►
Anaplasma marginale infects cattle for years by parasitizing red blood cells. ► Red blood cells only circulate for about four months and then are phagocytized. ► Prior
in vitro work suggested that ...endothelial cells might also be infected. ►
In vivo endothelial cell infection was investigated by multiple techniques. ► No conclusive proof of
A. marginale endothelial cell infection
in vivo was found.
Continuous culture of
Anaplasma marginale in endothelial cells and the potential implications for vaccine development heightened interest in determining the importance of endothelial cells in the
A. marginale life cycle.
A. marginale-infection trials were performed to determine if endothelial cells are an
in vivo host cell in cattle and if
A. marginale from
in vitro endothelial cells were infective to cattle. Adult, immunocompetent steers were infected by tick-feeding transmission and were euthanized at different points in the parasitemic cycle. Based on quantitative PCR, the tissue distribution of
A. marginale DNA during peak and trough parasitemia was variable with higher quantities observed in spleen, lung, hemal nodes, and abomasum.
A. marginale was not conclusively identified in tissue endothelial cells from the steers’ tick-bitten dermis or post-mortem tissues using three microscopy techniques (dual indirect immunofluorescence, transmission electron microscopy, and
in situ DNA target-primed rolling-circle amplification of a padlock probe). Intravenous inoculation of spleen-intact or splenectomized calves with endothelial cell culture-derived VA isolate
A. marginale did not cause seroconversion or clinical anaplasmosis regardless of whether the endothelial culture-derived bacteria were inoculated as host cell-free organisms or within endothelial cells and regardless of the type of endothelial cell culture used – RF/6A primate endothelial cells or primary bovine testicular vein endothelial cells. Data presented here suggest that endothelial cells are likely not a pivotal component of the
A. marginale life cycle
in vivo.
Abstract Background Available studies vary in their estimated prevalence of attention deficit/hyperactivity disorder (ADHD) in substance use disorder (SUD) patients, ranging from 2 to 83%. A better ...understanding of the possible reasons for this variability and the effect of the change from DSM-IV to DSM-5 is needed. Methods A two stage international multi-center, cross-sectional study in 10 countries, among patients form inpatient and outpatient addiction treatment centers for alcohol and/or drug use disorder patients. A total of 3558 treatment seeking SUD patients were screened for adult ADHD. A subsample of 1276 subjects, both screen positive and screen negative patients, participated in a structured diagnostic interview. Results Prevalence of DSM-IV and DSM-5 adult ADHD varied for DSM-IV from 5.4% (CI 95%: 2.4–8.3) for Hungary to 31.3% (CI 95%:25.2–37.5) for Norway and for DSM-5 from 7.6% (CI 95%: 4.1–11.1) for Hungary to 32.6% (CI 95%: 26.4–38.8) for Norway. Using the same assessment procedures in all countries and centers resulted in substantial reduction of the variability in the prevalence of adult ADHD reported in previous studies among SUD patients (2–83% → 5.4–31.3%). The remaining variability was partly explained by primary substance of abuse and by country (Nordic versus non-Nordic countries). Prevalence estimates for DSM-5 were slightly higher than for DSM-IV. Conclusions Given the generally high prevalence of adult ADHD, all treatment seeking SUD patients should be screened and, after a confirmed diagnosis, treated for ADHD since the literature indicates poor prognoses of SUD in treatment seeking SUD patients with ADHD.
To determine blood cell morphologic characteristics and hematologic and plasma biochemical reference ranges for iguanas housed in a warm indoor and outdoor environment with regular exposure to direct ...sunlight.
Original study.
51 clinically normal iguanas (18 males, 25 females, and 8 juveniles) housed in 3 Florida locations.
Blood was collected from the coccygeal or ventral abdominal vein. Any samples that had obvious hemolysis or clot formation were not used. Leukocyte counts were determined manually; other hematologic values were obtained by use of a commercially available cell counter. Plasma biochemical values were determined by use of a spectrophotometric chemistry analyzer. Blood smears were stained with Wright-Giemsa and cytochemical stains for morphologic and cytochemical evaluation.
Hematologic ranges were generally higher in this study than previously reported. Thrombocytes were variable in appearance between individuals and sometimes difficult to distinguish from lymphocytes on a Wright-Giemsa preparation. Concentrations of calcium, phosphorus, total protein, globulins, and cholesterol were significantly higher, and the albumin:globulin ratio was significantly lower, in healthy gravid females than in male or nongravid female iguanas. Nongravid females had significantly higher calcium and cholesterol concentrations, compared with males. The calcium:phosphorus ratio was > 1 in all iguanas. Gravid females had a calcium phosphorus product ranging between 210 and 800. Intracytoplasmic inclusions were identified within the erythrocytes of some iguanas.
Hematologic ranges for iguanas in this study are higher than those reported for iguanas. Sex and age of the iguana should be considered when evaluating biochemical values. Healthy ovulating and gravid females may have significantly increased electrolyte and protein concentrations, but maintain a calcium:phosphorus ratio > 1.
