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PubMed2004

Mechanism and use of the commercially available viability stain, BacLight.

BACKGROUND: BacLight (Molecular Probes, Eugene, OR, USA) is a popular fluorescence-based two-component stain for determining bacterial cell viability. The main purpose of this work was to fully elucidate the mechanism and to determine why it is sometimes reported that cells stain simultaneously live and dead. METHODS: Solutions of DNA were stained with the two components, propidium iodide (PI) and SYTO9, in different combinations, and fluorescence spectra were collected. RESULTS: K(PI) and K(SYTO9) were approximately 3.7 x 10(5)/M and 1.8 x 10(5)/M. SYTO9 emissions were stronger and overlapped those of PI. Fluorescence resonance energy transfer from SYTO9 to PI was observed. It was, even under normal conditions, possible for DNA bound SYTO9 to have a component in the red region equal to that of DNA bound PI. Potentially confusing emissions were also found to occur when PI was not in sufficient excess to saturate nucleic acid (>0.4 M PI to 1 M DNA base pairs). CONCLUSIONS: The mechanism is a combination of displacement of SYTO9 by PI and quenching of SYTO9 emissions by fluorescence resonance energy transfer. Confusing results can occur if the relative intensities of the stains or the concentration of PI relative to nucleic acid are not properly accounted for.

Cell Survival↗
PubMed1995

Polarization of mitochondria in the unfertilized mouse oocyte.

Maturation of an immature oocyte into one capable of being fertilized involves tightly choreographed movements of chromosomes and organelles. The localization of mitochondria during maturation was studied in live mouse oocytes by confocal laser scanning microscopy (CLSM). Mitochondria were labeled with rhodamine 123 or Mitotracker (Molecular Probes, Eugene, OR) both of which are cell permeant and accumulate in mitochondria; acridine orange was used to mark chromatin. Prior to maturation, oocytes appeared to be radially symmetrical with no evident polarity; fully mature oocytes exhibited obvious polarity marked by the position of the metaphase II spindle in the cortex. CLSM revealed several interesting features of mitochondrial distribution: 1) A cortical clump of mitochondria was seen approximately 30-45 degrees to one side of the metaphase II spindle and marked the region of polar body I extrusion. 2) Large foci of mitochondria (7-14 microM) were frequently found around the central region of the mature oocyte, while the central region often exhibited markedly fewer mitochondria. 3) Small mitochondrial foci (3 microM) in the cortex and near the GV characterized several oocytes which failed to mature. 4) Non-spindle-associated mitochondria were not uniformly distributed in the mature oocyte but were concentrated in the hemisphere containing the metaphase II spindle. 5) The distal margins of this mitochondrial hemisphere were sharply demarcated at the cortex. These findings should help us understand organelle localization during mammalian oocyte maturation, and may give insights into possible causes of infertility and into early events of preimplantation development.

1-Methyl-3-isobutylxanthine↗
PubMed1993

How much do we know about spontaneous human mutation rates?

The much larger number of cell divisions between zygote and sperm than between zygote and egg, the increased age of fathers of children with new dominant mutations, and the greater evolution rate of pseudogenes of the Y chromosome than of those on autosomes all point to a much higher mutation rate in human males than in females, as first pointed out by Haldane [Ann Eugen 13:262-271, 1947] in his classical study of X-linked hemophilia. The age of the father is the main factor determining the human spontaneous mutation rate, and probably the total mutation rate. The total mutation rate in Drosophila males of genes causing minor reduction in viability is at least 0.4 per sperm, and may be considerably higher. The great mutation load implied by a rate of approximately 1 per zygote can be greatly ameliorated by quasi-truncation selection. Corresponding data are not available for the human population. The evolution rate of pseudogenes in primates suggests some 10(2) new mutations per zygote. Presumably the overwhelming majority of these are neutral, but even the approximate fraction is not known. Statistical evidence in Drosophila shows that mutations with minor effects cause about the same heterozygous impairment of fitness as those that are lethal when homozygous. The magnitude of heterozygous effect is such that almost all mutant genes are eliminated as heterozygotes before ever becoming homozygous. Although quantitative data in the human species are lacking, anecdotal information supports the conclusion that partial dominance is the rule here as well. This suggests that if the human mutation rate were increased or decreased, the effects would be spread over a period of 50-100 generations.

