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Biomedical subjects

M C Morris

Publications and source records attributed to M C Morris.

At least 37 records · Page 2Linked to original sources
PubMed2001

A peptide carrier for the delivery of biologically active proteins into mammalian cells.

The development of peptide drugs and therapeutic proteins is limited by the poor permeability and the selectivity of the cell membrane. There is a growing effort to circumvent these problems by designing strategies to deliver full-length proteins into a large number of cells. A series of small protein domains, termed protein transduction domains (PTDs), have been shown to cross biological membranes efficiently and independently of transporters or specific receptors, and to promote the delivery of peptides and proteins into cells. TAT protein from human immunodeficiency virus (HIV-1) is able to deliver biologically active proteins in vivo and has been shown to be of considerable interest for protein therapeutics. Similarly, the third alpha-helix of Antennapedia homeodomain, and VP22 protein from herpes simplex virus promote the delivery of covalently linked peptides or proteins into cells. However, these PTD vectors display a certain number of limitations in that they all require crosslinking to the target peptide or protein. Moreover, protein transduction using PTD-TAT fusion protein systems may require denaturation of the protein before delivery to increase the accessibility of the TAT-PTD domain. This requirement introduces an additional delay between the time of delivery and intracellular activation of the protein. In this report, we propose a new strategy for protein delivery based on a short amphipathic peptide carrier, Pep-1. This peptide carrier is able to efficiently deliver a variety of peptides and proteins into several cell lines in a fully biologically active form, without the need for prior chemical covalent coupling or denaturation steps. In addition, this peptide carrier presents several advantages for protein therapy, including stability in physiological buffer, lack of toxicity, and lack of sensitivity to serum. Pep-1 technology should be extremely useful for targeting specific protein-protein interactions in living cells and for screening novel therapeutic proteins.

3T3 Cells↗
PubMed2001

Is bacterial vaginosis a sexually transmitted infection?

OBJECTIVES: To determine whether the risk factors associated with bacterial vaginosis (BV) are consistent with it being a sexually transmitted infection (STI) by comparing the characteristics of women with BV with those of women with infections recognised as being sexually and non-sexually transmitted. METHODS: A prospective cross sectional study was conducted among female patients presenting for diagnosis at a genitourinary medicine clinic in Sheffield between January 1996 and September 1998. Demographic and behavioural characteristics were reported from patient records and a standardised questionnaire was administered. Risk factor models for BV, two STIs, and two non-STIs were compiled using a multivariable logistic regression analysis. RESULTS: Among the 8989 females under 45 years eligible for analysis, the prevalence of BV was 12.9%. Risk factors associated with BV included some in common with gonorrhoea and Chlamydia trachomatis and some that were not associated with these two STIs. Risk factors common to BV and the STIs included having had more than one sexual partner in the past 3 months, having a history of a bacterial STI, being of black Caribbean ethnicity, and living in a deprived area. However, BV had a contrasting age profile, being most prevalent among those over the age of 30. BV was also more common in those who were divorced. CONCLUSIONS: BV is associated with some factors related to the acquisition of gonorrhoea and Chlamydia trachomatis. However, infection is not only determined by those factors and therefore factors other than sexual activity may be important in the development of the condition.

Adolescent↗
PubMed2000

An essential phosphorylation-site domain of human cdc25C interacts with both 14-3-3 and cyclins.

Human cdc25C is a dual-specificity phosphatase involved in the regulation of cell cycle progression in both unperturbed cells and in cells subject to DNA damage or replication checkpoints. In this study, we describe the structure-function relationship of an essential domain of human cdc25C that interacts with 14-3-3 proteins. We show that this domain is a bi-functional interactive motif that interacts with cyclins primarily through their P-box motif in addition to 14-3-3 proteins. Characterization of the structural features of this domain by NMR and circular dichroism reveals two distinct alpha helical moieties interconnected by a loop carrying the 14-3-3 binding site. Moreover, the helical folding is induced upon binding to 14-3-3, suggestive of a conformational regulation of this domain of cdc25C through interactions with partner proteins in vivo. Combining our structural and biochemical data, we propose a detailed model of the molecular mechanism of cdc25C regulation by differential association with 14-3-3 and cdc2-cyclin B.

