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J Rossi

Publications and source records attributed to J Rossi.

At least 19 recordsLinked to original sources
PubMed2006

Typical and atypical trafficking pathways of Ad5 penton base recombinant protein: implications for gene transfer.

The adenovirus (Ad) penton base protein facilitates viral infection by binding cell surface integrins, triggering receptor-mediated endocytosis and mediating endosomal penetration. Given these multiple functions, recombinant penton base proteins have been utilized as non-viral vehicles for gene transfer by our lab and others. Although we have previously demonstrated that penton base-derived vectors undergo integrin-specific binding and cell entry, less than desirable levels of gene expression have led us to re-evaluate the recombinant penton base as an agent for gene delivery. To do so, we have examined here the intracellular trafficking of an Ad serotype 5 (Ad5) recombinant penton base protein (PB). Here, we not only observed that PB utilizes a similar, typical trafficking pathway of whole Ad, but also found that PB entered HeLa cells through pathways not yet identified as contributing to cell entry by the whole virus. We show by high-resolution confocal microscopy and biochemical methods that binding to alphav-integrins is a requirement for cell entry, but that early internalization stages did not substantially pass through clathrin-positive and early endosomal compartments. Moreover, a subpopulation of internalized protein localized with caveolin-positive compartments and Golgi markers, suggesting that a certain percentage of proteins pass through non-clathrin-mediated pathways. Similar to the virus, trafficking toward the nucleus was affected by disruption of microtubules and dynein. The majority of penton base molecules avoided the lysosome while facilitating early vesicle release of low molecular weight dextran molecules. In further support of a vesicle escape capacity, a subpopulation of internalized penton base appeared to enter the nucleus, as observed by high-resolution confocal microscopy and cell fractionation. As a confirmation of these findings, we demonstrate that a recombinant penton base facilitated cytosolic entry of an siRNA molecule as observed by RNA interference of a marker gene. Based on our findings here, we suggest that whereas soluble penton base proteins may enter cells through clathrin- and non-clathrin-mediated pathways, vesicle escape and nuclear delivery appear to be supported by a clathrin-mediated pathway. As our previous efforts have focused on utilizing recombinant penton base proteins as delivery agents for therapeutics, these findings allow us to evaluate the use of the penton base as a cell entry and intracellular trafficking agent, and may be of interest concerning the development of vectors for efficient delivery of therapeutics to cells.

Capsid Proteins↗
PubMed2005

[Not Available].

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Journal Article↗
PubMed2005

Physical activity staging distribution: establishing a heuristic using multiple studies.

The purpose of this study was to identify the population prevalence across the stages of change (SoC) for regular physical activity and to establish the prevalence of people at risk. With support from the National Institutes of Health, the American Heart Association, and the Robert Wood Johnson Foundation, nine Behavior Change Consortium studies with a common physical activity SoC measure agreed to collaborate and share data. The distribution pattern identified in these predominantly reactively recruited studies was Precontemplation (PC) = 5% (+/- 10), Contemplation (C) = 10% (+/- 10), Preparation (P) = 40% (+/- 10), Action = 10% (+/- 10), and Maintenance = 35% (+/- 10). With reactively recruited studies, it can be anticipated that there will be a higher percentage of the sample that is ready to change and a greater percentage of currently active people compared to random representative samples. The at-risk stage distribution (i.e., those not at criteria or PC, C, and P) was approximately 10% PC, 20% C, and 70% P in specific samples and approximately 20% PC, 10% C, and 70% P in the clinical samples. Knowing SoC heuristics can inform public health practitioners and policymakers about the population's motivation for physical activity, help track changes over time, and assist in the allocation of resources.

Behavioral Research↗
PubMed2004

[Irreducible knee dislocation by medial capsulo-ligament incarceration].

