The Encapsulation of Squid Diisopropylphosphorofluoridate-Hydrolyzing Enzyme within Mouse Erythrocytes

This study describes the entrapment of squid-type diisopropylphosphorofluoridate-hydrolyzingenzyme (DFPase) within mouse red blood cells. These erythrocytesthereby gain the ability to rapidly hydrolyze alkylphosphatecholinesterase (ChE) inhibitors such as diisopropyl fluorophosphate(DFP). DFPase rapidly hydrolyzes DFP to diisopropyl phosphate.Resealed erythrocytes provide a stable carrier system that canpreserve the activity of encapsulated enzymes against otherwiserapid in vivo degradation; thus, ChE inhibitors can be degradedto relatively nontoxic metabolites by these erythrocyte carriers.Squid DFPase was purified from the hepatopancreas of Atlanticsquid and DFPase activity was determined by measuring changesin fluoride ion concentration using a fluoride ion selectiveelectrode. Mouse erythrocytes in suspension with excess squidDFPase were dialyzed against hypotonic buffer to allow the encapsulationof the enzyme to occur. Cells were then resealed by returningthe suspension to isosmotic with saline. Rate of DFP hydrolysisobserved with these cells was much greater than the rate ofnonenzymatic hydrolysis and was directly proportional to theamount of the erythrocyte suspension added to the assay solution.The rate of hydrolysis was first order in substrate. Erythrocytecontrols showed no endogenous DFPase activity. These resultssuggest that enzyme entrapment may be developed as a methodto prevent and antagonize organophosphate poisoning.     

Multicenter Evaluation of Fixed-Wire PTCA Catheters with Modified Distal Tips: Initial and 2-Month Follow-Up Results

Lesions in the distal portion of coronary arteries are technological challenges with currently available percutaneous transluminal coronary angioplasty (PTCA) equipment. A study was performed at multiple centers to determine if new "fixed-wire" devices with a modified distal tip (a 2-cm flexible tip and 1-cm standard tip) would be useful in these situations. Three hundred twenty-three patients were included in the study, 75% male, mean age 63 ± 11 years. Four hundred ninety-three lesions were attempted; the overall success rate was 85%. Forty-six (14.2%) clinical complications occurred including: 5 deaths (1.1%); 2 nonfatal myocardial infarctions (0.6%); and 6 emergency coronary artery bypass grafting procedures (2.6%). A device malfunction was noted in 32 patients (9.9%), and included: 8 balloon ruptures or perforations (2.0%); 6 bent or damaged tips (1.9%); and 4 shaft/manifold leaks (1.2%). Two-month follow-up assessment for restenosis was done in 352 of 371 (95%) patients and at that time 89% were asymptomatic, had a negative functional test, or patent PTCA site by angiography. The new fixed-wire PTCA catheters with 1- and 2-cm flexible tips hold promise for dilating lesions in distal locations with an acceptable rate of complications. Additional investigation is needed to determine their role in the treatment of coronary artery disease with PTCA.     

Structure-activity studies of hydrophobic amino acid replacements at positions 9, 11 and 16 of glucagon

We have designed and synthesized eight compounds 2-9 which incorporate neutral, hydrophobic amino acid residues in positions 9, 11 and 16 of the glucagon molecule: (2) [desHis¹,Va1⁹,11e^11,16] glucagon amide, (3) [desHis¹,Val^9,11,16]glucagon amide, (4) [desHis¹,Va1⁹,Leu^11,16]glucagon amide, (5) [desHis¹,Nle⁹,11e^11,16]glucagon amide, (6) [desHis¹,Nle⁹,Val^11,16]glucagon amide, (7) [desHis¹,Nle⁹,Leu^11,16]glucagon amide, (8) [desHis¹,Val⁹,Leu^11,16,Lys^17,18,Glu²¹]glucagon amide and (9) [desHis¹,Nle⁹,Leu^11,16,Lys^17,18,Glu²¹]glucagon amide. The effect of neutral, hydrophobic residues at positions 9, 11 and 16 led to good binding to the glucagon receptor. Compared to glucagon (IC₅₀= 1.5 nM), analogues 2-9 were found to have IC₅₀ values of 6.0, 6.0, 11.0, 9.0, 2.5, 2.8, 6.5 and 7.0 nM, respectively. When these compounds were tested for their ability to block adenylate cyclase (AC) activity, they were found to be antagonists having no stimulation of adenyl cyclase, with PA₂, values of 6.15, 6.20, 6.30, 7.25, 6.10, 7.30, 6.25 and 7.25, respectively. © Munksgaard 1997.     

