Evaluation of the stability of furosemide in tablet form during six-month storage in spaceflight and peculiarities of its pharmacokinetics and pharmacodynamics under conditions of anti-orthostatic hypokinesia.

OBJECTIVES: The present study investigated the stability of furosemide under space-flight conditions on board the International Space Station, as well as its pharmacokinetics and pharmacodynamics under conditions simulating exposure to some space-flight factors. METHODS: Quantitative analysis of furosemide tablets by HPLC was performed before spaceflight (background), then after six months storage under normal ground conditions (control) and under spaceflight conditions (SF). The pharmacokinetics and pharmacodynamics of furosemide were studied in six healthy volunteers after a single oral dose of 40 mg under normal conditions (background) and under anti-orthostatic hypokinesia (ANOH). RESULTS: Quantitative content of furosemide in tablets before SF was 40.19 ± 0.28 mg (100.47 ± 0.71%), after 6 months storage: under normal conditions (control) - 39.9 ± 0.39 mg (99.73 ± 0.98%), under SF - 39.24 ± 0.72 mg (98.11 ± 1.80%), which was within the prescribed limits. Studying basic hemodynamic parameters showed that in ANOH conditions 6 h after furosemide administration there was a statistically significant increase of the stroke volume (SV) (+36.5 Δ%), a tendency for increasing of the stroke index (SI) (+36.5 Δ%) and decreasing of the total peripheral resistance (TPR) (-21.9 Δ%) compared to baseline study. CONCLUSIONS: It has been established that various factors of space flight (overloading, excessive vibration, microgravity, etc.) do not negatively influence the stability of furosemide in tablet form during storage for 6 months on board the International Space Station.

Quantitative chromogenic immunohistochemical image analysis in cellprofiler software.

Visual grading of chromogenically stained immunohistochemical (IHC) samples is subjective, time consuming, and predisposed to considerable inter- and intra-observer variations. The open-source digital analysis software, CellProfiler has been extensively used for fluorescently stained cells/tissues; however, chromogenic IHC staining is routinely used in both pathological and research diagnostics. The current investigation aimed to compare CellProfiler quantitative chromogenic IHC analyses against the gold standard manual counting. Oral mucosal biopsies from patients with chronic graft-versus-host disease were stained for CD4. Digitized images were manually counted and subjected to image analysis in CellProfiler. Inter-observer and inter-platform agreements were assessed by scatterplots with linear regression and Bland-Altman plots. Validation comparisons between the manual counters demonstrated strong intra-observer concordance (r2 = 0.979), particularly when cell numbers were less than 100. Scatterplots and Bland-Altman plots demonstrated strong agreement between the manual counters and CellProfiler, with the number of positively stained cells robustly correlating (r2 = 0.938). Furthermore, CellProfiler allowed the determination of multiple variables simultaneously, such as area stained and masking to remove any nonstained tissue and white gaps, which also demonstrated reliable agreement (r2 = >0.9). CellProfiler demonstrated versatility with the ability to assess large numbers of images and allowed additional parameters to be quantified. CellProfiler allowed rapid high processing capacity of chromogenically stained chronic inflammatory tissue that was reliable, accurate, and reproducible and highlights potential applications in research diagnostics.

Milk processing quality of suckled/milked goats: effects of milk accumulation interval and milking regime.

Milk with a high concentration of fat and casein is required for cheese production, and these components have a major impact for both quality and yield of the curd. Recent observations have shown that suckling can elevate milk fat concentration in goats and our aim was therefore to check the hypothesis that animal welfare and cheese-processing properties of goat milk could be optimised by appropriate management of suckled/milked goats. Twelve Swedish dairy goats were kept together with one kid each in 4 different mixed management-systems (milking combined with partial suckling) in a cross-over design. Two milk accumulation intervals were tested; Short = dams and kids were together for 16 h (T16) and Long = ; dams and kids were together for 8 h (T8 h). In addition, two milking regimes were used; Suckled Before Milking = S and Milked Before Suckling = M. Milk accumulation interval referred to how long dams and kids were separated. The milk yield available for processing (milk offtake), was weighed and analysed from each milking occasion and the suckled milk yield was estimated by a weigh-suckle-weigh method (WSW) in combination with observing the suckling behaviour during the free suckling periods. Milking managements, such as 'suckling before milking (S)', increased milk fat concentration compared to milking before suckling (M) and 'Short accumulation treatments (T16)' gave higher milk fat, casein concentration and individual curd yield (%) compared to the 'Long accumulation treatment (T8)'. The total individual curd yield (g) was the same despite treatment, but the animal welfare was most likely higher in T16 where dams and kids spent more time together.

