The behavioral variant of Alzheimer's disease does not show a selective loss of Von Economo and phylogenetically related neurons in the anterior cingulate cortex.

BACKGROUND: The neurobiological origins of the early and predominant behavioral changes seen in the behavioral variant of Alzheimer's disease (bvAD) remain unclear. A selective loss of Von Economo neurons (VENs) and phylogenetically related neurons have been observed in behavioral variant frontotemporal dementia (bvFTD) and several psychiatric diseases. Here, we assessed whether these specific neuronal populations show a selective loss in bvAD. METHODS: VENs and GABA receptor subunit theta (GABRQ)-immunoreactive pyramidal neurons of the anterior cingulate cortex (ACC) were quantified in post-mortem tissue of patients with bvAD (n = 9) and compared to typical AD (tAD, n = 6), bvFTD due to frontotemporal lobar degeneration based on TDP-43 pathology (FTLD, n = 18) and controls (n = 13) using ANCOVAs adjusted for age and Bonferroni corrected. In addition, ratios of VENs and GABRQ-immunoreactive (GABRQ-ir) pyramidal neurons over all Layer 5 neurons were compared between groups to correct for overall Layer 5 neuronal loss. RESULTS: The number of VENs or GABRQ-ir neurons did not differ significantly between bvAD (VENs: 26.0 ± 15.3, GABRQ-ir pyramidal: 260.4 ± 87.1) and tAD (VENs: 32.0 ± 18.1, p = 1.00, GABRQ-ir pyramidal: 349.8 ± 109.6, p = 0.38) and controls (VENs: 33.5 ± 20.3, p = 1.00, GABRQ-ir pyramidal: 339.4 ± 95.9, p = 0.37). Compared to bvFTD, patients with bvAD showed significantly more GABRQ-ir pyramidal neurons (bvFTD: 140.5 ± 82.658, p = 0.01) and no significant differences in number of VENs (bvFTD: 10.9 ± 13.8, p = 0.13). Results were similar when assessing the number of VENs and GABRQ-ir relative to all neurons of Layer 5. DISCUSSION: VENs and phylogenetically related neurons did not show a selective loss in the ACC in patients with bvAD. Our results suggest that, unlike in bvFTD, the clinical presentation in bvAD may not be related to the loss of VENs and related neurons in the ACC.

[Preconception care: an essential part of the care for mother and child]. / Preconceptiezorg: een onlosmakelijk onderdeel van de zorg voor moeder en kind.

Despite the modernisation of antenatal care in the Western world, the incidence of a number of adverse pregnancy outcomes, such as birth defects, low birth weight and preterm birth, has not decreased over the past few decades. Since its inception at the beginning of the last century, the concept of antenatal care has not changed. The first antenatal visit typically starts at the end of the first trimester. By this stage of pregnancy organogenesis and early placentation have been completed making it almost impossible to reverse any unfavourable exposure on the foetus. Preconception care addresses risk factors that are present prior to pregnancy. By either eliminating or altering risk factors during this period, pregnancy outcome may improve. The goal of preconception care is to optimise the quality of foetal, newborn and infant life through primary prevention. With regard to genetic conditions the aims of preconception care are more nuanced. The principle components of preconception care include (a) risk assessment, (b) information and advice on health promotion, (c) specific counselling and (d) intervention. The effectiveness of preconception care has been demonstrated in women who are at increased risk of adverse pregnancy outcome. In women at low-risk, however, the usefulness of preconception care has yet to be established. The concept of preconception care is relatively simple, logical and promises much. For preconception care to be a success, it is crucial to make this form of preventive care available to all prospective parents. The Internet could play a major part in the dissemination of information that is relevant to a successful outcome of pregnancy.

Alkyl-polyacrylate esters are strong mucosal adjuvants.

