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PubMed2004

Long-lived retrograde fluorescent labeling of corticospinal neurons in the living animal.

For pathophysiological studies, it is advantageous to label specific neuronal populations in living animals. This study aimed to establish a method for stable and long-lasting fluorescent labeling of corticospinal neurons in the living animal. The two fluorescent dyes Fluoro-Red and Fluoro-Green were injected in the cervical spinal cord of anesthetized newborn rats. After a recovery period, treated rats were returned to the mother. After 24 h and 14 days, fixed brain sections revealed wide-spread fluorescence in elongated or pyramidal-shaped cell profiles in a discrete internal cortical layer, consistent with layer V pyramidal cells. Labeled neurons displayed spontaneous synaptic activity using the slice patch clamp method. These results suggest that these dyes are effective tools for pathophysiological and slice patch clamp studies focused on specific neuron groups.

Action Potentials↗
PubMed1998

Influence of feeding on metabolite excretion evidenced by urine 1H NMR spectral profiles: a comparison between subjects living in Rome and subjects living at arctic latitudes (Svaldbard).

Urines from 25 normal subjects living in Rome and 25 normal subjects living in Ny-Alesund (Svaldbard) were analysed by 1HNMR spectroscopy. The observed differences in the concentration of the major metabolites were correlated to the composition of the diet. It was found that a diet rich of carbohydrates, such as the Italian diet, is responsible for an increased excretion of citrate, lactate, alanine, and glycine. Thus, a correct diagnostical interpretation of urinary metabolites needs to consider feeding habits.

Adult↗
PubMedDate not supplied

The prevalence of functional disability in activities of daily living and instrumental activities of daily living among elderly Beijing Chinese.

In order to assess the prevalence of the functional disability defined by activity of daily living (ADL) and instrumental activity of daily living (IADL) and associated factors in elderly Chinese, a population-based cross-sectional study was performed in urban, plain rural and mountain rural regions of Beijing. Of the 3440 subjects, 1707 are males and 1733 are females, with mean age of 71.4+/-7.7 years. Demographic, socio-economic and health aspects were obtained by trained interviewers. The results showed functional disability prevalence was 6.5% on ADL and 7.9% on IADL. Among the three representative areas in Beijing, the plain rural had the highest disability rate, increasing with the progression of age. Bathing and doing heavy housework were the two most difficult functional tasks. The functional disabilities were associated with gender and marital status. Our data suggest that plain rural elderly are most likely to generate functional disability, and bathing and doing heavy housework are two promising predictors to monitor the development of functional disability in the elderly.

Journal Article↗
PubMed2003

Rapid onset of protection following vaccination of calves with multivalent vaccines containing modified-live or modified-live and killed BHV-1 is associated with virus-specific interferon gamma production.

The objective of this study was to determine the effect of vaccination with commercially-available multivalent vaccines containing either modified-live (MLV) bovine herpesvirus-1 (BHV-1) (Bovishield) or MLV plus killed (MLV + K) BHV-1 (Reliant Plus) on protection against challenge at 5 days after a single vaccination. An additional objective was to determine whether cell-mediated immunity as measured by virus-specific interferon gamma (IFN-gamma) production by peripheral blood mononuclear cells (PBMC) was associated with any early protection induced by vaccination. Clinical signs, serum neutralizing (SN) titers, and nasal virus isolation (VI) titers were also measured. The 12-16-week-old dairy cross-calves seronegative for antibodies to BHV-1 were vaccinated with a multivalent vaccine containing MLV BHV-1 (n = 19), a multivalent vaccine containing MLV + K BHV-1 (n = 19), or a control multivalent vaccine not containing BHV-1 (n = 10) on day 0 and challenged intranasally on day 5. PBMC were isolated on days 0, 3, 5, 8, 10, 14 and 19. PBMC were incubated in vitro with spent media, live BHV-1, or heat-inactivated BHV-1 for 72 h. Supernatants were assayed for bovine IFN-gamma by ELISA. Bovine herpesvirus-1-specific IFN-gamma production was expressed as percent of the kit positive control, with value for spent media subtracted. Clinical signs were monitored daily. Serum VN titers were measured on days 0-5 and 19. Nasal VI titer was measured every other day from days 5 to 19. Interferon gamma production was higher on day 5, and was significantly increased post-challenge, in both vaccine groups compared to controls. There was no difference between vaccine groups on any day. There was no significant difference in SN titer among groups on any day. Virus isolation titer was significantly higher in controls on days 6 and 8 compared to both vaccine groups. Temperatures were significantly higher and nasal discharge was present more often post-challenge in controls compared to vaccine groups. Vaccination 5 days prior to challenge with commercially-available vaccine containing MLV or MLV + K BHV-1 was associated with increased BHV-1-specific IFN-gamma production, decreased viral shedding, lower temperatures and less nasal discharge post-challenge. Cell mediated immune responses as measured by IFN-gamma production are stimulated rapidly following BHV-1 vaccination of calves.

