A splice acceptor variant in RGS6 associated with intellectual disability, microcephaly, and cataracts disproportionately promotes expression of a subset of RGS6 isoforms.

Intellectual disability (ID) is associated with an increased risk of developing psychiatric disorders, suggesting a common underlying genetic factor. Importantly, altered signaling and/or expression of regulator of G protein signaling 6 (RGS6) is associated with ID and numerous psychiatric disorders. RGS6 is highly conserved and undergoes complex alternative mRNA splicing producing ~36 protein isoforms with high sequence similarity historically necessitating a global approach in functional studies. However, our recent analysis in mice revealed RGS6 is most highly expressed in CNS with RGS6L(+GGL) isoforms predominating. A previously reported genetic variant in intron 17 of RGS6 (c.1369-1G>C), associated with ID, may provide further clues into RGS6L(+GGL) isoform functional delineation. This variant was predicted to alter a highly conserved canonical 3' acceptor site creating an alternative branch point within exon 18 (included in a subset of RGS6L(+GGL) transcripts) and a frameshift forming an early stop codon. We previously identified this alternative splice site and demonstrated its use generates RGS6Lζ(+GGL) isoforms. Here, we show that the c.1369-1G>C variant disrupts the canonical, preferred (>90%) intron 17 splice site and leads to the exclusive use of the alternate exon 18 splice site, inducing disproportionate expression of a subset of isoforms, particularly RGS6Lζ(+GGL). Furthermore, RGS6 global knockout mice do not exhibit ID. Thus, ID caused by the c.1369-1G>C variant likely results from altered RGS6 isoform expression, rather than RGS6 isoform loss. In summary, these studies highlight the importance of proper RGS6 splicing and identify a previously unrecognized role of G protein signaling in ID.

Protein Profiling of RGS6, a Pleiotropic Gene Implicated in Numerous Neuropsychiatric Disorders, Reveals Multi-Isoformic Expression and a Novel Brain-Specific Isoform.

A metanalysis identified regulator of G-protein signaling 6 (RGS6) as one of 23 loci with pleiotropic effects on four or more human psychiatric disorders. This finding is significant as it confirms/extends the findings of numerous other studies implicating RGS6 in CNS function and pathology. RGS6 is a highly conserved member of the RGS protein family whose cellular roles are likely affected by mRNA splicing and alternative domain inclusion/exclusion. Indeed, we previously identified multiple RGS6 splice variants predicted to produce 36 distinct protein isoforms containing either long (RGS6L) or short (RGS6S) N-terminal domains, an incomplete or intact GGL domain, and nine alternative C termini. Unfortunately, sequence similarities between the isoforms have made it difficult to confirm their individual existence and/or to determine their unique functions. Here, we developed three RGS6-specific antibodies that recognize all RGS6 protein isoforms (RGS6-fl), the N-terminus of RGS6L isoforms (RGS6-L), and an 18-amino acid alternate C-terminal sequence (RGS6-18). Using these antibodies, we demonstrate that RGS6L(+GGL) isoforms, predominating in both mouse (both sexes) CNS and peripheral tissues, are most highly expressed in the CNS. We further identify three novel RGS6 protein bands that are larger (61, 65, and 69-kDa) than the ubiquitously expressed 53- to 57-kDa RGS6L(+GGL) proteins. Importantly, we show that the 69-kDa protein is a brain-specific dephospho form of the 65-kDa band, the first identified phosphorylated RGS6 isoform. Together, these data begin to define the functional significance behind the complexity of RGS6 gene processing and further clarifies RGS6's physiological roles by resolving tissue-specific RGS6 protein expression.

Effect of high-fat diet on body composition and hormone responses to glucose tolerance tests.

To determine potential hormonal mediators of the effect of high-fat diets on the development of insulin resistance, blood leptin, growth hormone (GH), glucose, and insulin responses to a 2 g/kg BW oral glucose challenge were evaluated in weanling female Sprague Dawley rats that were randomly assigned to a high-fat (HF, 39% of calories, 20% fat by weight; n = 10) and moderate-fat (MF, 22% of calories, 10% fat by weight; n = 10) diets. Oral glucose challenges were administered following 5, 7, and 9 wk on the feeding trial. Animals were provided diet in excess of their requirements for growth. Body mass analysis was conducted by dual X-ray absorptiometry (DXA) on the 6th, 8th, and 10th weeks of the trial. HF animals gained more weight after 7 wk, had greater body fat than the MF animals, and similar glucose responses to the oral glucose challenges. HF rats secreted more insulin and leptin compared to MF animals. Lean body mass and serum GH and IGF-I concentrations were not different between the groups. Results of this study demonstrate that leptin but not GH or IGF-I is involved in the development of insulin resistance in growing rats as a result of excess energy intake in the form of dietary fat.