Synthetic polymers were examined for their potency to enhance mucosal immune responses to inactivated antigens. Aqueous solutions of polyacrylic acid with a MW of 450 kDa (p[AA]) or an butyl-ester thereof with 16% esterification (Butyl16-p[AA]) plus antigen were administered twice intranasally in mice with a 2 week interval. The frequency of IgA-antibody secreting cells (ASCs) in lung cell suspensions was determined 1 week after the second immunisation. Both polymers significantly enhanced the IgA response against inactivated Newcastle disease virus (iNDV), inactivated influenza virus strain MRC-11 (iMRC-11), haemagglutinin/neuraminidase subunits of influenza virus strain A/Texas (HA/NA) and bovine serum albumin (BSA). Butyl16-p(AA) was significantly more effective than non-derivatised p(AA), cholera toxin B subunit (CTB) or liposomes. The factor of increase in IgA-ASCs varied from <10- to >100-fold and depended on the type of antigen, the dose of antigen and the adjuvant. Extremely high responses of about 10,000 IgA-ASCs per million lung cells were detected after immunisation with 5 microg HA/NA plus 50 microg Butyl16-p(AA). Intranasal immunisation with Butyl16-p(AA) resulted in high IgA responses, not only in the lungs, but also in the spleen and in high IgG responses in these organs. We concluded that alkyl-esters of polyacrylate are an interesting, novel category of mucosal adjuvants.

Sulfolipo-cyclodextrin in squalane-in-water as a novel and safe vaccine adjuvant.

Previously, we described synergistic adjuvanticity of combinations of synthetic sulfolipo(SL)-derivatives of polysaccharide (SL-polysaccharides) and squalane-in-water emulsions (squalane/W). In this paper, effects of type of polysaccharide and nature of oil on adjuvanticity, reactogenicity and stability are described. SL-derivatives of the following polysaccharides were synthesised: synthetic polysucroses with weight-average molecular weight (MW) of 400,000 (Ficoll400), 70,000 (Ficoll70) and 39,000 Da (Ficoll39), polyfructose of 5,000 Da (inulin), linear polyglucose of 1,200 Da (maltodextrin) and cyclic polyglucose of 1,135 Da (beta-cyclodextrin). The number of sulphate groups per monosaccharide of the different SL-polysaccharides varied between 0.15 and 0.23 and the number of lipid groups per monosaccharide between 1.15 and 1.29. Adjuvant formulations were prepared by incorporating these SL-polysaccharides into oil-in-water emulsions of either squalane, hexadecane, soya oil or mineral oil. Adjuvanticity of the formulations obtained for humoral responses to inactivated pseudorabies virus (PRV) and inactivated influenza virus strains A/Swine (A/Swine) and MRC-11 (MRC-11) in pigs and MRC-11 and ovalbumin (OVA) in mice depended on the type of oil (squalane = mineral oil > hexadecane = soya oil) but not on the type of polysaccharide backbone of the SL-derivative. Reactogenicity assessed by local swelling in mice decreased with decreasing MW (SL-Ficoll400 = Ficoll70 = Ficoll39 > SL-inulin = SL-maltodextrin > SL-cyclodextrin) when combined with squalane and decreased with the type of oil in the following order: squalane > mineral oil > hexadecane > soya oil when combined with SL-Ficoll400. Stability of the SL-polysaccharide/squalane/W emulsions at elevated temperature increased with decreasing MW of the SL-polysaccharide (SL-Ficoll400 < SL-Ficoll70 = SL-Ficoll39 < SL-inulin = SL-maltodextrin = SL-cyclodextrin). SL-cyclodextrin/squalane/W remained stable for > 2.5 years at 4 degrees C, > 18 weeks at 37 degrees C and > 10 days at 60 degrees C. We concluded that reactogenicity and stability but not adjuvanticity of SL-polysaccharide/squalane/W formulations depended on the MW of SL-polysaccharide and that SL-cyclodextrin/squalane/W is a promising non-mineral oil adjuvant as it combines strong adjuvanticity (i.e. better than the mineral oil-based adjuvant presently applied) with low reactogenicity and good stability.

Effect of various adjuvants on secondary immune response in chickens.

Stimulatory effects of several types of adjuvants on secondary antibody response to inactivated Newcastle disease virus (iNDV) were examined in chickens. For this purpose, animals were primed with iNDV without adjuvant resulting in a low but significant antibody response, boosted with iNDV plus adjuvant 3 weeks later, and analysed for specific antibody titres in serum 3 weeks after the booster. Water-in-mineral oil emulsion (W/O) caused significant increase in antibody titres measured in an indirect enzyme-linked immunosorbent (ELISA), haemagglutination inhibition (HI), and virus neutralisation (VN) assay. The adjuvants tested included three oil-in-water emulsions (i.e. mineral oil-in-water, sulpholipo(SL)-Ficoll400/squalane-in-water and sulpholipo-cyclodextrin/squalane-in-water), three negatively-charged polymers with high molecular weight (i.e. polyacrylate, polystyrenesulphonate and sulpho(S)-Ficoll400) and two surface-active agents (i.e. dimethyldioctadecylammonium bromide (DDA) and Quil A). These adjuvants enhanced significantly the secondary immune response but none reached the titre obtained with W/O. Combinations of adjuvants with distinct physicochemical properties, i.e. polyacrylate and DDA revealed only slight, beneficial effects. We concluded that the various types of adjuvants tested can stimulate secondary immune responses in primed animals but that W/O is superior.