Animals↗
PubMed2004

Effects of Soy-derived diets on plasma and liver lipids, glucose tolerance, and longevity in normal, long-lived and short-lived mice.

We examined the effects of diets based on a low isoflavone or a high isoflavone soy protein isolates in normal, growth-hormone receptor knockout and Ames dwarf, and Prop 1 (df) mice that are hypoinsulinemic, insulin-sensitive, and exceptionally long-lived, as well as in growth hormone transgenic mice that are hyperinsulinemic, insulin-resistant, dyslipidemic, and short-lived. Soybean diets tended to normalize plasma cholesterol levels in dwarf and transgenic mice, while low isoflavone diet reduced plasma triglycerides in most of the examined genotypes. The effects of low isoflavone and high isoflavone diets on the levels of free and esterified cholesterol in the liver were strongly genotype-dependent. Fasting blood glucose levels were reduced and glucose tolerance improved by both low isoflavone and high isoflavone diets in growth hormone-transgenic mice and in their normal siblings. Glucose tolerance was also improved by high-isoflavone diet in growth hormone receptor knockout mice. Lifespan was increased by low isoflavone diet in normal mice from two of the examined stocks. High isoflavone diet increased lifespan in normal animals from one line, but reduced lifespan of normal mice from a different line. We conclude that dietary soy protein intake can improve plasma and hepatic lipid profiles, reduce fasting glucose, enhance capacity for glucose tolerance, and prolong life, but all of these effects are strongly genotype-dependent.

Animals↗
PubMed1995

The marital perceptions of elderly persons living or having a spouse living in a long-term care institution in Canada.

The focus of this study is on the way that older married persons living, or having a spouse living, in a long-term care institution perceive their marriages following relocation. A qualitative analysis of 161 interviews was conducted. For noninstitutionalized spouses (n = 74), four themes emerged from the data--"marriage as a memory," the "illusory marriage," the "changed marriage," and the "continuing marriage." For institutionalized spouses (n = 87), three themes emerged, including the "happy marriage," the "detached marriage," and the "altered marriage." Relocation did not appear to have a direct impact on the way respondents perceived their marriages. Continuity theory was helpful in understanding these marital perceptions.

Aged↗
PubMed1993

Kinetics of B cell subpopulations in peripheral lymphoid tissues: evidence for the presence of phenotypically distinct short-lived and long-lived B cell subsets.

A small proportion of the sIg+ B lymphocytes in peripheral lymphoid organs [22% in spleen and 6% in lymph node (LN)] in rat carries the Thy-1 antigen. These Thy-1+ B cells represent newly formed bone marrow (BM) derived (or immature) B cells. In this study we investigated the kinetic behavior of Thy-1+ and Thy-1- B cells in various lymphoid tissues. The renewal rates of these B cells in young adult rats were determined by continuous administration of 5-bromo-2-deoxyuridine (BrdU) for up to 6 weeks. At several time intervals, Thy-1+ and Thy-1- B cell subpopulations in BM, blood, spleen and popliteal LN were analyzed for the presence of incorporated BrdU, using three-color immunocytology on cytospin preparations. In all tissues studied, the Thy-1+ B cells were rapidly renewed with a rate that varied between 58% (BM) and 22% (LN) per day. Virtually all Thy-1+ B cells were labeled by BrdU within a period of 8-16 days, indicating that all these cells are relatively short-lived. By contrast, the replacement of Thy-1- B cells in these tissues was 30-40 times lower, and ranged between 2.0 and 0.8% per day. In absolute numbers we estimate that 57 million Thy-1+ B cells are renewed per day in the BM whereas only 10 million Thy-1- B cells are replaced in the pool of long-lived peripheral B cells. This implicates a cell loss of 80% at the transition of the Thy-1+ to the Thy-1- B cell stage. The few B cells that actually become incorporated into the pool of mature B cells are most probably selected on the basis of the specificity of their sIg.

Animals↗
PubMed1994

Passive protection of mice against lethal Francisella tularensis (live tularemia vaccine strain) infection by the sera of human recipients of the live tularemia vaccine.

