This paper presents the results of the research aiming at determining content of selected anions in the samples containing precipitated water (rainwater and snowfall water) by isotachophoretic method. This work is a continuation of the complex pollution monitoring in the areas of Siedlce and Olecko (near Suwałki). The samples were collected from November 2006 to May 2007. Days of sampling depended on atmospheric conditions. Sulfate and hydrogen carbonate turned up to be the dominant anions in samples which were collected in city centers. Higher contents were observed in Siedlce. The content of nitrites in both cities was low. The highest concentration of sulfates and chlorides in rainwater was obtained in the samples collected near the Company of Thermal Energy (CTE) in Siedlce. Similar results of sulfate concentration were observed in Olecko. In samples of snowfall water (collected near to the CTE) sulfates and sulfites were the dominating anions.

In this paper we present the numerical simulation-based design of a new microfluidic device concept for electrophoretic mobility and (relative) concentration measurements of dilute mixtures. The device enables stationary focusing points for each species, where the locally applied pressure driven flow (PDF) counter balances the species’ electrokinetic velocity. The axial location of the focusing point, along with the PDF flowrate and applied electric field reveals the electrokinetic mobility of each species. Simultaneous measurement of the electroosmotic mobility of an electrically neutral specie can be utilized to calculate the electrophoretic mobility of charged species. The proposed device utilizes constant sample feeding, and results in time-steady measurements. Hence, the results are independent of the initial sample distribution and flow dynamics. In addition, the results are insensitive to the species diffusion for large Peclet number flows (Pe > 400), enabling relative concentration measurement of each specie in the dilute mixture.

We measured the bone-specific alkaline phosphatase (ALP) isoenzyme activity in 67 plasma samples from 14 newborn Holstein calves using both a conventional method (featuring heat inactivation) and a commercial agarose gel electrophoresis (AGE) kit; the relevant isoenzymes were termed bone-specific ALP (BAP) and ALP isoenzyme 3 (ALP3). We explored whether the AGE kit afforded reliable data when used to analyze samples from Holstein calves. The blood was collected from the jugular vein of each calf immediately prior to the first colostrum feeding (pre-feeding), 20 and 40 h after pre-feeding, and on days 4 and 7; whereas three samples (from three calves) were not obtained. The total plasma ALP activity varied widely, exceeding the ranges of reference values. On electrophoresis, 52 of 67 plasma samples (77.6 %) clearly contained both ALP isoenzyme 2 and ALP3, as did control human serum. The total ALP activity of the 52 samples ranged from 166–1989 U/L (median: 1013 U/L), whereas the values for the other 15 samples (22.4%) exhibiting abnormal isoenzyme fractionation ranged from 1014–5118 U/L (median: 1780 U/L). In the 52 plasma samples exhibiting clearly separated isoenzymes, ALP3 and BAP activities were strongly positively correlated as revealed by Deming regression (y = 0.93x + 22.6, p<0.0001) and Bland-Altman analysis (ALP3/BAP activities limit of agreement: −5.1%). Thus, the AGE kit yields useful information on newborn calves, and can replace the conventional method when the total plasma ALP activity is less than approximately 1000 U/L.

A recent study found that an agarose gel electrophoresis (AGE) method yielded two distinct major bands corresponding to the hepatic and bone ALP isoenzymes (ALP2 and ALP3, respectively) in bovine serum treated with protease and neuraminidase (PN-treatment), although there were concerns that the intestinal ALP isoenzyme (ALP5) often overlapped with ALP3 in human serum treated with neuraminidase. Because ALP5 was separated from ALP3 in bovine serum treated with protease alone (P-treatment), we used a modified method employing both P- and PN-treated bovine sera to measure the activities of the three ALP isoenzymes in 53 lactating Holstein cows: 24 primiparous and 29 multiparous. Upon electrophoresis, 51 of 53 samples (96.2%) subjected to P-treatment yielded a distinct fraction corresponding to ALP5, as did the control serum. All PN-treated sera yielded a definite ALP2 fraction. The ALP3 fraction was calculated as the remainder after excluding ALP2 and ALP5. The activities of total ALP (t-ALP) and ALP3 in primiparous cows were higher than those in multiparous cows (p < 0.001) at early-to-peak [10–110 days in milk (DIM)] and mid (111–220 DIM) lactation. In the multiparous cows, the ALP3 activity at late lactation (221−477 DIM) was significantly higher than that at early-to-peak lactation. Thus, the modified AGE method described here is able to discriminate three fractions of ALP isoenzymes in the sera of lactating cows. The AGE pattern of circulating ALP isoenzymes will contribute to the understanding of the physiological bone metabolism status in lactating cows.

This study investigated changes in serum levels of hepatic, bone, and intestinal alkaline phosphatase (ALP) isoenzymes (ALP2, ALP3, and ALP5, respectively) in Holstein cows around parturition. Tartrate-resistant acid phosphatase 5b (TRAP5b) activity and calcium (Ca) concen- trations were also measured. We analyzed blood samples from 11 late-pregnant heifers (primipa- rous group) and 13 multiparous (2-4 lactations; multiparous group) cows at 3 weeks (18-24 days prepartum; -3 weeks), 2 weeks (17-11 days prepartum; -2 weeks), and 1 week (10-4 days prepar- tum; -1 weeks) before parturition; the day of calving (within 12 h post-calving; day 0); and 5 days postpartum (5 days). ALP3 activity was significantly higher in the primiparous group than in the multiparous group, whereas the activities decreased significantly in both groups after 5 days. ALP2 and ALP5 activities did not change, whereas ALP2 activity was significantly higher in the primiparous group than in the multiparous group. TRAP5b activity was significantly higher in the primiparous group than in the multiparous group and showed a transient significant increase at day 0. Ca concentration significantly decreased at day 0 in both groups; the Ca level at day 0 was significantly higher in the primiparous group than in the multiparous group. These data show that ALP3 activity in serum may indicate a change in osteoblastic bone forma- tion after calving, but further study is needed to determine the clinical application for measuring ALP isoenzymes in bovine medicine.