Crystallization during volume reduction of solutions with a composition corresponding to that in the collecting duct: the influence of hydroxyapatite seed crystals and urinary macromolecules

To examine the effect of hydroxyapatite (HAP) seed crystals and urinary macromolecules on the crystallization under conditions similar to those in the collecting duct, we evaporated 100 ml samples of salt solutions with an ion composition assumed to correspond to that in the collecting duct without and with HAP seed crystals. The crystallization in seeded solutions was assessed both with and without dialysed urine (dU). After evaporation the number and volume of crystals were recorded in a Coulter Multisizer and the crystal morphology examined with scanning electron microscopy (SEM) and X-ray crystallography. Addition of HAP crystals was apparently followed by an approximately 15–20% increase in heterogeneous nucleation of calcium oxalate (CaOx). In these experiments SEM and X-ray crystallography showed a high percentage of CaOx in the precipitate. In samples reduced to 40–69 ml, addition of dU to the collecting duct solution containing HAP seed resulted in a greater mean (SD) number of crystals; 3895 (1841) in samples with dU and 1785 (583) in samples without. This was mainly explained by an increased mean (SD) number of small crystals. The mean crystal volume was 17.8 (1.1) and 34.3 (9.1) in samples reduced to 40–69 ml with and without dU, respectively. This might reflect the inhibitory effect of dU on the growth and/or aggregation of the CaOx-CaP precipitate or a promoted nucleation resulting in a large number of small crystals. It is concluded that calcium phosphate formed above the collecting duct might induce heterogeneous nucleation of CaOx at lower levels of the renal collecting system, and that urinary macromolecules are powerful modifiers of these processes. Received: 8 July 1998 / Accepted: 12 March 1999     

Interpreting the effectiveness of a national antibiotic policy and comparing antimicrobial use between countries

Rubella infection in early pregnancy is associated with severe consequences in the developing fetus. In Hong Kong, 8-11% of women of child-bearing age are still susceptible to rubella infection. Therefore, rubella immune status of healthcare workers who may have contact with pregnant women is of particular concern. Rubella immunity of healthcare workers in a Department of Obstetrics and Gynaecology was analysed. In the one and a half years of study, 134 healthcare workers voluntarily submitted blood samples for immunity determination and 16.4% of them were susceptible to rubella infection. A substantial proportion of healthcare workers of childbearing age (14%) was negative for rubella antibody. Susceptible healthcare workers have a risk of acquiring and subsequently transmitting the potentially teratogenic rubella infection to their patients. There is a need to review the rubella immunization policy for healthcare workers in Hong Kong.     

Maternal mortality in Guinea-Bissau: the use of verbal autopsy in a multi-ethnic population

