A single booster dose of diphtheria vaccine is effective for travelers regardless of time interval since previous doses.

Our study showed the immune response before and after a booster against diphtheria given within the 20-year interval recommended in Sweden or after a prolonged interval. Of 40 travellers, 10/13 in recommended interval group were immune before booster and 19/27 with a delayed interval. After booster, 13/13 versus 26/27 were protected. One booster was sufficient to achieve immunity regardless of the interval.

Pleistocene genetic legacy suggests incipient species of Sebastes mentella in the Irminger Sea.

To investigate a possible speciation event within the redfish (Sebastes mentella) complex in the Irminger Sea, we examined genetics, traditional morphology, geometric morphometrics and meristics of individuals sampled throughout the Sea. Tissue samples from 1901 fish were collected in 1995 and 1996 and from 1999 to 2002, and the fish were genotyped at nine microsatellite loci, two of which were developed for this study. Individual-based genetic analyses showed that two different gene pools exist in the Irminger Sea. Although these groups overlap extensively geographically, they segregate according to depth: those above and below 550 m. This signal of genotype distinction with depth was evident in both the earlier and later sampling. Historical imprints in the genetic data indicated that the redfish in the Irminger Sea are likely to represent a case of an incipient speciation event that began in allopatry during the Pleistocene glaciations followed by secondary contact. Although hybridization was observed between groups, an analysis of traditional and geometric morphometrics and of meristic variables suggested that restricted gene flow between the currently parapatric deep- and shallow-mesopelagic incipient species may be maintained by ecological isolation mechanisms.

The genetic spectrum of a population-based sample of familial hemiplegic migraine.

Familial hemiplegic migraine (FHM) is a rare subtype of migraine with aura and transient hemiplegia. FHM mutations are known in three genes, the CACNA1A (FHM1) gene, the ATP1A2 (FHM2) and the SCN1A (FHM3) gene and seem to have an autosomal-dominant mode of inheritance. The aim of this study was to search for FHM mutations in FHM families identified through a screen of the Danish population of 5.2 million people. FHM patients were diagnosed according to the International Classification of Headache Disorders and all FHM patients had a physical and neurological examination by a physician. A total of 147 FHM patients from 44 different families were identified; 43 FHM families participated in this study. Linkage analysis of these families shows clear linkage to the FHM locus (FHM1) on chromosome 19, supportive linkage to the FHM2 locus whereas no linkage was found to the FHM3 locus. Furthermore, we sequenced all exons and promoter regions of the CACNA1A and ATP1A2 genes and screened for the Q1489K mutation in the SCN1A gene. CACNA1A gene mutations were identified in three of the FHM families, two known FHM mutations, R583Q and T666M and one novel C1369Y mutation. Three FHM families were identified with novel mutations in the ATP1A2 gene; a family with a V138A mutation, a family with a R202Q mutation and a family with a R763C mutation. None of the Danish FHM families have the Q1489K mutation in the SCN1A gene. Our study shows that only 14% (6/42) of FHM families in the general Danish population have exonic FHM mutations in the CACNA1A or ATP1A2 gene. The families we identified with FHM mutations in the CACNA1A and ATP1A2 genes were extended, multiple affected families whereas the remaining FHM families were smaller. The existence of many small families in the Danish FHM cohort may reflect less bias in FHM family ascertainment and/or more locus heterogeneity than described previously.

Genome-wide linkage screen of a consanguineous multiple sclerosis kinship.

Multiple sclerosis (MS), like Alzheimer's disease (AD) and Parkinson's disease (PD), is a common neurological disorder thought to be caused by the interaction of several genes with unknown environmental factors. In both AD and PD the identification of disease forms inherited in a classic Mendelian fashion has helped investigators elucidate pathogenetic mechanisms. In this study a whole-genome screen, with an average of 608 successful genotypes per person, was performed on nine members of a consanguineous family: the index case, three of her siblings and her daughter, all of whom have been diagnosed with definite MS; as well as the parents of the index case (first cousins), one of her five healthy siblings and her husband (who is also her first cousin). Nonparametric linkage analysis was performed on genotyping data. Based on the presence of consanguinity, the a priori hypothesis was that the disease is transmitted in an autosomal recessive fashion in the pedigree. Linkage analysis revealed a suggestive logarithm of odds (LOD) score of 2.29 on the long arm of chromosome 9. Four of five affected family members were identically homozygous for a haplotype under this peak, spanning approximately 43 cM, while the fifth affected subject and all unaffected family members were heterozygous for the haplotype.

