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Biomedical subjects

M Stanley

Publications and source records attributed to M Stanley.

At least 19 recordsLinked to original sources
PubMed2004

Genomic imbalances in 70 snap-frozen cervical squamous intraepithelial lesions: associations with lesion grade, state of the HPV16 E2 gene and clinical outcome.

Host genomic abnormalities may determine the natural history of cervical squamous intraepithelial lesions (SILs). We undertook comparative genomic hybridisation analysis of epithelium carefully microdissected from 70 cervical SILs, the largest series to date. In contrast to previous studies, we used frozen sections for optimal DNA quality and examined whether patterns of DNA copy number imbalance (CNI) are characteristic of SIL grade, human papillomavirus (HPV) status and postoperative recurrence. We identified more CNIs in cervical SIL than previously described, with more CNIs per case in high-grade squamous intraepithelial lesion (HG-SIL) than in low-grade squamous intraepithelial lesion (LG-SIL) (P=0.04). While some CNIs were seen at similar frequencies in HG-SIL and LG-SIL, others, including gain on 1q, 3q and 16q, were found frequently in HG-SIL but not in LG-SIL. There were significantly more CNIs per case in HG-SILs showing loss of the HPV16 E2 gene (a repressor of viral oncogene transcription) (P=0.026) and in HG-SILs that subsequently recurred (P=0.04). Our data are consistent with sequential acquisition of CNIs in cervical SIL progression. Higher frequency of CNI in association with E2 gene loss supports in vitro evidence that high-risk HPV integration is associated with genomic instability. Further investigation of the clinical value of specific host genomic abnormalities in cervical SIL is warranted.

DNA-Binding Proteins↗
PubMed2001

Early genetic events in HPV immortalised keratinocytes.

Cancer of the uterine cervix (CaCx) is the second most common cancer in women worldwide. More than 99% of all cervical cancers contain high-risk human papillomaviruses (HPVs), with type 16 predominating. HPV infection alone is not sufficient for neoplastic progression; the HPV-infected cell must undergo additional genetic changes. Cytogenetic analysis of CaCx has been limited due to difficulties in obtaining good-quality banded chromosome preparations. Oncogenic HPVs immortalise primary genital keratinocytes in vitro, and evidence suggests that the molecular genetic and cytogenetic abnormalities observed in HPV immortalised cells reflect the in vivo changes. Therefore, these lines represent suitable models for HPV-induced carcinogenesis. We have used both spectral karyotyping (SKY) and multiplex-FISH (M-FISH) analysis to identify karyotypic changes in HPV-16 immortalised keratinocyte cell lines and established CaCx lines. SKY and M-FISH identified chromosomal abnormalities in all cell lines examined, with a translocation of chromosome 10 or i(10q) occurring in 9 of the 12 cell lines investigated. Further studies with chromosome 10 band-specific probes identified the translocation event as involving 10q with the breakpoint at 10p11.2 in some cell lines or 10q11.2 in others. The pericentric region of chromosome 10 is known to contain duplicated sequences flanking the centromeric satellites. The duplicated sequences contain many zinc finger transcription factor encoding genes and disruption of these in HPV immortalised cell lines may alter the transcription with consequences for both cellular and viral gene expression.

Aneuploidy↗
PubMed2000

Association of aggressive behavior with altered serotonergic function in patients who are not suicidal.

OBJECTIVE: The purpose of this study was to determine whether aggression and serotonergic dysfunction are related in the absence of a history of suicidal behavior. Although serotonergic dysfunction has been implicated in aggressive and impulsive behavior, most studies of such behavior have included individuals with a history of suicide attempts. Low concentrations of CSF 5-hydroxyindoleacetic acid (5-HIAA) have been consistently associated with suicidal behavior, presenting a potential confound in the link between aggression and serotonergic dysfunction. METHOD: The authors examined the association between aggression and CSF 5-HIAA concentrations in a group of 64 patients who had different DSM-III-R axis I diagnoses and no past suicidal behavior. Aggressive (N=35) and nonaggressive (N=29) groups were defined by a median split on a six-item history of adulthood aggressive behavior. RESULTS: The aggressive group had significantly lower CSF 5-HIAA concentrations than the nonaggressive group. Aggressive individuals also scored significantly higher on self-report measures of hostility, impulsiveness, and sensation seeking. CSF 5-HIAA concentrations, however, did not correlate with self-reported hostility and impulsivity. CONCLUSIONS: There is an association between aggressive behavior and serotonergic dysfunction independent of suicidal behavior in patients with axis I disorders who exhibit relatively milder forms of aggressive behavior. Analogous to findings with suicidal behavior, a low concentration of CSF 5-HIAA is related to aggressive behavior but does not show the same relationship to the continuum of aggressive feelings and thoughts.

