Validating and optimizing spot sampling of urine to estimate urine output with creatinine as a marker in dairy cows.

An experiment was conducted to validate and optimize the procedure of spot urine sampling with urinary creatinine as a marker to estimate urine outputs of dairy cows. Twelve lactating cows were used in a randomized complete block design. Cows were grouped and randomly assigned to 2 experimental diets: a corn silage-based diet and an alfalfa silage-based diet with supplemental potassium. The experiment lasted for 21 d and total collection (TC) of urine was conducted for the last 3 d. Twelve spot samples of urine from individual cows were collected over a 3-d period during TC to represent every 2-h sampling in a 24-h cycle. Creatinine excretion rate (mg/kg of body weight per d) was variable among cows from 16.7 to 34.5 with an average of 27.3. Creatinine concentrations of spot samples within cow were averaged to simulate urine samples obtained from various spot sampling frequencies (equally spaced 12, 6, 4, and 2 time points starting at feeding: 12TP, 6TP, 4TP, and 2TP, respectively). Large diurnal variation of urinary creatinine concentration was observed within cow. Creatinine concentration was greater (75 vs. 65 mg/dL) for 12TP compared with TC, resulting in underestimating (29.8 vs. 32.6 kg/d) urine outputs. When compared among 12TP, 6TP, 4TP, and 2TP, creatinine concentrations were different and urine outputs tended to be different for 2TP compared with 12TP, 6TP, and 4TP. In addition, despite underestimation of urine output, a regression analysis indicated strong linear relationships between 12TP, 6TP, or 4TP and TC, suggesting that this technique can successfully identify the differences in urine outputs altered by dietary treatments. However, 4TP failed to detect statistical differences in urine outputs between a corn silage-based diet and the alfalfa silage-based diet with supplemental potassium, indicating that a spot urine sampling frequency of at least 6 was required to identify dietary effects on urine outputs. According to the pattern of diurnal changes in urinary creatinine concentration, a spot sample at about 10 h after feeding may have potential to obtain a urine sample that is more representative (i.e., creatinine concentration) of TC urine compared with urine from multiple sampling frequencies. Overall, urinary creatinine as a marker with spot sampling of urine underestimated urine output. However, 12TP and 6TP were successful in identifying changes in urine outputs by dietary treatments.

Sulfated hyaluronan/collagen I matrices enhance the osteogenic differentiation of human mesenchymal stromal cells in vitro even in the absence of dexamethasone.

Glycosaminoglycans (GAG) are multifunctional components of the extracellular matrix (ECM) involved in different steps of the regulation of cellular differentiation. In this study artificial extracellular matrices (aECM) consisting of collagen (Col) I and different GAG derivatives were used as a substrate for human mesenchymal stromal cells (hMSC) to study osteogenic differentiation in vitro. hMSC were cultured on aECM containing col and hyaluronan sulfates (HyaS) with increasing degrees of sulfation (DS(S)) and were compared with aECM containing col and the natural GAG hyaluronan or chondroitin 4-sulfate. hMSC were analyzed for osteogenic differentiation markers such as calcium phosphate deposition, tissue non-specific alkaline phosphatase (TNAP) and expression of runt-related transcription factor 2 (runx2), osteocalcin (ocn) and bone sialoprotein II (bspII). Compared with aECM containing Col and natural GAG all Col/HyaS-containing aECM induced an increase in calcium phosphate deposition, TNAP activity and tnap expression. These effects were also seen in the absence of dexamethasone (an established osteogenic supplement). The expression of runx2 and ocn was not altered and the expression of bspII was diminished on the col/HyaS-containing aECM. The impact of the Col/HyaS-containing aECM on hMSC differentiation was independent of the DS(S) of the HyaS derivatives, indicating the importance of the primary (C-6) hydroxyl group of N-acetylglucosamine. These results suggest that Col/HyaS-containing aECM are able to stimulate hMSC to undergo osteogenic differentiation even in the absence of dexamethasone, which makes these matrices an interesting tool for hMSC-based tissue engineering applications and biomaterial functionalizations to enhance bone formation.

