Reimbursement for drugs -- a register study comparing economic outcome for five healthcare centres in areas with different socioeconomic conditions.

AIMS: Previous studies have indicated the negative effects of socioeconomic deprivation on health status and morbidity. Nevertheless, the economic assignment systems for pharmaceutical benefits in Sweden do not take socioeconomic status (SES) into account. The aim of the study was, therefore, to compare reimbursement for subsidized drugs at primary healthcare centres (HCCs) with differing socioeconomic conditions in relation to real costs. The word reimbursement is used to denote economic compensation to the HCCs from the county council for drug benefit costs. METHODS: The numbers of individuals dispensed drugs, total costs and reimbursement at five HCCs with different socioeconomic conditions were compared. A socioeconomic index was calculated for each HCC on the basis of information from the municipality registries on income (with negative sign), assistance allowance, education, foreign background, and unemployment. Register data on drug benefit costs were retrieved from the National Corporation of Pharmacies (Apoteket AB) and the Swedish Prescribed Drug Register at the National Board of Health and Welfare. Data on listed and unlisted citizens at the Kalmar County Council and on public statistics from registers at the HCC municipalities where the HCCs were situated were retrieved. RESULTS: There was an almost inverse linear relationship between total cost compensation and the socioeconomic index (n = 5; r =-0.99; p = 0.001). The HCCs with the lowest SES received lower cost compensation. CONCLUSIONS: HCCs responsible for citizens with lower SES appeared to be disadvantaged by the prevalent reimbursement system in Sweden, thereby increasing differences in the state of health of the citizens. This, in turn, hampers health preventing programmes and lifestyle interventions. An HCC-specific standardized summary of socioeconomic burden is presented.

Hydrothermal treatment and malting of barley improved zinc absorption but not calcium absorption in humans.

OBJECTIVE: To study whether hydrothermal treatment or malting of barley (cv. Blenheim) improves zinc and calcium absorption in humans. DESIGN: : Two groups of 10 and 12 healthy subjects, respectively, were in a period of 2 months in a fasting state, served two single meals each containing porridge or breakfast cereals prepared from processed or unprocessed (control) barley (60 g). The meals included 200 g of milk, extrinsically labelled with (65)Zn and (47)Ca. Whole-body retention of both minerals was measured. SETTING: The study was carried out at the Department of Radiation Physics, Sahlgrenska University Hospital, Göteborg. SUBJECTS: The subjects were recruited among students at the Göteborg University. None dropped out. INTERVENTIONS: The activities of (65)Zn and (47)Ca were measured by whole-body counting four to five times over a 4-week period after each meal. RESULTS: Zinc absorption from hydrothermally treated barley porridge, containing 28 mg P as inositol tri- to hexaphosphates (InsP(3)-InsP(6)), was significantly higher (P<0.001) than from control porridge containing 111 mg P as InsP(3)-InsP(6), 25.2+/-6.9 vs 11.0+/-2.5% (n=12). Calcium absorption did not differ (P>0.05), 21.1+/-6.8 vs 19.5+/-4.7% (n=12). Zinc absorption from breakfast cereals of malted barley with phytase activity and containing 70 mg P as InsP(3)-InsP(6,) was significantly higher (P<0.05) than from flakes of barley, containing 108 mg P as InsP(3)- InsP(6) and no phytase activity, 22.9+/-5.8 vs 14.8+/-4.6% (n=10). The calcium absorption was 21.3+/-6.5 vs 18.5+/-4.3% (n=10) and did not differ significantly (P>0.05). CONCLUSION: Improvements of zinc absorption in breakfast meals can be achieved by optimised hydrothermal treatment or malting of barley. Calcium absorption was not influenced in the meals in this study. SPONSORSHIP: Supported by Semper AB, Sweden, Oy Lahden Polttimo, Finland, the SL-Foundation, Sweden, Swedish National Board for Industrial and Technical Development (NUTEK), the Nordic Industrial Foundation, Swedish Council for Forestry and Agricultural Research (SJFR, project no 50.0306/97).