The relative role that humoral immunity plays in protection against infection with the intracellular bacterium, Francisella tularensis, remains controversial. Cellular immunity is thought to play the major and perhaps only role. The authors, in this article, investigate the immunologic and protective properties of immune serum collected from human recipients of the live tularemia vaccine (LVS). Sera of recipients of the vaccine demonstrated reactivity with the vaccine strain by enzyme-linked immunosorbent assay and Western blot analysis. This reactivity appeared to be directed primarily against the lipopolysaccharide of LVS and demonstrated complete cross-reactivity with fully virulent F. tularensis (Schu4). Pooled immune sera protected mice fully against a 10,000 LD50 challenge with the LVS strain relative to non-immune sera. The protection was abrogated by dilution or preadsorption with the LVS strain but not by preadsorption with Escherichia coli, which suggests specificity of protection. The authors conclude that antibodies to the LVS strain of F. tularensis are generated by live vaccination in humans and play a significant role in protection of mice against lethal challenge with the same organism. These antibodies crossreact completely with fully virulent F. tularensis, but whether they play a role in protection against fully virulent human tularemia strains requires further experimentation.

Animals↗
PubMed1993

Bacterial clearance capability of living skin equivalent, living dermal equivalent, saline dressing, and xenograft dressing in the rabbit.

Two new skin substitutes, Living Skin Equivalent (LSE) and Living Dermal Equivalent (DE), have recently been developed. In this experiment, the ability of the LSE and DE preparations to function as biological dressings in an acute wound model was tested. Forty full-thickness wounds were made in New Zealand White rabbits. Each wound was inoculated with 5 x 10(5) Staphylococcus aureus organisms. Twenty-four hours later, one of the following four dressings was applied: saline gauze, porcine-derived xenograft, LSE, or DE. Daily dressing changes and wound biopsies for bacterial counts were performed. At 96 hours after inoculation, split-thickness autograft was applied to all wounds. Skin graft take was assessed 5 days later. In all treatment groups, bacterial counts decreased over time (p = 0.02). At 72 and 96 hours after inoculation, wounds dressed with LSE or DE had significantly lower mean bacterial counts than wounds treated with xenograft dressing (p < 0.01). No significant differences were found among the LSE-, DE-, or saline-treated groups. Skin grafts took well in LSE- and DE-treated wounds. In conclusion, the LSE and DE were more effective than xenograft in reducing bacterial wound contamination in this model, thereby demonstrating their potential application as biological dressing materials.

Animals↗
PubMed1992

An appraisal of pediatric liver transplantation from living relatives. Initial clinical experiences in 20 pediatric liver transplantations from living relatives as donors.

The authors performed 20 liver transplantations from living related donors between June 1990 and July 1991. The 20 pediatric patients (14 biliary atresia, two Budd-Chiari syndrome, one liver cirrhosis after hepatitis C viral infection (HCV hepatitis), 1 progressive intrahepatic cholestasis, 1 liver cirrhosis, 1 protoporphyria) were transplanted with 11 left lobes, eight left lateral segments, and one right lobe. The choice of donors was restricted to the parents of the recipients. The immunosuppressive treatment consisted of FK 506 and steroids. Seventeen recipients are alive, 15 of whom are well and at home. Two recipients, who underwent emergency transplantation, died of postoperative complications. Another recipient died of accidental asphyxia at 6 months after the transplantation. All 20 donors had uneventful postoperative courses and were able to resume their normal social lives. The arterial ketone body ratio (AKBR) increased to above 1.0 within 2 days after the transplantation in all cases. Relatively mild rejection episodes were encountered in only two cases transplanted with ABO-compatible grafts, and these were treated successfully with steroids and FK 506.

Adolescent↗
PubMed2004

Individual thyroid dose estimation for a case-control study of Chernobyl-related thyroid cancer among children of Belarus-part I: 131I, short-lived radioiodines (132I, 133I, 135I), and short-lived radiotelluriums (131MTe and 132Te).

Large amounts of radioiodines were released into the atmosphere during the accident at the Chernobyl nuclear power plant on 26 April 1986. In order to investigate whether the thyroid cancers observed among children in Belarus could have been caused by radiation exposures from the Chernobyl accident, a team of Belarusian, Russian, and American scientists conducted a case-control study to compare cases and controls according to estimated thyroid dose. The primary purpose of this paper is to present detailed information on the estimated thyroid doses, due to intakes of 131I, that were used in the case-control study. The range of the 131I thyroid doses among the 107 cases and the 214 controls was found to extend from 0.00002 to 4.3 Gy, with medians of approximately 0.2 Gy for the cases and 0.07 Gy for the controls. In addition, the thyroid doses resulting from the intakes of short-lived radioiodines (132I, 133I, and 135I) and radiotelluriums (131mTe and 132Te) were estimated and compared to the doses from 131I. The ratios of the estimated thyroid doses from the short-lived radionuclides and from I for the cases and the controls range from 0.003 to 0.1, with median values of approximately 0.02 for both cases and controls.

