In-home walking speeds and variability trajectories associated with mild cognitive impairment.

OBJECTIVE: To determine whether unobtrusive long-term in-home assessment of walking speed and its variability can distinguish those with mild cognitive impairment (MCI) from those with intact cognition. METHODS: Walking speed was assessed using passive infrared sensors fixed in series on the ceiling of the homes of elderly individuals participating in the Intelligent Systems for Assessing Aging Change (ISAAC) cohort study. Latent trajectory models were used to analyze weekly mean speed and walking speed variability (coefficient of variation [COV]). RESULTS: ISAAC participants living alone included 54 participants with intact cognition, 31 participants with nonamnestic MCI (naMCI), and 8 participants with amnestic MCI at baseline, with a mean follow-up of 2.6 ± 1.0 years. Trajectory models identified 3 distinct trajectories (fast, moderate, and slow) of mean weekly walking speed. Participants with naMCI were more likely to be in the slow speed group than in the fast (p = 0.01) or moderate (p = 0.04) speed groups. For COV, 4 distinct trajectories were identified: group 1, the highest baseline and increasing COV followed by a sharply declining COV; groups 2 and 3, relatively stable COV; and group 4, the lowest baseline and decreasing COV. Participants with naMCI were more likely to be members of either highest or lowest baseline COV groups (groups 1 or 4), possibly representing the trajectory of walking speed variability for early- and late-stage MCI, respectively. CONCLUSION: Walking speed and its daily variability may be an early marker of the development of MCI. These and other real-time measures of function may offer novel ways of detecting transition phases leading to dementia.

Mucosal T-cell responses to HIV: responding at the front lines.

Mucosal surfaces of the body serve as the major portal of entry for human immunodeficiency virus (HIV). These tissues also house a majority of the body's lymphocytes, including the CD4(+) T cells that are the major cellular target for HIV infection. Mucosal surfaces are defended by innate and adaptive immune mechanisms, including secreted antibodies and CD8(+) cytotoxic T cells (CTL). CTL in mucosal lymphoid tissues may serve to limit viral replication, decreasing the host's viral burden as well as reducing the likelihood of sexual transmission to a naïve host. This review summarizes recent literature on HIV-specific T-cell responses in mucosal tissues, with an emphasis on the gastrointestinal tract.

An unobtrusive in-home monitoring system for detection of key motor changes preceding cognitive decline.

The existing paradigm of ongoing or posttreatment monitoring of patients through periodic but infrequent office visits has many limitations. Relying on self-report by the patient or their family is equally unreliable. We propose an alternative paradigm in which continuous, unobtrusive monitoring is used to observe changes in physical behavior over time. We highlight the use of this technique for monitoring motor activity that may be predictive of early cognitive changes in the elderly. Initial results using a system of low-cost wireless sensors are presented, together with a discussion of appropriate analyses and interpretation of such data. Using low-cost wireless sensor network coupled with algorithms to detect changes in relevant patterns of behavior, we are able to detect both acute and gradual changes that may indicate a need for medical intervention.

Effects of intravenous lidocaine overdose on cardiac electrical activity and blood pressure in the horse.

This study aimed to identify blood serum lidocaine concentrations in the horse which resulted in clinical signs of intoxication, and to document the effects of toxic levels on the cardiovascular and cardiopulmonary systems. Nineteen clinically normal mature horses of mixed breed, age and sex were observed. Lidocaine administration was initiated in each subject with an i.v. loading dose of 1.5 mg/kg bwt and followed by continuous infusion of 0.3 mg/kg bwt/min until clinical signs of intoxication were observed. Intoxication was defined as the development of skeletal muscle tremors. Prior to administration of lidocaine, blood samples for lidocaine analysis, heart rate, mean arterial blood pressure, systolic blood pressure, diastolic blood pressure, respiratory rate and electrocardiographic (ECG) data were collected. After recording baseline data, repeat data were collected at 5 min intervals until signs of intoxication were observed. The range of serum lidocaine concentrations at which the clinical signs of intoxication were observed was 1.85-4.53 microg/ml (mean +/- s.d. 3.24 +/- 0.74 microg/ml). Statistically significant changes in P wave duration, P-R interval, R-R interval and Q-T interval were observed in comparison to control values, as a result of lidocaine administration. These changes in ECG values did not fall outside published normal values and were not clinically significant. Heart rate, blood pressures and respiratory rates were unchanged from control values. This study establishes toxic serum lidocaine levels in the horse, and demonstrates that there were no clinically significant cardiovascular effects with serum lidocaine concentrations less than those required to produce signs of toxicity.

