MAP2 IHC detection: a marker of antigenicity in CNS tissues

Immunohistochemistry (IHC) is used to detect antibody-specific antigens in tissues; the results depend on the ability of the primary antibodies to bind to their antigens. Therefore, results depend on the quality of preservation of the specimen. Many investigators have overcome the deleterious effects of over-fixation on the binding of primary antibodies to specimen antigens using IHC, but if the specimen is under-fixed or fixation is delayed, false negative results could be obtained despite certified laboratory practices. Microtubule-associated protein 2 (MAP2) is an abundant microtubule-associate protein that participates in the outgrowth of neuronal processes and synaptic plasticity; it is localized primarily in cell bodies and dendrites of neurons. MAP2 immunolabeling has been reported to be absent in areas of the entorhinal cortex and hippocampus of Alzheimer’s disease brains that were co-localized with the dense-core type of amyloid plaques. It was hypothesized that the lack of MAP2 immunolabeling in these structures was due to the degradation of the MAP2 antigen by the neuronal proteases that were released as the neurons lysed leading to the formation of these plaques. Because MAP2 is sensitive to proteolysis, we hypothesized that changes in MAP2 immunolabeling may be correlated with the degree of fixation of central nervous system (CNS) tissues. We detected normal MAP2 immunolabeling in fixed rat brain tissues, but MAP2 immunolabeling was decreased or lost in unfixed and delayed-fixed rat brain tissues. By contrast, two ubiquitous CNS-specific markers, myelin basic protein and glial fibrillary acidic protein, were unaffected by the degree of fixation in the same tissues. Our observations suggest that preservation of various CNS-specific antigens differs with the degree of fixation and that the lack of MAP2 immunolabeling in the rat brain may indicate inadequate tissue fixation. We recommend applying MAP2 IHC for all CNS tissues as a pre-screen to assess the quality of the tissue preservation and to avoid potentially false negative IHC results.     

Prevalence of risk factors for cardiovascular disease in Canadians 55 to 74 years of age: results from the Canadian Heart Health Surveys, 1986-1992

BACKGROUND: By 2016, the proportion of Canadians older than 65 years of age will increase to 16%, and there will be an increase in the absolute number of cases of cardiovascular disease in older Canadians. The Canadian Heart Health Surveys database provides information about this population upon which health policy related to cardiovascular disease can be based. This paper presents for the first time population-based data on the risk factors for cardiovascular disease in older Canadians. METHODS: Canadians from all 10 provinces participated in surveys of cardiovascular risk factors; health insurance registries were used as sampling frames. In each province, probability samples of 2200 adults 18 to 74 years old not living in institutions, on reserves or in military camps were asked to participate in interviews and to undergo testing at clinics for major risk factors for cardiovascular disease. RESULTS: A total of 2739 men (response rate 70%) and 2617 women (response rate 66%) aged 55 to 74 years participated in the survey and also provided follow-up clinical measurements at the clinic. Overall, 52% of participants were hypertensive, 26% had isolated systolic hypertension, and 30% had a total blood cholesterol level of 6.2 mmol/L or greater. Rates of current smoking were lower in women than men (17% v. 22%). Overall, 87% of men and 78% of women who were current smokers smoked at least 10 cigarettes per day. Only slightly more than half of participants exercised at least once a week for at least 15 minutes, and almost half had a body mass index of 27 or greater. In only 4% was no major risk factor for cardiovascular disease detected. INTERPRETATION: Significant numbers of older Canadians have one or more major risk factors for cardiovascular disease. Many of these risk factors are amenable to modification.     

Controlling harmful algal blooms through clay flocculation

The potential use of clays to control harmful algal blooms (HABs) has been explored in East Asia, Australia, the United States, and Sweden. In Japan and South Korea, minerals such as montmorillonite, kaolinite, and yellow loess, have already been used in the field effectively, to protect fish mariculture from Cochlodinium spp. and other blooms. Cell removal occurs through the flocculation of algal and mineral particles, leading to the formation of larger aggregates (i.e. marine snow), which rapidly settle and further entrain cells during their descent. In the U.S., several clays and clay-rich sediments have shown high removal abilities (e.g. > 80% cell removal efficiency) against Karenia brevis, Heterosigma akashiwo, Pfiesteria piscicida and Aureococcus anophagefferens. In some cases, the removal ability of certain clays was further enhanced with chemical flocculants, such as polyaluminum chloride (PAC), to increase their adhesiveness. However, cell removal was also affected by bloom concentration, salinity, and mixing. Cell mortality was observed after clay addition, and increased with increasing clay concentration, and prolonged exposure to clays in the settled layer. Mesocosm, field enclosure, and flume experiments were also conducted to address cell removal with increasing scale and flow, water-column impacts, and the possible benthic effects from clay addition. Results from these studies will be presented, especially those in regards to water quality, seawater chemistry, bottom erodibility and faunal impacts in the benthos. At this time, clay dispersal continues to be a promising method for controlling HABs and mitigating their impacts based on existing information and experimental data.     

