Short communication: Repeatability of β-hydroxybutyrate measurements in capillary blood obtained from the external vulvar skin

The primary objective of this study was to evaluate the repeatability of measurements of β-hydroxybutyrate (BHB) concentrations in capillary blood obtained from minimal-invasive puncturing of different sites at the external vulva skin of dairy cows. A total of 240 multiparous cows sampled once within 21 d in milk were enrolled in the study. Capillary blood samples were obtained from the right and left labium of the external vulva skin and additionally from a site close to the first puncture. Samples were analyzed by using an electronic hand-held device [FreeStyle Precison Neo (FSP-Neo), Abbott GmbH & Co., Wiesbaden, Germany)]. Additionally, samples from a coccygeal vessel were analyzed as reference in a laboratory. Mean deviations (± standard deviation) between the 3 capillary sampling sites ranged from 0.01 ± 0.14 (left vs. right labium) to 0.02 ± 0.13 mmol/L (left or right labium vs. additional site). The BHB concentrations in capillary blood compared with the reference showed a mean deviation of 0.14 ± 0.18 mmol/L. The Spearman correlation coefficient between BHB concentrations in capillary blood and coccygeal blood was 85.4%. Mixed model analyses showed no significant effect of sampling site or squeezing the skin during sampling procedures on the differences toward the reference. Based on laboratory BHB concentrations of ≥1.2 and ≥1.4 mmol/L, respectively, the prevalence of hyperketonemia was 10.4 (n = 25) and 5.0% (n = 12). Thresholds to detect hyperketonemia with the FSP-Neo were optimized by receiver operating characteristic analyses. Based on a laboratory BHB concentration of ≥1.2 mmol/L thresholds were determined as 1.1 (left labium, 2nd puncture) and 1.0 mmol/L (right labium), with sensitivities between 94 and 99% and specificities between 89 and 97%. Based on a serum BHB concentration of 1.4 mmol/L, receiver operating characteristic analyses resulted in optimized cutoffs of 1.4 mmol/L for all sampling sites at the vulva, with sensitivities of 100% and specificities between 96 and 99%. In conclusion, vulvar capillary blood is suitable for accurate measuring of BHB concentrations by using the FSP-Neo. After adjusting the thresholds, the FSP-Neo provides very good test characteristics independent of the capillary sampling site or squeezing the skin and allows a practical and reliable screening tool for dairy cows.     

Wettability of Surface Oxyfluorinated Polypropylene Fibres and Its Effect on Interfacial Bonding with Cementitious Matrix

The surface of high molecular weight polypropylene monofilament fibre was modified using a oxyfluorination method. The oxyfluorination treatment level was varied and a hydrolysis post-treatment was also applied. Contact angles of oxyfluorinated, hydro-lyzed oxyfluorinated and unmodified polypropylene fibres were obtained by dynamic contact angle (DCA) measurement using three liquids of known dispersion, acid and base surface free energy components. The surface free energies were then calculated according to the acid-base theory developed by Good, van Oss and Chaudhury. Surface oxyfluorination largely increased the acid and base components of the fibres' surface free energy compared with unmodified polypropylene fibres. The oxyfluorinated and unmodified polypropylene fibre surfaces were observed by Scanning Electronic Microscopy and Photoacoustic Infrared Spectroscopy. It was found that the surface oxyfluorination largely increases the roughness of the polypropylene surfaces and the carbonyl group content increases as the treatment level increases. The interfacial shear bond strengths between the cementitious matrix and the polypropylene fibres treated under various conditions were determined by embedded fibre pull-out tests. Results showed that the fibre surface oxyfluorination treatments increase the interfacial bond strengths. The correlations between the shear bond strengths and surface free energy components were established. Results showed that fibre/concrete interfacial bonding was best correlated with the acid component of surface free energy of polypropylene fibres.     

Fibroblast Growth Factor 21 Response in a Preclinical Alcohol Model of Acute-on-Chronic Liver Injury

Background and Aims: Fibroblast growth factor (FGF) 21 has recently been shown to play a potential role in bile acid metabolism. We aimed to investigate the FGF21 response in an ethanol-induced acute-on-chronic liver injury (ACLI) model in Abcb4^−/− mice with deficiency of the hepatobiliary phospholipid transporter. Methods: Total RNA was extracted from wild-type (WT, C57BL/6J) and Abcb4⁻/⁻ (KO) mice, which were either fed a control diet (WT-Cont and KO-Cont groups; n = 28/group) or ethanol diet, followed by an acute ethanol binge (WT-EtOH and KO-EtOH groups; n = 28/group). A total of 58 human subjects were recruited into the study, including patients with alcohol-associated liver disease (AALD; n = 31) and healthy controls (n = 27). The hepatic and ileal expressions of genes involved in bile acid metabolism, plasma FGF levels, and bile acid and its precursors 7α- and 27-hydroxycholesterol (7α- and 27-OHC) concentrations were determined. Primary mouse hepatocytes were isolated for cell culture experiments. Results: Alcohol feeding significantly induced plasma FGF21 and decreased hepatic Cyp7a1 levels. Hepatic expression levels of Fibroblast growth factor receptor 1 (Fgfr1), Fgfr4, Farnesoid X-activated receptor (Fxr), and Small heterodimer partner (Shp) and plasma FGF15/FGF19 levels did not differ with alcohol challenge. Exogenous FGF21 treatment suppressed Cyp7a1 in a dose-dependent manner in vitro. AALD patients showed markedly higher FGF21 and lower 7α-OHC plasma levels while FGF19 did not differ. Conclusions: The simultaneous upregulation of FGF21 and downregulation of Cyp7a1 expressions upon chronic plus binge alcohol feeding together with the invariant plasma FGF15 and hepatic Shp and Fxr levels suggest the presence of a direct regulatory mechanism of FGF21 on bile acid homeostasis through inhibition of CYP7A1 by an FGF15-independent pathway in this ACLI model. Lay Summary: Alcohol challenge results in the upregulation of FGF21 and repression of Cyp7a1 expressions while circulating FGF15 and hepatic Shp and Fxr levels remain constant both in healthy and pre-injured livers, suggesting the presence of an alternative FGF15-independent regulatory mechanism of FGF21 on bile acid homeostasis through the inhibition of Cyp7a1.     