Comparison of the hydrolytic stability of S-(N,N-diethylaminoethyl) isobutyl methylphosphonothiolate with VX in dilute solution.

The stability of S-(N,N-diethylaminoethyl) isobutyl methylphosphonothiolate--a V-type nerve agent developed by the former Soviet Union--in the environment is an important parameter in threat assessment analysis and for the determination of use, production, testing and storage of this chemical warfare agent. S-(N,N-Diethylaminoethyl) isobutyl methylphosphonothiolate is a structural isomer of the nerve agent VX developed by the USA and the UK and will be referred to as VXA (VX analog) in this presentation. Because VXA and VX differ structurally, even though they do have the same molecular formula, it is expected that their physical and chemical properties would be different. This preliminary investigation was undertaken to determine the relative hydrolysis rate of VXA compared with VX. The hydrolysis of each compound at approximately 1 mg x ml(-1) in unbuffered water at pH 7 was determined side-by-side. The half-lives for VXA and VX were determined to be 12.4 days and 4.78 days, respectively. Agent VXA hydrolyzed 2.6 times more slowly than VX, and each agent followed second-order hydrolysis kinetics. These results imply that VXA is more persistent in the environment and therefore poses a greater threat. These results also imply that VXA is more likely to be detected, if present, during an inspection in support of the Chemical Weapons Convention.

Adenoviral vectors targeted to CD40 enhance the efficacy of dendritic cell-based vaccination against human papillomavirus 16-induced tumor cells in a murine model.

Dendritic cells (DCs) represent a unique junction from which to initiate antigen-specific immunity. One of the most challenging obstacles for DC-based immunotherapy has been the means by which to convey tumor antigen-encoding genes to DCs. In this study, we show that adenoviral (or adenovirus, Ad) vectors targeted to CD40 by means of bispecific antibodies can enhance gene transfer to murine DCs. Moreover, we illustrate that this vector initiates phenotypic changes characteristic of DC maturation. To explore the in vivo potential of this strategy, we coupled this targeting approach with an Ad vector carrying the gene for a tumor antigen. In particular, the human papillomavirus (HPV) E7 antigen represents an attractive target for antigen-specific immunity of cervical cancer. Relative to DCs infected by untargeted Ad, DCs infected by AdE7 targeted to the receptor CD40 enhanced protection against HPV-16-induced tumor cells in a murine model. We have further established that this protection was both antigen specific and CD8+ T-cell dependent. Illustrating that Ad-modified DCs may be used in repeated vaccination, we report that preimmunization of animals with Ad infected DCs prior to E7 vaccination only moderately reduced vaccine efficacy. Finally, we have observed that CD40-targeted AdE7 can initiate partial therapeutic immunity in mice bearing established tumors. These findings suggest that gene-based vaccination of DCs with tumor antigens can elicit productive antitumoral immunity and that enhancements in gene transfer efficacy and/or DC maturation may facilitate this process.

Efficacy of the topical skin protectant in advanced development.

A topical skin protectant (TSP) (ICD 2289) is being developed to protect service members from exposure to chemical warfare agents (CWA). The TSP is designed for use on the skin at the overgarment closures and other vulnerable areas to enhance protection. The TSP, which is in phase II clinical studies, is a cream containing two chemically inert substances: perfluoroalkylpolyether and polytetrafluoroethylene. Animal data showed that the TSP was effective against percutaneous penetration of a blister agent, sulfur mustard (HD), by reducing the size of skin lesions and against T-2 mycotoxin by preventing the development of erythema and edema. The insect repellent N,N-diethyl-m-toluamide (DEET) reduced the TSP protection against HD regardless of the order of application on rabbit skin prior to dosing of HD. The protection was sustained when DEET was removed with a dry gauze prior to TSP application. The TSP was also effective against percutaneous exposure of nerve agents-thickened (with 5% methyl methacrylamide) soman (TGD) and VX (O-ethyl-S-[2-(diisopropylamino)ethyl]methylphosphonothioate )-by reducing the mortality rate and protecting the red blood cell acetylcholinesterase activity. The TSP was effective against VX when DEET was applied prior to TSP application. Because human efficacy studies using CWA cannot be conducted, the efficacy will be demonstrated by the level of protection against poison ivy (urushiol) contact dermatitis in humans.

