Autoradiographic detection of animal cell membrane mutants altered in phosphatidylcholine synthesis.

We have screened approximately 20,000 colonies of Chinese hamster ovary cells immobilized on filter paper [Esko, J.D. & Raetz, C.R.H. (1978) Proc Natl. Acad. Sci. USA 75, 1190-1193] for strains unable to incorporate [methyl-14C]-choline into trichloroacetic acid-precipitable phospholipid at 40 degrees C. Mutant 58, identified in this way, was specifically defective in choline incorporation, and other isolates were also blocked in thymidine and leucine incorporation into DNA and protein, respectively. Further analysis of mutant 58 revealed that the strain grew almost normally at 33 degrees C, the permissive temperature, but divided only once at 40 degrees C, the restrictive temperature. After a 20-hr incubation at 40 degrees C, the phosphatidyl-choline level dropped from 41% to 20% in the mutant whereas other phospholipids, including sphingomyelin, continued to accumulate. Wild-type cells contained approximately 50% phosphatidylcholine at both temperatures. Anion-exchange chromatography of the water-soluble choline metabolites extracted from mutant 58 revealed that phosphorylcholine accumulation increased from 6 nmol/mg of protein at 33 degrees C to 42 nmol/mg of protein at 40 degrees C whereas CDP-choline decreased from 0.42 nmol to less than 0.07 nmol per mg of protein. Phosphorylcholine also increased in wild-type cells shifted from 33 degrees C to 40 degrees C (from 1.8 nmol to 16 nmol per mg of protein), but the level of CDP-choline was not altered (from 0.52 nmol to 0.58 nmol per mg of protein). Enzymatic assays of extracts prepared from mutant and wild-type cells revealed a reduction of CTP: phosphorylcholine cytidylyltransferase (EC 2.7.7.15) activity (CDP-choline synthetase) in the mutant to 1/40th that in the wild type, and mixing experiments excluded the production of antagonists to CDP-choline synthesis in the mutant. Thus, the inability of the mutant to generate normal amounts of phosphatidylcholine in vivo was correlated with an enzymatic lesion in the biosynthesis of CDP-choline in vitro.     

Placebo-controlled double-blind trial of pirprofen and an ergotamine tartrate compound in migraine attacks

Sixty-one patients, 16 with classic and 45 with common migraine, were treated during three subsequent attacks with pirprofen, a new inhibitor of prostaglandin synthesis; an ergotamine tartrate compound; or placebo, in a randomized, double-blind multicentre study. Pain relief after a single dose and reduction of the attack intensity occurred most often with pirprofen in patients who needed more than one dose. Among them, however, the duration of attack was shortest with ergotamine. Working ability was well preserved with pirprofen, especially among patients with common migraine, and this treatment was ranked highest by the patients. However, no statistically significant differences were found between pirprofen and ergotamine. No serious side effects were observed with pirprofen. This study establishes the usefulness of pirprofen in the treatment of acute migraine.     

Endothelial heparan sulfate deficiency impairs L-selectin- and chemokine-mediated neutrophil trafficking during inflammatory responses

Here we have studied the involvement of endothelial heparan sulfate in inflammation by inactivating the enzyme N-acetyl glucosamine N-deacetylase-N-sulfotransferase-1 in endothelial cells and leukocytes, which is required for the addition of sulfate to the heparin sulfate chains. Mutant mice developed normally but showed impaired neutrophil infiltration in various inflammation models. These effects were due to changes in heparan sulfate specifically in endothelial cells. Decreased neutrophil infiltration was partially due to altered rolling velocity correlated with weaker binding of L-selectin to endothelial cells. Chemokine transcytosis across endothelial cells and presentation on the cell surface were also reduced, resulting in decreased neutrophil firm adhesion and migration. Thus, endothelial heparan sulfate has three functions in inflammation: by acting as a ligand for L-selectin during neutrophil rolling; in chemokine transcytosis; and by binding and presenting chemokines at the lumenal surface of the endothelium.     

Impact of childhood obesity treatment on body composition and metabolic profile

Background  

Childhood obesity is associated with adverse changes in cardiometabolic risk factors. A family-oriented group program stressing a health-promoting lifestyle has been more effective than routine counselling in the treatment of obesity in school children. The aim of the present study was to compare the impact of group program and routine councelling on body composition and metabolic profile, and to evaluate the associations of changes in adiposity with levels of cardiometabolic risk factors.     

Resident care needs and work stressors in special care units versus non-specialized long-term care units

Differences in how elderly residents' care needs affect staff's experiences of work stressors between special care units (SCUs) for dementia and psychiatric residents and non-SCUs were investigated. The data were drawn from 390 staff members in 38 long-term care SCUs, and 587 staff in 53 non-SCUs in Finland. Residents' care needs were based on the Resident Assessment Instrument (RAI) system measured by the Minimum Data Set 2.0. Work stressors (time-pressure and role-conflicts) were assessed with a staff survey questionnaire. Multiple-group regression analysis showed that residents' dependency in activities of daily living (ADL) was related to increased work stressors only in SCUs. A high proportion of behavioral problems was related to fewer work stressors for SCU staff, but more for non-SCU staff. Work stressors may be reduced by specializing, so that residents with similar care needs are placed together and care is focused.     

