Maternal lineages and Alzheimer disease risk in the Old Order Amish

Old Order Amish, founded by a small number of Swiss immigrants, exist in culturally isolated communities across rural North America. The consequences of genetic isolation and inbreeding within this group are evident by increased frequencies of many monogenic diseases and several complex disorders. Conversely, the prevalence of Alzheimer disease (AD), the most common form of dementia, is lower in the Amish than in the general American population. Since mitochondrial dysfunction has been proposed as an underlying cause of AD and a specific haplogroup was found to affect AD susceptibility in Caucasians, we investigated whether inherited mitochondrial haplogroups affect risk of developing AD dementia in Ohio and Indiana Amish communities. Ninety-five independent matrilines were observed across six large pedigrees and three small pedigrees then classified into seven major European haplogroups. Haplogroup T is the most frequent haplogroup represented overall in these maternal lines (35.4%) while observed in only 10.6% in outbred American and European populations. Furthermore, haplogroups J and K are less frequent (1.0%) than in the outbred data set (9.4–11.2%). Affected case matrilines and unaffected control lines were chosen from pedigrees to test whether specific haplogroups and their defining SNPs confer risk of AD. We did not observe frequency differences between AD cases compared to controls overall or when stratified by sex. Therefore, we suggest that the genetic effect responsible for AD dementia in the affected Amish pedigrees is unlikely to be of mitochondrial origin and may be caused by nuclear genetic factors.     

An analysis of genetic variation across the MBL2 locus in Dutch Caucasians indicates that 3′ haplotypes could modify circulating levels of mannose-binding lectin

Mannose-binding protein (MBL) is a critical component of innate immunity and provides first-line protection against pathogens. Both circulating MBL serum levels and functional activity have been correlated with common genetic variants in the MBL2 gene. Associations between MBL deficiency and severe infections have been reported in immuno-incompetent patients and for autoimmune disorders; however, measured MBL serum levels do not fully correlate with the ‘secretor haplotypes’. Previously, the MBL2 locus was resequenced and determined that a recombination hotspot divides MBL2 into two haplotype blocks. It was sought to investigate whether additional variants, in either block structure could associate with MBL serum levels. Therefore, 31 common variants were analysed across the locus in 212 DNA samples of healthy Caucasian individuals with known MBL serum concentrations. The additional 5′ variants were in strong linkage to the elements of the ‘secretor haplotypes’; functional alleles B, C and D also lie on restricted haplotypes. Four variants in the 3′ block (Ex4-1483T>C, Ex4-1067G>A, Ex4-901G>A and Ex4-710G>A) are components of a distinct haplotype block. The results of this study suggest that additional 5′ variants as well as markers of distinct 3′ haplotype blocks in MBL2 may contribute to circulating protein levels, but further studies are required to confirm these observations. Last, there could be a selective advantage for diversification of the 3′ region of the gene.Electronic Supplementary Material Supplementary material is available for this article at     

Extreme hemodilution with PEG-hemoglobin vs. PEG-albumin

Isovolemic hemodilution to 11% systemic hematocrit was performed in the hamster window chamber model using 6% dextran 70 kDa (Dx 70) and 5% human serum albumin (HSA). Systemic and microvascular effects of these solutions were compared with polyethylene glycol (PEG)-conjugated 5% albumin (MPA) and PEG-conjugated 4.2% Hb (MP4). These studies were performed for the purpose of comparing systemic and microvascular responses of PEG vs. non-PEG plasma expanders and similar oxygen-carrying vs. noncarrying blood replacement fluids. Mean arterial blood pressure was statistically significantly reduced for all groups compared with baseline (P < 0.05), HSA, MPA, and MP4 higher than Dx 70 (P < 0.05). MP4 and MPA had a significantly higher cardiac index than HSA and Dx 70, in addition to a positive base excess. Microvascular blood flow and capillary perfusion were significantly higher for the PEG compounds compared with HSA and Dx 70. Intravascular PO2 for MP4 and MPA was higher in arterioles (P < 0.05) compared with HSA and Dx 70, but there was no difference in either tissue or venular PO2 between groups. Total Hb in the MP4 group was 4.8 +/- 0.4 g/dl, whereas the remaining groups had a range of 3.6-3.8 g/dl. The hemodilution results showed that PEG compounds maintained microvascular conditions with lower concentrations than conventional plasma expanders. Furthermore, microvascular oxygen delivery and extraction in the window chamber tissue were significantly higher for the PEG compounds. MP4 was significantly higher than MPA (P < 0.05) and was not statistically different from baseline, an effect due to the additional oxygen release to the tissue by the Hb MP4.     