Evaluation of neutralized chemical agent identification sets (CAIS) for skin injury with an overview of the vesicant potential of agent degradation products.

Vesication and skin irritation studies were conducted in hairless guinea-pigs to determine the vesicant and skin irritation potential of chemically-neutralized Chemical Agent Identification Sets (CAIS). The CAIS are training items that contain chemical warfare-related material--sulfur mustard (HD), nitrogen mustard (HN) or lewisite (L)--and were declared obsolete in 1971. Animals were dosed topically with 'test article'--neat HD, 10% agent/chloroform solutions or product solutions (waste-streams) from neutralized CAIS--and evaluated for skin-damaging effects (gross and microscopic). Product solutions from the chemical neutralization of neat sulfur mustard resulted in microvesicle formation. All agent-dosed (HD or agent/chloroform solutions) sites manifested microblisters as well as other histopathological lesions of the skin. Waste-streams from the neutralization of agent (agent/chloroform or agent/charcoal) were devoid of vesicant activity. Cutaneous effects (erythema and edema) were consistent with the skin-injurious activity associated with the neutralizing reagent 1,3-dichloro-5,5-dimethylhydantoin (DCDMH). Chemical neutralization of CAIS was effective in eliminating/reducing the vesicant property of CAIS containing agent in chloroform or agent on charcoal but was inefficient in reducing the vesicant potential of CAIS containing neat sulfur mustard.

Magnopyramidal neurons in the anterior motor speech region. Dendritic features and interhemispheric comparisons.

OBJECTIVE: To test the hypothesis that the larger somal size of layer III magnopyramidal neurons in left than in right Brodmann area 45 is associated with greater measures of dendritic arborization in the left hemisphere neurons. DESIGN: A case series involving postmortem human brain specimens was used to compare dendritic parameters of Golgi-impregnated layer III pyramidal neurons in Brodmann area 45 in the left and right hemispheres. SUBJECTS: A convenience sample consisting of 9 subjects with no known neurologic or psychiatric disorders was obtained at autopsy. MAIN OUTCOME MEASURES: Dendritic parameters of the 10 largest Golgi-impregnated layer III pyramidal neurons were measured in each hemisphere of each brain using a eutectic neuron tracing system. The measures examined were somal size, neuron depth, mean segment length, number of branch points, maximal branch order, combined dendritic diameter, total dendritic length, horizontal extent of the dendritic field, and spine density. The Golgi-impregnated slides were also compared with Nissl-stained slides from adjacent blocks to determine the laminar location of the sampled neurons. RESULTS: Although the mean somal size was greater in the left than in the right hemisphere, none of the dendritic parameters examined was larger for the left hemisphere neurons. As expected, total dendritic length was positively correlated to somal size (r = 0.43, P < 001) in the left hemisphere. However, there was no correlation between these parameters in the right hemisphere (r = 0.02, P = .76). CONCLUSIONS: The difference between left and right Brodmann area 45 magnopyramidal neurons in the correlation of somal size and dendritic length provides additional evidence of anatomical differences between these 2 populations of neurons. However, the lack of interhemispheric differences in measures of dendritic arborization suggests that additional factors contribute significantly to the marked difference in somal size between the large magnopyramidal neurons in the left and right hemispheres.

Anatomical specialization of the anterior motor speech area: hemispheric differences in magnopyramidal neurons.