The effect of temperature on non-specific defence parameters of three strains of juvenile Atlantic halibut (Hippoglossus hippoglossus L.).

The effect of temperature (8, 12, 15 and 18 degrees C) on a variety of non-specific defence and haematological parameters was examined in three geographically distinct reared strains (Canadian, Icelandic, Norwegian) of Atlantic halibut. The results indicate that temperature exerts a considerable influence on some blood parameters (packed cell volume and the percentage population of leucocytes in peripheral blood) and on some humoral parameters (serum lysozyme activity and serum protein levels) of halibut. A high temperature of 18 degrees C caused a decrease in the number of circulating blood cells and an increase in serum lysozyme levels; effects consistent with those reported within the literature for stress. The different strains of halibut exhibited differing responses with respect to differential counts of peripheral blood lymphocytes and thrombocytes, and to serum protein concentrations, serum lysozyme activity, serum iron content, unsaturated iron binding capacity of serum and O2- production by kidney macrophages.

Assessment of alginic acid molecular weight and chemical composition through capillary electrophoresis.

The polydispersity of alginic acids regarding distributions in molecular weight and chemical composition is analyzed by capillary electrophoresis. Low concentrations of linear polyacrylamide in the electrolyte provide the means for separation of alginate fractions into the megadalton range, while high-resolution polysaccharide mapping of structural variants could be achieved at higher LPAA concentrations. The fraction and block length of the structurally important guluronic acid constituent is determined through the selective complexation with calcium ions. The methodology presented here might serve as a general approach for the assessment of alginate composition and size and the relation to technical properties desired in medical and pharmaceutical applications.

Reconstitution of opsonizing activity by infusion of mannan-binding lectin (MBL) to MBL-deficient humans.

Mannan-binding lectin (MBL, previously named mannan-binding protein, MBP) is a serum collectin, which activates complement upon binding to microbial carbohydrates. This results in opsonization of the microorganisms as well as direct complement-mediated killing. Clinical evidence indicates that MBL has an important role in the innate immune defence against various pathogens. Genetically determined MBL deficiency is associated with increased susceptibility to infections. We have infused two MBL-deficient individuals with clinical grade MBL, purified from pooled donor plasma, in doses sufficient to attain normal concentration of MBL in serum. This resulted in normalization of complement-mediated opsonization. An initial rapid decrease in the serum concentration of MBL was followed by a slower decline with an estimated half-life of about 6 days. No adverse effects were observed and anti-MBL antibodies could not be detected following several MBL infusions. One of the two MBL recipient, a two-year-old girl, who had been suffering from repeated infections from the age of 4 months, was given a total of six MBL infusions. She has subsequently remained healthy for more than three years.

High-Resolution Studies of Hyaluronic Acid Mixtures through Capillary Gel Electrophoresis.

Hyaluronic acid is a negatively charged polysaccharide with a high degree of polydispersity that makes the separation of its oligomers extremely difficult. Through the use of columns filled with a highly viscous polyacrylamide matrix, the unit resolution of hyaluronate oligomers could be achieved, up to at least 80 kDa of mass, through capillary electrophoresis. As analytical application examples, the fractions of enzymatically or ultrasonically degraded hyaluronates were monitored through this method. Because of the very high resolving power, peaks additional to the regular oligomers can be observed that are assumed to be conformers of this regular, unbranched biopolymer.

[Immunization against Haemophilus influenzae type b in Iceland. Results after six years use of PRP-D (ProHIBiT(R).].