PURPOSE OF THE STUDY: Irreducible dislocation of the knee is exceptional. Incarceration of the capsulo-ligamentary elements in the inter-condylar notch is a particular causal mechanism. We report four new cases and review the literature to describe the diagnostic and therapeutic approach. MATERIAL AND METHODS: Between January 2001 and January 2002, four patients underwent surgery after trauma (wind surf, ski accidents) producing an irreducible knee dislocation. Mean patient age was 53 years and mean follow-up was 16 months. The IKDC classification was used to assess outcome. RESULTS: Physical examination revealed characteristic signs: a groove facing the medial joint line with signs of cutaneous suffering just above and a fold line behind the medial condyle. Orthopedic reduction failed. Surgery was performed in all cases and revealed incarceration of the entire capsulo-ligamentary structure in the intercondylar notch with the medial condyle in an extra-articular position. After surgical reduction, healing was achieved in all cases without cutaneous necrosis. The mean subjective IKDC score was 53.9. DISCUSSION: Orthopedic reduction is impossible in about 4% of all cases of knee dislocation. Clinical signs are characteristic when the medial capsulo-ligamentary structures are incarcerated in the intercondylar notch; the anatomic injury is almost always the same. Vasculonervous complications are exceptional but an emergency arteriography should be obtained to rule out an infra-clinical intimal lesion. Cutaneous necrosis is a specific complication of irreducible dislocation. Early and complete reduction is required to prevent necrosis. Arthroscopic management raises the risk of compartment syndrome since peripheral lesions are not healed. Satisfactory functional recovery requires associated repair of the cruciate ligaments. CONCLUSION: This type of irreducible dislocation of the knee is a specific clinical entity. It usually results from postero-lateral rotation but may exceptionally occur after lateral dislocation. A groove associated with a fold line indicates the need for rapid surgical reduction to avoid the risk of cutaneous necrosis.

Adult↗
PubMed2003

[Video-nystagmography findings in healthy elderly subjects].

This study investigated 44 healthy elderly subjects aged between 64 and 87, who were analysed with videonystagmoscopy and quantitative videonystagmography, for establishing new standards for normal limits into this new diagnostic tool. 15.9% of the subjects were found to have spontaneous and provoked nystagmus at least in one position studied. Vertical nystagmus in head hanging position was the most frequent finding. In the Dix-Hallpike test we found one case of torsional nystagmus. No subject had seated position nystagmus. Nystagmus after head shaking and evoked nystagmus were not found.

Aged↗
PubMed2003

Phenotypic analysis of human BM T-cell depleted by soybean lectin agglutination and E rosetting with sheep RBC: relative enrichment of NK cells and a CD3(+),CD2(-dim) population.

BACKGROUND: T-cell depletion (TCD) of BM allows transplantation across HLA barriers. Although different methods are used throughout the world, the optimal application of TCD still remains unclear, partly due to the lack of thorough analyses of the cellular fractions eliminated or retained in each method, and their possible implications regarding GvHD, GvL, or engraftment. We have analyzed the phenotype of the successive fractions of 19 BM samples depleted by soybean lectin agglutination and sheep erythrocyte rosetting (elimination of T cells that form rosettes through CD2), focusing on the final fraction infused to patients. METHODS: Analysis was performed using three-color flow cytometry and strategies for optimal staining and individualism of the subsets of interest. RESULTS: The relative composition of the lymphoid population varied significantly along the successive steps in TCD: at the agglutination step, B cells and CD4 T cells are greatly reduced, while natural killer cells (NK) and TCRgammadelta+ T are augmented. The rosetting steps imply the relative enrichment of CD2-dim T cells, together with a further rise in the proportion of NK and double-negative T cells frequently TCRgammadelta+. DISCUSSION: The presence of minor subsets of CD2- TCRgammadelta+ and CD2- TCRalphabeta T cells has already been described in the peripheral blood of normal individuals. We report that, by using this method of TCD, CD2-dim T cells, frequently TCRgammadelta+, are retained in the grafts and infused in patients, together with NK cells as the main lymphoid population. We discuss the possible implications of these populations in the biology of the graft, regarding GvHD, GvL and engraftment.