The lateralization of motor speech function to the left hemisphere is supported by multiple lines of evidence, but relatively little is known about the anatomical basis of that specialization. In a preliminary study, we recently reported that area 45 of the left hemisphere (Broca's area) contained a subpopulation of magnopyramidal neurons which were significantly larger than any seen in the homotopic region of the right hemisphere (Hayes & Lewis, 1993a). In the present study we examined a larger sample of cases in order to determine how consistently this difference is present in the population, if it is specific to Broca's area or is a general feature of cortical regions mediating lateralized functions, and whether the subpopulation of large magnopyramidal neurons in left area 45 can be distinguished by their chemical phenotype. In Nissl-stained sections from 19 human brains, the mean (+/- SD) cross-sectional area of the largest layer III pyramidal neurons in area 45 was significantly (p < .0001) greater in the left hemisphere (522.1 +/- 128.3 microns2) than in the right (454.1 +/- 121.5 microns2). This interhemispheric difference appeared to be a unique characteristic of the largest neurons, since the mean size of all layer III pyramids in this area was not significantly different in the left (206.2 +/- 93.5 microns2) and right (213.3 +/- 103.9 microns2) hemispheres. In contrast to area 45, there was no interhemispheric difference in the mean cross-sectional area of the largest layer III pyramids in another lateralized region, primary motor cortex. In addition, in area 46, a region of prefrontal association cortex not known to be functionally lateralized, the mean somal size of the largest layer III pyramidal neurons was significantly (p < .001) smaller in the left hemisphere (402.4 +/- 84.9 microns2) than in the right (437.8 +/- 88.3 microns2). Finally, although the large layer III pyramids in area 45 were immunoreactive for nonphosphorylated neurofilament protein in both hemispheres, the mean cross-sectional area of the largest labeled neurons was significantly larger (p < .002) in the left hemisphere (525.2 +/- 149.0 microns2) than in the right (490.3 +/- 154.1 microns2). These findings demonstrate that layer III of Broca's area contains a distinctive subpopulation of neurons that may play an important role in the specific functional architecture of this region.

Heavy ion induced changes in small intestinal parameters.

The effects on 17 different structural parameters of mouse small intestine three days after treatment with three types of heavy ion (neon, iron and niobium) are compared, the first two being of particular relevance to space flight. The data for niobium are given in full, showing that changes after niobium ion treatment are not standard and are concentrated in the epithelial compartment, with few of the parameters having a response which is dose dependent. When comparisons are made for the three types of heavy ion, the damage is greatest after neon ion irradiation, implying that the additional non-epithelial damage produced as LET rises from X rays through neutrons to neon ions is not necessarily maintained as LET continues to rise. Further understanding is therefore needed of the balance between changes affecting the vascular and absorptive components of the organ. Variation from group to group is also important, as is variation of strain or gastrointestinal status. All such factors are important in the understanding of changes in multicellular organs after exposure to heavy ion radiation.

Hemispheric differences in layer III pyramidal neurons of the anterior language area.

OBJECTIVE: The major objective of this study was to determine if the lateralization of motor speech functions may be associated with hemispheric differences in the size of layer III pyramidal neurons in Brodmann's area 45. DESIGN: A case series design involving postmortem human specimens was used to compare the cross-sectional area of NissI-stained layer III pyramidal neurons of Brodmann's area 45 from the left and right hemispheres. SUBJECTS: A convenience sample consisting of seven cases with no known neurological or psychiatric disorders was obtained at autopsy. MAIN OUTCOME MEASURES: The cross-sectional area of layer III pyramidal neurons in both left and right hemispheres was measured in four fields per hemisphere per brain using a computerized image analysis system. Measurements of both the largest layer III pyramids and of all layer III pyramids were done. RESULTS: The largest layer III pyramidal neurons of area 45 were significantly larger in the left than in the right hemisphere in both an unblinded and a blinded series of measurements. However, this hemispheric difference appeared to be restricted to the largest neurons, since the mean size of all layer III pyramids in this area was not significantly different in the left and right hemispheres. CONCLUSIONS: The presence of a unique population of large pyramidal neurons in left Brodmann's area 45 may be related to the involvement of this region in the circuitry that mediates motor speech functions.

Dopamine and the neural circuitry of primate prefrontal cortex: implications for schizophrenia research.