Age Factors↗
PubMed1989

Alternative genetic models for the inheritance of the phenylthiocarbamide taste deficiency.

Pedigree segregation analysis was used to examine several one- and two-locus models of the inheritance of phenylthiocarbamide (PTC) taste deficiency that extend the traditional one-locus recessive model by the addition of either another allele or another locus, and in some cases predict two types of nontasters. These models allow nontaster by nontaster matings to produce taster offspring, consistent with our data and several previous studies which use the Harris and Kalmus [Annals of Eugenics 15:24-32, 1949] dilution method. The models fit our data set of 1,152 individuals from 120 families significantly better than the one-locus recessive model. The best fit was obtained with a two-locus model in which one locus controls PTC tasting and the other locus controls a more general taste ability. This model is consistent with research on the physiology of PTC tasting and with results from genetic linkage studies. Further study is suggested to evaluate better the accuracy of the proposed model.

Alleles↗
PubMed2026

(Re)imagining the Future of Genetic Counseling: A Reflexive Qualitative Analysis of Sociopolitical Power, Cultural Safety, Systemic Racism, and Comparative Practice in the United Kingdom, Aotearoa New Zealand and, Australia.

Genetic counseling is undergoing a rapid transformation as genomic medicine becomes embedded within mainstream healthcare systems. At the same time, the profession is being challenged to respond to systemic racism, colonial legacies, technological change, and evolving expectations regarding equity and justice. Historically, genetic counseling emerged within twentieth-century medical genetics and was influenced by political, social, scientific, and medical forces that included eugenic ideology, values, and practices. The profession has since evolved substantially toward psychosocial, patient-centered, and non-directive models of care. Contemporary debates regarding "newgenics" or "neugenics" further demonstrate how concerns regarding equity, reproductive ethics, disability, and genomic stratification continue to shape genomic healthcare discourse. This qualitative reflexive practice paper explores how systemic racism, colonial legacy, cultural safety and structural power shape genetic counseling practice in the United Kingdom (UK), Aotearoa New Zealand and Australia, and how these forces continue to reshape the profession's future identity. A reflexive, narrative, and comparative qualitative approach was employed, grounded in the authors' lived professional experiences across UK and Australasian contexts and informed by purposively selected policy, professional and scholarly literature relating to cultural safety, dignity, anti-racism, and Human Rights-Based Decision-Making. Through iterative reflexive dialogue, comparative analysis, and thematic synthesis, four interrelated themes were developed examining sociopolitical context, systemic racism, cultural safety and technologization within contemporary genetic counseling practice. Comparative analysis identified substantial differences in how culturally responsive practice is conceptualized and operationalized across settings. In Aotearoa, cultural safety is strongly shaped by Te Tiriti o Waitangi, bicultural accountability, and Māori sovereignty frameworks. In Australia, culturally safer genomic care has increasingly developed through Indigenous-led initiatives and workforce reform, including the Australian Alliance for Indigenous Genomics (ALIGN). In contrast, UK practice remains largely situated within equality, diversity, and inclusion (EDI) frameworks that may insufficiently address systemic racism and structural power within increasingly diverse populations. Reflexive clinical examples demonstrated how inequities may emerge through undocumented patient values, standardized pathways, assumptions regarding autonomy, and misinterpretation of culturally specific communication styles. Re-imagining the future of genetic counseling requires more than just technological advancement. It requires reflexive engagement with dignity, inequity, and the sociopolitical realities of the populations served. These insights re-imagine a culturally grounded, socially responsive future for genetic counseling in an era shaped by genomic mainstreaming, digital transformation, artificial intelligence and workforce reform and one in which the profession remains ethically anchored, relationally attuned, and committed to justice-oriented practice.

Humans↗
PubMed2007

Schizophrenia as split personality/Jekyll and Hyde: the origins of the informal usage in the English language.

The entry into the English language of the informal usage of schizophrenia as split personality/Jekyll and Hyde is traced and commented upon. The metaphor of split personality is followed from Eugene Bleuler via his translators and the wider psychiatric community into the present day. It, and to a lesser extent the Jekyll-Hyde personality, is found to be as much a product of the psychological professions as a product of lay misinterpretation. The informal definition of schizophrenia as split personality has outlived the scientific theory with which it was initially associated.