14-3-3 Proteins↗
PubMed2000

Translocating peptides and proteins and their use for gene delivery.

A dramatic surge in the development of peptides for gene delivery in vitro and in vivo has been witnessed in the past decade. A better understanding of the structural and mechanistic properties of peptides has been an important step for the rational design of optimal peptide-based gene delivery systems. Research has focused on the design of short synthetic peptides that overcome both extracellular and intracellular limitations of other gene delivery systems by binding reversibly and condensing DNA, specifically targeting cells and/or tissues, rapidly releasing plasmids into the cytoplasm and mediating efficient nuclear translocation.

DNA↗
PubMed2000

The cross-sectional association between blood pressure and Alzheimer's disease in a biracial community population of older persons.

BACKGROUND: The relation of blood pressure to Alzheimer's disease (AD) is complex because both an association of high blood pressure with increased risk of the disease and lower blood pressure as a consequence of the disease are possible. METHODS: We examined the cross-sectional association of blood pressure and AD in the Chicago Health and Aging Project (CHAP), a study of a geographically defined, biracial community. After in-home interviews with 6.162 residents > or =65 years, a stratified random sample of 729 participants was clinically evaluated; 709 had blood pressures measured, and 243 were diagnosed with AD. RESULTS: In logistic regression models adjusted for age, sex, education, and race there was no association between blood pressure measured as a continuous variable and Alzheimer's disease. In categorical analyses, however, prevalence of Alzheimer's disease was significantly higher among persons with low systolic pressure (<130 mmHg) compared with the referent group of 130-139 mmHg (odds ratio [OR] = 2.2, 95% confidence interval [CI]: 1.2,4.1), and with low diastolic pressure (<70 mmHg) compared to the referent of 70-79 mmHg (OR = 1.8, 95% CI: I. 1,3.1). High systolic and diastolic categories were not statistically different from the referent group, although there was some evidence that the associations differed by race. The odds ratios changed little with further adjustment for apolipoprotein E genotype, antihypertensive medications, body mass, stroke, diabetes, and heart disease. CONCLUSION: These findings are consistent with previous studies showing associations between low blood pressure and AD, but longitudinal studies are needed to characterize cause-and-effect associations.

Aged↗
PubMed2000

The prevalence of dizziness and its association with functional disability in a biracial community population.

BACKGROUND: Information regarding the prevalence of dizziness and its association with functional disability among African American and white residents from defined community populations is limited. METHODS: A total of 6,158 persons 65 years and older (78.8% of age-eligible persons) completed in-home interviews that included three common measures of self-reported disability: the Katz Activities of Daily Living (ADL) Scale, the Rosow-Breslau Functional Health Scale, and the Nagi Physical Disability Scale. A stratified random sample of 729 persons underwent a detailed evaluation that included questions on the frequency and severity of dizziness, which was defined as having an episode of dizziness or lightheadedness at least once a month. RESULTS: The overall prevalence of dizziness in this population was 9.6% (95% confidence interval [CI] 7.2-12.0). It increased with age, from 6.6% in those 65-74 years old, to 11.6% in those 75-84 years old, and to 18.4% in those persons > or =85 years old. It was more common in women (odds ratio [OR] 2.03, 95% CI 0.99-4.19) but was not associated with race. After adjusting for age, sex, and race, dizziness was associated with greater disability on the Rosow-Breslau (OR 2.29, 95% CI 1.18-4.46) and Nagi (OR 2.54, 95% CI 1.48-4.36) measures but not on the Katz ADL Scale (OR 1.18, 95% CI 0.64-2.20). CONCLUSIONS: Dizziness is common among older persons and is associated with functional disability.

Activities of Daily Living↗
PubMed2000

Comparison of informal caregiving by black and white older adults in a community population.