Animals↗
PubMed2002

Gene modulation in total brain induced by exposure to the bicyclic phosphorus ester trimethylolpropane phosphate (TMPP).

The effect of a single subconvulsive dose of the GABAergic convulsant trimethylolpropane phosphate (TMPP) on gene expression in total rat brain was examined using cDNA array analysis. Using threshold criteria that reduce the number of false positives to <1 gene per 3551 actively transcribed genes on the cDNA array, 41 genes/EST sequences were reproducibly modulated in response to 0.25 mg/kg TMPP. Several genes that were consistent with epileptogenesis and/or neuronal damage and repair mechanisms, such as trkB, alphaB-crystallin, and decorin, were modulated by TMPP exposure in the absence of clinical convulsions. Previous research indicates that rats exposed to subconvulsive doses of TMPP exhibit both "absence-like" EEG paroxysms and persisting central nervous system (CNS) sensitization, as evidenced by increased susceptibility to audiogenic seizures (AGS). Results of this study suggest that cDNA arrays can be used to identify gene modulation events induced by low-level exposure to a chemical convulsant in a reproducible manner.

Animals↗
PubMed2001

Effects of repeated exposure to JP-8 jet fuel vapor on learning of simple and difficult operant tasks by rats.

Groups of 16 Sprague-Dawley rats each were exposed by whole-body inhalation methods to JP-8 jet fuel at the highest vapor concentration without formation of aerosol (1,000 +/- 10% mg/m3); to 50% of this concentration (500 +/- 10% mg/m3); or to treated room air (70 +/- 81 L/min) for 6 h/d, 5 d/wk, for 6 wk (180 h). Although two subjects died of apparent kidney complications during the study, no other change in the health status of exposed rats was observed, including rate of weight gain. Following a 65-d period of rest, rats were evaluated for their capacity to learn and perform a series of operant tasks. These tasks ranged in difficulty from learning of a simple food-reinforced lever pressing response, to learning a task in which subjects were required to emit up to four-response chains of pressing three different levers (e.g., press levers C, R, L, then C). It was shown that repeated exposure to 1,000 mg/m3 JP-8 vapor induced significant deficits in acquisition or performance of moderately difficult or difficult tasks, but not simple learning tasks, as compared to those animals exposed to 500 mg/m3. Learning/performance of complex tasks by the 500-mg/m3 exposure group generally exceeded the performance of control animals, while learning by the 1,000-mg/m3 group was nearly always inferior to controls, indicating possible "neurobehavioral" hormesis. These findings appear consistent with some previously reported data for operant performance following acute exposure to certain hydrocarbon constituents of JP-8 (i.e., toluene, xylenes). There has, however, been little previously published research demonstrating long-term learning effects for repeated hydrocarbon fuel exposures. Examination of regional brain tissues from vapor-exposed rats indicated significant changes in levels of dopamine in the cerebral cortex and DOPAC in the brainstem, measured as long as 180 d postexposure, as compared to controls.

Administration, Inhalation↗
PubMed2001

Interactions between yeasts and bacteria in the smear surface-ripened cheeses.

In the initial phase of ripening, the microflora of bacterial smear surface-ripened cheeses such as Limburger, Taleggio, Brick, Münster and Saint-Paulin and that of surface mould-ripened cheeses such as Camembert and Brie may be similar, but at the end of the ripening, bacteria such as Brevibacterium spp., Arthrobacter spp., Micrococcus spp., Corynebacterium spp. and moulds such as Penicillium camemberti are, respectively, the dominant microorganisms. Yeasts such as Candida spp., Cryptococcus spp., Debaryomyces spp., Geotrichum candidum, Pichia spp., Rhodotorula spp., Saccharomyces spp. and Yarrowia lipolytica are often and variably isolated from the smear surface-ripened cheeses. Although not dominant within the microorganisms of the smear surface-ripened cheeses, yeasts establish significant interactions with moulds and especially bacteria, including surface bacteria and lactic acid bacteria. Some aspects of the interactions between yeasts and bacteria in such type of cheeses are considered in this paper.