Genetic transformation of novel isolates of chicken Lactobacillus bearing probiotic features for expression of heterologous proteins: a tool to develop live oral vaccines

Background  

The use of lactic acid bacteria as vehicles to delivery antigens to immunize animals is a promising issue. When genetically modified, these bacteria can induce a specific local and systemic immune response against selected pathogens. Gastric acid and bile salts tolerance, production of antagonistic substances against pathogenic microorganisms, and adhesive ability to gut epithelium are other important characteristics that make these bacteria useful for oral immunization.     

High genetic and morphological diversity despite range contraction in the diploid Hieracium eriophorum (Asteraceae) endemic to the coastal sand dunes of south‐west France

Endemic plants inhabiting coastal sand dunes show augmented extinction risks due to the dynamic nature of dunes and strong human pressure on coastal areas. To investigate the survival strategies and threats to long‐term survival of such species, we combined genetic, morphological and biogeographical approaches, using the example of Hieracium eriophorum (Asteraceae) and its putative cryptic sister species H. prostratum, which are endemic to the longest coastal sand dune in Europe. An analysis of amplified fragment length polymorphism revealed high within‐population genetic variability, and slight isolation by distance was the only indication of genetic population structure. Thus, no signs of genetic threats to survival were found. Furthermore, genetic and morphometric data provided no evidence for the existence of two species. Therefore, we propose to synonymize H. prostratum with H. eriophorum and provide a nomenclatural overview with typification. Finally, an analysis of historical distribution records showed that, during the last 100 years, the species was lost from its range margins, where its habitat became fragmented. Taken together, our results suggest that one successful survival strategy of narrow endemics may be the achievement of large local population sizes on a small geographical scale, thus avoiding the genetic problems inherent to small and fragmented populations. Dune management policies should thus take care that the current tendencies to allow more erosion will not result in too severe fragmentation of the remaining continuous stretches of dune habitat. © 2012 The Linnean Society of London, Botanical Journal of the Linnean Society, 2012, 169 , 365–377.     

Irisin immunohistochemistry in gastrointestinal system cancers

Cancer is the leading cause of morbidity and mortality worldwide. Some studies have shown that high heat kills cancer cells. Irisin is a protein involved in heat production by converting white into brown adipose tissue, but there is no information about how its expression changes in cancerous tissues. We used irisin antibody immunohistochemistry to investigate changes in irisin expression in gastrointestinal cancers compared to normal tissues. Irisin was found in human brain neuroglial cells, esophageal epithelial cells, esophageal epidermoid carcinoma, esophageal adenocarcinoma and neuroendocrine esophageal carcinoma, gastric glands, gastric adenosquamous carcinoma, gastric neuroendocrine carcinoma, gastric signet ring cell carcinoma, neutrophils in vascular tissues, intestinal glands of colon, colon adenocarcinoma, mucinous colon adenocarcinoma, hepatocytes, hepatocellular carcinoma, islets of Langerhans, exocrine pancreas, acinar cells and interlobular and interlobular ducts of normal pancreas, pancreatic ductal adenocarcinoma, and intra- and interlobular ducts of cancerous pancreatic tissue. Histoscores (area × intensity) indicated that irisin was increased significantly in gastrointestinal cancer tissues, except liver cancers. Our findings suggest that the relation of irisin to cancer warrants further investigation.     