Adult↗
PubMed2000

An outcome survey of psychology residency training program graduates of the University of Texas Houston Medical School.

A survey of 71 graduates of the psychology residency program at The University of Texas Houston Medical School and the Texas Research Institute of Mental Science indicated that two-thirds of the respondents were involved in private practice, conducting primarily individual therapy, with assessment also playing a major role in their practice. Managed care had the effect of placing limits on the numbers of sessions available to patients, reducing income, and increasing paperwork associated with practice. Implications for training during residency are that individual therapy, using focused short-term approaches, and assessment should continue to be the primary clinical experiences for trainees, and that there should also be training in the business aspect of practice, including marketing.

Adult↗
PubMed1999

An SCL 3' enhancer targets developing endothelium together with embryonic and adult haematopoietic progenitors.

The SCL gene encodes a basic helix-loop-helix transcription factor which is expressed in early haematopoietic progenitors throughout ontogeny and is essential for the normal development of blood and blood vessels. Transgenic studies have characterised spatially distinct 5' enhancers which direct lacZ expression to subdomains of the normal SCL expression pattern, but the same elements failed to produce appropriate haematopoietic expression. We now describe an SCL 3' enhancer with unique properties. It directed lacZ expression in transgenic mice to extra-embryonic mesoderm and subsequently to both endothelial cells and to a subset of blood cells at multiple sites of embryonic haematopoiesis including the yolk sac, para-aortic splanchnopleura and AGM region. The 3' enhancer also targeted expression to haematopoietic progenitors in both foetal liver and adult bone marrow. Purified lacZ(+ )cells were highly enriched for clonogenic myeloid and erythroid progenitors as well as day-12 spleen colony forming units (CFU-S). Within the total gated population from bone marrow, 95% of the myeloid and 90% of the erythroid colony-forming cells were contained in the lacZ(+) fraction, as were 98% of the CFU-S. Activation of the enhancer did not require SCL protein. On the contrary, transgene expression in yolk sacs was markedly increased in an SCL-/- background, suggesting that SCL is subject to negative autoregulation. Alternatively the SCL-/- environment may alter differentiation of extra-embryonic mesoderm and result in an increased number of cells capable of expressing high levels of the transgene. Our data represents the first description of an enhancer that integrates information necessary for expression in developing endothelium and early haematopoietic progenitors at distinct times and sites throughout ontogeny. This enhancer provides a potent tool for the manipulation of haematopoiesis and vasculogenesis in vivo.

Animals↗
PubMed1998

Expression of human papillomavirus type 16 L1 protein in Escherichia coli: denaturation, renaturation, and self-assembly of virus-like particles in vitro.

Major capsid protein L1 of HPV16 was produced in a fused form in Escherichia coli using an inducible expression system. The protein formed insoluble aggregations (inclusion bodies) and the yield was more than 10% of total cell proteins. The inclusion bodies were isolated and solubilised with 8 M urea and the L1 proteins were purified by chromatographic separation. Following removal of the urea by gradual dialysis, the denatured L1 proteins spontaneously renatured and subsequently assembled into polymorphologic aggregations in vitro. Electron microscopy showed that the assembled material included structures resembling native empty capsids as well as incompletely formed capsids. After separation from the pool of polymorphologic structures by sucrose gradient sedimentation, the correctly formed virus-like particles (VLE. coliPs) were recognised by a HPV16 type-specific, conformational-dependent monoclonal antibody in an ELISA. This system offers not only a model for investigation of the intrinsic interactions that occur during L1 assembly, but also a potential route for convenient manufacture of highly purified VLP vaccines.

