Modelling Germline Mosaicism and Different New Mutation Rates Simultaneously for Appropriate Risk Calculations in Families with Duchenne Muscular Dystrophy

For several genetic diseases two biological phenomena have been recognised as important: germline mosaicism; and different new mutation rates in males and females depending on mutation type. Both principles have been investigated separately and their influence on risk estimation in families has been exemplified in the literature. The aim of this paper is to present a general model that includes mosaicism and different new mutation rates. Mosaicism is introduced by defining additional alleles at the disease locus in combination with adapted segregation rules. Taking Duchenne muscular dystrophy as an example, we derive the conditions which have to be fulfilled for a population in mutation selection equilibrium. Our approach describes the model at the population level and not in individual subjects. This has the advantage of being able to use well known algorithms for the calculation of likelihoods in pedigrees, and to include additional diagnostic information such as marker genotypes and carrier deletion test results. We demonstrate the impact of the new model on a typical pedigree. In families where the patient is not available, the distinction between point mutations and deletions is important, since often molecular diagnostic tests for females can only screen for deletions. Negative deletion test results can now be included in the risk calculations.     

Health care use among young children in day care. Results in a randomized trial of early intervention

Exposure of young children to group day-care settings increases the risk of illness and may result in higher use of medical care. These observations raise concerns that the use of such settings for early intervention programs for low-birth-weight infants may increase the already high burden of medical care costs incurred by these children and their families. To address the question of medical care use associated with center-based care, we examined the hospital-based and ambulatory care reported for participants of the Infant Health and Development Program. This project is a multisite randomized trial of an early intervention program for preterm low-birth-weight infants with an intervention including 2 years of center-based care. The Intervention group did not differ in hospital-based care and averaged only two more physicians' visits over the 3-year observation period than the comparison group. We conclude that early intervention programs involving high-quality group care are not accompanied by substantial increases in health care use.     

Preparation of protected peptidyl thioester intermediates for native chemical ligation by Nα‐9‐fluorenylmethoxycarbonyl (Fmoc) chemistry: considerations of side‐chain and backbone anchoring strategies,and compatible protection for N‐terminal cysteine*,†

Abstract: Native chemical ligation has proven to be a powerful method for the synthesis of small proteins and the semisynthesis of larger ones. The essential synthetic intermediates, which are C‐terminal peptide thioesters, cannot survive the repetitive piperidine deprotection steps of N^α‐9‐fluorenylmethoxycarbonyl (Fmoc) chemistry. Therefore, peptide scientists who prefer to not use N^α‐t‐butyloxycarbonyl (Boc) chemistry need to adopt more esoteric strategies and tactics in order to integrate ligation approaches with Fmoc chemistry. In the present work, side‐chain and backbone anchoring strategies have been used to prepare the required suitably (partially) protected and/or activated peptide intermediates spanning the length of bovine pancreatic trypsin inhibitor (BPTI). Three separate strategies for managing the critical N‐terminal cysteine residue have been developed: (i) incorporation of N^α‐9‐fluorenylmethoxycarbonyl‐S‐(N‐methyl‐N‐phenylcarbamoyl)sulfenylcysteine [Fmoc‐Cys(Snm)‐OH], allowing creation of an otherwise fully protected resin‐bound intermediate with N‐terminal free Cys; (ii) incorporation of N^α‐9‐fluorenylmethoxycarbonyl‐S‐triphenylmethylcysteine [Fmoc‐Cys(Trt)‐OH], generating a stable Fmoc‐Cys(H)‐peptide upon acidolytic cleavage; and (iii) incorporation of N^α‐t‐butyloxycarbonyl‐S‐fluorenylmethylcysteine [Boc‐Cys(Fm)‐OH], generating a stable H‐Cys(Fm)‐peptide upon cleavage. In separate stages of these strategies, thioesters are established at the C‐termini by selective deprotection and coupling steps carried out while peptides remain bound to the supports. Pilot native chemical ligations were pursued directly on‐resin, as well as in solution after cleavage/purification.     

Use of a computer-based health risk appraisal by older adults

BACKGROUND. A health risk appraisal (HRA) is a tool for health promotion. Conversational microcomputer-based HRAs may be more cost effective than other HRA formats. The acceptability of conversational HRAs, however, has not been demonstrated for older adults. METHODS. We studied the acceptability of a conversational microcomputer-based HRA in a sample of 247 adults at the Minnesota State Fair and the Senior Options Exposition. All users were offered the appraisal via mouse or keyboard interface. Acceptability was measured in terms of user-reported helpfulness, intent to change, time of use, and willingness to view HRA health recommendations. Data on completion time and willingness to view HRA recommendations were collected for Senior Exposition users only. Regression analyses were used to examine the combined impact of interface (mouse or keyboard), location (State Fair or Senior Exposition), age, and sex on user acceptability. Results. Interface and location had no effect on helpfulness or change ratings. Older users rated the appraisal more helpful (P less than .007). Both older and female users reported more intent to change behavior (P = .016, both). Time to use the appraisal was related to interface, age, and sex. Mouse users (P less than .0001), older users (P less than .0001) and female users (P less than .05) took significantly longer to use the appraisal. Significantly more mouse users declined to see recommendations (P less than .02). CONCLUSIONS. Older users can derive as much or more value from conversational health risk appraisals as younger users; however, a mouse interface may be less effective for this age group.     