Bacteria↗
PubMed2001

Serum cleaved Tau protein and neurobehavioral battery of tests as markers of brain injury in experimental bacterial meningitis.

Brain injury due to bacterial meningitis affects multiple areas of the brain with a heterogeneous distribution generating a challenge to assess severity. Tau proteins are microtubular binding proteins localized in the axonal compartment of neurons. Brain injury releases cleaved Tau proteins (C-tau) into the extracellular space where they are transported to the cerebral spinal fluid. We hypothesized that C-tau crosses the blood-brain barrier during inflammation and that it can be detected in serum. The correlation between serum C-tau levels and the extent of the meningitic insult was examined. Furthermore, we studied whether the use of a subset of neurobehavioral tasks can assess the extent of brain injury after meningitis. The tests were chosen primarily for their ability to detect deficits in the acoustic system, low brain, reflexive responding, as well as for impaired motor coordination and the higher brain functions of learning and memory. A rat model of group B streptococcal meningitis with variable severity was utilized. At five days after bacterial inoculation followed by antibiotic therapy neurobehavioral tests were performed and serum C-tau and histologic samples of the brain were obtained. Our study shows that during meningitis C-tau appears in serum and reflects the extent of neurologic damage. Neurobehavioral performance was altered after bacterial meningitis and could be correlated with histologic and biochemical markers of neurologic sequelae. We conclude that serum C-tau and a composite of neurobehavioral tests could become useful markers for assessing the severity of neurological damage in experimental bacterial meningitis.

Animals↗
PubMed2001

Effects of repeated exposure of rats to JP-5 or JP-8 jet fuel vapor on neurobehavioral capacity and neurotransmitter levels.

The U.S. Naval Service is anticipating transition from the nearly exclusive use of JP-5 jet fuel to predominant use of JP-8, consistent with the primary utilization by the U.S. Army, U.S. Air Force, and the militaries of most NATO countries. To compare the relative risk of repeated exposure to JP-5 versus JP-8 vapor, groups of 32 male Sprague-Dawley rats each were exposed for 6 h/d, 5 d/wk for 6 wk (180 h) to JP-8 jet fuel vapor (1,000 +/- 10% mg/m3), IP-5 vapor (1,200 +/- 10% mg/m3), or room air control conditions. Following a 65-d rest period, rats completed 10 tests selected from the Neurobehavioral Toxicity Assessment Battery (NTAB) to evaluate changes in performance capacity. Repeated exposure to JP-5 resulted in significant effects on only one test, forelimb grip strength (FGS), while exposure to JP-8 vapor resulted in a significant difference versus controls on appetitive reinforcer approach sensitization (ARAS). Rats were further evaluated for concentrations of major neurotransmitters and metabolites in five brain regions and in the blood serum. Levels of dopamine, the dopamine metabolite dihydroxyphenylacetic acid (DOPAC), and the serotonin metabolite homovanillic acid (HVA) were significantly modulated in various brain regions, as measured 85+ d postexposure. Similarly, serum levels of the serotonin metabolite 5-hydroxyindoleacetic acid (5-HIAA) were differentially modulated following JP-8 or JP-5 exposure. Results are compared to previously published research evaluating the neurotoxicity of repeated exposure to other hydrocarbon fuels and solvents.

3,4-Dihydroxyphenylacetic Acid↗
PubMed2001

Behavioral sensitization following exposure to low doses of trimethylolpropane phosphate.