BACKGROUND: In developing countries with scanty resources it is very important to have reliable data to establish priorities for the health sector; e.g. to reduce maternal mortality it is necessary to determine the most important causes. The majority of deaths, however, occur without previous contact with the health system and consequently conventional analyses of death certificates are not feasible. Instead, studies have been carried out in some developing countries with various forms of post-mortem interviews, the so-called verbal autopsies (VA). METHODS: We developed a structured interview with filter questions, which was applied to all deaths of women of fertile age in a cohort of 10,000 women living in 100 clusters in Guinea-Bissau and followed over a period of 6 years. The cause of death was ascertained by means of a series of diagnostic algorithms for the most common causes of maternal mortality, including postpartum haemorrhage, antepartum haemorrhage, puerperal infection, obstructed labour, eclampsia, abortion, and ectopic pregnancy. RESULTS: Of the 350 deaths of women of fertile age, 32% were maternal and it seems unlikely that a significant proportion of maternal deaths have not been classified correctly. Using the diagnostic algorithm 70% could be given a specific diagnosis, the most important causes being postpartum haemorrhage (42% [29/69]), obstructed labour (19% [13/69]), and puerperal infection (16% [11/69]). We attempted to identify the factors that are critical for obtaining sufficient information to reach a diagnosis. In the univariate analyses, it was important whether the respondent had been present during the last illness (P = 0.04) and whether the death occurred more than one week after delivery (P = 0.04). The husband was a better respondent than a co-wife (P = 0.08), and men in general provided more specific information than women (P = 0.08). Furthermore, information appeared to be better if the woman had died in the rainy season (P = 0.08). The length of the recall period, parity, age of woman, place of death, rural/urban residence, and ethnic group were not decisive. In the multivariate analysis sex and presence of respondent and time after delivery were significantly associated with the risk of not reaching a specific diagnosis. Women are less likely to provide adequate information for a diagnosis than men (odds ratio [OR] 3.1; 95% confidence interval [CI]: 1.2-8.1). Respondents that did not reside in the village during the departed woman's illness/delivery carried equal risk of not reaching a conclusion (OR 3.1; CI: 1.1-9.1). Deaths occurring more than one week after delivery were also less likely to be classified (OR 6.1; CI: 1.7-22.0). CONCLUSION: The VA described in the present paper left 30% of the maternal deaths unclassified without a specific diagnosis. Had all interviews been with husbands, only 14% would have remained unclassified. If we had only asked people who were present during the terminal phase of the victim's illness the proportion of classified deaths would have risen from 70% to 75%. It is likely that delayed maternal deaths have not been adequately covered by the present algorithms, but they may also simply be more difficult to describe due to the duration of the disease episode. In contrast to methods by which cause of death is established by a panel of medical experts, the present VA should be economically and technically viable in areas where health workers have only minimal training.     

Detection of polycyclic aromatic hydrocarbon metabolites by high-pressure liquid chromatography after purification on immunoaffinity columns in urine from occupationally exposed workers

Objective: The objective in our study was to quantitate benzo[a]pyrene (B[a]P) metabolites by a combination of immunoaffinity chromatography and high-pressure liquid chromatography (HPLC) with fluorescence detection in urine from workers exposed to high levels of polycyclic aromatic hydrocarbons (PAH). Furthermore, by the simultaneous quantitation of 1-hydroxypyrene, the correlation between the B[a]P-tetrol and 1-hydroxypyrene would provide a means of evaluating the validity of 1-hydroxypyrene as a surrogate biomarker for occupational exposure to the potent carcinogen B[a]P in an electrode paste plant. Methods: The study was carried out at an electrode paste plant that produces electrode paste for Söderberg electrodes. A total of 34 pre- and post-shift urine samples and 17 personal air samples were collected from 17 workers during a normal work week. The concentration of 1-hydroxypyrene was measured in all urine samples. A recent method of quantitating B[a]P-r-7, t-8, t-9, c-10-tetrol in urine of humans exposed to low levels of PAH has been described. A modified version of this method involving purification of urine samples on immunoaffinity columns and HPLC analysis with fluorescence detection was used on urine samples from workers exposed to high levels of PAH. A monoclonal antibody (8E11) with binding affinity to B[a]P-tetrols was used. This antibody also binds several PAH-DNA adducts and metabolites, including 1-hydroxypyrene. Gas chromatography/mass spectroscopy (GC/MS) was also used for identification of metabolites isolated by HPLC fractionation. Results: From personal air sampling the mean exposure to particulate PAHs was 38?μg/m³. The mean concentration of urinary 1-hydroxypyrene was 3.9?μmol/mol creatinine in preshift samples and 10.2?μmol/mol creatinine in postshift samples. We could not identify detectable amounts of urinary B[a]P-tetrol by HPLC or fluorescence spectroscopy after purification on immunoaffinity columns. However, in the HPLC analysis we identified several hydroxyphenantrene metabolites that were detected at relatively high concentrations in all of the workers' urine samples. We could not separate 2- and 3-hydroxyphenanthrene (2?+?3-OH-Phe) in peak 1, and peak 2 contained both 1- and 9-hydroxyphenanthrene (1?+?9-OH-Phe). The phenanthrene metabolites were mainly conjugated to glucuronic acid and sulfate. There was a significant correlation between the 1-hydroxypyrene concentration and 2?+?3-OH-Phe (r?=?0.73) and 1?+?9-OH-Phe (r?=?0.64) in the urine samples. 1-Hydroxypyrene was measured in all post-shift urine samples but was not significantly correlated with workplace pyrene exposure, indicating that skin exposure is an important route of pyrene exposure in this factory. As with 1-hydroxypyrene, dermal PAH uptake may also account for the poor correlation between 2?+?3- and 1?+?9-OH-Phe and ambient phenanthrene. Discussion: Since dermal uptake is likely to be important in occupational PAH exposure in addition to inhalation, estimation of total PAH exposure is best achieved by quantitation of PAHs excreted into body fluids. However, it remains unclear whether there might be a difference in uptake and urinary excretion of 3-ring, 4-ring, or 5-ring PAHs and in the correlation between these metabolites and ambient-air PAH measurements. In summary, using immunaffinity chromatography, we did not find detectable amounts of B[a]P-tetrol in urine from workers occupationally exposed to PAH. However, by an HPLC/immunoaffinity method, relatively high amounts of 1-hydroxypyrene as well as 2?+?3- and 1?+?9-OH-Phe were quantitated in the urine samples, both of which are relevant as biomarkers of PAH exposure.     