Major surface protein 5 (Msp5) of Anaplasma marginale is highly conserved in the genus Anaplasma and the antigen used in a commercially available competitive enzyme-linked immunosorbent assay ...(cELISA) for serologic identification of cattle with anaplasmosis. This study analyzes the degrees of conservation of Msp5 among various isolates of Anaplasma phagocytophilum and the extent of serologic cross-reactivity between recombinant Msp5 (rMsp5) of Anaplasma marginale and A. phagocytophilum. The msp5 genes from various isolates of A. phagocytophilum were sequenced and compared. rMsp5 proteins of A. phagocytophilum and A. marginale were used separately in an indirect ELISA to detect cross-reactivity in serum samples from humans and dogs infected with A. phagocytophilum and cattle infected with A. marginale. Serum samples were also tested with a commercially available competitive ELISA that uses monoclonal antibody ANAF16C1. There were 100% sequence identities in the msp5 genes among all of the A. phagocytophilum isolates from the United States and a horse isolate from Sweden. Sheep isolates from Norway and dog isolates from Sweden were 99% identical to one another but differed in 17 base pairs from the United States isolates and the horse isolate. Serologic cross-reactivity was identified when serum samples from cattle infected with A. marginale were reacted with rMsp5 of A. phagocytophilum and when serum samples from humans and dogs infected with A. phagocytophilum were reacted with rMsp5 of A. marginale in an indirect-ELISA format. Serum samples from dogs or humans infected with A. phagocytophilum did not cross-react with rMsp5 of A. marginale when tested with the commercially available cELISA. These results suggest that rMsp5 of A. phagocytophilum is highly conserved among United States and European isolates and that serologic distinction between A. phagocytophilum and A. marginale infections cannot be accomplished if rMsp5 from either organism is used in an indirect ELISA.
The aim of this study was to compare three different enzyme-linked immunosorbant assays (recombinant major antigenic protein 2 (rMAP2)-ELISA, the Immunocomb
® (Biogal, Israel) and the Snap
® 3Dx ...assay (IDEXX Laboratories Inc., USA)) with the indirect immunofluorescent antibody test in detecting anti-
Ehrlichia canis immunoglobulin-G (IgG) antibodies. Samples tested were collected from dogs suspected to be naturally infected with
E. canis and from experimentally infected dogs.
When qualitative results (positive/negative) were compared, there was an overall agreement of 81% (54/67) between the indirect immunofluorescence antibody (IFA) test and the rMAP2-ELISA. An overall agreement of 94% (63/67) was found between the IFA test and the Immunocomb
®, and an overall agreement of 91% (61/67) was found between the IFA test and the Snap
® 3Dx assay. In 50 of 67 (74.6%) samples tested, complete agreement in the qualitative results was found in all four tests. Sixteen of 17 samples with disagreement in the qualitative results were found to have IFA titers of 1:320 or less. The sensitivities and specificities of the tests were found to be 0.71 and 0.85 for the rMAP2-ELISA, 0.86 and 0.98 for the Immunocomb
®, and 0.71 and 1.00 for the Snap
® 3Dx assay.
The tests performed in this study were found to be highly specific in detecting
E. canis antibodies. Their sensitivity was found to be low with sera having IFA titers of ≤1:320, while high with sera having titers greater than 1:320. Repeating the serological tests 1–2 weeks after the first antibody assay may overcome the sensitivity problem with titers of ≤1:320.
We analyzed the structure of the expression site encoding the immunoprotective protein MSP2/P44 from multiple Anaplasma phagocytophilum strains in the United States. The sequence of p44ESup1 had ...diverged in Ap-variant 1 strains infecting ruminants. In contrast, no differences were detected between A. phagocytophilum strains infecting humans and domestic dogs.
Objective The objective of the study was to create a predictive model for preterm birth (PTB) from available clinical data and serum analytes. Study Design Serum analytes and routine pregnancy ...screening plus cholesterol and corresponding health information were linked to birth certificate data for a cohort of 2699 Iowa women with serum sampled in the first and second trimester. Stepwise logistic regression was used to select the best predictive model for PTB. Results Serum screening markers remained significant predictors of PTB, even after controlling for maternal characteristics. The best predictive model included maternal characteristics, first-trimester total cholesterol, total cholesterol change between trimesters, and second-trimester alpha-fetoprotein and inhibin A. The model showed better discriminatory ability than PTB history alone and performed similarly in subgroups of women without past PTB. Conclusion Using clinical and serum screening data, a potentially useful predictor of PTB was constructed. Validation and replication in other populations, and incorporation of other measures that identify PTB risk, like cervical length, can be a step toward identifying additional women who may benefit from new or currently available interventions.
While anthropogenic emissions have dramatically elevated lead concentrations in the North Atlantic troposphere and surface waters by orders of magnitude above natural levels Murozumi et al., 1969; ...Schaule and Patterson, 1983; Boyle et al., 1986, it has been assumed that the relatively low lead levels in North Atlantic abyssal waters are not yet contaminated Schaule and Patterson, 1981; Flegal and Patterson, 1983. That misperception is redressed by the following stable lead isotopic composition data which reveal the advective transport of industrial lead into those deep basin waters through the formation of North Atlantic Deep Water (NADW). Additionally, spatial gradients in the isotopic signatures of anthropogenic lead within the North Atlantic abyss appear to serve as transient tracers of contaminant penetration rates.
Cholesteryl ester storage disease (CESD) is a rare autosomal recessive disease caused by mutations in LIPA. Here we describe two different clinical presentations of this disease: one case with a ...clear phenotype of familial hypercholesterolaemia and one case with hepatosplenomegaly from childhood onwards. These two cases exemplify the diversity of clinical phenotypes of patients with CESD. Knowledge on the phenotypic variability of the disease is of clinical relevance in light of enzyme replacement therapy (sebelipase alpha) for patients with mutations in LIPA, which is currently under development.
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