Bovine chromosomal regions affecting rheological traits in rennet-induced skim milk gels.

Optimizing cheese yield and quality is of central importance to cheese manufacturing. The yield is associated with the time it takes before the gel has an optimal consistency for further processing, and it is well known that gel formation differs between individual milk samples. By identifying genomic regions affecting traits related to rennet-induced gelation, the aim of this study was to identify potential candidate genes affecting these traits. Hence, rennet-induced gelation, including rennet coagulation time, gel strength, and yield stress, was measured in skim milk samples collected from 379 animals of the Swedish Red breed using low-amplitude oscillation measurements. All animals had genotypes for almost 621,000 segregating single nucleotide polymorphisms (SNP), identified using the Bovine HD SNPChip (Illumina Inc., San Diego, CA). The genome was scanned for associations, haplotypes based on SNP sets comprising highly associated SNP were inferred, and the effects of the 2 most common haplotypes within each region were analyzed using mixed models. Even though the number of animals was relatively small, a total of 21 regions were identified, with 4 regions showing association with more than one trait. A major quantitative trait locus for all traits was identified around the casein cluster explaining between 9.3 to 15.2% of the phenotypic variation of the different traits. In addition, 3 other possible candidate genes were identified; that is, UDP-N-acetyl-α-d-galactosamine:polypeptide N-acetylgalactosaminyl-transferase 1 (GALNT1), playing a role in O-glycosylation of κ-casein, and 2 cathepsins, CTSZ and CTSC, possibly involved in proteolysis of milk proteins. We have shown that other genes than the casein genes themselves may be involved in the regulation of gelation traits. However, additional analysis is needed to confirm these results. To our knowledge, this is the first study identifying quantitative trait loci affecting rennet-induced gelation of skim milk through a high-density genome-wide association study.

Genetic parameters for rennet- and acid-induced coagulation properties in milk from Swedish Red dairy cows.

Milk coagulation is an important processing trait, being the basis for production of both cheese and fermented products. There is interest in including technological properties of these products in the breeding goal for dairy cattle. The aim of the present study was therefore to estimate genetic parameters for milk coagulation properties, including both rennet- and acid-induced coagulation, in Swedish Red dairy cattle using genomic relationships. Morning milk samples and blood samples were collected from 395 Swedish Red cows that were selected to be as genetically unrelated as possible. Using a rheometer, milk samples were analyzed for rennet- and acid-induced coagulation properties, including gel strength (G'), coagulation time, and yield stress (YS). In addition to the technological traits, milk composition was analyzed. A binary trait was created to reflect that milk samples that had not coagulated 40min after rennet addition were considered noncoagulating milk. The cows were genotyped by using the Illumina BovineHD BeadChip (Illumina Inc., San Diego, CA). Almost 600,000 markers remained after quality control and were used to construct a matrix of genomic relationships among the cows. Multivariate models including fixed effects of herd, lactation stage, and parity were fitted using the ASReml software to obtain estimates of heritabilities and genetic and phenotypic correlations. Heritability estimates (h(2)) for G' and YS in rennet and acid gels were found to be high (h(2)=0.38-0.62) and the genetic correlations between rennet-induced and acid-induced coagulation properties were weak but favorable, with the exception of YSrennet with G'acid and YSacid, both of which were strong. The high heritability (h(2)=0.45) for milk coagulating ability expressed as a binary trait suggests that noncoagulation could be eliminated through breeding. Additionally, the results indicated that the current breeding objective could increase the frequency of noncoagulating milk and lead to deterioration of acid-induced coagulation through unfavorable genetic associations with protein content (0.38) and milk yield (-0.61 to -0.71), respectively. The outcome of this study suggests that by including more detailed compositional traits genetically associated with milk coagulation or by including milk coagulation properties directly within the breeding goal, it appears possible to breed cows that produce milk better suited for production of cheese and fermented products.