Developing leadership in healthcare administration: a competency assessment tool.

Despite the many graduate programs that specialize in health administration, healthcare leaders and practitioners have expressed concern about the quality of preparation of health administration graduates. The purpose of this study was to facilitate one part of an integrated approach to leadership development that spans academic and practitioner settings. The approach was to design a competency assessment tool for early careerists who have two to five years of postgraduate experience and who aspire to fill senior leadership positions in complex, provider-based healthcare organizations. Open-ended interviews with key informants and a comprehensive review of relevant literature were done to identify and categorize a set of competencies relevant to early careerists. Based on data from key informants, specific work experiences and academic courses were mapped to each competency, indicating where and how such competencies might be developed. A simple rating system was then added to assess each competency, which resulted in the completion of the tool. Finally, the tool was piloted in a practitioner setting at the Massachusetts General Hospital Administrative Fellowship Program and in an academic setting at the Yale University Health Management Program. The resulting tool includes 52 competencies categorized into four domains: (1) technical skills (operations, finance; information resources, human resources, and strategic planning/external affairs); (2) industry knowledge (clinical process and healthcare institutions); (3) analytic and conceptual reasoning; and (4) interpersonal and emotional intelligence. Early experience with the tool suggests that it can facilitate career planning among graduate students, early careerists, and their mentors. Further, the tool can help directors of both academic and practitioner programs identify strengths and gaps in their existing curricula or training programs. By offering specific competencies linked to work experiences and graduate courses, the tool is an initial step toward promoting collaborative efforts between academic and practitioner programs.

A survey of pediatric nurses' knowledge about breastfeeding.

The purpose of this study was to obtain baseline data on pediatric nurses' knowledge about breastfeeding, to inform an education program being developed in a large Melbourne pediatric teaching hospital. A pediatric breastfeeding questionnaire was developed. A random sample of 278 nurses in a variety of clinical units was selected. The questionnaire return rate was 54%. Overall, the difference in the mean percentage knowledge scores between the most experienced and the least experienced nurses was small. The result indicate that an ongoing lactation education program and a Breastfeeding Day-Stay Unit would benefit nursing staff, mothers, and babies.

Maternal and fetal insulin-like growth factor system and embryonic survival during pregnancy in rats: interaction between dietary chromium and diabetes.

Chromium (Cr) depletion may exacerbate hyperglycemia and negative outcomes of pregnancy in the streptozotocin (STZ) diabetic pregnant rat model through the regulation of the insulin-like growth factor (IGF) system. To test this hypothesis, 40 female rats, all fed a low Cr diet (i.e., 70 microgram Cr/kg diet ) from 21 d of age, were randomly assigned one of four treatments, applied on Day 1 of pregnancy, in a 2 x 2 factorial design: 1) very low Cr diet (40 microgram Cr/kg diet) + citrate buffer injection, 2) very low Cr diet + STZ injection (30 mg STZ/kg body wt in citrate buffer), 3) adequate Cr diet (2 mg Cr [from added CrK(SO4)2]/kg diet) + citrate buffer injectionand 4) adequate Cr diet + STZ injection. Blood and tissues were collected on Day 20 of pregnancy. Chromium depletion increased (P < 0.05) urinary hydroxyproline excretion, 22-kDa IGF binding protein (IGFBP) concentration and litter size but decreased (P < 0. 05) placental wt, percentage of protein per fetus, and fetal IGF-I and -II concentrations. Chromium had no effect (P > 0.10) on maternal hormones, 32-kDa IGFBP, glucose, or placental and fetal hydroxyproline concentrations. Diabetes decreased (P < 0.05) maternal wt gain, embryonic survival, litter size, mean pup wt and maternal insulin concentrations, increased (P < 0.05) maternal blood glucose, IGF-I concentrations and maternal hydroxyproline excretion but did not affect fetal concentrations of hormones, IGFBP, glucose or hydroxyproline. Interaction between chromium and diabetes tended (P < 0.10) to affect maternal IGF-II concentrations, but had no effect on other maternal or fetal variables. In conclusion, maternal chromium depletion did not exacerbate hyperglycemia or pregnancy outcome in STZ-induced diabetic rats, but may negatively affect fetal protein content by decreasing fetal IGF-II concentrations. Diabetes may negatively affect fetal growth through its effect on maternal glucose, insulin and IGF-I.