OBJECTIVES: To examine the prevalence of informal caregiving and demographic factors associated with caregiving time in older community residents and compare caregiving prevalence and time spent providing care by black and white residents. DESIGN: A cross-sectional, population-based study. SETTING: The study was conducted as part of the Chicago Health and Aging Project (CHAP) in a geographically defined community of black and white residents aged 65 and older. PARTICIPANTS: Participants were 5,924 community residents (61.4% black; 38.6% white) who answered questions about informal caregiving responsibilities during a structured interview about a broad range of health and social factors. METHODS: Data were collected during an in-home interview. Multiple logistic and linear regression models were used to examine the association between caregiving and race, gender, age, marital status, and education. RESULTS: More than 16% of residents had provided care to others during the previous 12 months, and 10.3% were currently providing care. Compared with whites, blacks were 30% more likely to be caregivers, spent almost 13 more hours each week in caregiving activities, and were more likely to assist friends. The probability of caregiving increased significantly with age for married persons, decreased with age for unmarried persons, and was lower for men compared with women. The time spent providing care each week increased significantly with age for married persons and did not differ between men and women. CONCLUSIONS: Although physicians and other healthcare providers typically view older people as the recipients of informal care, individuals older than age 65 provide a substantial amount of care to others with health problems and disability. Most research has focused on the needs of young and middle-aged caregivers, and little is known about the needs of these older caregivers. Future research should use sampling strategies that provide adequate numbers of white and non-white participants for meaningful comparisons. This will permit identification of racial and cultural differences in caregiving so that interventions can be tailored to specific groups.

Black or African American↗
PubMed1999

The thumb domain of the P51-subunit is essential for activation of HIV reverse transcriptase.

The biologically relevant and active form of human immunodeficiency virus reverse transcriptase is a heterodimer produced in a two-step dimerization process. Dimerization involves first the rapid association of the two subunits, followed by a slow conformational change yielding a fully active form. In the present study, we demonstrate that the interaction between the thumb domain of p51 and the RNase-H domain of p66 plays a major role in an essential conformational change required for proper folding of the primer/template and the tRNA-binding site, for maturation and for activation of heterodimeric reverse transcriptase. A synthetic peptide derived from the sequence within the thumb domain of p51, which forms the interface with the RNase-H domains of p66, binds heterodimeric reverse transcriptase with an apparent dissociation constant in the nanomolar range and selectively inhibits activation of heterodimeric reverse transcriptase with an inhibition constant of 1.2 microM. A detailed study of the mechanism of inhibition reveals that this peptide does not require dissociation of heterodimeric RT for efficient inhibition and does not affect subunit association, but interferes with the conformational change required for activation of heterodimeric reverse transcriptase, resulting in a decrease in the affinity of reverse transcriptase for the tRNA and an increase in the stability of the primer/template/reverse transcriptase complex. We have previously proposed that the dimeric nature of reverse transcriptase represents an interesting target for the design of antiviral agents. On the basis of this work, we propose that the conformational changes involved in the activation of reverse transcriptase similarly represent an important target for the design of novel antiviral compounds.

Amino Acid Sequence↗
PubMed1999

A novel potent strategy for gene delivery using a single peptide vector as a carrier.

We have shown previously that a peptide, MPG, derived from the hydrophobic fusion peptide of HIV-1 gp41 and the hydrophilic nuclear localisation sequence of SV40 large T antigen, can be used as a powerful tool for the delivery of oligonucleotides into cultured cells. Now we extend the potential of MPG to the delivery of nucleic acids into cultured cells. In vitro, MPG interacts strongly with nucleic acids, most likely forming a peptide cage around them, which stabilises and protects them from degradation in cell culture media. MPG is non-cytotoxic, insensitive to serum and efficiently delivers plasmids into several different cell lines in only 1 h. Moreover, MPG enables complete expression of the gene products encoded by the plasmids it delivers into cultured cells. Finally, we have investigated the potential of MPG as an efficient delivery agent for gene therapy, by attempting to deliver antisense nucleic acids targeting an essential cell cycle gene. MPG efficiently delivered a plasmid expressing the full-length antisense cDNA of human cdc25C, which consequently successfully reduced cdc25C expression levels and promoted a block to cell cycle progression. Based on our results, we conclude that MPG is a potent delivery agent for the generalised delivery of nucleic acids as well as of oligonucleotides into cultured cells and believe that its contribution to the development of new gene therapy strategies could be of prime interest.

3T3 Cells↗
PubMed1999

A new potent HIV-1 reverse transcriptase inhibitor. A synthetic peptide derived from the interface subunit domains.

