S-CMC-Lys protective effects on human respiratory cells during oxidative stress.

The mucoactive drug S-carbocysteine lysine salt monohydrate (S-CMC-Lys) stimulates glutathione (GSH) efflux from respiratory cells. Since GSH is one of the most important redox regulatory mechanisms, the aim of this study was to evaluate the S-CMC-Lys effects on GSH efflux and intracellular concentration during an oxidative stress induced by the hydroxyl radical (xOH). Experiments were performed on cultured human respiratory WI-26VA4 cells by means of patch-clamp experiments in whole-cell configuration and of fluorimetric analyses at confocal microscope. xOH exposure induced an irreversible inhibition of the GSH and chloride currents that was prevented if the cells were incubated with S-CMC-Lys. In this instance, the currents were inhibited by the specific blocker CFTR(inh)-172. CFT1-C2 cells, which lack a functional CFTR channel, were not responsive to S-CMC-Lys, but the stimulatory effect of the drug was restored in LCFSN-infected CFT1 cells, functionally corrected to express CFTR. Fluorimetric measurements performed on the S-CMC-Lys-incubated cells revealed a significant increase of the GSH concentration that was completely hindered after oxidative stress and abolished by CFTR(inh)-172. The cellular content of reactive oxygen species was significantly lower in the S-CMC-Lys-treated cells either before or after xOH exposure. As a conclusion, S-CMC-Lys could exert a protective function during oxidative stress, therefore preventing or reducing the ROS-mediated inflammatory response

Prolongation of survival of dogs with oral malignant melanoma treated by en bloc surgical resection and adjuvant CSPG4-antigen electrovaccination

Reported post-surgery 1-year survival rate for oral canine malignant melanoma (cMM) is around 30%; novel treatments are needed as the role of adjuvant chemotherapy is unclear. This prospective study regards adjuvant electrovaccination with human-CSPG4-encoded plasmid in 23 dogs with resected II/III-staged CSPG4-positive oral cMM compared with 19 dogs with resected only II/III-staged CSPG4-positive oral cMM. Vaccination resulted in 6/12/18/24-month survival rate of respectively 95.6/73.9/47.8/30.4% (MST 684 days, range 78-1694, 8/23 dogs alive) and 6/12/18/24-month DFI rate of respectively 82.6/47.8/26.1/17.4% (DFI 477 days, range 50-1694). Non-vaccinated dogs showed 6/12/18/24-month survival rate of respectively 63.2/26.3/15.8/5.3% (MST 200 days, range 75-1507, 1/19 dogs alive) and 6/12/18/24-month DFI rate of respectively 52.6/26.3/10.5/5.3% (DFI 180 days, range 38-1250). Overall survival and DFI of vaccinated dogs was longer in those <20 Kg. In vaccinated and non-vaccinated dogs local recurrence rate was respectively 34.8% and 42% while lung metastatic rate was respectively 39% and 79%

Unknown Renal Impairment: A Rare Case of Inhaled Tobramycin Induced Acute Kidney Injury in a Cystic Fibrosis Patient

Acute kidney injury is a reversible medical condition commonly caused by nephrotoxic agents. The infrequency that a nebulized medication elicits a renal insult presents a rare diagnostic challenge. Within this case, we report a 57-year-old cystic fibrosis patient with chronic kidney disease (CKD) Stage G3b (baseline 1.5–1.6 mg/dL) who developed an acute kidney injury (AKI) with a serum creatinine elevation to 4.08 mg/dL and associated worsening vestibular dysfunction related to twice-daily nebulized tobramycin inhalation solution (TIS). The patient was found to have a tobramycin serum level of 4.2 μg/mL 2.5 h after TIS dosing, with elevation remaining present at 1.1 μg/mL 24 h after discontinuation of therapy. Laboratory values at one month continued to show elevated creatinine levels at 2.1 mg/dL, suggesting progression of his baseline CKD. This case supports the benefit of obtaining tobramycin serum levels and vestibular/audiology function testing when evaluating patients on chronic nebulized TIS who present with acute or chronic renal dysfunction. From these serum levels, adjustments to daily dosing, regular monitoring of tobramycin serum levels, or discontinuation of treatment should be made to prevent permanent renal damage in patients with CKD. Calculated Naranjo ADR Probability Scale: 9; Definite