Dopaminergic systems have been implicated in the pathophysiology of schizophrenia; the prefrontal cortex may be a site of dysfunction in this disorder. Until recently, however, relatively little was known about the organization of dopaminergic afferents, or the relationship of those axons to other elements of neural circuitry in the expanded and highly differentiated prefrontal cortex of primates. In this paper we review recent studies demonstrating that monkey prefrontal cortex is innervated by dopaminergic axons in a highly specific regional and laminar fashion. These findings are considered in the context of the organization of other neural systems in monkey prefrontal cortex and with regard to the extent to which they represent the organization of human prefrontal cortex. The resulting model of prefrontal cortical circuitry is then discussed in terms of the ways in which that circuitry might be disrupted in schizophrenia.

Nonphosphorylated neurofilament protein and calbindin immunoreactivity in layer III pyramidal neurons of human neocortex.

Subpopulations of pyramidal neurons in the neocortex have been shown to contain nonphosphorylated neurofilament protein (NPNFP) and calbindin D28K (Morrison et al., 1987; Campbell and Morrison, 1989; Hof et al., 1990; Kobayashi et al., 1990; Hof and Morrison, 1991; Mesulam and Geula, 1991). However, it is not known what relations, if any, exist between the pyramidal neurons containing each of these proteins. In this study, the expression of NPNFP and calbindin immunoreactivity was compared in six regions of human neocortex. Characteristic laminar patterns of immunoreactivity for each protein were seen in most regions examined, and both NPNFP- and calbindin-labeled pyramidal neurons were found in layer III. However, the pyramidal neurons labeled with NPNFP and calbindin differed in several respects. First, the sublaminar distribution of NPNFP-labeled pyramids within layer III differed across regions, ranging from an even distribution throughout the layer in a visual association region (area 18) to a predominance of labeled neurons in the deep half of that layer in a higher association region (area 20). The distribution of calbindin-immunoreactive pyramidal neurons also varied regionally, but in a different manner than that of the NPNFP-labeled neurons. Second, in every region examined, the average size of NPNFP-labeled layer III pyramids was greater than that of calbindin-immunoreactive pyramids. However, there was substantial regional heterogeneity in the extent to which the size distributions of neurons in each of the two populations overlapped. Third, in the regions in which NPNFP- and calbindin-immunoreactive neurons were most similar in size, the amount of colocalization (as identified by double-labeling studies) was also greatest. Similarly, in the regions in which there was minimal overlap in the size of the NPNFP- and the calbindin-immunoreactive neurons, there was minimal colocalization. These regional characteristics of NPNFP- and calbindin-immunoreactive layer III pyramidal neurons have implications for the involvement of these neuronal populations in Alzheimer's disease.

A comparative analysis of the distribution of prosomatostatin-derived peptides in human and monkey neocortex.

Comparative analyses were made of the immunohistochemical and biochemical distributions of three prosomatostatin-derived peptides (PSDP) in human, perfused monkey, and unperfused monkey neocortex. The PSDP we examined were the tetradecapeptide somatostatin 14 (SS14); the N-terminal extension of this peptide, somatostatin 28 (SS28); and somatostatin 28(1-12) (SS28(1-12)). In immunohistochemical experiments, numerous SS28-immunoreactive perikarya were located in both superficial and deep layers of perfused monkey cortex, but none were present in the cerebral cortex from unperfused monkey or autopsied human brains. In contrast, the number of SS28(1-12)-immunoreactive neurons was five times greater in the superficial cortical layers of unperfused monkey than of perfused monkey brain. Moreover, unperfused monkey and human cortex contained notably more SS14-immunoreactive processes than perfused monkey cortex. These data suggested that SS28 may have been converted into SS14 and SS28(1-12) in unperfused tissue during the post-mortem interval. This hypothesis was examined biochemically by measuring the levels of immunoreactivity of SS14, SS28, and SS28(1-12) in samples of unperfused monkey cortex frozen at different time intervals after removal from the brain. Samples frozen 10 minutes or longer after removal contained only 10-20% the level of SS28 immunoreactivity measured in samples frozen immediately or 1 minute after removal. The levels of SS14 and SS28(1-12) immunoreactivity did not demonstrate such reductions, and may instead have increased at early time points. To further characterize post-mortem effects on PSDP and to explore for species differences, we performed a detailed comparison of the regional, laminar, and cellular distribution of SS28(1-12) immunoreactivity under the three conditions. A progressive loss of immunoreactivity, particularly in radial fibers, was found at increasing post-mortem intervals in unperfused monkey neocortex, indicating that differences in density and distribution of immunoreactive fibers between human and perfused monkey may result from post-mortem peptide degradation in unperfused tissue. In contrast, the larger size of SS28(1-12)-immunoreactive white matter neurons in humans as compared to monkeys appeared partially due to a post-mortem effect but also reflected a species difference. In addition, the density of white matter neurons was found to be significantly greater in human than in perfused or unperfused monkey. These data indicate that any study of human autopsy material must be assessed in light of possible post-mortem effects.(ABSTRACT TRUNCATED AT 250 WORDS)