History, 19th Century↗
PubMed2003

Quantitative assessment of chondrocyte viability after laser mediated reshaping: a novel application of flow cytometry.

BACKGROUND AND OBJECTIVES: Lasers can be used to reshape cartilage by accelerating mechanical stress relaxation. In this study, fluorescent differential cell viability staining and flow cytometry were used to determine chondrocyte viability following laser heating. STUDY DESIGN/MATERIALS AND METHODS: Porcine septal cartilages were irradiated with an Nd:YAG laser (lambda = 1.32 microm, 25 W/cm(2)) while surface temperature, stress relaxation, and diffuse reflectance were recorded. Each slab received one, two, or three laser exposures (respective exposure times of 6.7, 7.2, 10 seconds). Irradiated samples were then divided into two groups analyzed immediately and at 5 days following laser exposure. Chondrocytes were isolated following serial enzymatic digestion, and stained using SYTO/DEAD Red (Molecular Probes, Eugene, OR). A flow cytometer was then used to detect differential cell fluorescence; size; granularity; and the number of live cells, dead cells, and post-irradiation debris in each treatment population. RESULTS: Nearly 60% of chondrocytes from reshaped cartilage samples isolated shortly after one irradiation, were viable while non-irradiated controls were 100% viable. Specimens irradiated two or three times demonstrated increasing amounts of cellular debris along with a reduction in chondrocyte viability: 31 and 16% after two and three exposures, respectively. In those samples maintained in culture medium and assayed 5 days after irradiation, viability was reduced by 28-88%, with the least amount of deterioration in untreated and singly irradiated samples. CONCLUSIONS: Functional fluorescent dyes combined with flow cytometric analysis successfully determines the effect of laser irradiation on the viability of reshaped cartilage.

Animals↗
PubMed2006

Sexual health care in persons with intellectual disabilities.

In the past, preventive health concerning sexuality of people with intellectual disabilities was addressed through surgical sterilization as part of nationwide eugenic programs in many countries. For more than 30 years now, it has come progressively to light in the scientific literature that, besides major ethical and legal problems, these programs also failed to assess many of the individual's needs in sexual health. The fact that an increasing number of people with intellectual disabilities live in the community rather than in institutions has heightened public awareness that these individuals have sexual expectancies, desires, and needs that must be supported through both education and health services. The emergence of AIDS, including descriptions of cases among people with intellectual disabilities, has further demonstrated that surgical sterilization cannot be considered a global option to achieve preventive sexual health. The aim of this paper is to review scientific studies that have assessed the expectancies and support needs of persons with intellectual disabilities in terms of sexual health. These needs vary widely from one individual to another, according to life milieu, level of disability, and potential comorbidity. From this review, it appears that hygiene management, global gynecological care, and prevention of unplanned pregnancy, sexually transmitted diseases, and abuse have been frequently identified as areas in which the presence of intellectual disability dictates specific support needs. Different approaches that have been evaluated to address these issues will also be discussed.

Acquired Immunodeficiency Syndrome↗
PubMed2002

Issues and concerns of couples presenting for preimplantation genetic diagnosis (PGD).

BACKGROUND: The use of preimplantation genetic diagnosis (PGD) to select genetically 'normal' human embryos and to transfer them to the uterus of a woman has generated considerable controversy. Debate has occurred over the implications of PGD, sex selection, safety of embryonic manipulation and eugenics. This study evaluates a range of social and moral concerns of couples towards PGD and assisted reproductive technologies (ART) prior to treatment to obtain unbiased authentic attitudes independent of the treatment cycle and the outcome. METHODS: A total of 121 subjects were administered a structured questionnaire after each couple's in vitro fertilization (IVF) or genetic counselling session. Group A consisted of 41 subjects presenting for PGD of single gene disorders (PGD-SG) and group B consisted of 48 subjects undertaking PGD for aneuploidy screening (PGD-AS). A control group consisted of 32 subjects that were about to commence their first IVF cycle. RESULTS AND DISCUSSION: All groups found PGD to be a highly acceptable treatment. They expressed little concern about its extension to testing non-disease states such as sex and they were strongly in favour of a shared decision-making model in which couples have considerable autonomy over decisions about the embryo(s) to transfer. Differences between the groups included issues surrounding the transfer of embryos, restrictions to PGD and the destruction of embryos.

