Microcephaly, intractable seizures and developmental delay caused by biallelic variants in TBCD: further delineation of a new chaperone-mediated tubulinopathy.

Microtubule dynamics play a crucial role in neuronal development and function, and several neurodevelopmental disorders have been linked to mutations in genes encoding tubulins and functionally related proteins. Most recently, variants in the tubulin cofactor D (TBCD) gene, which encodes one of the five co-chaperones required for assembly and disassembly of α/ß-tubulin heterodimer, were reported to underlie a recessive neurodevelopmental/neurodegenerative disorder. We report on five patients from three unrelated families, who presented with microcephaly, intellectual disability, intractable seizures, optic nerve pallor/atrophy, and cortical atrophy with delayed myelination and thinned corpus callosum on brain imaging. Exome sequencing allowed the identification of biallelic variants in TBCD segregating with the disease in the three families. TBCD protein level was significantly reduced in cultured fibroblasts from one patient, supporting defective TBCD function as the event underlying the disorder. Such reduced expression was associated with accelerated microtubule re-polymerization. Morpholino-mediated TBCD knockdown in zebrafish recapitulated several key pathological features of the human disease, and TBCD overexpression in the same model confirmed previous studies documenting an obligate dependency on proper TBCD levels during development. Our findings confirm the link between inactivating TBCD variants and this newly described chaperone-associated tubulinopathy, and provide insights into the phenotype of this disorder.

An affective disorder in zebrafish with mutation of the glucocorticoid receptor.

Upon binding of cortisol, the glucocorticoid receptor (GR) regulates the transcription of specific target genes, including those that encode the stress hormones corticotropin-releasing hormone (CRH) and adrenocorticotropic hormone. Dysregulation of the stress axis is a hallmark of major depression in human patients. However, it is still unclear how glucocorticoid signaling is linked to affective disorders. We identified an adult-viable zebrafish mutant in which the negative feedback on the stress response is disrupted, due to abolition of all transcriptional activity of GR. As a consequence, cortisol is elevated, but unable to signal through GR. When placed into an unfamiliar aquarium ('novel tank'), mutant fish become immobile ('freeze'), show reduced exploratory behavior and do not habituate to this stressor upon repeated exposure. Addition of the antidepressant fluoxetine to the holding water and social interactions restore normal behavior, followed by a delayed correction of cortisol levels. Fluoxetine does not affect the overall transcription of CRH, the mineralocorticoid receptor (MR), the serotonin transporter (Serta) or GR itself. Fluoxetine, however, suppresses the stress-induced upregulation of MR and Serta in both wild-type fish and mutants. Our studies show a conserved, protective function of glucocorticoid signaling in the regulation of emotional behavior and reveal novel molecular aspects of how chronic stress impacts vertebrate brain physiology and behavior. Importantly, the zebrafish model opens up the possibility of high-throughput drug screens in search of new classes of antidepressants.

Zebrafish arylalkylamine-N-acetyltransferase genes - targets for regulation of the circadian clock.

Daily rhythms of melatonin production are controlled by changes in the activity of arylalkylamine-N-acetyltransferase (AANAT). Zebrafish possess two aanats, aanat1 and aanat2; the former is expressed only in the retina and the latter is expressed in both the retina and the pineal gland. Here, their differential expression and regulation were studied using transcript quantification and transient and stable in vivo and in vitro transfection assays. In the pineal gland, the aanat2 promoter exhibited circadian clock-controlled activity, as indicated by circadian rhythms of Enhanced green fluorescent protein (EGFP) mRNA in AANAT2:EGFP transgenic fish. In vivo transient expression analyses of the aanat2 promoter indicated that E-box and photoreceptor conserved elements (PCE) are required for expression in the pineal gland. In the retina, the expression of both genes was characterized by a robust circadian rhythm of their transcript levels. In constant darkness, the rhythmic expression of retinal aanat2 persisted while the aanat1 rhythm disappeared; indicating that the former is controlled by a circadian clock and the latter is also light driven. In the light-entrainable clock-containing PAC-2 zebrafish cell line, both stably transfected aanat1 and aanat2 promoters exhibited a clock-controlled circadian rhythm, characteristic for an E-box-driven expression. Transient co-transfection experiments in NIH-3T3 cells revealed that the two, E-box- and PCE-containing, promoters are driven by the synergistic action of BMAL/CLOCK and orthehodenticle homeobox 5. This study has revealed a shared mechanism for the regulation of two related genes, yet describes their differential phases and photic responses which may be driven by other gene-specific regulatory mechanisms and tissue-specific transcription factor profiles.

Functional development of the zebrafish pineal gland: light-induced expression of period2 is required for onset of the circadian clock.

In zebrafish, the pineal gland is a photoreceptive organ that contains an intrinsic circadian oscillator and exhibits rhythmic arylalkylamine-N-acetyltransferase (zfaanat2) mRNA expression. In the present study, we investigated the role of light and of a clock gene, zperiod2 (zper2), in the development of this rhythm. Analysis of zfaanat2 mRNA expression in the pineal gland of 3-day-old zebrafish embryos after exposure to different photoperiodic regimes indicated that light is required for proper development of the circadian clock-controlled rhythmic expression of zfaanat2, and that a 1-h light pulse is sufficient to initiate this rhythm. Analysis of zper2 mRNA expression in zebrafish embryos exposed to different photoperiodic regimes indicated that zper2 expression is transiently up-regulated by light but is not regulated by the circadian oscillator. To establish the association between light-induced zper2 expression and light-induced clock-controlled zfaanat2 rhythm, zPer2 knock-down experiments were performed. The zfaanat2 mRNA rhythm, induced by a 1-h light pulse, was abolished in zPer2 knock-down embryos. These experiments indicated that light-induced zper2 expression is crucial for establishment of the clock-controlled zfaanat2 rhythm in the zebrafish pineal gland.

