Enhanced cytotoxicity of bleomycin, cisplatin, and carboplatin on equine sarcoid cells following electroporation-mediated delivery in vitro.

Electroporation is a method used to deliver poorly permeant chemotherapeutic drugs to tumor cells, potentiating the cytotoxic effects of drugs and overall clinical response. Despite existing evidence of the potential benefits of electroporation to enhance the antitumoral effects of drugs, there is a lack of understanding about the effects of electroporation on equine tumor cells. This study investigated the combined effects of electroporation and bleomycin, cisplatin, and carboplatin on an equine sarcoid cell line (EqS04b). The use of electroporation increases the cytotoxic effects of bleomycin, cisplatin, and carboplatin on the equine sarcoid cell line by 5-fold, 4-fold, and 3-fold, respectively. These very promising findings demonstrate proof of principle for future preclinical studies on different tumor cells to investigate the in vivo effects of electroporation in sarcoid tumors.

Hepatic manifestations of tuberous sclerosis complex: a genotypic and phenotypic analysis.

Hepatic manifestations of tuberous sclerosis complex: a genotypic and phenotypic analysis. A retrospective review of the clinical records and radiological images of 205 patients with tuberous sclerosis complex (TSC) was performed to evaluate the prevalence and progression of hepatic lesions; examine the association of hepatic phenotype with genotype, age, and gender; and investigate the relationships between hepatic, renal, and pulmonary involvement. Hepatic angiomyolipomas (AML), cysts, and other benign lesions were identified in 30% of the cohort, and some lesions grew significantly over time. However, no patient had clinical symptoms or complications from hepatic lesions. TSC2 patients exhibited a higher frequency of AML compared to TSC1 patients (p = 0.037), and patients with no mutation identified exhibited a higher frequency of cysts compared to TSC2 patients (p = 0.023). Age was positively correlated with frequency of hepatic involvement (p < 0.001), whereas hepatic phenotype was independent of gender. Presence of hepatic AML was associated with presence of renal AML (p = 0.001). These findings confirm a high rate of asymptomatic hepatic lesions in TSC and further characterize the TSC phenotype.

Herpes simplex virus-1 thymidine kinase mutants created by semi-random sequence mutagenesis improve prodrug-mediated tumor cell killing.

Cancer suicide gene therapy affords the prospect of using the most optimal genes available because the source of the therapeutic gene is often irrelevant. Currently, there are numerous preclinical and clinical trials to develop tumor ablative therapies that use viral, yeast, or bacterial genes. One such gene, the herpes simplex virus type 1 (HSV-1) thymidine kinase (TK) is widely used as a suicide gene in combination with ganciclovir. In the study reported here, a restricted set of random sequences (semi-random) was introduced into the active site of HSV-1 TK, and the resulting variants were selected on the basis of their ability to confer increased ganciclovir or acyclovir sensitivity to Escherichia coli. Sequence analysis demonstrated that functional mutants contained three to five amino acid substitutions that are unique and novel combinations. On the basis of enzyme assay results, three mutants were identified for further analysis in vitro. These three mutants conferred substantial increased sensitivity to both ganciclovir and acyclovir when compared with IC50s of wild-type TK expressing rat C6 glioma cells. One mutant, SR39, was further evaluated in a xenograft tumor model in nude mice. Expression of SR39 in tumors was shown to prevent tumor growth at prodrug dosages that did not affect wild-type HSV-1 TK-expressing tumors. The use of any of these mutants as a suicide gene should provide a more effective and safer alternative to wild-type TK, because lower, less immunosuppressive doses of ganciclovir will be necessary for tumor ablation, and the use of acyclovir may now be possible.

Enhanced ganciclovir killing and bystander effect of human tumor cells transduced with a retroviral vector carrying a herpes simplex virus thymidine kinase gene mutant.