The biologically relevant and active forms of human immunodeficiency viruses type 1 and 2 reverse transcriptase found in infectious virions are heterodimers produced in a two-step dimerization process. Dimerization involves first the rapid association of the two subunits, followed by a slow conformational change yielding a fully active form. We have shown that the dimeric nature of reverse transcriptase represents a important target for the design of a new class of antiviral agents. In this work, we propose a new strategy for its inhibition by targeting protein/protein interactions during viral formation in infected cells. From the screening of peptides derived from the tryptophan cluster at the interface of the connection subdomain, we have designed a short peptide (10 residues) corresponding to residues 395-404, which can block dimerization of reverse transcriptase in vitro and in infected cells. This peptide is highly efficient in abolishing the production of viral particles, without any adverse toxic side effects, when transduced into human immunodeficiency virus type 1-infected cells together with a new peptide carrier.

Amino Acid Sequence↗
PubMed1999

Methodological issues in the study of cognitive decline.

In recent years, epidemiologists have given increased attention to cognition, especially to the dementing illnesses that occur in old age. Central to this study is the measurement of change in cognition as opposed to cognition measured at a single point in time. This article addresses conceptual and methodological issues in the study of changes in cognitive function, including: 1) difficulties encountered with the use of single measurements of cognition and the importance of measuring changes in cognition; 2) sources of measurement variation and its potential effects; 3) the importance of careful modeling of age and education; 4) considerations in categorizing outcomes or combining the results of cognitive tests; and 5) the benefits of using multiple-outcome statistical models.

Aged↗
PubMed1999

Effects of phosphorylation of threonine 160 on cyclin-dependent kinase 2 structure and activity.

We have prepared phosphorylated cyclin-dependent protein kinase 2 (CDK2) for crystallization using the CDK-activating kinase 1 (CAK1) from Saccharomyces cerevisiae and have grown crystals using microseeding techniques. Phosphorylation of monomeric human CDK2 by CAK1 is more efficient than phosphorylation of the binary CDK2-cyclin A complex. Phosphorylated CDK2 exhibits histone H1 kinase activity corresponding to approximately 0.3% of that observed with the fully activated phosphorylated CDK2-cyclin A complex. Fluorescence measurements have shown that Thr160 phosphorylation increases the affinity of CDK2 for both histone substrate and ATP and decreases its affinity for ADP. By contrast, phosphorylation of CDK2 has a negligible effect on the affinity for cyclin A. The crystal structures of the ATP-bound forms of phosphorylated CDK2 and unphosphorylated CDK2 have been solved at 2.1-A resolution. The structures are similar, with the major difference occurring in the activation segment, which is disordered in phosphorylated CDK2. The greater mobility of the activation segment in phosphorylated CDK2 and the absence of spontaneous crystallization suggest that phosphorylated CDK2 may adopt several different mobile states. The majority of these states are likely to correspond to inactive conformations, but a small fraction of phosphorylated CDK2 may be in an active conformation and hence explain the basal activity observed.

Adenosine Triphosphate↗
PubMed1999

Characterization of the interactions between human cdc25C, cdks, cyclins and cdk-cyclin complexes.

We have overexpressed and purified human dual-specificity phosphatase cdc25C from a prokaryotic expression system at high levels and in a soluble, active form, and have studied and quantified its potential to interact with cdks, cyclins and preformed cdk-cyclin complexes by fluorescence spectroscopy and size-exclusion chromatography. Our data indicate that human cdc25C forms stable complexes, through hydrophobic contacts, with cdk and cyclin monomers, as well as with preformed cdk-cyclin complexes. In vitro, cdc25C interacts with cyclin monomers with high affinity, with tenfold less affinity with cdks, and with intermediate affinity with cdk-cyclin complexes. Moreover, changes observed in the intrinsic fluorescence of cdks, cyclins and cdk-cyclin complexes upon interaction with cdc25C are indicative of concomitant conformational changes within cdks and cyclins. From our results, we propose that in vitro, in the presence of monomeric cdks and cyclins, cdc25C forms stable ternary complexes, first through a high affinity interaction with a cyclin, which may then help target cdc25C towards a cdk. We discuss the biological relevance of our results and propose that a similar, two-step mechanism of interaction between cdc25C and cdk-cyclin complexes may occur in vivo.