Field workers' records and a microcomputer: monitoring child health in Nepal

A district-level project for monitoring uptake of family planning,child health and immunization services was started in Sindhupalchok,Nepal, in 1983. The project entailed setting up skill-basedin-service training for health workers and their supervisors,and organizing collection of data from field workers' routinereports at quarterly intervals after training. A microcomputerand dBase II software were used to analyse the data, producingreports for different management purposes. Data from each fieldworker area were aggregated and then analysed with respect tothe number of occasions on which households were contacted successfully.Despite geographical variations in the availability of immunizationfacilities, the uptake of immunization was rapid in all areascovered. Family planning services were universally availablein the project area. However, the increase in the percentageof couples practising family planning was more gradual. Theseresults support the linkage between visible improvements inchild health services (including home visiting) and uptake offamily planning. Data on morbidity and nutritional status showedno overall improvements among any regularly-visited childrenwithin the first 2 years of the project. Although the trainingprovided in the project was effective, and the information systemwas felt to be appropriate and useful, the quality of supervisionof field workers in the government Mother and Child/Family PlanningDivision was not adequate to ensure follow-up and reportingin areas remote from the district centre. However the informationyielded through the project does reveal the potential benefitsof home visiting services for children's health in Nepal. Theinformation also permits the setting of targets for achievementsby field workers responsible for promoting child health in thehills of Nepal.     

Alcohol misuse prevention for young people: a systematic review reveals methodological concerns and lack of reliable evidence of effectiveness

In a systematic review we assessed the methodological quality of evaluations of alcohol misuse prevention programmes for young people, and recorded evidence of effectiveness. Studies were identified through systematic searches of electronic databases; hand searches of all obtained papers, existing reviews and several journals; and mailshots to key organizations, conferences and individuals. Relevant papers were checked and cross-checked by members of the review team, and only those studies with an experimental or quasi-experimental design and positive attributes on a number of other quality criteria were included in the review. After pre-screening over 500 papers which reported prevention programmes, information was systematically abstracted from 155 papers. Only 33 studies merited inclusion in the review, and most of these had some methodological shortcomings. Twenty-one studies reported some significant short- and medium-term reductions in drinking behaviour. Of two studies which carried out longer-term evaluations, only one reported a significant longer-term effect, with small effect sizes. No factors clearly distinguished partially effective from ineffective or harmful prevention programmes. In conclusion, the lack of reliable evidence means that no one type of prevention programme can be recommended. In particular there is a need to carry out well-designed scientific evaluations of the effectiveness of current or new prevention efforts which target young people's alcohol misuse.     

Determination of the ratios of the aromatic amino acid residues by first- or second-derivative UV spectrometry for a simple characterization of peptides

A method for the simultaneous determination of the ratios of the three aromatic amino-acid residues in peptides was set up in acidic conditions. Binary and ternary mixtures of these amino acids were prepared, and first- and second-derivative spectra then calculated from their 0.1 nm resolution spectra between 240 and 320 nm. Certain spectral bands were chosen to differentiate tyrosine from tryptophan on the first-derivative spectra, and phenylalanine from tyrosine and tryptophan on the second-derivative spectra. Variation of the amplitude of the chosen bands was shown to be a linear function of the ratio of the aromatic amino acids in the mixture. This technique was validated with peptides whose sequence was known. The difference between theoretical and experimentally determined ratios was lower than 10%. Since the results are obtained as ratios, neither the concentration nor the nature of the peptide has to be known. The feasibility of application using a photodiode array detector with high resolution in reversed-phase high-performance liquid chromatography is discussed. © Munksgaard 1995.     

Change Processes in Psychotherapy

Efforts to understand the processes of change in psychotherapy have typically been based on either clinicians' experiences or on empirical studies somewhat removed from the day-to-day work and concerns of psychotherapists. This paper reviews selected examples of insights and issues derived from each of these sources. It is suggested that our understanding of change processes in psychotherapy will be enhanced by the growing convergence between the two approaches.     

A Subset of Patients With Recurrent Spontaneous Abortion Is Deficient in Transforming Growth Factor β-2-Producing “Suppressor Cells” in Uterine Tissue Near the Placental Attachment Site

PROBLEM : To determine if patients with unexplained recurrent miscarriage have a deficiency of decidual immunosuppressor cells that produce transforming growth factor β type 2, as has been found in mice with abortion due to rejection and/or trophoblast failure. METHODS : Decidual biopsy specimens were taken as near to the placental attachment site as possible under ultrasound guidance from first trimester legal termination (control) patients with recurrent miscarriage and non-viable pregnancy, and from patients with sporadic missed abortion. The tissue was tested for TGFβ-2⁺ suppressor cells by in situ hybridization, immunohistochemistry, and analysis of supernatants. RESULTS : TGFβ-2-related suppressor molecules similar but not identical to those identified in pregnant mice were released by decidual lymphoid cells. Fifty percent of 14 recurrent miscarriage patients showed a lack of suppressor cells and 59% were subnormal in comparison to 20 controls and 5 sporadic miscarriage patients, where 80–85% of the patients had detectable suppressor cells. CONCLUSIONS : Suppressor cell deficiency is compatible with a role for rejection and/or trophoblast failure in some patients with recurrent miscarriage. Presence of suppressor cells in most patients with missed abortion (4/5) is compatible with an alternative cause of fetal death, similar to findings reported in genetic fetal death mice.