Gene transfer of the herpes simplex virus thymidine kinase (TK) gene associated with ganciclovir (GCV) treatment can lead to death of TK-expressing cells, and of neighboring TK- cells because of the bystander effect. Thus, a small proportion of TK+ cells in a tumor can lead to its complete regression after GCV treatment. However, a lack of efficacy of gene transfer into tumors associated with low GCV sensitivity and poor bystander effect of human cancer cells currently limit the clinical use of this suicide gene therapy approach. To increase the potency of suicide gene therapy, we have tested the GCV sensitivity and the bystander effect of TK mutants that have been previously described. After retroviral transduction of the TK mutants into human tumor cells of various origins, we have found a strong killing effect of GCV with cells expressing the mutants TK30 or TKF161C. The GCV sensitivity of several human tumor cell types expressing TK30 was 9- to 500-fold higher than cells containing wild-type TK. Furthermore, TK30-expressing cells were able to kill bystander cells much more efficiently than TK-expressing cells. Thus, TK30 mutant is a promising candidate for suicide gene therapy clinical trials.

Targeted delivery of DNA encoding cytotoxic proteins through high-affinity fibroblast growth factor receptors.

Nonviral DNA delivery strategies for gene therapy have generally been limited by a lack of specificity and efficacy. However, ligand-mediated endocytosis can specifically deliver DNA in vitro to cells bearing the appropriate cognate receptors. Similarly, in order to circumvent problems related to efficacy, DNA must encode proteins with high intrinsic activities. We show here that the ligand basic fibroblast growth factor (FGF2) can target FGF receptor-bearing cells with DNA encoding therapeutic proteins. Delivery of genes encoding saporin, a highly potent ribosomal inactivating protein, or the conditionally cytotoxic herpes simplex virus thymidine kinase, a protein that can kill cells by activating the prodrug ganciclovir, is demonstrated. The saporin gene was codon optimized for mammalian expression and demonstrated to express functional protein in a cell-free assay. FGF2-mediated delivery of saporin DNA or thymidine kinase DNA followed by ganciclovir treatment resulted in a 60 and 75% decrease in cell number, respectively. Specificity of gene delivery was demonstrated in competition assays with free FGF2 or with recombinant soluble FGF receptor. Alternatively, when histone H1, a ligand that binds to cell surface heparan sulfate proteoglycans ("low-affinity" FGF receptors), was used to deliver DNA encoding thymidine kinase, no ganciclovir sensitivity was observed. These findings establish the feasibility of using ligands such as FGF2 to specifically deliver genes encoding molecular chemotherapeutic agents to cells.

Expression vector pT7:TKII for the synthesis of authentic biologically active RNA encoding vaccinia virus thymidine kinase.

A transcription vector, pT7: TKII, was constructed by a novel application of the polymerase chain reaction. Chimeric oligodeoxynucleotides were used to direct the synthesis of a DNA fragment which consisted of a truncated bacteriophage T7 promoter element fused to the vaccinia virus (VV) thymidine kinase gene (tk). This fragment was cloned into a pUC118 plasmid and sequenced to ensure no mutations had occurred during its synthesis. When linearized at the 3' end of the VV tk gene at the BamHI site located in the polylinker region of the vector, pT7:TKII was efficiently transcribed by T7 RNA polymerase into a 595 nucleotide transcript whose 5' end was identical to that found on authentic nascent VV tk mRNA. When translated in a rabbit reticulocyte lysate system, the synthetic VV tk RNA was shown to be biologically active in that it directed the synthesis of a 20-kDa protein which assembled into an enzymatically active 80-kDa tetrameric complex which was indistinguishable from VV thymidine kinase (TK) enzyme isolated from VV-infected cells. The pT7:TKII vector provides a powerful approach with which: (i) to investigate the translational and posttranslational regulation of the VV tk gene; (ii) to use directed genetics to identify potential cis-acting regulatory sequences or structures present within the VV tk RNA; and (iii) to apply protein engineering procedures to identify the catalytic, allosteric and subunit interactive domains of the VV TK enzyme. As an example, the translational effects of adding a m7G cap structure to the pT7:TKII-derived VV tk RNA are presented.

Selection of new biologically active molecules from random nucleotide sequences.