Exhalation flow and pressure-controlled reservoir collection of exhaled nitric oxide for remote and delayed analysis.

BACKGROUND: Expiratory flow rate, soft palate closure, and dead space air may influence exhaled levels of nitric oxide (NO). These factors have not been evaluated in the reservoir collection of NO. METHODS: Exhaled NO was collected into a reservoir during a single flow and pressure controlled exhalation. RESULTS: NO collected in a reservoir containing silica gel was stable for 24 hours. Nasally delivered 4.8% argon measured by mass spectrometry did not contaminate exhaled argon levels (0.1 (0.02)%) in five volunteers during exhalation against a resistance (10 (0.5) cmH2O), hence proving an effective soft palate closure. Exhaled NO in the reservoir was 11 (0.2) ppb, 8.6 (0.1) ppb, 7.1 (0.6) ppb, and 6.6 (0.4) ppb in five normal subjects and 48.3 (18) ppb, 20.3 (12) ppb, 16.9 (0.3) ppb and 10.1 (0.4) ppb in 10 asthmatic subjects at four studied expiratory flows (5-6, 7-8, 10-11, and 12-13 l/min, respectively), with NO levels equal to direct measurement (7.3 (0.5) ppb and 17.4 (0.5) ppb for normal and asthmatic subjects respectively, p < 0.05) at the flow rate 10-11 l/min. Elimination of dead space proved necessary to provide NO levels comparable to the direct measurement. Exhaled NO collected into the reservoir without dead space during flow controlled exhalation against mild resistance provided close agreement (mean (SD) difference -0.21 (0.68), coefficient of variation 4.58%) with direct measurement in 74 patients (NO range 1-69 ppb). CONCLUSIONS: Flow and pressure controlled collection of exhaled NO into a reservoir with silica gel provides values identical to the direct measurement and may be used to monitor asthma at home and where analysers are not on site.

Risky business: pre-powdered gloves or powder-free gloves in the operating suite?

Two of the emerging issues for the health-care sector in the 1990s are occupational health and safety, and iatrogenic issues. Both of these issues are implicated in the use of pre-powdered latex gloves. Hospital health-care workers are exposed to latex in many ways: gloves, intravenous sets, ventilator circuits, dental products, resuscitation equipment, anaesthetic equipment. Post-operative complications, delayed wound healing, scar formation, and the potential for misdiagnosis, in the presence of starch powder, have been well documented in the literature with the need for through glove rinsing prior to surgery. Another route for glove powder to enter wounds is through a barrier breach. For an institution to ensure it provides the most durable and effective barrier for healthcare worker protection and patient safety, knowledge is needed regarding the various factors which lead to glove barrier failure. The primary aim of the study was to evaluate the in-use durability of the surgical gloves in current use against powder-free gloves. Descriptive statistics were used to analyse the data, in addition a cost analysis was calculated. The results of this study demonstrated clinically important differences between existing glove products in terms of barrier quality.

A non-invasive system to evaluate diaphragmatic strength in ventilated patients.

In this paper we describe a system that allows an indicator of diaphragmatic strength to be determined non-invasively in mechanically ventilated patients. The system is comprised of an occlusion device that can be incorporated into the ventilator tubing and an occlusion control unit to operate the occlusion. The system electronically coordinates the timing of airway occlusion at the mouthpiece, application of a magnetic phrenic nerve stimulus and rapid removal of the occlusion once the measurement has been made. The system therefore permits measurement of the change in airway pressure produced by a stimulated diaphragm contraction, without disconnection from the ventilator. Other important respiratory measurements such as twitch mouth pressure, simulated cough and P0.1 can also be performed on non-ventilated patients in a repeatable and systematic manner.

Lasers in dermatology.

The use of dermatologic laser therapy is rapidly expanding. Thirty years of experience has produced advances in the technology, techniques, and therapeutic efficacy of dermatologic lasers. The original lasers have been improved and modified, and new types of lasers have expanded the dermatologist's therapeutic repertoire. Extensive research has provided a greater understanding of the skin's clinical and histologic response to laser treatment. This has allowed dermatologists to expand their therapeutic options and techniques and to improve clinical outcome.