Behavioral sensitization is commonly studied within the context of drugs known to directly increase activity in the brain's dopamine system, particularly drugs of abuse. However, the present research suggests such behavioral changes can also be observed following exposure to other compounds that indirectly affect the dopamine system. One such compound is trimethylolpropane phosphate (TMPP), a bridged organophosphate that can be produced by the partial pyrolysis of certain synthetic lubricants used on military ships and aircraft. Although TMPP is a potent convulsant, it has been demonstrated that treatment with doses below seizure threshold results in long-term behavioral sensitization. The effect has been demonstrated with a number of neurobehavioral endpoints, particularly those assessing appetitive responding. More specifically, sensitization has been observed in acquisition of schedule-induced polydipsia (SIP), appetitive reinforcer approach sensitization (ARAS) and social interaction as measured in neonatal ultrasonic vocalizations, juvenile play and adult conspecific approach. Overall, the rats demonstrated a heightened appetitive response pattern. More specifically, TMPP reliably reduced the number of SIP sessions necessary to induce asymptotic drinking level and increased the time spent investigating (sniffing) a food reinforcer as measured in the ARAS task. Specific effects of TMPP on social interaction were an increase in ultrasonic vocalizations when the neonate was isolated from the dam and littermates and an increase in both measures of juvenile play (pins and dorsal contacts). A complex set of interactions emerged for the measures of adult social investigation where the drug effect was modulated by such factors as sex and neutral vs. stress-inducing experiences coincident with the drug treatment. In contrast to the above results, no behavioral changes were recorded for measures in the elevated plus maze and open field exploration. These results suggest that TMPP produces neurophysiological changes that persist much longer than the pharmacological effect of the compound, particularly in the neural correlates for appetitive behavior.

Aging↗
PubMed2001

Multivalent anti-CCR ribozymes for stem cell-based HIV type 1 gene therapy.

HIV-1 infection of susceptible cells is mediated by the specific interaction of viral envelope glycoproteins with the cell surface CD4 receptor and a chemokine coreceptor, CCR5 or CXCR4. Individuals with a CCR5 genetic defect show resistance to HIV-1 infection, indicating that downregulation of CCR5 expression on target cells can prevent viral infection. In previous studies we demonstrated the utility of an anti-CCR5 ribozyme targeted to a single cleavage site in downregulating CCR5 expression and consequently providing resistance to viral infection. To improve on the level of downregulation we designed a construct containing an anti-CCR5 ribozyme heterotrimer (R5RbzTM) targeted to three different cleavage sites in CCR5 mRNA. In vitro tests showed that the anti-CCR5 ribozyme heterotrimer could effectively cleave the CCR5 RNA substrates to yield products of the expected sizes. This construct was introduced into various retroviral vectors for stable gene transduction. HOS.CD4/R5 cells stably transduced with this anti-CCR5 heterotrimer showed a marked reduction in the surface expression of CCR5 and a concomitant 70% reduction in macrophage-tropic viral infection. In addition, a retroviral vector containing the anti-CCR5 ribozyme heterotrimer and an anti-HIV-1 tat-rev ribozyme heterodimer was constructed. This construct also showed a similar inhibition of CCR5 surface expression and reduced infectability by the macrophage-tropic HIV-1 vector in HOS.CD4/R5 cells. The trimeric and multimeric ribozyme constructs were transduced into CD34+ hematopoietic progenitor cells to determine their effects on lineage-specific differentiation. We show that multivalent ribozyme gene-transduced hematopoietic progenitors differentiated normally into mature macrophages that bear CD14 and CD4 surface markers. Macrophages containing the transgenes expressed ribozymes, and showed resistance to M-tropic HIV-1 infection. These results provide strong support for the use of the trimeric anti-CCR5 ribozyme approach in a gene therapy setting for the treatment of HIV infection.

CCR5 Receptor Antagonists↗
PubMed2001

Trimethylolpropane phosphate induces epileptiform discharges in the CA1 region of the rat hippocampus.