Haemophilus influenzae type b (Hib) causes meningitis bacteremia and epiglottitis, dangerous infections, which occur mainly in children under five years of age. Incidence of Hib meningitis in that age group in Iceland was 43/100.000 1974-1988. In the fall of 1988 Icelandic health authorities decided to offer infant immunisation against Hib with PRP-D (ProHIBiT(R)) vaccine, product of Connaught Ltd, Canada. Results are presented of this immunisation programme which has been running since spring 1989. The vaccine is administered at the age of three, four, six and 14 months. During the first year of the immunisation programme one dose was offered to children aged 15 months up to end of third year. During the 10 years 1980-1989, 92 children had Hib meningitis, 61 Hib bacteremia or arthritis and 21 acute epiglottitis. During the five years 1990-1994 no child had Hib meningitis or epiglottitis but three had Hib bacteremia. Hib strains were 10-16% of Haemophilus influenzae strains isolated from surface swabs from 0-5 years old children at different periods until spring 1991 but became very scarse after that. Anti-PRP antibodies in blood measured <0.15 u.g/ ml in 20% of children after three doses of vaccine but >1.0j.ig/ml in 95% after four doses. No fully immunized child has had invasive Hib disease, but one had meningitis and two bacteremia after one dose of vaccine and one bacteremia after three doses. In 1993 21 fully immunized three to four years old children received a booster dose of PRP-D. Geometric Mean Titer of anti-PRP was 1.11 jig/ml before and 137.11u.g/ml after the dose. Mean antibodies against diphtheria were 0.37 IU before and 11.69 IU after the dose. It remains uncertain how long anti-PRP will last in vaccinees when Hib strains disappear.

[Enuresis amongst schoolchildren in an Icelandic town.].

Enuresis is defined as uncontrolled voiding during day or night after normal control is expected. It is a common disorder, which can be divided into primary and secondary. We looked at the health files from children born between 1986-1988, written when they started school. There were 62 children which were said to have enuresis matched with 62 controls of same sex and from the same class. Questionnaires were sent to the parents of each child. That way five more children were found. A total of 101 took part in the study, 52 patients and 49 controls. The prevalence of enuresis was 9.8%, boys being almost two thirds. 69% had primary enuresis, 56% only/also at daytime. Over 44% of patients had a parent with history of enuresis and 48% other relatives too. 88% said that the disorder influenced their children's lives. Many parents worried and sought advice. Urinary infection was commoner among patients but not other diseases. Many children were evaluated and therapy attempted in 54%, mostly drugs and/or alarm device. Permanent results were disappointing. Our results are similar to other studies. Most of our patients still had enuresis when the study took place but 15-17% should spontaneously cure each year. A few still might have this problem in adulthood.

Maltooligosaccharides as chiral selectors for the separation of pharmaceuticals by capillary electrophoresis.

Complexation between the linear maltodextrin oligosaccharides and certain enantiomeric compounds of pharmaceutical interest in buffered solutions can lead to an analytically desirable chiral recognition. Different maltodextrins were assessed in their capacity to cause enantiomeric separations under various conditions of capillary electrophoresis. The mechanism of chiral recognition has been probed through electrophoretic mobility and selectivity measurements for different buffer solutions and organic solvent additives. A differential interaction of chiral solutes with the maltodextrin helical entities emerges as the basis of such enantioselectivity. This notion is further supported by 1H- and 13C-NMR experiments. Optimized separations of simendan, ibuprofen, warfarin, and ketoprofen enantiomers are demonstrated together with a chiral determination of ibuprofen in a blood serum sample at the therapeutic level.

Modification of the electrophoretic mobility of neutral and charged polysaccharides.

Suitable buffer additives can either induce or reduce the electrophoretic mobility of certain polysaccharides during capillary electrophoresis. With water-soluble cellulose derivatives, adsorption of detergents on the analyte molecules governs the migration rates. Different types of adsorption sites, including multilayer formation, were found to be present on the cellulose-derived polymers. For highly charged polysaccharides (heparins), the electrophoretic mobility can be controlled by the use of ion-pairing reagents. The concentration and number of cationic charges appear useful in optimizing the separation selectivity. Migration models based on secondary thermodynamic equilibria are presented and evaluated for different polysaccharides.

Resolution of the branched forms of oligosaccharides by high-performance capillary electrophoresis.

The heterogeneous nature of most polysaccharides found in nature includes distribution in molecular weight, primary sequence, and branching. The analytical methodology used in the characterization of these structural aspects must ensure high separation efficiency and selectivity. This paper reports on the high-performance capillary electrophoresis (HPCE) separation of branched forms of oligosaccharides as well as some variants in the primary structure. Oligosaccharide maps were obtained after selective debranching using isoamylase, laminarinase, and cellulase enzymes. The samples investigated were alpha-D-glucans (amylose, amylopectin, and pullulan) and beta-D-glucans (exemplified by lichenan). The solutes were separated as fluorescent derivatives with 8-aminonaphthalene-1,3,6-trisulfonate (ANTS) and detected by laser-induced fluorescence at 514 nm using a He/Cd laser (excitation at 325 nm). The number of theoretical plates was in excess of one million per meter. Baseline resolution of oligosaccharides with a degree of polymerization approximately 70 was obtained within 15 min using borate buffer as the electrolyte.