Genetic diversity can be achieved in vitro by inserting random nucleotide (nt) sequences into cloned genes. In the case of enzymes, subsequent genetic complementation can be used to select for new mutants that exhibit different substrate specificities, altered catalytic activities, or altered temperature sensitivities. Using this technique, one can also analyze the contribution of different amino acid residues to the structure and function of enzyme. Selecting biologically active DNA sequences from large random populations provides a new method for identifying nt sequences with unique functions. Analogous random sequence selection techniques have been applied to determine the consensus sequence of the Escherichia coli promoters, DNA and RNA sequences that bind specific protein(s), DNA regulatory sequences, ribozyme(s) and ligand-specific RNA(s). In this manuscript, we will consider recent data obtained in our laboratory as a result of inserting random sequences into the putative nucleoside-binding site of herpes simplex virus type 1 (HSV-1) thymidine kinase (TK). We have obtained over 2000 new mutant HSV-1 TKs, some of which are stable at higher temperatures or have altered substrate specificity and/or catalytic rates when compared to those of the wild-type enzyme.

Psychiatric disorder in patients with advanced breast cancer: prevalence and associated factors.

The prevalence of psychiatric disorder and associated factors has been examined in 139 women with advanced breast cancer. Patients completed a self-report assessment of mood, the Hospital Anxiety and Depression Scale (HAD). They were also interviewed to obtain sociodemographic details, UICC performance status and past psychiatric history. Overall, 35 (25%) scored 11 or above (out of a maximum of 21) on either the anxiety or the depression subscales, or both, of the HAD and were therefore probable cases of anxiety and/or depression. These patients are likely to benefit from psychosocial intervention. Clinical anxiety was unrelated to any sociodemographic or disease related factors. Clinical depression was significantly more prevalent amongst patients in the lower socioeconomic classes (P = 0.01) and those with poor performance status (P = 0.007). Depression can be difficult to detect in patients with advanced breast cancer and these factors may be useful indicators to clinicians of patients at high risk of this disorder.

In vitro evaluation of mutant HSV-1 thymidine kinases for suicide gene therapy.

Herpes Simplex Virus type 1 (HSV-1) thymidine kinase (TK) is currently the most widely used suicide agent for gene therapy of cancer. Tumor cells that express HSV-1 thymidine kinase are rendered sensitive to prodrugs due to preferential phosphorylation by this enzyme. While ganciclovir (GCV) is the prodrug of choice for use with TK, this approach is limited in part by the toxicity of this prodrug. From a random mutagenesis library of over a million mutant thymidine kinases, ten thymidine kinase variants were identified on the basis of activity towards ganciclovir and acyclovir (Black ME, Newcomb TG, Wilson H-MP and Loeb LA: Proc. Natl. Acad. Sci. U.S.A. 93: 3525-3529, 1996). Six mutants described here contain three to six amino acid changes and render mammalian cells more sensitive to acyclovir (ACV) including one that demonstrates an 8.5-fold reduction in IC50 compared to wild-type TK. These novel enzymes could provide benefit to ablative gene therapy by now making it feasible to use the relatively non-toxic acyclovir at nanomolar concentrations.

Older women and mammography screening behavior: do possible selves contribute?

This study sought to explore the contribution of the self-concept to older women's adherence to regular mammography screening behavior. The PRECEDE and health belief model concepts were incorporated with a measure of the women's future selves to determine whether the self-concept adds to our ability to predict screening. A self-administered questionnaire was completed by 210 community-dwelling women ages 50 to 75 years, recruited from urban and rural women's groups. Logistic regression analyses revealed that predictors of adherence were clinical breast examination, physician recommendation, age, barriers, benefits, feared health-related possible self, and self-efficacy in the feared domain. The addition of the self measures significantly improved the overall fit of the model. Implications for theory development, practice, and future research are discussed.

What did popular women's magazines from 1929 to 1949 say about breast cancer?

Despite the increasing risk of breast cancer with age, older women are less likely to engage in breast cancer screening behaviors than are younger women. In order to examine articles written on the topic of breast cancer and cancer, a historical survey was conducted of women's magazines popular between 1929 and 1949. This survey was an attempt to identify the messages of that period and compare them with the beliefs and knowledge prevalent among women who are today 75 to 84 years of age. There were striking similarities found between the messages in the early magazines and in older women's beliefs today. This literature was also compared with the medical literature of the same time to identify the role of the nurse and common themes. This article outlines specific mass-media strategies that nurses can use to design appropriate messages for women of different ages as well as for policy makers and the public at large through popular media such as magazines.

Evaluation of the reorganization of a public health nursing division.