Reactions of healthy persons and persons suffering from allergic rhinitis when exposed to office dust

OBJECTIVES: Reactions to airborne office dust among healthy subjects and subjects suffering from allergic rhinitis were investigated. METHODS: Twelve healthy and 11 subjects suffering from allergic rhinitis were exposed to clean air [17 (SD 2) microg/m3] and office dust [439 (SD 68) microg/m3] for 245 minutes. The effect measurements included subjective sensations (questionnaire and potentiometer ratings), mood scale, peak flow, bronchial provocation with histamine using forced expiratory volume in 1 second as the effect measure, nasal mucosal swelling, tear film stability, epithelial damage, foam formation in the eye canthus, threshold for eye irritation with carbon dioxide, eye redness, cellular content of conjunctival fluid, and an addition test for distraction. As many investigations were made and as many statistical analyses (including subgroup analyses) were carried out, the risk of mass significance appeared. This problem was dealt with using the Bonferroni correction for multiple significance tests. RESULTS: The mean ratings of the potentiometer were higher (the subjects showed more irritation) during the dust exposure. The objective investigations showed only indications of effects of dust exposure, and some of the indications were in biologically unexplainable directions. No difference in the reactions to dust was observed between the healthy subjects and the subjects suffering from allergic rhinitis. CONCLUSIONS: Dust does not seem to have objective or subjective effects on humans, as only indications of dust effects were found. Subjects suffering from allergic rhinitis do not appear to be a risk group in relation to dust exposure.     

Outbreak of meningococcal disease in western Norway due to a new serogroup C variant of the ET-5 clone: effect of vaccination and selective carriage eradication

A new sulphonamide resistant (SR) C: 15:P1.7,16 meningococcal strain, a variant of the ET-5 clone, dominated in an outbreak of 22 cases in western Norway commencing in 1995. The first eight patients were 15-21 years old from the Nordhordland area, initiating a carrier study in the local high schools. Carriage of SR serogroup C meningococci was detected by routine methods and treated with a single dose of ofloxacin 400 mg. Of 20 treated carriers, 14 harboured the outbreak strain C: 15:P1.7,16. Vaccination of 4000 children, adolescents and close contacts of patients was also performed. After the intervention, 14 additional cases of meningococcal disease occurred, 8 due to the outbreak strain. However, incidence rates dropped from 180 to 30 per 100000 per year in the student population, but increased from 0 to 13 in the rest of the population in Nordhordland. Carriage eradication is not generally recommended in Norway. However, tracing and treating meningococcal carriage may have reduced transmission and disease in this outbreak situation.     

Decreased glutamate metabolism in cultured astrocytes in the presence of thiopental.