Capsid Proteins↗
PubMed1998

Energy intake in early infancy and childhood fatness.

OBJECTIVE: To investigate whether aspects of infant energy intake are related to fatness in early childhood. DESIGN: Longitudinal investigation of infants studied at 12 weeks and 2-3.5 y. SUBJECTS: 20 healthy infants, breast-fed or formula-fed, from the general population. MEASUREMENTS: Milk volume intake (MVI) by deuterium turnover, estimated energy intake, weaning status and body composition in infancy, body composition in childhood. RESULTS: MVI was not related to infant skinfolds or percentage fat. Weaning was inversely related to MVI (P < 0.04) at 12 weeks, and inversely related to skinfolds (P = 0.055) and fat mass (P = 0.020) in childhood. MVI and total energy intake were not related to childhood fatness. CONCLUSIONS: Early weaning was associated with a moderate reduction in childhood fatness. Two possible mechanisms are discussed. However, early infant energy intake was not an important determinant of later fatness in this population.

Aging↗
PubMed1997

Investigation of the relationship between infant temperament and later body composition.

OBJECTIVE: To investigate the ability of maternally-rated infant temperament to predict fatness and activity patterns in early childhood. DESIGN: Longitudinal investigation of infants studied at 12 weeks and followed up at 2-3.5 y of age. SUBJECTS: Thirty healthy full-term infants from the general population. MEASUREMENTS: Body composition, behavioural activity and temperament at 12 weeks; anthropometry, body composition, diet and behavioural activity at follow-up. RESULTS: Infant temperament predicted later behaviour and fatness. Easily soothable infants had leaner childhood skinfold thicknesses (P < 0.02) and were more active in childhood (P < 0.025). Infant distress was also related to childhood diet composition. CONCLUSIONS: Infant temperament can predict later body composition and behaviour. Both energy intake and energy expenditure may be mechanisms by which the relationship develops.

Age Factors↗
PubMed1997

Current trends in the clinical management of an old enemy: congestive heart failure in the elderly.

Much has been learned in the past decade, through quality medical and nursing research, regarding the pathophysiology and management of congestive heart failure. Primary care practitioners are challenged to apply the latest data to the clinical management of these patients when there is clear evidence of improvement in the control of symptoms and quality of life. Because of the complexities of the disease process, older adults with congestive heart failure require a comprehensive approach that is particularly well suited to the knowledge and skills of the advanced practice nurse.

Aged↗
PubMed1997

Abnormal expression and mutation of p53 in cervical cancer--a study at protein, RNA and DNA levels.

OBJECTIVES: The objectives of this study are to document the status of p53 expression and mutation in cervical cancer at protein, RNA and DNA levels and to relate this to the presence of HPV. MATERIALS AND METHODS: Biopsy specimens from one hundred and three squamous cell carcinoma of the cervix and histologically normal ectocervix were analysed. Fresh tissues were extracted for protein, RNA and DNA and flash frozen tissue cryostat sectioned for immunohistochemical staining. HPV DNA status was determined by PCR using L1 consensus primers and typed for HPV 16 and 18 with E6 specific primers. p53 expression was determined at the protein level by Western blotting on protein extracts and at RNA level by Northern blotting. RESULTS: There was no p53 overexpression or mutation detectable in the protein extracts. Three of 65 (4.6%) of the carcinomas were positive for p53 by immunostaining with the polyclonal antibody CM1. Overexpression at the RNA level was detected in 2 of 32 (6.3%) carcinomas. p53 mutation was screened for by PCR/SSCP (single strand conformation polymorphism) followed by sequencing to define the site of mutation. Two of the cervical cancers (2.0%) showed mutation in p53 in exons 7 or 8. The mutation rate in HPV positive tumours was 1.2% (1/81) and in HPV negative tumours was 5.2% (1/19). CONCLUSION: p53 overexpression or mutation does not seem to play a significant role in cervical carcinomas.

Blotting, Northern↗
PubMed1996

Comparison of cerebrospinal fluid monoamine metabolite levels in dominant-aggressive and non-aggressive dogs.