The actions of trimethylolpropane phosphate (TMPP), an ethyl bicyclophosphate convulsant produced during the partial pyrolysis of some phosphate ester-based lubricants, were tested on CA1 neurons of rat hippocampal slices using intracellular recording techniques. Bath application of TMPP (0.1-100 microM) induced spontaneous paroxysmal depolarizing shifts and the associated spontaneous epileptiform bursts followed by after-hyperpolarizations in 63% of neurons tested. The TMPP-induced epileptiform bursts were blocked by muscimol, a gamma-aminobutyric acid A (GABA(A)) receptor agonist, diazepam (DZP), a GABA(A)-benzodiazepine ionophore complex agonist, or baclofen, a GABA(B) receptor agonist. While bath application of muscimol, DZP, or baclofen suppressed spontaneous activity in CA1 neurons not previously exposed to TMPP, subsequent application of TMPP (10 microM) reversed the actions of muscimol and diazepam, but not baclofen. TMPP (0.1-100 microM) also induced membrane hyperpolarization associated with an increase in peak input resistance and inward rectification in 33% of neurons tested or membrane depolarization associated with an increase in input resistance in 17% of neurons tested. In summary, TMPP induced epileptiform activities in hippocampal CA1 neurons. The epileptogenic effects of TMPP are consistent with its interaction with GABA(A)-benzodiazepine receptors.

Action Potentials↗
PubMed2001

Acute neurobehavioral effects in rats from exposure to HFC 134a or CFC 12.

1,1,1,2-Tetrafluoroethane (HFC 134a), a chlorine-free hydrofluoroalkane, is internationally replacing billions of pounds of dichlorodifluoromethane (CFC 12) for coolant, refrigerant and aerosol propellant applications. The ALC50 for HFC 134a in rats is 567,000 ppm for 4 h; its potential for cardiac epinephrine sensitization in beagle dogs is acceptable (75,000 ppm); and its capacity to induce carcinogenicity or developmental disorders in animals is minimal. HFC 134a, with a serum half life estimated at 4-11 min, has been accepted for use as a propellant in metered-dose inhalant products, implying a low human toxicity risk from periodic brief exposures. There has been little published human or animal research evaluating possible neurobehavioral toxicity from longer HFC 134a exposures, as may be expected to occur in operational scenarios. In this study, male Wistar rats were exposed to various concentrations of HFC 134a or CFC 12 for up to 30 min while performing in either a rotarod/motorized running wheel apparatus or in an operant chamber The relative neurobehavioral toxicity of CFC 12 and its ozone-depleting substance replacement HFC 134a was assessed by comparing both gross motor system incapacitation and more subtle changes in ability to perform an operant discrimination task. It was shown that exposure to HFC 134a or CFC 12 concentrations from 40,000 to 470,000 ppm, for up to 30 min, induced neurobehavioral deficits in every subject, ranging from reduced operant efficiency to apparent anesthesia. For neurobehavioral endpoints examined in these experiments, HFC 134a inhalation was shown to induce deficits more rapidly, and at lower concentrations when compared to CFC 12 exposure.

Administration, Inhalation↗
PubMed2001

Reduction of motor seizures in rats induced by the ethyl bicyclophosphate trimethylolpropane phosphate (TMPP).

1. Trimethylolpropane phosphate (TMPP) is a potent cage convulsant, reported to act through binding to the picrotoxinin and/or benzodiazepine receptor sites of the gamma-aminobutyricA (GABA(A)) ionophore complex. 2. Adult male Fischer-344 rats were pretreated by intraperitoneal (i.p.) injection with either diazepam (DZP) [0.5-5.0 mg/kg], Phenobarbital (PB) [5-20 mg/kg], dizocilpine maleate (MK-801) [0.5-3.0 mg/kg], Tiagabine (TGB) [0.5-5.0 mg/kg], 6,7-dinitro-quinoxaline-2,3-dione (DNQX), [5-20 mg/kg], or scopolamine [SCP] (0.25-1.0 mg/kg) 30 min prior to i.p. injection with a convulsive dose of TMPP (0.6 mg/kg). 3. Rats were rated for occurrence of convulsive activity for 120 min post-injection. Time from TMPP injection to observation of subclinical seizures, generalized (tonic-clonic) seizures, and lethality was rated for each pretreatment group. 4. In general, DZP = PB > TGB in reduction of TMPP subclinical and/or clinical seizures. MK-801, at dose levels inducing near sedation, was also effective in modulation of TMPP-induced seizures. SCP or DNQX were generally ineffective in reducing or eliminating TMPP-induced seizures.

Animals↗