Protein phosphorylation/dephosphorylation in the inner membrane of potato tuber mitochondria.

Inside-out inner mitochondrial membranes free of matrix proteins were isolated from purified potato tuber (Solanum tuberosum L.) mitochondria and incubated with ¿gamma-(32)PATP. Proteins were separated by SDS-PAGE and visualized by autoradiography. Phosphorylation of inner membrane proteins, including ATPase subunits, was strongly inhibited by the phosphoprotein phosphatase inhibitor NaF. We propose that an inner membrane phosphoprotein phosphatase is required for activation of the inner membrane protein kinase. When prelabelled inner membranes were incubated in the absence of ¿gamma-(32)PATP, there was no phosphoprotein dephosphorylation unless a soluble matrix fraction was added. This dephosphorylation was inhibited by NaF, but not by okadaic acid. We conclude that the mitochondrial matrix contains a phosphoprotein phosphatase that is responsible for dephosphorylation of inner membrane phosphoproteins.

Determination of the retention of 47Ca by whole-body counting.

Retention of intravenously or orally administered 47Ca in the human body are described by a two-parameter function. It is then sufficient to make only a few whole-body measurements to determine the retention function, avoiding faeces sampling and stool markers. Seven days after intake the non-absorbed calcium was excreted and the model agreed with the measured relative retention. Absorption of calcium could then, in some cases (e.g. comparative studies), be described by relative retention at the 7th day after intake.

Two subunits of the F0F1-ATPase are phosphorylated in the inner mitochondrial membrane.

Inside-out submitochondrial particles from potato tuber mitochondria were incubated with [gamma-32P]ATP. More than 16 phosphorylated polypeptides were detected by autoradiography on an SDS-gel. Two phosphoproteins, migrating at 22 and 28 kDa, were excised from the SDS-gel, electroeluted, and purified further by anion chromatography. The phosphoproteins were N-terminally sequenced. Over the regions sequenced, the 22 and 28 kDa phosphoproteins had 100% sequence identity with potato proteins identified as the delta'-subunit of the F1-ATPase and the b-subunit of the F0-ATPase, respectively. We suggest that phosphorylation of these proteins may control the interaction between F1 and F0 and regulate energy coupling in oxidative phosphorylation.

A study of some important vitamins and antioxidants in a blackcurrant jam with low sugar content and without additives.

Blackcurrant (Ribes nigrum) jam was manufactured with the aim of producing a jam with a low sugar content, and without any additives. Four temperatures were investigated, namely 60 degrees C, 76 degrees C, 92 degrees C and 97 degrees C. Processing time varied between 1-20 min. After processing, the highest content of ascorbic acid was found in the jam processed at 97 degrees C for 1 min, which contained 63.3 +/- 2.6 mg ascorbic acid/100 g jam. At all combinations investigated more than 60% of the original amount of ascorbic acid was retained after manufacturing and packaging. The jam made at 92 degrees C was stored in a shelf-life study for 13 months. The jam was then stored at 8 degrees C, ambient temperature and at 37 degrees C. At ambient temperature the jam was stored both in dark and in daylight, at 8 degrees C and at 37 degrees C the jam was stored in dark. After 13 months of storage, at 8 degrees C, 60% of the amount of ascorbic acid and 29% of the amount of anthocyanins were retained. In the jam stored at higher temperatures less of both was retained. The beta-carotene in the jam was found to be stable throughout the whole shelf-life study. Exposure to light did not have any effect on any of the components studied. The degradation of anthocyanins was best described by a second-order reaction and the activation energy was determined to be 90 kJ/mol. A jam of blackcurrant may be considered as a good source of vitamins and antioxidants after one year, if certain precautions concerning manufacture and storage conditions are taken.