Effects of breed and casein genetic variants on protein profile in milk from Swedish Red, Danish Holstein, and Danish Jersey cows.

In selecting cows for higher milk yields and milk quality, it is important to understand how these traits are affected by the bovine genome. The major milk proteins exhibit genetic polymorphism and these genetic variants can serve as markers for milk composition, milk production traits, and technological properties of milk. The aim of this study was to investigate the relationships between casein (CN) genetic variants and detailed protein composition in Swedish and Danish dairy milk. Milk and DNA samples were collected from approximately 400 individual cows each of 3 Scandinavian dairy breeds: Swedish Red (SR), Danish Holstein (DH), and Danish Jersey (DJ). The protein profile with relative concentrations of α-lactalbumin, ß-lactoglobulin, and α(S1)-, α(S2)-, κ-, and ß-CN was determined for each milk sample using capillary zone electrophoresis. The genetic variants of the α(S1)- (CSN1S1), ß- (CSN2), and κ-CN (CSN3) genes for each cow were determined using TaqMan SNP genotyping assays (Applied Biosystems, Foster City, CA). Univariate statistical models were used to evaluate the effects of composite genetic variants, α(S1)-ß-κ-CN, on the protein profile. The 3 studied Scandinavian breeds differed from each other regarding CN genotypes, with DH and SR having similar genotype frequencies, whereas the genotype frequencies in DJ differed from the other 2 breeds. The similarities in genotype frequencies of SR and DH and differences compared with DJ were also seen in milk production traits, gross milk composition, and protein profile. Frequencies of the most common composite α(S1)-ß-κ-CN genotype BB/A(2)A(2)/AA were 30% in DH and 15% in SR, and cows that had this genotype gave milk with lower relative concentrations of κ- and ß-CN and higher relative concentrations of αS-CN, than the majority of the other composite genotypes in SR and DH. The effect of composite genotypes on relative concentrations of the milk proteins was not as pronounced in DJ. The present work suggests that a higher frequency of BB/A(1)A(2)/AB, together with a decrease in BB/A(2)A(2)/AA, could have positive effects on DH and SR milk regarding, for example, the processing of cheese.

The occurrence of noncoagulating milk and the association of bovine milk coagulation properties with genetic variants of the caseins in 3 Scandinavian dairy breeds.

Substantial variation in milk coagulation properties has been observed among dairy cows. Consequently, raw milk from individual cows and breeds exhibits distinct coagulation capacities that potentially affect the technological properties and milk processing into cheese. This variation is largely influenced by protein composition, which is in turn affected by underlying genetic polymorphisms in the major milk proteins. In this study, we conducted a large screening on 3 major Scandinavian breeds to resolve the variation in milk coagulation traits and the frequency of milk with impaired coagulation properties (noncoagulation). In total, individual coagulation properties were measured on morning milk collected from 1,299 Danish Holstein (DH), Danish Jersey (DJ), and Swedish Red (SR) cows. The 3 breeds demonstrated notable interbreed differences in coagulation properties, with DJ cows exhibiting superior coagulation compared with the other 2 breeds. In addition, milk samples from 2% of DH and 16% of SR cows were classified as noncoagulating. Furthermore, the cows were genotyped for major genetic variants in the αS1- (CSN1S1), ß- (CSN2), and κ-casein (CSN3) genes, revealing distinct differences in variant frequencies among breeds. Allele I of CSN2, which had not formerly been screened in such a high number of cows in these Scandinavian breeds, showed a frequency around 7% in DH and DJ, but was not detected in SR. Genetic polymorphisms were significantly associated with curd firming rate and rennet coagulation time. Thus, CSN1S1 C, CSN2 B, and CSN3 B positively affected milk coagulation, whereas CSN2 A(2), in particular, had a negative effect. In addition to the influence of individual casein genes, the effects of CSN1S1-CSN2-CSN3 composite genotypes were also examined, and revealed strong associations in all breeds, which more or less reflected the single gene results. Overall, milk coagulation is under the influence of additive genetic variation. Optimal milk for future cheese production can be ensured by monitoring the frequency of unfavorable variants and thus preventing an increase in the number of cows producing milk with impaired coagulation. Selective breeding for variants associated with superior milk coagulation can potentially increase raw milk quality and cheese yield in all 3 Scandinavian breeds.