Few electron limit of n-type metal oxide semiconductor single electron transistors.

We report the electronic transport on n-type silicon single electron transistors (SETs) fabricated in complementary metal oxide semiconductor (CMOS) technology. The n-type metal oxide silicon SETs (n-MOSSETs) are built within a pre-industrial fully depleted silicon on insulator (FDSOI) technology with a silicon thickness down to 10 nm on 200 mm wafers. The nominal channel size of 20 × 20 nm(2) is obtained by employing electron beam lithography for active and gate level patterning. The Coulomb blockade stability diagram is precisely resolved at 4.2 K and it exhibits large addition energies of tens of meV. The confinement of the electrons in the quantum dot has been modeled by using a current spin density functional theory (CS-DFT) method. CMOS technology enables massive production of SETs for ultimate nanoelectronic and quantum variable based devices.

Prognosis of central retinal artery occlusion: local intraarterial fibrinolysis versus conservative treatment.

BACKGROUND AND PURPOSE: Local intraarterial fibrinolysis (LIF) is one of several methods used in treating central retinal artery occlusion (CRAO). We investigated whether LIF is more effective than conservative methods in the treatment of CRAO. METHODS: In this retrospective study, a total of 178 patients (125 men and 53 women) with CRAO were treated at the Eye Hospital of the University of Freiburg from 1980 to 2000. The average age of the patients was 66.8 years (SD, 12 years). In group I, 116 patients were treated conservatively by anterior chamber paracentesis, massage of the globe, isovolemic hemodilution, acetazolamide, Pentoxifyllin, acetylsalicylic acid, and reduction of arterial hypertension. Some combination but not all of the mentioned conservative methods were used in the conservatively treated patients. In group II, 62 patients receiving LIF received local injection of urokinase or recombinant tissue plasminogen activator into the proximal part of the ophthalmic artery. In case of ipsilateral carotid artery occlusion or high grade stenosis (14 of 62 patients), the thrombolytic agent was administered into the internal maxillary artery. RESULTS: Among 178 patients, the CRAO was subtotal in 130 (73.0%), incomplete in 39 (21.9%), and total in nine (5.1%). Statistical calculations showed a significantly better visual acuity in group II patients, who were treated with LIF, in comparison with group I patients, who were treated conservatively (P =.0022). CONCLUSION: For patients with CRAO, LIF is superior to conservative treatment.

Rat hepatic sinusoidal endothelial cells in monolayer culture. Biochemical and ultrastructural characteristics.

Sinusoidal endothelial cells were isolated by collagenase-pronase digestion of rat livers followed by centrifugal elutriation. The main endothelial cell fraction consisted of more than 85% endothelial cells as shown by electron microscopy and enzyme histochemistry. Contamination by Kupffer cells was less than 5%. The endothelial cells formed a coherent stable monolayer on dishes coated with collagen type IV in the presence of an RPMI 1640 medium supplemented with 4% Ultroser. Fc receptors were undetectable immediately after elutriation but reappeared after 12 h in culture. Von Willebrand factor (formerly factor VIII-related antigen) could not be detected unequivocally by immunofluorescence. Unchallenged endothelial cells did not produce eicosanoids. In the presence of free arachidonate, however, prostaglandins D2 and E2 as well as thromboxane B2 and 6-keto-prostaglandin F1 alpha were detected by radioimmunoassay and by high-performance liquid chromatography analysis of [3H]arachidonate-exposed cells. Cells treated with the Ca2+ ionophore A23187 produced the same spectrum of immunologically measured prostanoids. In contrast to Kupffer cells in primary culture, eicosanoid formation by endothelial cells was neither triggered by phagocytotic stimuli nor suppressed by pretreatment with dexamethasone.