Adult↗
PubMed1997

Immunofluorescence assay for the quantitative and qualitative evaluation of intracellular interleukin-8 in microtiter plates.

In order to monitor the effects of drugs on interleukin-8 (IL-8) production by cells, a microtiter plate assay that determines four parameters simultaneously was established (i) levels of secreted IL-8 (supernatant ELISA), (ii) levels of intracellular IL-8 (cell ELISA), (iii) intracellular localization (fluorescence microscopy), and (iv) the amount of cellular protein (colorimetric assay). The quantitative and qualitative determination of intracellular IL-8 was achieved by immunofluorescence using the ELF-detection system (Molecular Probes, Eugene, OR), which is approximately 10 times more sensitive than conventional immunofluorescence detection systems. Thus, a 32x objective magnification (without immersion oil) is sufficient to precisely assess the subcellular localization of IL-8. Experiments were carried out with human umbilical vein endothelial cells. Drugs interfering with transcription or translation and inhibitors of protein kinase C inhibited both production and secretion of IL-8. Brefeldin A (BFA), colchicine, and the HMGCoA-reductase inhibitor fluvastatin disrupted the characteristic Golgi localization of IL-8, but only BFA inhibited its secretion. This assay can therefore be used to distinguish drugs that inhibit both IL-8 synthesis and secretion from those that inhibit IL-8 secretion only. It can easily be adapted to other cellular proteins for which a sensitive detection method is required.

Brefeldin A↗
PubMed1995

A confocal scanning laser microscope for quantitative ratiometric 3D measurements of [Ca2+] and Ca2+ diffusions in living cells stained with Fura-2.

A confocal scanning laser microscope (CSLM) for observation and quantitative ratiometric measurements of the intracellular dynamics of Ca2+ ions in living neurons has been developed. The instrument consists of a UV-enhanced CSLM, an optical arrangement providing simultaneous excitation at two wavelengths, an electronic arrangement for processing the simultaneous fluorescence response, and software for computing the absolute Ca2+ concentrations, ([Ca2+]). The instrument can be used for any excitation ratiometric measurements, provided that the dye substance used is excitable by wavelengths between 334 nm and 750 nm (such as, e.g. Fura-2). The spatial resolution of the CSLM, as well as a temporal resolution of 20 ms per line (maximum sampling rate) for dynamic measurements are provided by the instrument. Using Fura-2 in calibrated Ca2+ buffer solutions, the instrument measures [Ca2+] between 0 and 1.35 mumol.l-1 with an error of less than 1%. The capability of the instrument to measure absolute [Ca2+] was verified by recording fluorescence images of test solutions with well defined [Ca2+] values (Molecular Probes, Eugene, Ore., USA, C-3009 calibration solutions). In order to verify the dynamic capability of the instrument in real biological specimens, fluorescence changes of Fura-2 that were due to an intracellular flux of Ca2+ ions, and to an increase of [Ca2+]i (the intracellular Ca2+ concentration) have been recorded in Fura-2-loaded cultured cells of the line TE 671.

Calcium↗
PubMed1985

The conceptual neutering of gender and the criminalization of sex.

Thirty years ago the term gender was borrowed from philology for use in sexological psychology in a paper on hermaphroditism (Money, 1955). As originally defined, gender role consists of both introspective and the extraspective manifestations of the concept. In general usage, the introspective manifestations soon became separately known as gender identity. The acronym, G-I/R, being singular, restores the unity of the concept. Without this unity, gender role has become a socially transmitted acquisition, divorced from the biology of sex and the brain. Sex and gender have been partitioned between body and mind, respectively. The desexualization of gender is in accord with the Zeitgeist of contemporary sexual politics together with victimology and an expanding criminalization of sex. The funding of sexological research is being diverted to victimology, which is, de facto, a branch of law enforcement. Victimologists--and sexological professionals among them--are vulnerable to a backlash of being themselves criminalized. This happens as a result of false accusations of various types of malpractice, including sexual abuse of clients, especially children. Under Hitler, there was an historical parallel when the destruction of sexology was effected by the application of the theory of social eugenics and racial purity with sexologists had endorsed. They were among the first of Hitler's victims.