Minimally Invasive Coronary Artery Bypass Grafting: A New Method Using an Anterior Mediastinotomy

The benefit of internal mammary artery (IMA) grafting as a long-lasting intervention for coronary artery disease is well recognized. However, largely because they are less invasive, catheter based alternatives are frequently chosen, particularly to treat single or double vessel disease. To retain the advantages of the IMA graft, and to offset the invasiveness of conventional coronary artery bypass grafting, we developed a new minimally invasive method using an anterior mediastinotomy for treating left anterior descending (LAD) or right coronary artery disease, or both. Feasibility studies using 16 pigs and a human cadaver led to approval by the Institutional Review Board for use of this procedure to treat six patients (four men, two women; mean age, 63.8 ± 13.6 [SD] yrs) who granted informed consent. Pedicle dissection of the IMA, using video assisted thoracoscopy if necessary, was made through a 2-to 3-inch horizontal anterior mediastinotomy. The underlying LAD artery was grafted during femoral vessel cardiopulmonary bypass, with cooling to 30°C, induced ventricular fibrillation, and left ventricular venting if required. Transesophageal echocardiography performed after bypass showed that two patients maintained normal wall motion and four had improvement from the original impairment. One patient suffered a recurrence of angina 4 weeks after the procedure; recatheterization showed an acutely angled IMA, subsequently corrected by balloon angioplasty. The results of follow-up dobutamine echocardiographic stress tests were negative in all patients. With this minimally invasive approach, the procedure should provide the benefits of IMA grafting with shorter hospital stay, more rapid recovery, and less overall cost.     

Laser placement in noncontact Nd:YAG cyclophotocoagulation

We treated 40 eyes of 40 patients by using noncontact transscleral Nd:YAG cyclophotocoagulation to determine whether treatment 1.5 or 3.0 mm posterior to the corneoscleral lumbus was preferable. Patients were randomly assigned to one of the treatment groups, and all other variables, including power, number, and distribution of laser applications, were kept constant. Six months postoperatively, those treated 1.5 mm posterior to the corneoscleral limbus had a lower intraocular pressure (P = .0047) than those treated 3.0 mm from the corneoscleral limbus. The 1.5-mm group also required fewer retreatment (P = .017) and had a slightly lower occurrence of visual acuity loss after this procedure. We believe it may be advantageous to locate the laser application approximately 1.5 mm posterior to the corneoscleral limbus, rather than further posteriorly, when performing noncontact transscleral Nd:YAG cyclophotocoagulation.     

MAO-A inhibition in brain after dosing with esuprone, moclobemide and placebo in healthy volunteers: in vivo studies with positron emission tomography

Objective: The aim of the study was to investigate whether or not esuprone binds substantially to MAO-A in the human brain. Methods: In a randomised double-blind placebo-controlled study 16 male healthy volunteers were examined␣with positron emission tomography (PET) with [¹¹C]harmine. Eight of the volunteers were given daily doses of 800 mg esuprone, four were given bi-daily doses of 300 mg moclobemide, and four volunteers were given placebo tablets. PET was performed before initiation of a 7-day treatment period. On day 7, one investigation was made immediately before administration of the drug, representing 23 h after the previous day's treatment for esuprone and 11 h after the last tablets of moclobemide. Further investigations were made 4 h and 8 h after the morning dose on day 7. Results: PET showed a high degree of binding of [¹¹C]harmine, a high-affinity ligand for MAO-A, before the start of treatment, and a marked and similar reduction after treatment with esuprone and moclobemide. A slight tendency for normalisation of enzyme binding was observed at the last time point. In the placebo group no change was observed. Plasma kinetics of esuprone showed a rapid elimination with a half-life of about 4 h. Conclusion: The study demonstrates that esuprone was comparable to moclobemide in its effect on MAO-A inhibition in the brain at the doses given. This is an illustration of the potential of PET to monitor drug effects directly on target biochemical systems in the brain in human volunteers, and the possibility of using these data, rather than pharmacokinetic data, for the determination of dosing intervals. Received: 21 August 1996 / Accepted in revised form: 22 November 1996