Collared crypts in irradiated small intestine.

Crypts of Lieberkuhn are radiosensitive: the technique of crypt counting is an established method of assessing radiation induced changes in the small intestine. However, there has been little work done on the surface contours of the crypts, as they open into the intervillous cleft. The current paper describes the structure of control mouse crypt mouths as unobtrusive openings approximately 5 microns in diameter. After radiation with heavy ion particles, the crypt mouths are substantially larger (up to 10 microns in diameter) with a marked collar which is similar to that sometimes seen in coeliac disease. The shape and incidence of the collared crypts is described for specimens irradiated with neon, silicon and iron ions, with treatment with iron producing the most marked collars: it is suggested that the size and incidence of the collared crypts may be related to the LET of the beam used. It is of interest that the abnormal crypts are not produced after single doses of X-irradiation. The consideration of the structure of the collared crypts may require a redefinition of the terms crypt and villus with priority being given to the position of subepithelial vessels rather than surface shape. Finally, although the collared crypts can not be directly equated with 'tunnel' or 'channel' lesions, it is pointed out that they do represent localised damage with a specific position and shape.

A specimen holder for low-temperature scanning electron microscopy.

A miniature vise built into a copper stub is described that holds bulk, pre-frozen, hydrated biological specimens during examination under the electron beam of the scanning electron microscope.

Internal structure of the intestinal villus: morphological and morphometric observations at different levels of the mouse villus.

The internal structure of the mouse jejunal villus was examined from apex to base in transverse serial sections. The villi were arbitrarily divided into apical (1-160 microns), middle (161-320 microns) and basal (321-480 microns) thirds. Histological and ultrastructural morphometry was performed on villous epithelium, stroma, blood vessels and lacteals at the three levels. The cross sectional area and the number of cells/section increased apico-basally in both the villous epithelium and the stroma, but the increase was greater in the latter. The enterocytes in the villous basal third were narrow and had oblong nuclei. The enterocytes gradually became wider and their nuclei attained a more round or oval shape towards the villous apex. The microvilli of these cells also increased in length from the villous base to apex. Although the number of capillaries/section was approximately the same at the three levels examined, it appeared that the apical third was relatively more vascular if the overall apico-basal increase in the villous size was taken into consideration. The villi had a single lacteal, whose cross sectional area generally increased towards the villous base. The results of this study indicate that the internal structure of the intestinal villus is not uniform along its length. Therefore, it is important that when studying normal and abnormal villi, like should be compared with like so as to ensure that the normally occurring variations in the villous morphology are not confused with those induced by disease or experimental processes.

In vitro interactions between pulmonary macrophages and respirable particles.

Pulmonary alveolar macrophage cells (PAM) are an important component of the pulmonary response to particles deposited in the deep lung. To more fully characterize the interactions between macrophages and particulate materials, a correlative light and electron microscopic technique was developed that allowed light microscopic identification of individual cell viability after in vitro particle exposure, followed by scanning and transmission electron microscopic analyses of specific PAM, including surface morphology, X-ray microanalytic evaluation of particle content, and internal cellular structure. Individual cell viability, particle content, and morphologic alterations were evaluated for three particle types: Ni3S2, TiO2, and glass beads. Cell death and stages of cell disruption including bleb cluster formation, loss of surface features, formation of membrane tears and holes, and cell degranulation correlated with Ni3S2 and TiO2 content. Glass beads were not associated with cell disruption or viability reduction. Correlative microscopic techniques were essential in describing particle-dependent effects on an individual cell basis.