Adult↗
PubMed1989

[Plato: forerunner of demographic thinking?].

Since Malthus, several commentators have noted quantitative considerations in Plato's work, regarding him as a forerunner of demographic thinking. But how to explain several demographic contradictions between the Laws and the Republic? This paper brings out the consistency of Plato's thought, which cannot be demonstrated at a purely demographic level. His fascination for mathematics and the influence of the Pythagorean school should 1st be taken into account. However, the City, as an ideal concept and as a concrete social organization is the key to Plato's thought. Confronted with a basic problem of political philosophy, that of [power and justice, Plato proposed to restore the harmony between the City as a political body and the individual citizens. Hence policy measures which we regard unduly as eugenic and demographic, are in fact based on a conception of man quite different from ours.

Demography↗
PubMed1984

[Recent developments in medical law].

The most spectacular aspect is the extremely rapid expansion of medical law. Even if there is a close connection between developments in medicine and in law, the question must be asked as to what extent new discoveries and advances in medicine play a dominant role here, and to what extent the emphasis is on the further development of law. How advances in medicine can give rise to new legal problems was most impressively demonstrated some time ago by the discussion about cerebral death. In view of the progress made in the field of re-animation and intensive care, the current question is whether or not the physician's duties and rights to maintain life should be limited in hopeless cases when patients are incapable of making decisions themselves. This is demonstrated in particular by the discussion about the binding character of "patient testaments" in which healthy subjects declare that they do not want treatment under such circumstances. The decisive factor will continue to be the presumptive will of the patient at the respective time, and this will have to be ascertained considering all circumstances prevailing at that time. New questions with regard to the ethical and legal limitation of the technically feasible also arise from the possibility of culturing embryos from legal abortions or extracorporally fertilized ova to obtain transplants, and from the possibility of implanting extracorporally fertilized ova into the uterus, perhaps that of a "hired childbearing wet-nurse." In addition to ethical and legal problems, questions of parentage would arise here similar to those already of current interest in connection with artificial heterologous insemination. For physicians practicing these methods, questions concerning liability and the limitation of professional secrecy vis-à-vis the semen donor might become the issue of law suits in the near future. Current problems of "unsuccessful sterilization" and nonperformance of an abortion through the physician's fault although abortion was indicated for eugenic reasons are, on the other hand, primarily due to the fact that the law--possibility even for acceptable reasons--establishes legal obligations for the physician which, in the last analysis, aim at preventing human life from coming into being.(ABSTRACT TRUNCATED AT 400 WORDS)

Germany, West↗
PubMed1989

Meeting report: human fetal tissue transplantation research panel.

On September 14 through 16, 1988, a meeting on the use of human fetal tissue in transplantation was held at the National Institutes of Health, Bethesda Maryland, USA. The meeting sponsored by NIH for the Human Fetal Tissue Transplantation Research Panel, a consultant group to the Advisory Committee to the Director. The consultant group was convened to deal with the scientific, judicial and moral questions associated with research involving transplantation of human fetal tissue obtained after induced abortions. The first day of the meeting was devoted to presentations addressing scientific issues. Included among the speakers was Dr. Lars Olson, Professor of Neurobiology, Karolinska Institute, Stockholm, who described the use of transplanted human fetal tissue in the treatment of patients with Parkinson's disease and Dr. Eugene Redmond, Professor of Psychiatry, Yale University School of Medicine, who showed results of work with transplantation of tissue to correct induced Parkinson-like disease in monkeys. Other speakers addressed the present, past or potential use of fetal tissue in the treatment of diabetes, immune disorders, and other diseases, as well as the use of fetal cells in the production of biologicals. At the conclusion of the meeting the panel did not recommend that research be halted on fetal tissue within the context discussed, although the recommendation of the committee is not binding, and an additional assembly of the panel will probably occur before the final recommendation to an NIH advisory committee is made in November. Other meetings on this subject include a meeting on the use of fetal tissue sponsored by the American Association of Tissue Banks, March 6-7, 1989, in Washington D. C. (Crystal City) and a meeting June 10, 1989, the day before the annual meeting of the Tissue Culture Association, USA, in Orlando, Florida, on fetal cells and ownership of cultured cells and products derived from clinical specimens. Following are statements to the Human Fetal Tissue Transplantation Research Panel presented September 14, 1988, by Dr. David Barnes, Associate Professor of Biochemistry and Biophysics in the Environmental Health Sciences Center at Oregon State University, USA, who was asked to address for the panel recent advances in cell culture related to fetal tissue, and Dr. Robert E. Stevenson, Director of the American Type Culture Collection, President of the Tissue Culture Association, USA, and Chairman of the Committee on Cells and Tumors of the American Association of Tissue Banks.