Maintenance of quantal size and immediately releasable granules in rat chromaffin cells by glucocorticoid

Glucocorticoid is reported to regulate catecholamine synthesis and storage. However, it is not clear whether the actual amount of catecholamine released from individual granules (quantal size, Q) in mature chromaffin cells is affected by glucocorticoid. Using carbon fiber amperometry, we found that dexamethasone did not affect mean cellular Q or the proportional release from different populations of granules in rat chromaffin cells cultured for 1 day in a serum-free defined medium. After two extra days of culture in the defined medium, there was a rundown in mean cellular Q, and it was associated with a shift in the proportional release from the different granule populations. This phenomenon could not be rescued by serum supplementation but could be prevented by dexamethasone via an action that was independent of changes in voltage-gated Ca²⁺ channel (VGCC) density. Using simultaneous measurements of membrane capacitance and cytosolic Ca²⁺ concentration, we found that for cells cultured in defined medium dexamethasone enhanced the exocytotic response triggered by a brief depolarization (50 ms) without affecting the VGCC density or the fast exocytotic response triggered via flash photolysis of caged Ca²⁺. Thus glucocorticoid may regulate the number of immediately releasable granules that are in close proximity to a subset of VGCC. Because chromaffin cells in vivo are exposed to high concentrations of glucocorticoid, our findings suggest that the paracrine actions of glucocorticoid maintain the mean catecholamine content in chromaffin cell granules as well as the colocalization of releasable granules with VGCCs. catecholamines; paracrine action; exocytosis; calcium channels     

fecB, a gene potentially involved in iron transport in Mycobacterium avium, is not induced within macrophages

FecB is a protein involved in the transport of iron from ferric citrate in Escherichia coli and is present in the Mycobacterium tuberculosis genome sequence. Since the ability to retrieve iron from the host is crucial and may be related to virulence, we characterized the gene fecB from Mycobacterium avium, strain 101. An E. coli-mycobacterial shuttle plasmid with a fecB-promoter green fluorescence protein (gfp)-fusion was transformed into M. avium strain 104 to study the fecB-regulation. In vitro, the fecB expression in M. avium weakly correlated with the amount of iron present in the medium but the expression was maximal when there was no iron in the culture medium. In macrophages, M. avium fec B was not induced during the early phase of infection, suggesting that the iron concentration in the mycobacterial phagosome is not sufficiently low to stimulate the expression of fecB in M. avium.     

Color enhances orthonasal olfactory intensity and reduces retronasal olfactory intensity

The effect of color on orthonasal and retronasal odor intensities was investigated. When odorants were smelled orthonasally (i.e., through the nostrils), color enhanced odor intensity ratings, consistent with previous reports. However, when odorants were smelled retronasally (i.e., the odorous solution was put in the mouth), color reduced odor intensity ratings. These different effects of color on odor intensity (i.e., enhancement orthonasally and suppression retronasally) appear to be the result of route of olfactory stimulation rather than of any procedural artifact. This supports previous reports that retronasal and orthonasal odors are perceived differently.     

Receptor activator of NF-kappaB ligand protein expression in UMR-106 cells is differentially regulated by parathyroid hormone and calcitriol

Expression of the cytokine, receptor activator of NF-kappaB ligand (RANKL), is stimulated by both parathyroid hormone (PTH) and calcitriol in osteoblasts. Most studies have examined the effects on RANKL mRNA, and less information is available on the protein products. We have determined the effects of PTH, the adenylate cyclase stimulator forskolin, and calcitriol, alone and in combination, on endogenous RANKL protein expression in UMR-106 rat osteoblastic osteosarcoma cells by Western blotting and enzyme immunoassay (EIA). PTH and forskolin dose dependently increased a approximately 52 kDa band in whole cell lysates that was detected by both C- and N-terminal directed RANKL antibodies. Calcitriol treatment produced little or no expression of this approximately 52 kDa band, but markedly increased the expression of a approximately 32 kDa band that was only detected with an antibody directed to the N-terminus of RANKL. An EIA based on RANKL binding to OPG detected a large increase in RANKL expression following calcitriol treatment, and much smaller increases with PTH or forskolin. The combination of PTH and calcitriol or forskolin and calcitriol elicited effects similar to those of PTH and forskolin alone, as detected by both Western blotting and EIA. In contrast to the effects on protein, all agents increased RANKL mRNA expression, with the greatest effects seen with the co-treatments. The results indicate that PTH, likely through effects on cyclic AMP, has a different effect on RANKL processing than calcitriol. The approximately 52 and approximately 32 kDa RANKL products appear to interact differently with OPG, which could affect responses to the agents in target cells.     

Doubling the cloak of (in)competence in client/therapist interactions

Cultural competence is used (often implicitly) to make decisions in human service settings. When therapists make decisions about whether or not a particular service will be offered, they place themselves in a position where their own competence can be judged. Using narrative data on independence and the elderly, we apply Edgerton's idea of the cloak of competence to demonstrate this doubling effect.