From 1986 to 1988, the Nursing Division of the Hamilton-Wentworth Department of Public Health Services undertook an analysis of our present organizational structure, incorporating knowledge from current literature, community trends and staff perceptions. From this analysis, a division-wide reorganization was implemented from the current practice in which the Public Health Nurse (PHN) delivered nursing care across all ages and stages (generalist role) to one organized by target populations. These events were described in an earlier article. As part of the plan for reorganization, the nurse management team implemented an evaluation process which included five components: 1) an internal staff survey; 2) an external agency survey; 3) a program-specific activity analysis; 4) a program-specific measure of knowledge change and; 5) a longitudinal job satisfaction survey. This article describes this evaluation and presents implications for the future.

Personal, home and community safety promotion with community-dwelling elderly persons: response to a public health nurse intervention.

Public health clients aged 65 years and older were randomly allocated to receive an intervention by a public health nurse during a home visit, promoting either safety behaviours or influenza immunization. Sixty-eight clients reported making a total of 103 safety changes, with almost equal proportions making changes in the safety and influenza groups (21.9% vs. 18.3%). A higher percentage of males (25.0%) than females (17.3%) reported making safety changes following the intervention. Further research should focus on the effectiveness of safety promotion strategies which are expanded to include seniors participation, policy development and community planning and design.

Attitudes of diploma student nurses toward adult clients.

This descriptive study was conducted to determine the attitudes of diploma nursing students towards adult clients of different ages in various health situations. The present study has attempted to fill a gap in nursing research using a new approach: "the person perception paradigm." One hundred college students in the last year of their nursing program completed a questionnaire. Students in this study rated eight photos of male clients, ranging in ages from 20 to 90 years, in various health situations. The rating scales were set up with 12 pairs of bipolar adjectives which reflected three dimensions: competence, benevolence, and health. The second part of the questionnaire consisted of seven questions pertaining to the particular respondent. The subject data sheet required the respondents to state their age, sex, current status in the diploma program, previous education, experiences with the elderly, and preference of work upon completion of the diploma nursing program. A multivariate repeated measures analysis of variance indicated that nursing students demonstrated positive attitudes toward the elderly as compared to younger clients for competence, benevolence, and health. In particular, the student nurses rated the 60- and 80-year-old men more positively than younger clients in both success and failure conditions. The magnitude of the differences for the success and failure ratings was much greater for the 20- and 60-year-old men than the magnitude of the ratings for the 40-year-old men or 80-year-old men. The success and failure interaction did not clearly support the expectation of a subtle age bias with the 60- and 80-year-old men.(ABSTRACT TRUNCATED AT 250 WORDS)

Collapsing health care in Serbia and Montenegro.

Serbia and Montenegro together form the Federal Republic of Yugoslavia. As well as the Serb majority this includes the mixed province of Vojvodina, the mainly Albanian population in Kosovo, and the large Muslim minority in Sandzak. Since the start of war in 1991 the attention and sympathies of the world have focused on Bosnia and Croatia. The United Nations imposed economic sanctions on the federal republic in 1992, although in theory medical supplies and aid are exempt. The economy has now collapsed under the triple burden of war, loss of trade between the republics, and UN sanctions. A major public health catastrophe is unfolding in the federal republic.

Specialist outreach clinics in general practice.

OBJECTIVES: To establish the extent and nature of specialist outreach clinics in primary care and to describe specialists' and general practitioners' views on outreach clinics. DESIGN: Telephone interviews with hospital managers. Postal questionnaire surveys of specialists and general practitioners. SETTING: 50 hospitals in England and Wales. SUBJECTS: 50 hospital managers, all of whom responded. 96 specialists and 88 general practitioners involved in outreach clinics in general practice, of whom 69 (72%) and 46 (52%) respectively completed questionnaires. 122 additional general practitioner fundholders, of whom 72 (59%) completed questionnaires. MAIN OUTCOME MEASURES: Number of specialist outreach clinics; organisation and referral mechanism; waiting times; perceived benefits and problems. RESULTS: 28 of the hospitals had a total of 96 outreach clinics, and 32 fundholders identified a further 61 clinics. These clinics covered psychiatry (43), medical specialties (38), and surgical specialties (76). Patients were seen by the consultant in 96% (107) of clinics and general practitioners attended at only six clinics. 61 outreach clinics had shorter waiting times for first outpatient appointment than hospital clinics. The most commonly reported benefits for patients were ease of access and shorter waiting times. CONCLUSIONS: Specialist outreach clinics cover a wide range of specialties and are popular, especially in fundholding practices. These clinics do not seem to have increased the interaction between general practitioners and specialists.