Cell Cycle Proteins↗
PubMed1999

Current and remote blood pressure and cognitive decline.

CONTEXT: Previous studies raise the possibility that blood pressure (BP) in middle age predicts later cognitive decline. OBJECTIVE: To examine prospectively the relationship of BP with level of and change in cognitive function in the elderly. DESIGN: Longitudinal, population-based study comprising subjects enrolled in the East Boston component of the Established Populations for the Epidemiologic Study of the Elderly (EPESE) (1982-1983) and the Hypertension Detection and Follow-Up Program (HDFP) (1973-1974). SETTING: East Boston, Mass. PARTICIPANTS: Of the 3657 participants in the EPESE with baseline BP measurements, 2068 also participated in the HDFP. Subjects were aged 65 to 102 years at baseline in the EPESE and had mental status and memory assessed at baseline and 3 and 6 years. MAIN OUTCOME MEASURES: Numbers of errors on the Short Portable Mental Status Questionnaire and the East Boston Memory Test and rates of change in these numbers of errors. Subjects had BP measured both at baseline in the EPESE and 9 years before, as part of the HDFP. RESULTS: In analyses adjusted for age, sex, and education, there was no strong linear association between BP and cognition. The associations found were fairly small in magnitude, and varied according to which test was used to measure cognition. There was little evidence for an effect of BP on change in cognitive function with either test, or for an effect on level of function on the memory test. In analyses of level of mental status questionnaire performance, however, elevated systolic BP (> or =160 mm Hg) 9 years before baseline was associated with a 14% (95% confidence interval [CI], 4%-25%) increase in error rate, relative to the referent (130-139 mm Hg). Baseline systolic BP had a U-shaped association with the number of errors; error rates were 9% higher compared with the referent group among those with systolic BP lower than 130 mm Hg (95% CI, 1%-17%) and 7% greater (95% CI, 0%-15%) among those with elevated systolic BP. Diastolic BP 9 years before baseline also had a U-shaped association with errors on the mental status questionnaire. CONCLUSION: The findings do not suggest a linear association of BP with cognitive decline, but they are consistent with a more complex relationship between BP and cognition than previously appreciated.

Aged↗
PubMed1999

Design and synthesis of a peptide derived from positions 195-244 of human cdc25C phosphatase.

We have designed, synthesized and purified a 51 amino acid peptide derived from an essential domain of human cdc25C phosphatase. In vivo, differential phosphorylation of this domain regulates either the induction of mitotic processes, or the checkpoint arrest of eukaryotic cells in response to DNA damage. Peptide synthesis was achieved using the stepwise Fmoc strategy and resulted in an important yield of highly pure peptide. The final peptide was identified by amino acid analysis, electrospray mass spectrometry and nuclear magnetic resonance, which revealed that one of the two methionines within the peptide was oxidized into its sulphoxide derivative We investigated whether this 51 amino acid peptide folded into secondary structures in solution by circular dichroism and observed the formation of alpha helices in TFE. Finally, we verified that this peptide could bind to its biologically relevant 14-3-3 partner in vitro by fluorescence spectroscopy.

Amino Acid Sequence↗
PubMed1999

Cognitive activity in older persons from a geographically defined population.

Patterns of cognitive activity, and their relation to cognitive function, were examined in a geographically defined, biracial population of persons aged 65 years and older. Persons (N = 6,162) were given cognitive performance tests and interviewed about their participation in common cognitive activities, like reading a newspaper. Overall, more frequent participation in cognitive activities was associated with younger age, more education, higher family income, female gender, and White race; participation in activities judged to be more cognitively intense was not strongly related to age, but was associated with more education, higher family income, male gender, and White race. Substantial heterogeneity in activity patterns remained after accounting for demographic factors, however. In an analysis controlling for demographic variables, level of cognitive function on performance tests was positively related to composite measures of the frequency and intensity of cognitive activity. Longitudinal studies are needed to assess the relation of cognitive activity patterns to stability and change in cognitive function in older persons.

Aged↗