Cutaneous fibrous dysplasia: an incomplete form of the McCune-Albright syndrome.

A number of genetic disorders have been described in limited form. We report a patient with precocious puberty, large irregularly shaped café-au-lait spots since birth and a diffuse scalp alopecia showing the cutaneous changes of fibrous dysplasia. Histologically, the hair follicles were replaced by convolutions of fibrous tissue. This is to our knowledge the first patient reported with an apparently localized cutaneous form of this syndrome, a pattern predicted by the recently described somatic mutation of the Gs alpha gene. This is the second patient reported to our knowledge with the cutaneous McCune-Albright syndrome and scalp alopecia, and the first with diffuse scalp alopecia, the latter being the presenting sign. We believe that the differential diagnosis of both localized and diffuse alopecia should include the McCune-Albright syndrome.

Infection control and human immunodeficiency virus: perceptions of risk among nurses and hospital domestic workers.

In December 1993 the first case of patient-to-patient transmission of human immunodeficiency virus (HIV), at a doctor's surgery in New South Wales, was documented. In an environment of heightened anxiety about HIV transmission and the adequacy of infection-control measures taken by health providers, it is important to explore perceptions of occupational risk of exposure to infection among hospital workers, reasons why hospital domestic workers sometimes depart from standard procedure in infection control, and how they regard the patients who have infectious diseases. In this study, at an infectious diseases hospital where there is an acute awareness of such issues, nurses had accurate knowledge about control of infection, including HIV, but had limited trust of that knowledge. They gave rationales for why they sometimes departed from infection-control procedures. They had low levels of fear of homosexuals and of acquired immune deficiency syndrome. The hospital domestic workers had lower levels of accurate knowledge about infection control, including HIV, and less trust of that knowledge and of protection by health provider from occupational exposure to infection. They had low levels of fear of homosexuals and HIV. Both groups sought regular, small-group, interactive education programs on infection control and HIV to allow them to discuss their concerns. Participatory education of workers should include eliciting concerns of participants, and should discuss concerns regarding administrators' and educators' interests in their safety and wellbeing.

Ecchymotic Kaposi's sarcoma.

We report two patients with ecchymotic patches that did not suggest the diagnosis of Kaposi's sarcoma. The principal complaint of one patient was facial and periorbital edema with bilateral periorbital ecchymosis. On his trunk were patches resembling pityriasis rosea-like Kaposi's sarcoma. Both types of lesions proved to be Kaposi's sarcoma on histologic examination. The second patient had scattered ecchymotic lesions, with typical lesions of Kaposi's sarcoma elsewhere on his body. The ecchymotic lesions showed a large amount of extravasated red blood cells and no evidence of amyloid. This variant has not been described previously to our knowledge.

Drug discrimination training with low doses: maintenance of discriminative control.

Procedures are reported that maintain control by the drug cue during and after drug discrimination training with lower doses that yield predominantly vehicle-appropriate choices. Twelve pigeons were trained to discriminate chlordiazepoxide (CDP) from saline using two-key (drug vs. vehicle) drug discrimination procedures. Intermixed within each block of 30 sessions were nine sessions of training with 8.0 mg/kg CDP, nine with one of seven lower training doses (4.0, 2.8, 2.0, 1.4, 1.0, 0.7, or 0.5 mg/kg CDP), and 12 with saline. The lower training dose was decreased across blocks. The three lowest training doses (1.0, 0.7, and 0.5 mg/kg CDP) yielded predominantly saline-appropriate choices but had no effect on discrimination of 8.0 mg/kg CDP or saline. Three doses (2.0, 1.4, and 1.0 mg/kg CDP) were retrained, and each yielded percentages of drug-appropriate choices nearly identical to those obtained during previous training. This drug discrimination procedure maintains control by the drug cue during and after training with vehicle-like doses of the training drug and may allow for repeated assessment of effects of low training doses.

Sclerosing basal cell carcinoma: management with electron beam therapy.

There are two types of sclerosing basal cell carcinoma (BCC), one entirely with this histology and a second with a prominent sclerosing component. Sclerosing BCC and nodular BCC with a sclerosing component often represent a difficult diagnostic and therapeutic challenge.