Analysis of oral uptake of a synthetic adjuvant by an immunoassay with monoclonal antibodies.

A blocking ELISA with specific antibodies against a synthetic sulpholipo-polysucrose (SL-Ficoll400) was developed to quantify this vaccine adjuvant component in faeces with the purpose of estimating the uptake thereof by the gastrointestinal tract after oral administration. Specific monoclonal antibodies (MABs) prepared against SL-Ficoll400 did not cross-react with components in murine faeces. SL-Ficoll400 inhibited the reactivity of the MABs in a dose-dependent fashion in the blocking ELISA and the concentration causing 50% inhibition (IC50) was about 1 microg SL-Ficoll400 per milliliter. Derivatives of Ficoll400 lacking sulphate or lipid groups or both were unable to block the MABs (IC50 > 1000 microg/ml) which confirmed the specificity of the test system. Quantification of SL-Ficoll400 in extracts of faeces of untreated mice spiked with SL-Ficoll400 revealed that more than 90% could be recovered. The blocking ELISA was used to quantify SL-Ficoll400 in the faeces of mice given this compound orally. One, two and three days after oral administration, the cumulative recovery of SL-Ficoll400 in faeces was 30, 40 and 92%, respectively. From these data, we concluded that the MABs obtained were specific for SL-Ficoll400, that SL-Ficoll400 in murine faeces could be quantified by a blocking ELISA with MABs and that SL-Ficoll400 was poorly absorbed by the gastrointestinal tract.

Quantitative analysis of a synthetic adjuvant by an immunoassay.

Antibodies against a sulpholipo-derivative of synthetic polysucrose (SL-Ficoll) were prepared with the purpose of developing a detection system for this adjuvant component. SL-Ficolls with polysucrose backbones of 400 kDa (SL-Ficoll400) or 22 kDa (SL-Ficoll22) were not immunogenic. However, SL-Ficoll22 conjugated to bovine serum albumin (SL-Ficoll22-BSA) induced high levels of specific antibodies against SL-Ficoll in mice as determined by an indirect ELISA with either SL-Ficoll400 or SL-Ficoll22 as coating antigen. The specificity of the antibodies was analysed further in a blocking ELISA with SL-Ficoll400 as coat. Dose-dependent inhibition of the ELISA titre was obtained with SL-Ficoll22 and SL-Ficoll400 and the concentration causing 50% inhibition (IC50) was < 1 microg/ml. Non-derivatised polysucrose (Ficoll400) and compounds lacking either lipid or sulphate were not recognised (IC50 > 1000 microg/ml). The reaction of SL-Ficoll400-derivatives with antibodies increased with increasing sulphate and lipid content and maximal inhibition was produced by compounds with a composition similar to the SL-Ficoll22 used for immunisation. Quantitative analysis of SL-Ficoll400 in adjuvant formulations comprising SL-Ficoll400 incorporated in a squalane-in-water emulsion revealed high recovery and sufficient precision. From these data, we conclude that specific antibodies are generated against the synthetic SL-Ficoll400 and that this component of a novel adjuvant formulation can be quantified by an immunoassay.

Alkyl-esters of polyacrylic acid as vaccine adjuvants.

