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Background: Given the cost and complexity of home subcutaneous biologic use, a centralized specialty medication management (CSMM) service was developed utilizing clinical pharmacist practitioners (CPPs)., Objective: To determine the implementation feasibility of the CSMM service., Practice Description: The CSMM service was implemented in a Veterans Health Administration (VHA) hospital. The VHA does not use dedicated specialty pharmacies., Practice Innovation: The CSMM service was developed utilizing CPPs who operated as an advance practice provider under a national VHA scope of practice. The CPP staffing the CSMM service performed patient education, screened for medication appropriateness, and monitored for safety and efficacy via videoconference or telephone. All patients newly starting a home subcutaneous biologic were offered the service from allergy, cardiology, dermatology, gastroenterology, and rheumatology clinics, as appropriate., Evaluation Methods: A retrospective chart review was completed through the first year the service was offered, which included the recruitment and retention of patients, number of follow-up appointments, and interventions made., Results: Of the 68 patients offered clinic enrollment, 54 were enrolled in the service (79.4%). Of the 44 who had completed an initial appointment with the CPP, 42 had either received an initial follow-up or were scheduled for a follow-up (95.5%). A total of 161 clinical outcomes have been performed by the CSMM CPP including administrative assistance (33.5%), initial patient education (17.4%), technique correction (7.5%), and medication changes or discontinuations (6.8%)., Conclusion: Given the high rate of enrollment and retention, the implementation of the CSMM service at a VA hospital was feasible. The service contributed to safe and effective medication use for enrolled patients and continues to grow in both patient enrollment and services offered., (Copyright © 2022 American Pharmacists Association®. All rights reserved.)

In this report, we have developed a simple approach using single-detector fluorescence autocorrelation spectroscopy (FCS) to investigate the Förster resonance energy transfer (FRET) of genetically encoded, freely diffusing crTC2.1 (mTurquoise2.1-linker-mCitrine) at the single molecule level. We hypothesize that the molecular brightness of the freely diffusing donor (mTurquoise2.1) in the presence of the acceptor (mCitrine) is lower than that of the donor alone due to FRET. To test this hypothesis, the fluorescence fluctuation signal and number of molecules of freely diffusing construct were measured using FCS to calculate the molecular brightness of the donor, excited at 405 nm and detected at 475/50 nm, in the presence and absence of the acceptor. Our results indicate that the molecular brightness of cleaved crTC2.1 in a buffer is larger than that of the intact counterpart under 405-nm excitation. The energy transfer efficiency at the single molecule level is larger and more spread in values as compared with the ensemble-averaging time-resolved fluorescence measurements. In contrast, the molecular brightness of the intact crTC2.1, under 488 nm excitation of the acceptor (531/40 nm detection), is the same or slightly larger than that of the cleaved counterpart. These FCS-FRET measurements on freely diffusing donor-acceptor pairs are independent of the precise time constants associated with autocorrelation curves due to the presence of potential photophysical processes. Ultimately, when used in living cells, the proposed approach would only require a low expression level of these genetically encoded constructs, helping to limit potential interference with the cell machinery., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2021 Kay, Aplin, Simonet, Beenken, Miller, Libal, Boersma, Sheets and Heikal.)

Living cells are complex, crowded, and dynamic and continually respond to environmental and intracellular stimuli. They also have heterogeneous ionic strength with compartmentalized variations in both intracellular concentrations and types of ions. These challenges would benefit from the development of quantitative, noninvasive approaches for mapping the heterogeneous ionic strength fluctuations in living cells. Here, we investigated a class of recently developed ionic strength sensors that consists of mCerulean3 (a cyan fluorescent protein) and mCitrine (a yellow fluorescent protein) tethered via a linker made of two charged α-helices and a flexible loop. The two helices are designed to bear opposite charges, which is hypothesized to increase the ionic screening and therefore a larger intermolecular distance. In these protein constructs, mCerulean3 and mCitrine act as a donor-acceptor pair undergoing Förster resonance energy transfer (FRET) that is dependent on both the linker amino acids and the environmental ionic strength. Using time-resolved fluorescence of the donor (mCerulean3), we determined the sensitivity of the energy transfer efficiencies and the donor-acceptor distances of these sensors at variable concentrations of the Hofmeister series of salts (KCl, LiCl, NaCl, NaBr, NaI, Na 2 SO 4 ). As controls, similar measurements were carried out on the FRET-incapable, enzymatically cleaved counterparts of these sensors as well as a construct designed with two electrostatically neutral α-helices (E6G2). Our results show that the energy transfer efficiencies of these sensors are sensitive to both the linker amino acid sequence and the environmental ionic strength, whereas the sensitivity of these sensors to the identity of the dissolved ions of the Hofmeister series of salts seems limited. We also developed a theoretical framework to explain the observed trends as a function of the ionic strength in terms of the Debye screening of the electrostatic interaction between the two charged α-helices in the linker region. These controlled solution studies represent an important step toward the development of rationally designed FRET-based environmental sensors while offering different models for calculating the energy transfer efficiency using time-resolved fluorescence that is compatible with future in vivo studies.

Inhibition of hepatic drug metabolizing enzymes is an important cause of clinically relevant drug interactions. A close correlation was found between influence of pentobarbital-induced sleeping time in outbred Swiss mice and hepatic drug metabolism in man for several established inhibitors, i.e. chloramphenicol, cimetidine, idrocilamide, isoniazid, metronidazole, miconazole and triacetyloleandomycin, and control drugs, i.e. baclofen, ranitidine and spiramycin. Provided that room temperature is carefully controlled, barbiturate sleeping time is a simple, inexpensive and reproducible predictive tool for the experimental detection of pharmacological agents likely to inhibit hepatic drug metabolism.

Pentobarbital-induced sleeping time was found to be significantly prolonged in mice within at least 4 days following either whooping cough, tetanus, rubella or poliomyelitis vaccination. By contrast, barbital-induced sleeping time remained unaffected, These findings provide further evidence of a correlation between inhibition of liver drug metabolizing enzymes and stimulation of the immune response.

Evidence suggests that stimulation of the immune response may be associated with impairment of hepatic drug metabolism. Antipyrine kinetics has therefore been determined in 12 patients on the fourth day following tetanus vaccination. Under these conditions, antipyrine plasma half-life was found to be 14.8 h. Despite the lack of control over antipyrine kinetics, these preliminary results indicate that tetanus vaccination is likely to impair the antipyrine metabolism.

To test the hypothesis that hepatic drug metabolism affected the pharmacokinetics of antipyrine, 12 persons who had received vaccination against influenza were given antipyrine on the fourth day following. Plasma antipyrine was assayed from venous blood samples taken 5, 11 and 24 h later. The results indicate that the immunization procedure transiently increased the hepatic drug metabolizing enzyme activity against antipyrine.