Parkinson's disease symptoms--patients' perceptions.

Self-assessment is significant for the accurate evaluation of patient' needs. This study examined the frequency and severity of symptoms reported by 39 patients with Parkinson's disease and compared them with symptoms suggested by the literature and by specialists as bothering Parkinson's patients. Four categories of symptoms were examined: (a) motor disability or activity loss, (b) mental change, (c) psychosocial difficulties, and (d) nonspecific symptoms. The findings show that there was correspondence between expert judgements and subjects' reports regarding symptoms such as dyskinesia/tremor as well as walking, freezing gait, and changing position. Symptoms such as dressing self, getting in/out of bed, morning stiffness and deficit in cognitive sequencing, which experts described as characteristic of Parkinson's disease patients, bothered subjects less. In general, patients mental and psychosocial symptoms were higher in their frequencies and perceived severity than problems of performing activities of daily living.

The effect of an alphanumeric paging system on patient compliance with medicinal regimens.

There are numerous actual and potential impediments to patient compliance with medicinal regimens. The implications of poor compliance include worsening disease states or symptoms, with resultant increased costs of care. Using a population of hospice patients at high risk for noncompliance, this study evaluated the use of an alphanumeric paging system as a memory prosthetic, finding that compliance rose from a mean of 56 percent to 96 percent when the system was used. We conclude that such a program is of use in this setting, and that these findings warrant study and suggest applications in other populations.

The moderating effect of age on self-care.

The study examined the moderating effect of age on the relations between symptom severity, self-care and others' care, patients' perception of the extent of problem solution, their satisfaction with the solution, and their perception of control over their health. Participating in the study were 121 female and 167 male chronically ill patients. Results indicated that age did play a major role in the relations between self-care and others' care and self-care outcomes. The young population sought maximum relief of symptoms with minimum effort from the self. The elderly, in contrast, could be said fo maximize control because what was important for them was not to depend on others but to be self-sufficient in solving their health problems.

Ethical reasoning concerning the feeding of severely demented patients: an international perspective.

Structured interviews were held with 149 registered nurses in seven countries in America, Asia, Australia and Europe concerning the feeding of severely demented patients who do not accept food. The most common reasons for nurses being willing to change their decision to feed or not to feed were an order from the medical head, a request from the patient's husband and/or the staff meeting. There was a connection between the willingness to feed and the ranking of ethical principles. Nurses who were most prone to feed the patient most often gave a high rank to the ethical principle of sanctity of life, while those who primarily chose not to feed the patient gave a high rank to the ethical principle of autonomy. All nurses stressed the ethical principle of beneficence.

From diploma to degree: follow up of R.N.-B.A. graduates of Tel Aviv University.

The Tel Aviv University post-basic nursing program, established in 1968, was the first step of nursing academization in Israel. Since then, one-third of the nursing schools in Israel have been academized. The objectives of the present study were to document changes in the worklife of the graduates since completion of the program, to learn about the influence of the program, and to identify factors promoting or impeding study in the program. A mailed questionnaire to 709 graduates produced a 46.8% response. At the time of this study 93% were working, almost half of them in hospital. Data also showed upward mobility, with 38% employed at the institutional to national supervisor level. Recommendations include continuation of the post-basic program, expansion of the clinical content, and enhanced interaction between students, graduates and faculty.

Academia--a stimulus for change.

When the Department of Nursing at Tel Aviv University celebrated its 20th anniversary in 1988, interest was generated on this programme's impact on its graduates and on their contribution to nursing in Israel. Based on a previous study, a questionnaire was sent to 709 graduates, of which 46.8% responded, and a telephone survey was made of 20% of nonrespondents. Below, the study's revelations.

An evaluation of the internship program: the Jerusalem experience.

Concurrent evaluation of an internship which was part of a four year BSN programme, at the Haddassah School, of Nursing was conducted. Data for this evaluation included input from students, teachers and clinical preceptors who answered questionnaires regarding goals' accomplishments in this particular programme. The results consists of both statistical and content analysis of the evaluation. The ones which were evaluated are: students' achievements, preceptors' skills and abilities to surprise students, nursing teachers' abilities in accompanying the programme, students' grades were also evaluated in order to compare their performance during the school years and the internship programme. This evaluation allowed reviewing the intership's goals, at the Haddassah School of Nursing. Similar tools are being used at the present as part of a yearly prospective evaluation and are found to be of great value in improving the transition from students' to professional status.

Ethical reasoning associated with the feeding of terminally ill elderly cancer patients. An international perspective.

An international nursing research study examined the ethical decision-making of "good and experienced" registered nurses in eight countries. The subjects were asked about their decision to feed or not to feed a hypothetical terminally ill, mentally alert, elderly cancer patient who refuses to eat. Cultural variations were demonstrated in the decisions as well as differences in ethical justification. The majority of nurses who would not feed appeared to use the principle of autonomy, whereas nurses who would feed the patient used beneficence as justification. Conditions under which nurses would change their decision to either feed or not feed the patient against her will included doctor's orders and lack of peer support for the decision. The majority of nurses clearly experienced a dilemma.

Follow-up of Hadassah Nursing School's first seven graduate classes.

This article presents a study that evaluated the work performance of the first seven graduate classes of Hadassah Nursing School, The Hebrew University of Jerusalem, Israel. Most of the graduates were located and their fields of employment and future professional plans determined. Strengths and weaknesses in the graduates' practical work are discussed. While interviewing the head nurses at the graduates' workplaces, the absorption strategies of accepting the graduates were questioned as were the ways in which the graduates were evaluated.