Propofol in the treatment of refractory status epilepticus

Objectives To study prospectively the effects of propofol anesthesia on seizure control, hemodynamics and course of intensive care in patients with refractory status epilepticus.Design and setting Prospective observational study in the general intensive care unit in a tertiary university hospital.Patients Ten patients with refractory status epilepticus.Interventions Patients received propofol anesthesia aiming to burst suppression EEG pattern for 12 h.Measurements and results Dose of propofol, quality of burst suppression EEG, hemodynamics and the course of intensive care were recorded. Clinical and electrophysiological seizures terminated quickly, but maintaining burst suppression EEG pattern required incremental doses of propofol. Despite high doses of propofol, recovery from anesthesia was fast.Conclusions High doses of propofol are needed in the treatment of refractory status epilepticus. The maintenance of continuous-burst suppression is difficult, and vigilant titrating of dosage of propofol is necessary under continuous EEG monitoring.     

Mixed venous oxygen saturation cannot be estimated by central venous oxygen saturation in septic shock

Objective Central venous oxygen saturation (ScvO₂) in initial resuscitation is included in the Surviving Sepsis Campaign guidelines. ScvO₂ monitoring has also been suggested to be comparable to mixed venous oxygen saturation (SvO₂) for clinical purposes. The aim of our study was to assess the correlation and agreement of ScvO₂ and SvO₂ and compare ScvO₂–SvO₂ difference to lactate, oxygen-derived and hemodynamic parameters in early septic shock in ICU after initial resuscitation.Design and setting Prospective clinical study with 16 patients with septic shock at two university hospital ICUs. A dose of norepinephrine over 0.1 μg/kg/min was required for inclusion.Measurements and results Five paired ScvO₂ and SvO₂ samples at 6-h intervals, altogether 72 samples, were collected during 24 h. The mean SvO₂ was below the mean ScvO₂ at all time points. Bias of difference was 4.2% and 95% limits of agreement ranged from –8.1% to 16.5%. The difference correlated significantly to CI and DO₂.Conclusions The difference between paired ScvO₂ and SvO₂ varies highly. Therefore, SvO₂ may not be estimated on the basis of ScvO₂ in treatment of septic shock after resuscitation period in ICU.Electronic supplementary material The electronic reference of this article is . The online full-text version of this article includes electronic supplementary material. This material is available to authorised users and can be accessed by means of the ESM button beneath the abstract or in the structured full-text article. To cite or link to this article you can use the above reference.     

HLA genotyping is useful in the evaluation of the risk for coeliac disease in the 1st-degree relatives of patients with coeliac disease

OBJECTIVE: Coeliac disease (CD) is a common disease with a strong heredity. About 10-20% of 1st-degree relatives of probands develop CD. Relatives should be screened for CD, because if not treated, CD exposes patients to numerous complications. The heterogeneity of symptoms and the lifetime-spanning risk of CD render the timing of CD antibody and/or gastroscopy screenings difficult. As CD susceptibility has been shown to be strongly associated with the HLA alleles DQA1*0501 and DQB1*0201 (together encoding the DQ2 heterodimer) and DRB1*04 (associated with the DQ8 heterodimer), our aim was to investigate whether HLA genotyping might be useful in the identification of 1st-degree relatives of CD patients who do not need further screening for CD. MATERIAL AND METHODS: The study comprised 54 Finnish CD families including 54 CD probands and 382 living 1st-degree relatives. All subjects who were willing to participate were screened for CD (duodenal and skin biopsies; endomysial, reticulin and gliadin antibodies). The DQA1*0501, DQB1*0201 and DRB1*04 allele frequencies of CD patients and the 1st-degree relatives were determined. RESULTS: Altogether 17.6% (5.9% of the parents, 15.7% of the siblings, 25.8% of the offspring) of the investigated 1st-degree relatives (n = 245) did not carry any of the alleles studied. All of the CD patients (n = 136) with the exception of one (0.7%) carried at least one of the alleles investigated. CONCLUSIONS: By using the HLA genotyping a considerable proportion of 1st-degree relatives of CD probands could be excluded from further screening for CD.     

Gene dose effect of the DQB1*0201 allele contributes to severity of coeliac disease

OBJECTIVE: Coeliac disease (CD) susceptibility has been shown to be associated with the HLA alleles DQA1*0501 and DQB1*0201. This HLA-associated risk has been estimated to account for 29-40% of the genetic component of CD. Conflicting data have been published on the gene dose effect of these HLA alleles on the risk and severity of CD. In this study the aim was to investigate the association between the number of HLA risk alleles and the severity of CD. MATERIAL AND METHODS: Fifty-four Finnish CD families, including 144 CD patients mainly diagnosed in adulthood (94.4%), were enrolled in the study. The association between the number of DQA1*0501 and DQB1*0201 alleles and villous atrophy, symptoms and laboratory parameters at the time of diagnosis, and the association with villous atrophy after one year of treatment on a gluten-free diet were studied. RESULTS: The homozygosity for the DQB1*0201 allele was associated with a more severe form of CD assessed by more severe villous atrophy (p=0.011), younger age (p=0.036), more severe diarrhoea (p=0.048) and a lower level of blood haemoglobin at the time of diagnosis (p=0.010). Furthermore, the homozygosity for the DQB1*0201 allele was associated with a slower recovery of villous atrophy after a gluten-free diet (p=0.041). In contrast, the DQA1*0501 allele did not have a significant association with the severity of CD. CONCLUSIONS: Our results demonstrate a gene dose effect of the DQB1*0201 allele on the clinical heterogeneity of CD and on the rate of recovery from villous atrophy in patients on a gluten-free diet.