Effect of storage and separation of milk at udder quarter level on milk composition, proteolysis, and coagulation properties in relation to somatic cell count.

Coagulation properties of milk are altered by elevated somatic cell count (SCC), partly due to increased proteolytic and lipolytic activity in the milk and, thereby, degradation of protein and fat during storage. Milk is commonly stored on the farm at cooling conditions for up to 2 d before transport to the dairy for processing. This study evaluated the effects of storage on milk with altered composition due to high SCC and the effects of exclusion of milk from individual udder quarters with high SCC on milk composition, proteolysis, and coagulation properties. Udder-quarter milk and cow-composite milk samples from 13 cows having at least 1 quarter with SCC above 100,000 cells/mL were collected on 1 occasion. In addition, commingled milk from only healthy quarters (<100,000 cells/mL) of each cow was collected, representing a cow sample where milk with elevated SCC was excluded. The milk samples were analyzed for total protein content; protein content in the whey fraction; casein, fat, and lactose contents; SCC; proteolysis; curd yield; coagulation time; and total bacterial count, on the day of sampling and after 2 and 5 d of storage at +4°C. In addition to SCC, duration of storage and total bacterial count had an effect on milk quality. The content of total protein, fat and protein contents in the whey fraction, and curd yield were found to have different storage characteristics depending on the level of SCC at udder-quarter level. The exclusion of milk from udder quarters with elevated SCC decreased the content of total protein and protein content in the whey fraction and increased the content of lactose at cow level. However, the effect of separating milk at udder-quarter level needs to be further studied at bulk tank level to evaluate the effect on overall total milk quality.

Day-to-day variation in milk yield and milk composition at the udder-quarter level.

Automatic in-line measurement of milk composition and milk yield could be a useful tool in management of the dairy herd. Data on milk components and milk yield provide information on milk quality alterations and cow health status but are also useful in planning feeding and breeding. In automatic milking systems, udder quarters are milked individually, enabling analysis and recording at the udder-quarter level. Frequent records of components require knowledge about day-to-day variations. A component with greater day-to-day variation needs more frequent sampling when used as a diagnostic tool and for management decisions. Earlier studies have described the day-to-day variations in milk components for cow composite milk, but with the quarter milking technique and the possible sampling at the udder-quarter level, knowledge about day-to-day variations at the udder-quarter level is needed. In this study, udder-quarter and cow composite milk samples were collected from 42 consecutive milkings of 10 cows during 21 d. Milk yield was recorded and the milk was analyzed for total protein, whey protein, casein, fat, lactose, and somatic cell count. The results showed that the day-to-day variations and mean values for 4 healthy udder quarters within a cow were similar. In addition, different milk components had different levels of day-to-day variation, the least variation being found in lactose (0.9%) and the greatest in fat (7.7%). This suggests that repeated milk sampling and analysis at the udder-quarter level can be used to detect alterations in composition and cow health and would, thus, be helpful in the management of the dairy herd.

Evaluation of quality changes in udder quarter milk from cows with low-to-moderate somatic cell counts.