Aggression has been shown to be related to reduced serotonergic activity in humans and non-human primates, and in rodents. We now studied the relationship between cerebrospinal fluid (CSF) monoamine metabolites and canine aggression in 21 dominant-aggressive dogs (Canis familiaris) and 19 controls. The diagnosis of dominance-related aggression was based upon a history of biting family members in contexts associated with dominance challenges. Post-mortem CSF 5-HIAA, MHPG and HVA were measured by high-performance liquid chromatography using electrochemical detection. Concentrations of CSF 5-HIAA (P = 0.01) and HVA (P < 0.001) were lower in the aggressive group (median values: 5-HIAA 202.0 pmol/ml; HVA 318.0 pmol/ml) than in controls (5-HIAA 298.0 pmol/ml; HVA 552.0 pmol/ml). No differences were noted in CSF MHPG levels. Differences in 5-HIAA were maintained after controlling for breed and age of dogs, but HVA differences may have been breed-dependent. Lower levels of 5-HIAA (P = 0.02) and HVA (P = 0.04) were found in the subgroup of aggressive dogs with a history of biting without warning (5-HIAA 196.0 pmol/ml; HVA 302.0 pmol/ml) compared to dogs that warned (5-HIAA 244.0 pmol/ml; HVA 400.0 pmol/ml). This study suggests that reduced serotonergic function is associated with aggressive behavior and impaired impulse control in dogs, a finding that is consistent with observations in primates, and suggests that serotonin modulates aggressive behavior throughout mammals.

Aggression↗
PubMed1996

Attempted suicide characteristics and cerebrospinal fluid amine metabolites in depressed inpatients.

BACKGROUND: Serotonin abnormalities have been reported in the brain of suicide victims. Evidence of a serotonin deficiency in suicide attempters is less consistent. We hypothesized that a serotonin deficiency may be present in suicide attempters whose attempt behavior more closely approximates completed suicide. METHOD: Sixty-seven (67) drug-free depressed inpatients (46 suicide attempters, 21 nonattempters) underwent research clinical assessments and a lumbar puncture. Cerebrospinal fluid (CSF) monoamine metabolites were assayed. Degree of medical damage and intent of the most recent suicide attempt were rated. RESULTS: CSF amine metabolites did not differentiate suicide attempters as a group from nonattempters. However, reduced serotonergic activity, as indicated by lower levels of CSF-5-hydroxyindoleacetic acid (5-HIAA) was associated with a history of planned suicide attempts and with suicide attempts that resulted in greater medical damage. Other monoamine metabolites did not correlate with seriousness of suicidal behavior, except for low CSF homovanillic acid and higher medical damage. No correlation was found with violent method. CONCLUSIONS: Planned and more medically damaging suicide attempts appear to be associated specifically with low serotonergic activity and, therefore, resemble completed suicide both behaviorally and biochemically. It remains to be determined whether low levels of CSF 5-HIAA can predict greater medical damage in future suicide attempts.

Adult↗
PubMed1996

Sepsis in the elderly.

Sepsis in the elderly occurs frequently and carries a high rate of mortality. With increasing numbers of elderly patients being cared for in critical care units, the critical care nurse must have a thorough understanding of the unique aspects of this patient population. Nurses must be prepared to deliver expert nursing care that is knowledge-based and incorporates current research findings. Unit protocols that encompass the preventive measures needed by these special patients will assist in reducing the incidence of sepsis. When sepsis occurs, careful trending of data will signal the need for timely and precise interventions that may result in a good outcome for the elderly patient with sepsis.

Age Factors↗
PubMed1996

The relationship between components of infant energy expenditure and childhood body fatness.

OBJECTIVE: To investigate whether any component of infant energy expenditure is related to fatness in early childhood, and whether infant fatness is related to childhood variables. DESIGN: Longitudinal investigation of infants studied at 12 weeks and followed up at 2.5 to 3.5 years of age. SUBJECTS: 30 healthy full-term infants selected from the general population. MEASUREMENTS: Sleeping metabolic rate, total energy expenditure, anthropometry and behaviour at 12 weeks; anthropometry, body composition and behaviour in follow-up. RESULTS: Energy expenditure at 12 weeks (minimal metabolism, total energy expenditure, energy expended on physical activity, behaviour) showed no relationship with later fatness. Infant fatness (skinfold thicknesses and percentage fat) showed in contrast a strong relationship with childhood fatness. Infant fatness also predicted childhood behaviour. CONCLUSIONS: These data do not support the theory that reduced energy expenditure in early infancy is related to later fatness. However, infant fatness influences both later fatness and activity patterns.