Flocculation and removal of the brown tide organism, Aureococcus anophagefferens (Chrysophyceae), using clays

Previous attempts to remove the brown tideorganism, Aureococcusanophagefferens, through flocculation withclays have been unsuccessful, in spite ofadopting concentrations and dispersalprotocols that yielded excellent cellremoval efficiency (RE >90%) with otherspecies, so a study was planned to improvecell removal. Four modifications in claypreparation and dispersal were explored: 1)varying the salinity of the claysuspension; 2) mixing of the clay-cellsuspension after clay addition; 3) varyingof concentration of the initial clay stock;4) pulsed loading of the clay slurry. Theeffect of salinity was dependent on theclay mineral type: phosphatic clay (IMC-P2)had a higher RE than kaolinite (H-DP) whenseawater was used to disperse the clay, butH-DP removed cells more efficiently whensuspended in distilled water prior toapplication. Mixing after dispersalapproximately doubled RE for both clayscompared to when the slurry was layeredover the culture surface. Lowering theconcentration of clay stock and pulsing theclay loading increased RE, regardless ofmineral type. However, this increase wasmore apparent for clays dispersed inseawater than in distilled water. Ingeneral, application procedures thatdecrease the rate of self-aggregation amongthe clay particles and increase thecollision frequency between clay particlesand A. anophagefferens achieve highercell removal efficiency. These empiricalstudies demonstrated that clays might be animportant control option for the brown tideorganism, given the proper attention topreparation, dispersal methods,environmental impacts, and the hydrodynamicproperties of the system being treated. Implications for the treatment of browntides in the field are discussed.     

Osteogenesis imperfecta: the audiological phenotype lacks correlation with the genotype

Background

Neurofibromatosis 1 (NF1), a common autosomal dominant disorder, was shown in one study to be associated with a 15-year decrease in life expectancy. However, data on mortality in NF1 are limited. Our aim was to evaluate mortality in a large retrospective cohort of NF1 patients seen in France between 1980 and 2006.

Methods

Consecutive NF1 patients referred to the National French Referral Center for Neurofibromatoses were included. The standardized mortality ratio (SMR) with its 95% confidence interval (CI) was calculated as the ratio of observed over expected numbers of deaths. We studied factors associated with death and causes of death.

Results

Between 1980 and 2006, 1895 NF1 patients were seen. Median follow-up was 6.8 years (range, 0.4-20.6). Vital status was available for 1226 (65%) patients, of whom 1159 (94.5%) survived and 67 (5.5%) died. Overall mortality was significantly increased in the NF1 cohort (SMR, 2.02; CI, 1.6-2.6; P < 10^-4). The excess mortality occurred among patients aged 10 to 20 years (SMR, 5.2; CI, 2.6-9.3; P < 10^-4) and 20 to 40 years (SMR, 4.1; 2.8-5.8; P < 10^-4). Significant excess mortality was found in both males and females. In the 10-20 year age group, females had a significant increase in mortality compared to males (SMR, 12.6; CI, 5.7-23.9; and SMR, 1.8; CI, 0.2-6.4; respectively). The cause of death was available for 58 (86.6%) patients; malignant nerve sheath tumor was the main cause of death (60%).

Conclusions

We found significantly increased SMRs indicating excess mortality in NF1 patients compared to the general population. The definitive diagnosis of NF1 in all patients is a strength of our study, and the high rate of death related to malignant transformation is consistent with previous work. The retrospective design and hospital-based recruitment are limitations of our study. Mortality was significantly increased in NF1 patients aged 10 to 40 years and tended to be higher in females than in males.     

Removal of harmful algal cells (Karenia brevis) and toxins from seawater culture by clay flocculation

Harmful algal blooms (HABs) occur worldwide causing serious threat to marine life, and to public health through seafood-borne illnesses and exposure to toxin-containing marine aerosol. This study was undertaken to assess the ability of phosphatic clay to remove the toxic dinoflagellate, Karenia brevis, and the potent neurotoxins (brevetoxins) produced by this species. Results showed that the addition of an aqueous slurry of 0.75 g (dry weight) clay to 3 l of K. brevis culture, containing 5×10⁶ and 10×10⁶ cells/l, removed 97±4% of brevetoxins from the water column within 4 h after the addition of clay. Clay flocculation of extra-cellular brevetoxins, released from cells ruptured (lyzed) by ultrasonication, removed 70±10% of the toxins. Addition of the chemical flocculant, polyaluminum chloride (PAC), removed all of the extra-cellular toxins. A 14 day study was undertaken to observe the fate of brevetoxins associated with clay flocculation of viable K. brevis cells. At 24 h following the clay addition, 90±18% of the toxins were removed from the water column, along with 85±4% of the cells. The toxin content of clay diminished from 208±13 μg at Day 1, to 121±21 μg at Day 14, indicating that the phosphatic clay retained about 58% of the toxins throughout the 14-day period. These studies showed the utility of natural clay as a means of reducing adverse effects from HABs, including removal of dissolved toxins, in the water column, although considerable work clearly remains before this approach can be used on natural blooms in open waters.