Much emphasis has been put on evaluating alterations in milk composition caused by clinical and subclinical mastitis. However, little is known about changes in milk composition during subclinical mastitis in individual udder quarters with a low-to-moderate increase in milk somatic cell count (SCC). This information is needed to decide whether milk from individual udder quarters with a moderate-to-high increase in milk SCC should be separated or not. The aim of this study was to determine how milk composition in separate udder quarters is affected when cow composite milk has low or moderately increased SCC levels. Udder quarter and cow composite milk samples were collected from 17 cows on one occasion. Milk yield was registered and samples were analyzed for SCC, fat, total protein, whey proteins, lactose, citric acid, non-protein nitrogen (NPN), lactoferrin, protein profile, free fatty acids (FFAs), lactate dehydrogenase (LDH), proteolysis, sodium and potassium. Bacteriological samples were collected twice from all four quarters of all cows. The cows were divided into three groups depending on their SCC at udder quarter level. The first group comprised healthy cows with four udder quarters with low SCC, <50 000 cells/ml; composition was equal when opposite rear and front quarters were compared. In the second and the third groups, cows had one udder quarter with 101 000 cells/ml < SCC < 600 000 cells/ml and SCC > 700 000 cells/ml, respectively. The remaining udder quarters of these cows had low SCC (<100 000 cells/ml). Despite the relatively low average cow composite SCC = 100 000 cells/ml of Group 2, milk from affected udder quarters exhibited lower casein number, content of lactose and ß-casein (ß-CN), while the content of whey protein, sodium, LDH and α-lactoalbumin (α-la) were higher compared to healthy opposite quarters. In addition to these changes, milk from affected udder quarters in Group 3 also exhibited lower values of potassium and αs1-casein (αs1-CN) and higher values of lactoferrin when compared to milk from opposite healthy quarters. This indicates that even when the SCC in cow composite milk is low, there might exist individual quarters for which milk composition is changed and milk quality impaired.

Effects on blood pressure after treatment of obstructive sleep apnoea with a mandibular advancement appliance - a three-year follow-up.

Obstructive sleep apnoea (OSA) is a highly prevalent sleep disorder; it affects 4% of males and 2% of females. Hypertension has been shown to occur in 28-57% of OSA patients. There is a steady increase in evidence linking OSA to long-term cardiovascular morbidity including hypertension. The purpose of this study was to investigate whether mandibular advancement oral appliance (OA) treatment of OSA affects the patient's blood pressure (BP) in a 3-month and a 3-year perspective. Twenty-nine consecutive patients, with verified OSA defined as apnoea index (AI) >5 per hour and/or apnoea/hypopnoea index (AHI) > or =10 per hour, received an OA as treatment. BP was measured on three occasions; before treatment, after 3 months of treatment, and after 3 years of treatment. BP was measured with an electronic blood pressure monitor. The treatment effect of OA was measured after 3 months by repeated somnographic registration while the patient was wearing the OA. A treatment response was defined as AHI < 10; this was achieved in 25 of 29 patients (86%) at the 3-month evaluation. Significant reductions in blood pressure were attained between baseline and the 3-month evaluation (P < 0.001) and these changes remained at the 3-year follow-up in both systolic BP of -15.4 +/- 18.7 mm Hg and diastolic BP of -10.3 +/- 10.0 mm Hg. OA therapy reduced blood pressure in both a 3-month and a 3-year perspective in patients with OSA.

Udder quarter milk composition at different levels of somatic cell count in cow composite milk.