Curcumin-Induced Heme Oxygenase-1 Expression Prevents H2O2-Induced Cell Death in Wild Type and Heme Oxygenase-2 Knockout Adipose-Derived Mesenchymal Stem Cells

Mesenchymal stem cell (MSC) administration is a promising adjuvant therapy to treat tissue injury. However, MSC survival after administration is often hampered by oxidative stress at the site of injury. Heme oxygenase (HO) generates the cytoprotective effector molecules biliverdin/bilirubin, carbon monoxide (CO) and iron/ferritin by breaking down heme. Since HO-activity mediates anti-apoptotic, anti-inflammatory, and anti-oxidative effects, we hypothesized that modulation of the HO-system affects MSC survival. Adipose-derived MSCs (ASCs) from wild type (WT) and HO-2 knockout (KO) mice were isolated and characterized with respect to ASC marker expression. In order to analyze potential modulatory effects of the HO-system on ASC survival, WT and HO-2 KO ASCs were pre-treated with HO-activity modulators, or downstream effector molecules biliverdin, bilirubin, and CO before co-exposure of ASCs to a toxic dose of H₂O₂. Surprisingly, sensitivity to H₂O₂-mediated cell death was similar in WT and HO-2 KO ASCs. However, pre-induction of HO-1 expression using curcumin increased ASC survival after H₂O₂ exposure in both WT and HO-2 KO ASCs. Simultaneous inhibition of HO-activity resulted in loss of curcumin-mediated protection. Co-treatment with glutathione precursor N-Acetylcysteine promoted ASC survival. However, co-incubation with HO-effector molecules bilirubin and biliverdin did not rescue from H₂O₂-mediated cell death, whereas co-exposure to CO-releasing molecules-2 (CORM-2) significantly increased cell survival, independently from HO-2 expression. Summarizing, our results show that curcumin protects via an HO-1 dependent mechanism against H₂O₂-mediated apoptosis, and likely through the generation of CO. HO-1 pre-induction or administration of CORMs may thus form an attractive strategy to improve MSC therapy.     

Induction of Bax-alpha precedes apoptosis in a human B lymphoma cell line: potential role of the bcl-2 gene family in surface IgM-mediated apoptosis

The members of the bcl-2 gene family are major regulators of programmed cell death, but their role in sIg-triggered apoptosis remains unclear. Using sensitive and resistant variants of the human B cell line BL-41, we studied the expression of the bcl-2 gene family during surface IgM-mediated apoptosis. We found constitutive Bcl-2 and Bcl-x expression, which remained unaltered after sIg cross-linking, in both resistant and sensitive cells. This and other experiments suggest that constitutive expression of Bcl-2 or Bcl-x alone is not sufficient to protect from activation-induced cell death in B cells. We therefore investigated Bax-alpha, the death-promoting splice variant of Bax, and found strong induction of both mRNA and protein upon sIg stimulation in sensitive cells. However, resistant subclones showed only weak expression, which was not inducible by sIg cross-linking. We provide evidence that up-regulation of Bax-alpha and the resulting imbalance of Bcl-2/Bax might be a major regulator of sIg-mediated apoptosis. Additionally, we found strong constitutive expression of Bcl-xs, the death promoting variant of Bcl-x, in sensitive cells, whereas resistant cells showed only weak Bcl-xs expression. Thus, we observed a much stronger expression of the death-promoting proteins Bax-alpha (inducible) and Bcl-xs (constitutive) in sensitive cells than in resistant cells. We therefore propose a potential role of the novel bcl-2 gene family members bcl-x and bax in surface IgM-triggered apoptosis.     

Evaluation of drug concentration in tissues after percutaneous administration of non-steroidal antirheumatics]

Forty-two cases of medulloblastoma of posterior fossa in children are presented in this paper. Of the children, 28 were males and 14 females; their age ranged from 1.5 to 12 (mean 7.3) years. The foci found were in vermis (36) and cerebellar hemispheres (6). All of these children presented symptoms and signs of obviously increased intracranial pressure, and 31(77%) of them had the signs of cerebellar functional deficits. CT scan showed severe obstructive hydrocephalus in 39 cases. 42 children with posterior fossa medulloblastoma underwent surgical resection. Three died of postoperative prespiratory and circulatory failure, 39 made good recovery and received craniospinal radiotherapy and adjunct chemotherapy. Up to now, none died. Surgical intervention and operative procedures were emphasized particularly. Radiotherapy and chemotherapy were discussed also.