The effect of thiopental on glutamate metabolism was studied by 13C magnetic resonance spectroscopy. Cerebral cortical astrocytes were incubated with 0.5 mM [U-13C]glutamate for 2 hr in the presence of 0.5 or 1 mM thiopental. Labeled glutamate, glutamine, aspartate, and glutathione were observed in cell extracts, and glutamine, aspartate, and lactate in the medium. Not only present in the medium was uniformly labeled glutamate, but also glutamate derived from the tricarboxylic acid (TCA) cycle, and thus glutamate release could be detected. The amounts of [U-13C]glutamate and unlabeled glucose taken up by astrocytes were unchanged in the presence of 0.5 mM thiopental and decreased to about 50% and 80%, respectively when the concentration was increased to 1 mM. The amounts of most metabolites synthesized from [U-13C]glutamate were unchanged in the presence of 0.5 mM thiopental, but decreased [U-13C]glutamine, [U-13C]aspartate, and [U-13C]lactate were observed in the 1 mM group. Surprisingly, the amounts of [1,2,3-13C]glutamate, [2,3-13C]aspartate, and [3,4-13C]aspartate (2nd turn via the TCA cycle) were unchanged. However, this was not the case for [1,2-13C]lactate and [2,3-13C]lactate. Such variations indicate cellular compartmentation, possibly caused by a heterogeneous glutamate concentration within the cells affecting TCA cycle turnover rates differently.     

Improving the nicotinic pharmacophore with a series of (Isoxazole)methylene-1-azacyclic compounds: synthesis, structure-activity relationship, and molecular modeling

A series of (isoxazole)methylene-1-azacyclic compounds was prepared. The compounds were tested for affinity to central nicotinic acetylcholine receptors (nAChRs) and central muscarinic receptors. The compounds covered a broad range of affinities for the nAChRs (IC(50) = 0.32 to >1000 nM), with selectivities for the nAChRs over the muscarinic receptors in the range of 3-183. The high-affinity compound (Z)-26 (3-(4-methyl-5-isoxazolyl)methylene-1-azabicyclo[2.2. 2]octane, IC(50) = 3.2 nM) having only one energy minimum was used as the reference structure in a computational study. This ligand has enabled definition of an important distance parameter, and the existence of this parameter was supported by showing that other potent nicotinic ligands (for example, nicotine and epibatidine) fit the model.     

CYP2D6 polymorphism in systemic lupus erythematosus patients

Objectives: To determine whether patients with idiopathic systemic lupus erythematosus (SLE) are associated with impaired CYP2D6 activity and to gain insight into whether there is an association between particular CYP2D6 genotypes and susceptibility to SLE, and whether CYP2D6 polymorphism is linked to any specific clinical features of SLE. Methods: Debrisoquine sulfate (10 mg p.o.) was given to 159 healthy volunteers and 39 idiopathic SLE patients. Genotypic assay was carried out in 80 healthy volunteers and 32 patients. A 10-ml blood sample was drawn for genotypic assay. Debrisoquine and 4-hydroxydebrisoquine were determined in 8-h urine samples. Blood samples were analysed for the presence of mutations in the CYP2D6 gene, by using polymerase chain reaction (PCR) specific for CYP2D6*3 and CYP2D6*4 alleles. Results: The metabolic ratio of debrisoquine to 4-hydroxydebrisoquine ranged from 0.01 to 86.98 in healthy subjects and from 0.02 to 96 in SLE patients. We observed the poor metabolizer(PM) debrisoquine phenotype in three of 39 patients with idiopathic SLE (7.6%) and five of 159 healthy subjects (3.1%). There was no significant difference in the frequency of PM phenotypes between idiopathic SLE and healthy subjects (Fisher's exact test, P = 0.19). No significant difference in the distribution of overall genotypes and allele frequencies were observed between the two groups. No significant relationships were found between specific clinical features and the overall genotype. Conclusion: The results of this study confirm that CYP2D6 activity is not impaired in SLE and that there is no association between SLE and phenotypic CYP2D6 status. The results also showed that there was no difference in the frequency of CYP2D6A and CYP2D6B alleles between controls and patients with SLE. Received: 14 May 1998 / Accepted in revised form: 19 October 1998