Separation of complex oligosaccharide mixtures by capillary electrophoresis in the open-tubular format.

The fluorescent derivatives of complex oligosaccharide mixtures from different origins are separated in open capillaries. The number of negatively charged sulfonate groups in the tag molecule strongly affects separation efficiency and selectivity. Coating of the capillary surface with linear polyacrylamide has been essential to ensure fast and stable migration velocities. Efficiencies in excess of 1 million plates/m have been achieved, facilitating resolution of branched oligosaccharides. The effects of field strength and buffer composition on the apparent electrophoretic mobility of dextran and dextrin oligomers are discussed, with relation to borate complexation. Preliminary examples of applications to monitoring the action of hydrolytic and synthesizing enzymes are also described.

Separation of glucuronides from urine by coupled-column separation using underivatized silica as precolumn.

Glucuronides are separated from urine by coupled-column separations (CCSs). The fraction containing the glucuronide(s) is transferred on-line from a silica precolumn to the analytical column (octadecyl derivatized silica), enriched, and separated by ion-pair chromatography. The retention and selectivity on the precolumn are controlled by pH, buffer components, organic modifier, and ion-pair agent. After the injection of filtered urine samples, glucuronides with different chemical properties can be separated. The total analysis of morphine-3-glucuronide and morphine-6-glucuronide is accomplished in less than six minutes, with UV detection at 210 nm.

Direct coupled column separation and determination of the diastereomeric glucuronides of almokalant, a new class III antiarrhythmic drug, in human urine.

A reversed-phase coupled column separation (CCS) system for the analysis of two diastereomeric glucuronides of almokalant, a new class III antiarrhythmic drug, in human urine is described. After direct injection of urine samples (50 microliters) the glucuronides were isolated by complex formation on a terbium(III) loaded strong cation exchanger at alkaline pH. The solutes were eluted from the precolumn by an acidic mobile phase, enriched and separated on Hypercarb (porous graphitic carbon) as analytical column with 0.1 M acetic acid pH 2.8 and 30% acetonitrile as mobile phase. The calibration graph was linear (r2 = 0.9999) and the detection limits were in the low picomole (UV) or femtomole (fluorescence) range. Optimization of the analytical column revealed that elution order and selectivity for the glucuronides were dependent on the buffer agent and temperature used. By appropriate choice of mobile phase conditions all four diastereomers could be separated.

An unusual inhibition of an anti-P1.

Tests for unexpected antibodies using commercially prepared red blood cells demonstrated no agglutination even though crossmatching and testing with saline washed commercially prepared reagent red blood cells identified an anti-P1. This antibody was inhibited when tested with red blood cells suspended in a modified Alsever's solution and several commercially prepared reagent red blood cell diluents. The inhibitory substance was identified as inosine. Hypoxanthine, adenine, and thymine also inhibited the anti-P1. Partial inhibition was produced by uracil, 5-methylcytosine, and xanthine. No other example of anti-P1 has been found to exhibit this unusual inhibiting characteristic.

Co-trimoxazole and nitrofurantoin in urinary-tract infections: a controlled clinical study.

The clinical efficacies of co-trimoxazole and nitrofurantoin were compared in patients with urinary-tract infections associated with significant bacteriuria with coliform bacteria tested and found to be sensitive to both drugs. Of 27 patients treated with co-trimoxazole, 23 were cured after 10 days' treatment with 320 mg trimethoprim and 1600 mg sulphamethoxazole daily (85%). Of 18 patients treated with nitrofurantoin (200 mg/day for 10 days), 7 were cured (39%). Thus, cotrimoxazole was significantly superior (p less than 0.01) in this study. A significant (p less than 0.001) but temporary and modest rise of the serum-creatinine levels was noted after 5 days' treatment with co-trimoxazole. Such a rise was not observed in the nitrofurantoin group. Only a few minor side effects were noted in both groups.