Evaluation of Home Medication Reconciliation by Clinical Pharmacists for Adult and Pediatric Cystic Fibrosis Patients

Medication reconciliation is an important aspect of a patient&rsquo;s care process that is ideally performed by clinical pharmacists. Despite literature supporting this process in other patient populations, cystic fibrosis (CF) lacks research in this area. To address this, we designed a retrospective, multi-centered, non-controlled, cross-sectional study at four CF Foundation-accredited centers in the United States to evaluate the medication reconciliation process for adult and pediatric CF patients by documenting the number of home medications reconciled by clinical pharmacists and the number of patients with home medications that did not align with the current CF guidelines published in 2013. There were 105 adult patients and 72 pediatric patients included in the study analysis with a mean number of medications reconciled by clinical pharmacists of 17.4 (standard deviation (SD) 6.7) for adults and 13 (SD 4.6) for pediatric patients. The mean number of discrepancies from guidelines per patient was 1.61 (SD 1.2) for adult patients and 0.63 (SD 0.9) for pediatric patients. Pharmacists play an essential role in identifying and managing medication interactions and further research is necessary to investigate pharmacist impact on medication reconciliation

Antiestrogen- and tamoxifen-induced effects on calcium-activated chloride currents in epithelial cells carrying the ∆F508-CFTR point mutation

Abstract Background Although pharmacological treatment has increased the average life expectancy of patients with cystic fibrosis, the median survival of females is shorter than that of males. In vitro and in vivo studies have shown that estrogens play a relevant role in the disease progression. The aim of this study was to investigate the effects of 17β-estradiol and tamoxifen citrate (TMX) on calcium-activated chloride channel (CaCC) currents in human bronchial epithelial cells carrying the ΔPhe508-CFTR mutation both in homozygosis and in heterozygosis. Methods Perforated patch clamp experiments were performed on single cells of the immortalized cell lines CFBE and IB3–1. Gramicidin (10 or 20 μM) was added to the electrode solution to reach the whole cell configuration. The electrical stimulation protocol consisted of square voltages ranging from − 80 to + 80 mV, in steps of 20 mV and with a duration of 800 msec. Results The presence of 17β-estradiol significantly reduced the CaCC currents, both in basal conditions and in the presence of ATP (100 μM). The addition of TMX (10 μM) completely restored the currents abolished by 17β-estradiol, in basal conditions and after stimulation with ATP in both CFBE and IB3–1 cells. TMX had a strong, direct action on membrane current density, which significantly increased more than 4-fold in both cases. The membrane current stimulation produced by TMX was further enhanced by the addition of ATP. CFBE cells incubated for 24 h with 3 μM VX-809 (a CFTR corrector) and then acutely stimulated with VX-770 (a CFTR potentiator) in the presence of forskolin, showed an increase of chloride currents which were abolished by Inh-172. The chloride current density induced by TMX + ATP was, on average, greater than that obtained with VX-809 + VX-770 + forskolin. The currents elicited by TMX + ATP were abolished by the addition of NPPB, a CaCC inhibitor. The combined administration of TMX/ATP and VXs/FSK had an additional effect on chloride currents. Conclusions Our results show that TMX restores CaCC currents inhibited by 17ß-estradiol and directly activates the transmembrane chloride currents potentiated by ATP, an effect which is mutation independent. The combined effect of TMX with current used treatments for cystic fibrosis could be of benefit to patients

Surveys and investigations for sportfish management in lakes and rivers in Illinois Annual Report July 1, 2014 through June 30, 2015

Reports on progress and results of sportfish studies including sportfish enhancement, harvest, regulations, and tournaments, habitat restoration, recruitment of sportfish, and sportfish sampling efficiency.Illinois Department of Natural Resources, Division of Fisheries, Federal Aid Project F-185-R-2unpublishednot peer reviewedOpe