Cells, Cultured↗
PubMed2003

[Jakob Klaesi on his 120th birthday].

Jakob Klaesi was born on the 29th May 1883 in Glarus Canton (Switzerland) and was assistant and later head physician at the Psychiatric University Hospital in Zurich, directed by Eugen Bleuler. Klaesi directed the new Psychiatric Outpatient Department in Basel from 1923 to 1926 and later founded the Schloss Kronau private clinic in the Zurich Canton. In 1933 he became director of the Psychiatric University Hospital in Bern. His attitude toward somatic treatment methods was skeptical, although about 1920 he founded a psychiatric sleep cure with Somnifen. Klaesi was primarily a psychotherapist and interested in the psychodynamics of his patients. With his great empathy, he was able to understand especially well their expressive behaviour. This capacity for empathy and his philosophic orientation enabled him to develop a phenomenological analysis of expression. He died on the 17th August 1980. Differences and parallels to the thinking of Alfred Adler and Karl Jaspers are discussed.

History, 19th Century↗
PubMed2005

[Considerations on the work of the neuropsychiatrist Gabriel Anton (1858-1933)].

Gabriel Anton (1858-1933) attained fundamental clinical neuropathological results. His scientific career and research were highly influenced by Theodor Meynert. However, in contrast to Meynert, Anton sought not only descriptions of pathological changes but also their therapy. The surgical treatment of hydrocephalus was one of his most important achievements. With this, he gave important impetus to the still new science of neurosurgery. Nowadays, Anton is still known in medical nomenclature by Anton's syndrome. The detailed description and explanation of visual anosognosia and asomatoagnosia and furthermore his emphasis on their importance can be regarded as Anton's most notable contribution to science. However, his research on basal ganglia disorders is not as well-known, although Anton's thoughts mark the beginning of further systematic research on these disorders. In addition, he did preliminary work in the development of child and adolescent psychiatry. Today, some of his work in the field of eugenics is controversial.

Blindness, Cortical↗
PubMed2007

[Nephropexy in the course of time. Aspects of an historical surgical technique].

The first successful nephropexy was performed in the year 1881. From this time, surgical therapy of nephroptosis has always been a subject of discussion. A partly uncritical acceptance led to nephropexy being the most performed urological operation at the beginning of the 20th century, with up to 200 different surgical variations. As early as the 15th century, a first description of ren mobilis was made by Alessius de Pedemontanus. The first surgical intervention for the treatment of nephroptosis was performed by Gilmore in 1870. In 1877, the American Dowell from New Orleans tried a fixation of the kidney through a seton, however, this operation failed. Eventually in 1881, Eugen Hahn from Berlin was able to perform the first successful nephropexy, he named this method "nephroraphy". In 1882, the first modification was made by Bassini with sutures through the renal capsule. Finally, the gynaecologist George Edebohl led nephropexy into a great popularity and secured the method through numerous technical innovations. By 1936, approximately 170 different surgical methods existed for fixation of the kidney. An accurate diagnosis is imperative before performing nephropexy. There were times in which this operation was carried out much too often and, therefore, had a bad reputation. However, it is not correct to drop nephropexy altogether as some would prefer. The statement by Professor Voelcker from Halle in the year 1911 that for all those who have a urinary obstruction and those with a beginning dilation, nephropexy is still justified and may - when correctly performed provide many blessings". Nothing needs to be added to this.

Europe↗