Body Burden↗
PubMed2003

Telomeres shorten more slowly in long-lived birds and mammals than in short-lived ones.

We know very little about physiological constraints on the evolution of life-history traits in general, and, in particular, about physiological and molecular adjustments that accompany the evolution of variation in lifespan. Identifying mechanisms that underlie adaptive variation in lifespan should provide insight into the evolution of trade-offs between lifespan and other life-history traits. Telomeres, the DNA caps at the ends of linear chromosomes, usually shorten as animals age, but whether telomere rate of change is associated with lifespan is unknown. We measured telomere length in erythrocytes from five bird species with markedly different lifespans. Species with shorter lifespans lost more telomeric repeats with age than species with longer lifespans. A similar correlation is seen in mammals. Furthermore, telomeres did not shorten with age in Leach's storm-petrels, an extremely long-lived bird, but actually lengthened. This novel finding suggests that regulation of telomere length is associated not only with cellular replicative lifespan, but also with organismal lifespan, and that very long-lived organisms have escaped entirely any telomeric constraint on cellular replicative lifespan.

Aging↗
PubMed2006

Disparate patterns of age-related changes in lipid peroxidation in long-lived naked mole-rats and shorter-lived mice.

A key tenet of the oxidative stress theory of aging is that levels of accrued oxidative damage increase with age. Differences in damage generation and accumulation therefore may underlie the natural variation in species longevity. We compared age-related profiles of whole-organism lipid peroxidation (urinary isoprostanes) and liver lipid damage (malondialdehyde) in long living naked mole-rats [maximum lifespan (MLS) > 28.3 years] and shorter-living CB6F1 hybrid mice (MLS approximately 3.5 years). In addition, we compared age-associated changes in liver non-heme iron to assess how intracellular conditions, which may modulate oxidative processes, are affected by aging. Surprisingly, even at a young age, concentrations of both markers of lipid peroxidation, as well as of iron, were at least twofold (P < 0.005) greater in naked mole tats than in mice. This refutes the hypothesis that prolonged naked mole-rat longevity is due to superior protection against oxidative stress. The age-related profiles of all three parameters were distinctly species specific. Rates of lipid damage generation in mice were maintained throughout adulthood, while accrued damage in old animals was twice that of young mice. In naked mole-rats, urinary isoprostane excretion declined by half with age (P < 0.001), despite increases in tissue iron (P < 0.05). Contrary to the predictions of the oxidative stress theory, lipid damage levels did not change with age in mole-rats. These data suggest that the patterns of age-related changes in levels of markers of oxidative stress are species specific, and that the pronounced longevity of naked mole-rats is independent of oxidative stress parameters.

Aging↗
PubMed1970

Tumor immunity produced by the intradermal inoculation of living tumor cells and living Mycobacterium bovis (strain BCG).

The intradermal inoculation of mixtures containing living tumor cells and living Mycobacterium bovis (strain BCG) into unimmunized syngeneic guinea pigs results in an inflammatory reaction to the BCG, and there is no progressive tumor growth. In the absence of BCG the tumor grows progressively, metastasizes, and kills the animal. By conventional methods, it has not been possible to immunize syngeneic guinea pigs to the tumor used. Guinea pigs that receive mixtures of BCG and tumor cells, however, develop specific systemic tumor immunity as measured by delayed cutaneous hypersensitivity and by suppression of tumor growth.

Animals↗
PubMed1990

DNA probes show genetic variation in cyanobacterial symbionts of the azolla fern and a closer relationship to free-living nostoc strains than to free-living anabaena strains.