Automatic milking systems have made possible the separation of high- and low-quality milk at the udder quarter level during the milking process. The aim of this study was to investigate the composition and yield of milk from individual udder quarters to determine whether deteriorated milk composition occurs in udders that are assumed to be healthy and whether quarters with high-quality milk are found in udders with high milk somatic cell count (SCC). Milk samples were collected on one occasion from 90 cows at udder quarter level and cow composite level. The milk was analyzed for content of total protein, whey protein, casein, fat, lactose, citric acid and SCC; milk yield was registered. The cows were divided into three groups depending on the SCC of their composite milk. Cows in group 1, cow composite SCC < 100 000 cells/ml, were assumed to have healthy udders. However, instances of increased SCC and decreased milk quality were discovered in one or more udder quarters of approximately 30% of the group. Cows in group 2, cow composite SCC of 100 000 to 300 000 cells/ml, and group 3, cow composite SCC > 300 000 cells/ml, were assumed to have affected udders. However, the majority of these cows had one or more udder quarters in which increased SCC and deteriorated milk quality were not detected. Calculations of bulk-tank milk values, when separation of milk from affected udder quarters was performed, indicate that SCC changes to a much greater degree compared to the other milk components. These results show that milk from affected udder quarters suffers compositional changes, but calculations of simulated separation indicate that the compositional changes in bulk-tank milk are small. The effect of separation of milk from individual udder quarters on bulk-tank milk needs to be further studied.

Effect of minor milk proteins in chymosin separated whey and casein fractions on cheese yield as determined by proteomics and multivariate data analysis.

The objective of this work was to find regressions between minor milk proteins or protein fragments in the casein or sweet whey fraction and cheese yield because the effect of major milk proteins was evaluated in a previous study. Proteomic methods involving 2-dimensional gel electrophoresis and mass spectrometry in combination with multivariate data analysis were used to study the effect of variations in milk protein composition in chymosin separated whey and casein fractions on cheese yield. By mass spectrometry, a range of proteins significant for the cheese yield was identified. Among others, a C-terminal fragment of beta-casein had a positive effect on the cheese yield expressed as grams of cheese per 100 g of milk, whereas several other minor fragments of beta-, alpha(s1)-, and alpha(s2)-casein had positive effects on the transfer of protein from milk to cheese. However, the individual effect of each identified protein was relatively low. Therefore, further studies of the relations between different proteins/peptides in the rennet casein or sweet whey fractions and cheese yield are needed for advanced understanding and prediction of cheese yield.

Effect of beta-casein, kappa-casein and beta-lactoglobulin genotypes on concentration of milk protein variants.

Individual mid-lactation milk samples were collected from 116 cows of the Swedish Red and Swedish Holstein breeds with known genotypes of beta-casein, kappa-casein and beta-lactoglobulin. Detailed milk protein composition and allele-specific expression of beta-casein, kappa-casein and beta-lactoglobulin proteins in milk were analysed using reversed-phase high-performance liquid chromatography. Aggregate beta-/kappa-casein genotype was significantly associated with the kappa-casein concentration in milk. The lowest kappa-casein concentration was found in milk of cows with genotypes including kappa-casein E (A(1)A(2)/AE, A(1)A(1)/AE) and also A(2)A(2)/AA milk, whereas highest levels were associated with genotypes including kappa-casein B. Casein number was positively and concentration of beta-lactoglobulin negatively associated with the beta-lactoglobulin BB genotype. In heterozygote cows, beta-casein A(1) and beta-lactoglobulin A proteins were found at higher concentrations in milk compared with the protein variant encoded by the alternative allele at these loci, whereas kappa-casein A and B variants were found at similar concentrations in heterozygote AB cows.

Effect of protein composition on the cheese-making properties of milk from individual dairy cows.