K+ channel cAMP activated in guinea pig gallbladder epithelial cells

In guinea pig gallbladder epithelial cells, an increase in intracellular cAMP levels elicits the rise of anion channel activity. We investigated by patchclamp techniques whether K1 channels were also activated. In a cell-attached configuration and in the presence of theophylline and forskoline or 8-Br-cAMP in the cellular incubation bath, an increase of the open probability (Po) values for Ca21-activated K1 channels with a single-channel conductance of about 160 pS, for inward current, was observed. The increase in Po of these channels was also seen in an inside-out configuration and in the presence of PKA, ATP, and cAMP, but not with cAMP alone; phosphorylation did not influence single-channel conductance. In the inside-out configuration, the opioid loperamide (1025 M) was able to reduce Po when it was present either in the microelectrode filling solution or on the cytoplasmic side. Detection in the epithelial cells by RT-PCR of the mRNA corresponding to the a subunit of largeconductance Ca21-activated K1 channels (BKCa) indicates that this gallbladder channel could belong to the BK family. Immunohistochemistry experiments con- firm that these cells express the BK asubunit, which is located on the apical membrane. Other K1 channels with lower conductance (40 pS) were not activated either by 8-Br-cAMP (cell-attached) or by PKA 1 ATP 1 cAMP (inside-out). These channels were insensitive to TEA1 and loperamide. The data demonstrate that under conditions that induce secretion, phosphorylation activates anion channels as well as Ca21- dependent, loperamide-sensitive K1 channels present on the apical membrane

Surveys and investigations for sportfish management in lakes and rivers in Illinois Annual Report July 1, 2014 through June 30, 2015

Illinois Department of Natural Resources, Division of Fisheries, Federal Aid Project F-185-R-2unpublishednot peer reviewedOpe

Volume-regulated Cl- channels in human pleural mesothelioma cells

Anion channels in human mesothelial and mesothelioma cell lines were characterized by patch-clamp and biomolecular approaches. We found an outwardly rectifying anionic current which was inactivated at positive voltages and inhibited by extracellular adenosine 5P-triphosphate (ATP). Mesothelial and mesothelioma cells behaved differently concerning current inactivation properties. Inactivation is more pronounced and has a steeper onset in mesothelial cells. Different reversal potentials, in asymmetrical Cl- solutions, that could be attributed to a different selectivity of the channel, have been observed in the two cell lines. Mesothelioma cell single-channel analysis indicates that the number of the same active anion channel (3-4 pS)increased under hypoosmotic conditions. Immunocytochemistry experiments showed the presence of ICln protein in the cytosol and in the plasma membrane. Western blot analysis revealed an increase of ICln in the membrane under hypotonic conditions, an event possibly related to the activation of Cl- channels

Effects of Physiological and Pathological Urea Concentrations on Human Microvascular Endothelial Cells

Urea is the uremic toxin accumulating with the highest concentration in the plasma of chronic kidney disease (CKD) patients, not being completely cleared by dialysis. Urea accumulation is reported to exert direct and indirect side effects on the gastrointestinal tract, kidneys, adipocytes, and cardiovascular system (CVS), although its pathogenicity is still questioned since studies evaluating its side effects lack homogeneity. Here, we investigated the effects of physiological and pathological urea concentrations on a human endothelial cell line from the microcirculation (Human Microvascular Endothelial Cells-1, HMEC-1). Urea (5 g/L) caused a reduction in the proliferation rate after 72 h of exposure and appeared to be a potential endothelial-to-mesenchymal transition (EndMT) stimulus. Moreover, urea induced actin filament rearrangement, a significant increase in matrix metalloproteinases 2 (MMP-2) expression in the medium, and a significant up- or down-regulation of other EndMT biomarkers (keratin, fibrillin-2, and collagen IV), as highlighted by differential proteomic analysis. Among proteins whose expression was found to be significantly dysregulated following exposure of HMEC-1 to urea, dimethylarginine dimethylaminohydrolase (DDAH) and vasorin turned out to be down-regulated. Both proteins have been directly linked to cardiovascular diseases (CVD) by in vitro and in vivo studies. Future experiments will be needed to deepen their role and investigate the signaling pathways in which they are involved to clarify the possible link between CKD and CVD