Previously, we demonstrated that polyacrylic acid (PAA) augmented significantly the immune response to inactivated Newcastle disease virus (iNDV) in chickens, but that efficacy was insufficient to replace the water-in-mineral oil (W/O) adjuvant applied for boosting primed animals. Attempting to improve its adjuvanticity, PAA with weight-average molecular weight (Mw) of 450 kDa was grafted with alkyl-chains by esterifying the carboxylic groups with octanol and butanol. The butyl-PAA and octyl-PAA esters obtained varied in degree of esterification between 10% and 92%. Adjuvant activity of water-soluble esters for humoral responses to iNDV was examined in chickens primed previously with iNDV without adjuvant. The alkyl-PAA esters exhibited significantly higher responses than unmodified PAA and titres increased with increasing dose of adjuvant. At doses of 2 mg per animal, octyl- and butyl-PAA esters with a substitution rate of 16% (octyl16-PAA and butyl16-PAA, respectively) gave similar titres as W/O. In aged animals primed with live NDV at early age, butyl16-PAA and W/O elicited comparable antibody responses. Butyl16-PAA was also more effective than PAA in stimulating primary immune responses in mice which was accompanied by stronger local reaction determined by monitoring swelling at the site of injection. Reactogenicity of butyl16-PAA was less than of W/O. We concluded that alkyl-PAA esters are strong adjuvants for primary and secondary responses and that they are promising alternatives to the mineral oil-based adjuvants presently used in various veterinary vaccines.

Health care management in workers' compensation.

A high-performing, effective health care delivery system is critical to the recovery of injured workers within a workers' compensation insurance system. Timely and effective health care has the potential to minimize indemnity costs and therefore contribute to the insurer's financial state. While costs remain a concern to insurers, cost-containment initiatives within the health care arena have evolved from a strict "deep discount" approach to more sophisticated health care strategies that follow managed care-style models. In the future, health care strategies are likely to become more integrated within the business operations of workers' compensation insurance systems. The next evolution of health care strategy within workers' compensation will likely include consensus-based contracts with providers that stipulate the role and function of each party while reinforcing a continuous improvement mindset. It is probable that a component of this evolving system will include shared risk and reimbursement that is based on performance. Insurers who begin to evaluate the true impact of a comprehensive health care strategy will find it necessary and advantageous to modify their business relationship with health care providers. Those who are able to articulate a business strategy that capitalizes on the skills of the health care community are likely to gain a competitive advantage. Most importantly, this bridging of intellectual capacity across the insurance and health care domains will result in a delivery system that is valued by, and contributes to, its key participants--the employers and the injured workers.

Safe biotechnology. 8. Transport of infectious and biological materials. Working Party "Safety in Biotechnology" of the European Federation of Biotechnology.

The transport of infectious and biological material is regulated by a number of international organizations. This mini-review has been compiled to increase awareness within the scientific community of problems caused by differences in terminology (such as infectious materials/substances, biological products, diagnostic specimens, genetically modified microorganisms) and certain technical aspects of the main international guidelines, and to assist policy makers in the creation of harmonized guidelines. A list of relevant Internet resources has been compiled.

Effect of an 18-wk weight-training program on energy expenditure and physical activity.

The purpose of this study was to examine the effect of an 18-wk weight-training program on average daily metabolic rate (ADMR). Before the intervention and in weeks 8 and 18 (T0, T8, and T18, respectively) data on body composition, sleeping metabolic rate (SMR), food intake, energy cost of the weight-training program (EEex), and nontraining physical activity (accelerometer) were collected in the exercise group (EXER, n = 18 males). ADMR was determined in a subgroup (EX12, n = 12) by using doubly labeled water. At T0 and T18, data (except ADMR) were also collected in a control group (Con, n = 8). Body mass did not change in EXER or Con. Fat-free mass increased only in EXER with 2.1 +/- 1.2 kg, whereas fat mass decreased in EXER as well as Con (2.0 +/- 1.8 and 1.4 +/- 1.0 kg, respectively). Initial ADMR (12.4 +/- 1.2 MJ/day) increased at T8 (13.5 +/- 1.3 MJ/day, P < 0.001) with no further increase at T18 (13.5 +/- 1.9 MJ/day). SMR did not change in EXER (4.8 +/- 0.5, 4.9 +/- 0.5, 4.8 +/- 0.5 kJ/min) or Con (4.7 +/- 0.4, 4.8 +/- 0.4 kJ/min). Energy intake did not change in EXER (10.1 +/- 1.8, 9.7 +/- 1.8, 9.2 +/- 1.9 MJ/day) or Con (10.2 +/- 2.6, 9.4 +/- 1.8, 10.1 +/- 1.5 MJ/day) and was systematically underreported in EX12 (-21 +/- 14, -28 +/- 18, -34 +/- 14%, P < 0.001). EEex (0.47 +/- 0.20, 0.50 +/- 0.18 MJ/day) could only explain 40% of the increase in ADMR. Nontraining physical activity did not change in both groups. In conclusion, although of modest energy cost, weight-training induces a significant increase in ADMR.