Twenty-two isolates of Anabaena azollae derived from seven Azolla species from various geographic and ecological sources were characterized by DNA-DNA hybridization. Cloned DNA fragments derived from the genomic sequences of three different A. azollae isolates were used to detect restriction fragment length polymorphism among all symbiotic anabaenas. DNA clones were radiolabeled and hybridized against southern blot transfers of genomic DNAs of different isolates of A. azollae digested with restriction endonucleases. Eight DNA probes were selected to identify the Anabaena strains tested. Two were strain specific and hybridized only to A. azollae strains isolated from Azolla microphylla or Azolla caroliniana. One DNA probe was section specific (hybridized only to anabaenas isolated from Azolla ferns representing the section Euazolla), and five other probes gave finer discrimination among anabaenas representing various ecotypes of Azolla species. These cloned genomic DNA probes identified 11 different genotypes of A. azollae isolates. These included three endosymbiotic genotypes within Azolla filiculoides species and two genotypes within both A. caroliniana and Azolla pinnata endosymbionts. Although we were not able to discriminate among anabaenas extracted from different ecotypes of Azolla nilotica, Azolla mexicina, Azolla rubra and Azolla microphylla species, each of the endosymbionts was easily identified as a unique genotype. When total DNA isolated from free-living Anabaena sp. strain PCC7120 was screened, none of the genomic DNA probes gave detectable positive hybridization. Total DNA of Nostoc cycas PCC7422 hybridized with six of eight genomic DNA fragments. These data imply that the dominant symbiotic organism in association with Azolla spp. is more closely related to Nostoc spp. than to free-living Anabaena spp.

Journal Article↗
PubMed2000

In vivo characterization of the murine intranasal model for assessing the immunogenicity of attenuated Salmonella enterica serovar Typhi strains as live mucosal vaccines and as live vectors.

Attenuated Salmonella enterica serovar Typhi live vector vaccine strains are highly immunogenic in mice following intranasal but not orogastric inoculation. To elucidate the relationship between organs within which vaccine organisms are found and the induction of specific serum immunoglobulin G (IgG) antibodies, we examined the in vivo distribution of serovar Typhi vaccine strain CVD 908-htrA following intranasal administration. Vaccine organisms were cultured from the nasal lymphoid tissue (NALT), lungs, and Peyer's patches 2 min after intranasal inoculation. Vaccine organisms persisted longer in NALT than in other organs. By decreasing the volume of intranasal inoculum containing 10(9) CFU (from a single 30- or 10-microl dose to four 2.5-microl doses given over the course of 1 h), we were able to significantly reduce the number of vaccine organisms isolated from the lungs (P < 0.05) without reducing the number of vaccine organisms in NALT. Reducing the number of vaccine organisms in the lungs resulted in a significant decrease in the serum tetanus antitoxin response elicited by CVD 908-htrA expressing tetanus toxin fragment C under the control of the redox-responsive nir15 promoter. In contrast, a similar construct expressing tetanus toxin fragment C under control of the constitutive lpp promoter stimulated a strong serum IgG tetanus antitoxin response with both inoculation regimens. The data suggest that following intranasal inoculation, NALT is a sufficient inductive site for elicitation of an immune response against both the live vector and heterologous antigen and, as occurs following oral inoculation of humans, attenuated serovar Typhi vaccine organisms elicit serum IgG responses.

Administration, Intranasal↗
PubMedDate not supplied

Use of short-lived 18F and long-lived 14C in double tracer autoradiography for simultaneous measurement of LCBF and LCGU.

We have developed a quantitative autoradiographic technique for the simultaneous measurement of local cerebral blood flow (LCBF) and local cerebral glucose utilization (LCGU) using a combination of 14C-long-lived and 18F-short-lived radionuclides as labels. To obtain the LCGU image, a 50-fold greater radioactivity of 18F than of 14C was administered and the first exposure was done for 2 hours. Three days later, when most of the 18F had decayed, a second exposure was done for 5 to 6 days to obtain the LCBF image. 18F standards were prepared in each experiment. The technique provides, for the first time, the local glucose flow ratio (LGFR). LGFR, obtained by dividing the LCGU by the LCBF image, was expressed as percent mumol/ml (multiplied by 100). Measurement of these three values in the same animal is expected to prove useful in the investigation of the pathophysiology of the brain. The advantages of this method are that cross contamination is less than 4%, chemical or physical manipulation of the slices is unnecessary, and final results can be obtained within a week.

Animals↗
PubMed2002

Conditional live virus as a novel approach towards a safe live attenuated HIV vaccine.

To control the worldwide spread of HIV, a safe and effective prophylactic vaccine is urgently needed. Studies with the simian immunodeficiency virus demonstrated that a live attenuated virus can be effective as a vaccine, but serious concerns about the safety of such a vaccine virus have arisen. We propose a conditional live virus, of which the replication can be switched on and off at will, as a novel approach for an HIV vaccine.

AIDS Vaccines↗