for the pelvis, but only by 2.5 mm. The results indicate that dynamic hip range of movement and gender are significant factors in HJC quality. The findings also suggest that a rigid sacral cluster for HJC estimation is an acceptable alternative for relying solely on traditional pelvis markers.

Heparin-induced thrombocytopenia: laboratory studies.

This report describes studies into the pathophysiology of heparin-induced thrombocytopenia. The IgG fraction from each of nine patients with heparin-induced thrombocytopenia caused heparin-dependent platelet release of radiolabeled serotonin. Both the Fc and the Fab portions of the IgG molecule were required for the platelet reactivity. The platelet release reaction could be inhibited by the Fc portion of normal human or goat IgG, and patient F(ab')2, but not F(ab')2 from healthy controls. These results suggested that the Fab portion of IgG binds to heparin forming an immune complex and the immune complexes initiate the platelet release reaction by binding to the platelet Fc receptors. To directly challenge this hypothesis, we preincubated the serotonin-labeled platelets with the monoclonal antibody against the platelet Fc receptor (IV.3). This monoclonal antibody completely inhibited the release reaction caused by heparin and patient sera, as well as heat aggregated IgG, but did not block collagen or thrombin-induced platelet release. Heparin-dependent platelet release also could be inhibited in vitro by the addition of monocytes and neutrophils, but not by red cells, presumably because the Fc receptors on the phagocytic cells have a higher binding affinity for IgG complexes than do platelets. Platelets from patients with congenital deficiencies of specific glycoproteins Ib and IX (Bernard-Soulier syndrome) and IIb and IIIa (Glanzmann's thrombasthenia) displayed normal heparin-dependent release indicating that the release reaction did not require the participation of these glycoproteins. These studies indicate that heparin-induced thrombocytopenia is an IgG-heparin immune complex disorder involving both the Fab and Fc portion of the IgG molecule.

Quantitation of red cell-associated IgG using an immunoradiometric assay.

In this report, we describe a sensitive immunoradiometric assay (IRMA) for quantitating IgG on the surface of red cells. Washed red cells were prepared to a purity of greater than 99.9 percent. Varying dilutions of these cells were incubated with a fixed concentration of 125I-anti-IgG. After equilibrium was achieved, the unbound 125I-anti-IgG was measured by the addition of IgG covalently linked to agarose beads. The red cells were lysed by detergent, and the 125I-anti-IgG bound to the IgG-beads was measured. The amount of IgG on the red cells was determined by relating the concentration of test red cells causing 50 percent inhibition of binding of the 125I-anti-IgG to the IgG-beads to 50 percent inhibition of binding caused by the IgG standard. Using this assay, the red cell-associated IgG (RCA-IgG) of 20 healthy male and female controls with normal hemoglobin concentrations was 7.23 +/- 6.11 fg IgG per 10(3) cells (mean +/- 2 SD). The mean RCA-IgG on washed cells from 34 different tests performed on 19 anemic patients with clinically diagnosed autoimmune hemolytic anemia was 176.1 +/- 375.6 fg IgG per 10(3) cells. There was no correlation between the levels of RCA-IgG and the hemoglobin levels or reticulocyte counts in these patients.

The amount of platelet-bound albumin parallels the amount of IgG on washed platelets from patients with immune thrombocytopenia.

The biologic relevance of the increased platelet-associated IgG (PAIgG) on platelets from patients with idiopathic thrombocytopenic purpura (ITP) is unclear. Platelets from ITP patients are often larger than normal, and it is possible that the increased IgG is not specific but passively related to platelet size. The measurement of platelet-bound albumin could provide information concerning the specificity of the platelet-bound IgG, since albumin, like IgG, is a plasma protein, but unlike IgG, is not an active participant in immunologic reactions. Albumin is also a normal constituent of platelet membrane, and increased platelet albumin could indicate an increased platelet mass. Platelet-bound albumin, IgG, and total platelet protein were measured on both intact and disrupted platelets from healthy individuals (n = 25) and patients with ITP (n = 21). Platelet IgG and albumin were measured in an immunoradiometric assay using intact antisera and F(ab')2 fragments prepared from the same antisera. There was no relationship between platelet-bound IgG or albumin, and platelet size measured by either platelet protein or platelet volume, (r less than 0.3 for all interactions). In contrast, there was a significant correlation between platelet-bound albumin and platelet-bound IgG (r = 0.7, n = 21, p less than 0.001). Those patients with elevated platelet PAIgG also had elevated platelet albumin, and this relationship was irrespective of the total platelet protein content or mean platelet volume. It is possible that the increased platelet-bound IgG in ITP reflects an increase in platelet surface area or contaminating platelet fragments that are not manifested as an increase in platelet volume or total platelet protein. Alternatively, a platelet membrane abnormality may occur in ITP that results in the uptake of significant amounts of plasma proteins. Either possibility implies that not all of the IgG on platelets from patients with ITP is pathologic IgG.