Safe biotechnology. 7. Classification of microorganisms on the basis of hazard. Working Party "Safety in Biotechnology" of the European Federation Biotechnology.

The current systems for classifying human pathogens on the basis of hazard are well developed and their basic criteria are in general agreement one with another. Of more importance, the safety practices based on these classifications have generally been successful. They have enabled extensive research activities, medical practice and industrial production to be conducted on an ever-increasing scale, involving dangerous microorganisms (e.g. in vaccine production and treatment of infected patients) with a very low incidence of adverse effects on the workers involved and the general public. Although the EU has adopted a harmonised list of agents in groups 1-4 there is as yet no complete agreement among member states and individual microbiologists. The purpose of this paper is to present a historical survey and to discuss the current processes for identifying and classifying the hazards posed by the use of microorganisms in research and technology. This is essential in the design of appropriate methods of counteracting potential risks.

Safe biotechnology. Part 6. Safety assessment, in respect of human health, of microorganisms used in biotechnology.

The assessment of microorganisms in respect to human health is an important step for the introduction of new natural and genetically modified production strains to biotechnology. This report outlines the potential hazards posed by industrial microorganisms, important considerations related to pathogenicity, such as routes and portals of entry into the human body, mechanisms of spread of biological material and a definition of pathogenicity. Furthermore the most important steps in the assessment of pathogenicity of unknown strains are described. A short overview on characterization and in vitro and in vivo tests is presented. The hazard related to allergens and toxic metabolites is reviewed and the choice of methods and the handling of strains with unknown potential are discussed.

Safe biotechnology (5). Recommendations for safe work with animal and human cell cultures concerning potential human pathogens.

The benefits of using animal or human cell cultures have been clearly demonstrated in diagnostic and therapeutic research and in their application for manufacturing. Cell cultures serve as a tools for the production of vaccines, receptors, enzymes, monoclonal antibodies and recombinant DNA-derived proteins. They represent an integral part of drug development for which corresponding facilities, equipment and manufacturing processes are required. Although the cells themselves offer no particular risk to workers in laboratories and production areas or to the environment, the cell cultures may be contaminated with viruses, mycoplasma, bacteria, yeast and fungi or might contain endogenous viruses. The containment level for animal and human cells is therefore determined by the risk class of these agents. The history of animal and human cell cultures has proved that they can be handled safely. The recommendations in this publication concern the safe handling of cell cultures (tissue explants, primary cell cultures) and permanent cell lines of animal and human origin. A classification system of safety precautions has been elaborated according to the potential for contamination with the pathogenic agents involved.

Safe biotechnology (4). Recommendations for safety levels for biotechnological operations with microorganisms that cause diseases in plants.

The Working Party on Safety in Biotechnology of the European Federation of Biotechnology has proposed a classification of microorganisms that cause diseases in plants. In this paper appropriate safety levels are proposed for these classes of microorganisms in order to ensure that research, development and industrial fermentation work with plant pathogens will limit the risk of outbreaks of diseases in crops that could result from work with such microorganisms when they are cultivated in laboratories, glasshouses and biotechnology installations.

Salivation discordant with hunger.

In the present study, the authors tested whether an increase in salivation is associated with an increase in subjectively experienced hunger. After conditioning, subjects showed a significant increase in salivation flow. Hunger levels, however, were significantly decreased after conditioning. No correlation was found between salivation flow and hunger levels. It is argued that salivation responses and subjectively experienced hunger are loosely coupled systems. Salivation flow reflects the learning history of a subject which may sometimes be paralleled by a biological state which is called hunger, whereas, at other times, hunger may be absent. The authors conclude that conditioning of preparatory responses such as salivation depends on the probability relationship between exposure to cues (CSs) and food intake (US), as well as the intensity of the US.

Diazepam mixed micelle--comparison with diazepam in propylene glycol and midazolam.

A mixed micelle formulation of diazepam was compared with midazolam and diazepam in propylene glycol for evidence of venous intolerance following IV injection. The overall incidence of venous morbidity was 17% for diazepam mixed micelle, 26% for midazolam and 90% for diazepam in propylene glycol. Diazepam mixed micelle is suggested as a preferable alternative to the standard formulation.