Direct evidence for the presence of two external NAD(P)H dehydrogenases coupled to the electron transport chain in plant mitochondria.

Exogenous NADPH oxidation by purified mitochondria from both potato tuber and Arum maculatum spadix was completely and irreversibly inhibited by sub-micromolar diphenyleneiodonium (DPI), while exogenous NADH oxidation was inhibited to only a small degree. Addition of DPI caused the collapse of the membrane potential generated by NADPH oxidation, while the potential generated by NADH was unaffected. We conclude that there are two distinct enzymes on the outer surface of the inner membrane of plant mitochondria, one specific for NADH, the other relatively specific for NADPH, with both enzymes linked to the electron transport chain.

Purification and characterization of an NADH-hexacyanoferrate(III) reductase from spinach leaf plasma membrane.

Plasma membranes were purified from spinach (Spinacea oleracea L.) leaves by aqueous two-phase partitioning. The NADH-hexacyanoferrate(III) reductase was released from the membrane by Chaps solubilization and purified 360-fold by ion-exchange chromatography followed by affinity chromatography and size-exclusion chromatography on FPLC. A major band of 45 kDa and a minor contaminant of 66 kDa were detected by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). The band at 45 kDa cross-reacted with antibodies raised against an NADH-hexacyanoferrate(III) reductase from potato tuber microsomes. The native size of the enzyme was 160 kDa as determined by size-exclusion chromatography indicating that it is a tetramer. Two-dimensional gel electrophoresis, isoelectric focusing, followed by SDS-PAGE revealed three main bands of identical molecular weight with pI of 5.3-5.6. The enzyme contained about one flavin adenine dinucleotide (FAD) per 45-kDa subunit as determined by fluorescence spectroscopy, was specific for the beta-hydrogen of NADH, preferred NADH over NADPH as electron donor, and preferred hexacyanoferrate(III) as electron acceptor, e.g., it reduced Fe3+-EDTA, cytochrome c, oxygen, and duroquinone at < 10% of the rate with hexacyanoferrate(III). p-Chloromercurobenzoate, mersalyl, and dicumarol inhibited the activity by > 70% whereas FAD, flavin mononucleotide, duroquinone, and ubiquinone0 did not affect the activity.

Comparison of the Stereospecificity and Immunoreactivity of NADH-Ferricyanide Reductases in Plant Membranes.

The substrate stereospecificity of NADH-ferricyanide reductase activities in the inner mitochondrial membrane and peroxisomal membrane of potato (Solanum tuberosum L.) tubers, spinach (Spinacea oleracea L.) leaf plasma membrane, and red beetroot (Beta vulgaris L.) tonoplast were all specific for the [beta]-hydrogen of NADH, whereas the reductases in wheat root (Triticum aestivum L.) endoplasmic reticulum and potato tuber outer mitochondrial membrane were both [alpha]-hydrogen specific. In all isolated membrane fractions one or several polypeptides with an apparent size of 45 to 55 kD cross-reacted with antibodies raised against a microsomal NADH-ferricyanide reductase on western blots.

The outer membrane of plant mitochondria contains a calcium-dependent protein kinase and multiple phosphoproteins.

Highly purified mitochondria from potato (Solanum tuberosum L. cv. Bintje) tubers were subfractionated into a matrix fraction, an inner membrane fraction and an outer membrane fraction with minimal cross-contamination. When the matrix and inner membrane fractions were incubated with [gamma-32P]ATP only one and three prominent phosphoproteins were detected after SDS-PAGE and autoradiography, respectively. In contrast, more than 20 phosphoproteins could be labelled in the outer membrane fraction, the main ones at 12, 18, 26, 43, 58, 60, 65, 74 and 110 kDa. Only one band, at 18 kDa, was detectable when the labelling was done in the presence of EGTA. We conclude that the outer membrane of plant mitochondria contains at least one Ca(2+)-dependent protein kinase and more than 20 endogenous substrates.