The objective of this study was to evaluate the effect of variations in milk protein composition on milk clotting properties and cheese yield. Milk was collected from 134 dairy cows of Swedish Red and White, Swedish Holstein, and Danish Holstein-Friesian breed at 3 sampling occasions. Concentrations of alphaS1-, beta-, and kappa-casein (CN), alpha-lactalbumin, and beta-lactoglobulin (LG) A and B were determined by reversed phase liquid chromatography. Cows of Swedish breeds were genotyped for genetic variants of beta- and kappa-CN. Model cheeses were produced from individual skimmed milk samples and the milk clotting properties were evaluated. More than 30% of the samples were poorly coagulating or noncoagulating, resulting in weak or no coagulum, respectively. Poorly and noncoagulating samples were associated with a low concentration of kappa-CN and a low proportion of kappa-CN in relation to total CN analyzed. Furthermore, the kappa-CN concentration was higher in milk from cows with the AB genotype than the AA genotype of kappa-CN. The concentrations of alphaS1-, beta-, and kappa-CN and of beta-LG B were found to be significant for the cheese yield, expressed as grams of cheese per one hundred grams of milk. The ratio of CN to total protein analyzed and the beta-LG B concentration positively affected cheese yield, expressed as grams of dry cheese solids per one hundred grams of milk protein, whereas beta-LG A had a negative effect. Cheese-making properties could be improved by selecting milk with high concentrations of alphaS1-, beta-, and kappa-CN, with high kappa-CN in relation to total CN and milk that contains beta-LG B.

Immunohistochemical study of the ontogeny of prochymosin--and pepsinogen-producing cells in the abomasum of sheep.

The appearance and development of prochymosin- and pepsinogen-producing cells were investigated in the ovine abomasum from fetus to adult using immunohistochemistry. Prochymosin immunoreactivity appeared first in the proper gastric glands of the 100-day-old fetus. The intensity and distribution of prochymosin-immunoreactive cells increased gradually with the progress of gestation, and their most intense immunoreactivities and widest distribution were observed in 3-day-old lambs. They were subsequently reduced throughout postnatal growth. A few prochymosin-immunoreactive cells were scattered in the glands of adult sheep. Pepsinogen immunoreactivity appeared at first in a small number of cells in the base of some proper gastric glands of 120-day-old fetuses. After 130 days, pepsinogen-immunoreactive cells increased their staining intensities and distribution. They reached a peak in area at 21 days, which is comparable to adult sheep. In the pyloric glands, prochymosin- and pepsinogen-immunoreactive cells appeared from 100 and 130 days, respectively. Numbers were reduced in comparison to gastric glands and their occurrence was capricious. The results demonstrated that the ontogeny of prochymosin- and pepsinogen-immunoreactive cells in the abomasum of sheep is more similar to that in cattle than to that in the goat. The present data will contribute to the overall understanding of the development of ruminant gastric proteases.

[Psychosocial survey of drunken drivers within the KAPUBRA project. A new interview technique suitable for matching the right treatment with the right client]. / Psykosocial kartläggning av rattfyllerister inom KAPUBRA-projektet. Ny intervjumetod lämplig för att para ihop rätt behandling med rätt klient.

Three different programs for individuals convicted of drunken driving are being evaluated in a randomized design in collaboration between researchers at the Karolinska Institute and the Prison and Probation Service in Sweden. In the years 1996-1998, 912 clients were interviewed by means of a structured interview, the Addiction Severity Index (ASI), which covers seven problem areas (medical status, alcohol use, employment status, drug use, legal status, family/social and psychiatric status). So far about half of the subjects have been re-investigated two years after leave. Initially, the group being investigated had problems particularly in the areas of criminality and alcohol use. Two years later a positive trend in most of the problem areas could be observed for clients in follow-up.

Determination of silver speciation in natural waters. 1. Laboratory tests of Chelex-100 chelating resin as a competing ligand.

Batch and column experiments were performed to investigate the suitability and chemical characteristics of Chelex-100 for use as a competing ligand in ionic silver (Ag(I)) speciation determinations in natural waters. A conditional stability constant (Kcond) for Ag+ chelation by iminodiacetate groups on the surface of Chelex resin was determined by fitting results of batch and column experiments with an equilibrium speciation model. Results of experiments in which Chelex competed with cyanide ion and thiosulfate ion for aqueous Ag+ were fitted well by a model in which log Kcond(Ag-Chelex) was set to 7.2. This value is similar to literature equilibrium constants for a 1:1 Ag(+)-EDTA chelate. In batch experiments with Chelex, equilibration times of 24 h were found to be sufficient to bring samples close to equilibrium. Effects of resin counterion and total Ag(I) concentration on extent of Ag(I) chelation were found to be minor. Effect of pH on Ag(I) chelation was minor over a range of 6-10. Column experiments (detention time = 6 s, empty-column basis) in which thiosulfate competed with Chelex for Ag(I) gave similar results to batch experiments with thiosulfate. This implies that batch and column experiments could be compared to explore ligands in natural water systems with different rates of dissociation.