Adipose Tissue↗
PubMed1996

Different susceptibility of cervical keratinocytes containing human papillomavirus to cell-mediated cytotoxicity.

OBJECTIVE: To detect the factors responsible for the susceptibility of cervical keratinocytes infected with human papillomavirus (HPV) to non-specific lysis mediated by natural killer (NK) and lymphokine activated killer (LAK) cells. MATERIALS AND METHODS: Five cervical keratinocyte lines: CaSki, SiHa, HeLa (representing high grade squamous intraepithelial lesion (HSIL) ), W12 (representing low grade squamous intraepithelial lesion (LSIL) ) and NCx, (normal cervix) were used as target cells in the four-hour lactate dehydrogenase (LDH) release cytotoxicity assay. The effector cells were NK and LAK. The modulatory effects of interferon gamma (IFN gamma) and tumor necrosis factor alpha (TNF alpha) pretreatment of keratinocytes were investigated by adding IFN gamma or TNF alpha into the flasks of target cells 48 hours before the cytotoxicity assays. The blocking effects of anti-intercellular adhesion molecule-1 (ICAM-1) and anti-lymphocyte function-associated antigen-1 (LFA-1) monoclonal antibodies (Mabs) were also studied. RESULTS: All the 5 cervical keratinocytes were susceptible to LAK, but not to NK. The sensitivity varied among the cell lines. LAK had better killing effects on HSIL than on LSIL. Pretreatment of target cells with IFN gamma and TNF alpha increased the killing mediated by LAK, but had little effect on NK activity. Anti-ICAM-1 and anti-LFA-1 Mabs inhibited LAK-mediated cytotoxicity. CONCLUSIONS: All the HPV infected keratinocytes used in the experiments are NK-resistant and LAK-sensitive cells. IL-2, IFN gamma and TNF alpha play some critical roles in the regulation of the susceptibility of cervical keratinocytes, especially HSIL to LAK-mediated cytotoxicity in vitro.

Cells, Cultured↗
PubMed1995

Concordance of PCR and antibody results from HIV testing of injecting drug users.

Standard HIV-1 testing relies on the enzyme immunoassay (EIA) for detecting antibodies specific to HIV-1. This technique may misclassify persons as HIV-1-negative in instances where testing follows infection but precedes development of antibody to HIV-1. To evaluate the occurrence of HIV infection in the absence of positive antibody, polymerase chain reaction (PCR) for viral DNA in the blood has been applied. Research comparing these two testing techniques has generally focused on populations of homosexual and bisexual men. This study compares PCR and antibody testing of 337 injecting drug users recruited from street settings in San Francisco. Of 286 HIV-1 antibody-negative samples, 3 (1.0%) were PCR-positive. Of 49 HIV-1 antibody-positive samples, 1 (2.0%) was PCR-negative. Two samples were antibody-indeterminate and PCR-negative. This yielded an overall concordance of 331/335 (98.8%), excluding the indeterminate results. These results suggest that current antibody methodology is adequate. However, misclassification among recently infected individuals may occur, which is of concern in high-incidence groups.

Adult↗
PubMed1995

The interaction between human papillomavirus type 16 E1 and E2 proteins is blocked by an antibody to the N-terminal region of E2.

Replication of papillomavirus DNA requires two virally encoded proteins, E1 and E2. We expressed human papillomavirus (HPV) type 16 E1 and E2 in bacteria and showed that purified full-length E2 protein interacted directly with E1, in the absence of HPV16 DNA. It was established that the first 142 amino acids of E1 were not required for binding as E2 protein was able to interact with E1 devoid of this region. The interaction of E2 with E1 could be blocked by a monoclonal antibody that bound E2 in the region of amino acids 18-41 of E2 whereas a monoclonal antibody reactive with a nearby part of the molecule (amino acids 2-17) only partially blocked this interaction. These results suggest that a region in the N-terminus of E2 around amino acids 18-41 is a site of interaction with the E1 protein.

Adenosine Triphosphatases↗