Utilisation by homeless people of acute hospital services in London.

OBJECTIVES: To estimate the numbers and distribution of homeless people in London; to quantify the utilisation of acute inpatient services by homeless people in two health authorities; and to predict the total numbers of admissions in homeless people in district health authorities across London. DESIGN: Data were collected from various sources on the distribution of homeless people across London boroughs. All unplanned acute inpatient admissions during November 1990 to relevant hospitals were identified. SETTING: Bloomsbury and Paddington and North Kensington, two former inner London district health authorities. SUBJECTS: Homeless people in London residing in bed and breakfast and private sector leased accommodation, residing in hostels, and of no fixed abode. MAIN OUTCOME MEASURES: Number and cost of acute unplanned admissions in homeless people in two health authorities in November 1990; predicted number of such admissions each year in district health authorities in London. RESULTS: There were at least 60,000 homeless people in London in March 1990. The majority were housed in temporary accommodation (55,412). There were at least 3295 hostel dwellers and 651 people sleeping rough. Homeless people accounted for 105 (8%) of the 1256 acute unbooked admissions in residents of Bloomsbury and Paddington and North Kensington health authorities in November 1990. Considerable variations in the pattern of acute unplanned admissions in homeless people were observed in the two districts with respect to housing status and specialty of admission. The total number of acute unplanned admissions in homeless people across London each year was estimated at 7598, ranging from 38 in Bexley to 1515 in Parkside. CONCLUSIONS: The results have fundamental implications for resource allocation across London. Allocation must take better account of the heterogeneity, uneven distribution, and extra health needs of homeless people.

A control question test oriented towards students.

Control Question Tests were altered for 12 of 24 students who were examined with a polygraph about a mock crime which half of them had committed. The altered tests substituted control questions about students' cheating and plagiarism for the standard questions about crime issues. Responses to the altered tests were compared with those from tests using regular control questions which are usually about criminal issues. All tests were conducted by a professor. Detection scores derived from response magnitudes of skin resistance differed between innocent (M = 2.0) and guilty participants (M = -1.9). Guilt and innocence interacted with the type of test. Those examined with control questions oriented towards students scored as more innocent when actually innocent (M = 4.3) than guilty students examined with the student form (M = -3.0) or the crime form (M = -0.8) of the test and innocent students (M = -0.3) examined with control questions oriented towards crimes. The discussion is augmented by results from a direct analysis of magnitude of scores.

The Control Question Test in polygraphic examinations with actual controls for truth.

Subjects who were guilty of a mock crime, innocent and informed of the details of the crime or innocent, and uninformed of the details were examined on the polygraph with a modified version of a Control Question Test, which normally contains questions which are incriminating, ambiguous, and likely to be answered with a lie. We challenged the necessity of using incriminating, ambiguous control questions which may demand a lie as an answer. Instead, we created and tested control questions that were unambiguous and answered truthfully. The results, with the modified questions, showed correct identification of 86% of the guilty subjects and 83 to 89% of the innocent subjects.

Breasts, babies and universities--reflections of two lactating professors.

This is a transcript of a presentation made by Mary E Black and Amanda Sinclair at the "Winds of Change" conference in Sydney on July 12, 1998. This was an international conference looking at issues around women in universities. It is reprinted here as many of the issues are of relevance to the readers of this journal. Both Mary and Amanda are experienced nursing mothers who also have high profile careers and are strongly committed to their families. In a sense they are pioneers as they speak openly about the conflicts both internal and external that they encounter. They were both asked to fly to Sydney to present this paper but decided not to, citing family responsibilities. Instead they persuaded the conference organisers to fund them to produce a video, shot in both Cairns and Melbourne and later edited into a presentation. This was shown to acclaim at the conference and provoked a lively discussion. If organisations are creative, they can do things differently and thus ensure that nursing mothers can fully participate in professional life.