Determination of silver speciation in natural waters. 2. Binding strength of silver ligands in surface freshwaters.

Three competing ligand methods were compared to determine characteristics of Ag(I) complexation by dissolved and colloidal ligands present in three rivers and one sewage treatment plant effluent. Iminodiacetate groups on Chelex-100 resin (used in batch and column experiments) and diethyldithiocarbamate (DDC) were used as competing ligands. Results of batch Chelex and DDC competition experiments show good agreement with regard to relative extent of Ag binding by natural ligands among the three river systems. Results of both methods also show a possible correlation between extent of Ag(I) complexation and organic matter concentration and/or Fe concentration. Fraction of Ag(I) associated with Chelex in both batch and column Chelex experiments was similar in each of the four systems tested, indicating that lability of Ag complexes does not change significantly on time scales ranging from seconds to 24 h. Results of Chelex and DDC competition were compared using a model based on a hypothetical single natural ligand. Under the experimental conditions used, this model quantified Ag(I) complexes with log Kcond values from approximately 12 to 14. For the three rivers studied, ligands with silver-association characteristics similar to those of reduced sulfur groups (log K = 14-16) present at subnanomolar concentrations likely dominate Ag(I) speciation in these systems. A weaker ligand (e.g., log Kcond < 12) at concentrations > 0.7 nM dominated Ag(I) speciation in the treatment plant effluent. This may result from elevated concentration of metals that compete for reduced sulfur groups rather than from a lower total concentration of these groups.

Partitioning of total mercury and methylmercury to the colloidal phase in freshwaters.

Using tangential flow ultrafiltration, total mercury (HgT) and methylmercury (MeHg) concentrations in the colloidal phase (0.4 microm-10 kDa) were determined for 15 freshwaters located in the upper Midwest (Minnesota, Michigan, and Wisconsin) and the Southern United States (Georgia and Florida). Unfiltered concentrations were typical of those reported for freshwater and ranged from 0.9 to 27.1 ng L(-1) HgT and from 0.08 to 0.86 ng L(-1) MeHg. For some rivers, HgT and MeHg in the colloidal phase comprised up to 72% of the respective unfiltered concentration. On average, however, HgT and MeHg concentrations were evenly distributed between the particulate (>0.4 microm), colloidal, and dissolved (<10 kDa) phases. The pool of Hg in the colloidal phase decreased with increasing specific conductance. Results from experiments on freshwaters with artificially elevated specific conductance suggest that HgT and MeHg may partition to different subfractions of colloidal material. The colloidal-phase HgT correlation with filtered organic carbon (OC(F)) was generally poor (r2 < 0.14; p > 0.07), but the regression of MeHg with OC(F) was strong, especially in the upper Midwest (r2 = 0.78; p < 0.01). On a mass basis, colloidal-phase Hg concentrations were similar to those of unimpacted sediments in the Midwest. Mercury to carbon ratios averaged 352 pg of HgT/mg of C and 25 pg of MeHg/mg of C and were not correlated to ionic strength. The log of the partition coefficient (log K(D)) for HgT and MeHg ranged from 3.7 to 6.4 and was typical of freshwater values determined using a 0.4 microm cutoff between the particulate phase and the dissolved phase. Log K(D) calculated using the <10 kDa fraction as "dissolved" ranged from 4.3 to 6.6 and had a smaller standard deviation about the mean. In addition, our data support the "particle concentration effect" (PCE) hypothesis that the association of Hg with colloids in the filter-passing fraction can lower the observed log K(D). The similarity between colloidal and particulate-phase partition coefficients suggests that colloidal mass and not preferential colloidal partitioning drives the PCE.