Cofactor regeneration by a soluble pyridine nucleotide transhydrogenase for biological production of hydromorphone

We have applied the soluble pyridine nucleotide transhydrogenase of Pseudomonas fluorescens to a cell-free system for the regeneration of the nicotinamide cofactors NAD and NADP in the biological production of the important semisynthetic opiate drug hydromorphone. The original recombinant whole-cell system suffered from cofactor depletion resulting from the action of an NADP(+)-dependent morphine dehydrogenase and an NADH-dependent morphinone reductase. By applying a soluble pyridine nucleotide transhydrogenase, which can transfer reducing equivalents between NAD and NADP, we demonstrate with a cell-free system that efficient cofactor cycling in the presence of catalytic amounts of cofactors occurs, resulting in high yields of hydromorphone. The ratio of morphine dehydrogenase, morphinone reductase, and soluble pyridine nucleotide transhydrogenase is critical for diminishing the production of the unwanted by-product dihydromorphine and for optimum hydromorphone yields. Application of the soluble pyridine nucleotide transhydrogenase to the whole-cell system resulted in an improved biocatalyst with an extended lifetime. These results demonstrate the usefulness of the soluble pyridine nucleotide transhydrogenase and its wider application as a tool in metabolic engineering and biocatalysis

Report in the Grand River Eagle

A report in the Grand River Eagle that a committee was organized in Grand Rapids to help the worthy and enterprising pastor, the Rev. A. C. Van Raalte, and his people in their settlement. Some members of the committee were appointed to visit Holland. E. B. Bostwick was chairman and A. D. Rathbone, secretary.https://digitalcommons.hope.edu/vrp_1840s/1138/thumbnail.jp

A novel experiential work-based learning model in paediatric secondary care using entrustable professional activities to develop clinical knowledge and communication skills

\ua9 2024 The AuthorsBackground: Initial education and training standards for pharmacists in Great Britain require early clinical exposure to patients using experiential work-based learning. However, there is poor evidence of this approach in some settings, such as paediatric care. The aim of this study was therefore to explore a novel model of experiential work-based learning for student pharmacists in a paediatric setting. Methods: Fourth-year student pharmacists enrolled on a Master of Pharmacy programme were allocated five three-hour placement sessions at a paediatric hospital. Sessions consisted of a briefing, ward activities, scaffolded consultations with children and their carers, followed by a debriefing session with a clinical supervisor. Data were collected relating to the ward, patient details, student reported activities, learning outcomes and if follow up was required by a member of the clinical team. Data were cleaned, quality checked, then descriptive statistical analysis and inductive content analysis were conducted. Main findings: Seventy-four students took part in 28 individual sessions and 233 consultations were recorded. Consultations included a best-possible medical history (76%, n = 177), a satisfactory drug history (45%, n = 104), or discussed hospital discharge (11%, n = 26). Students were exposed to patients with diagnosed acute conditions (41%, n = 96) and chronic conditions (33%, n = 76), as well as children awaiting diagnosis (13%, n = 30). Students reported learning about the pathology, diagnosis and symptoms of paediatric conditions (48%, n = 81), medicines used in children (24%, n = 41), patient experiences of recieving care (15%, n = 25), carer experiences (2%, n = 3), the hospital environment (2%, n = 4), career progression (2%, n = 4), and experiences of social care (11%, n = 18). Findings were synthesised with existing entrustable professional activities from the literature to generate novel EPAs specific to paediatric settings. Conclusions: A paediatric setting offers a suitable environment to host experiential work-based learning in pharmacy education. Standards of initial education and training which require pharmacists to prescribe in Great Britain must recognise the importance of exposure to the health needs and experiences of children, young people\u27s and carers prior to graduation

Self, memory, and imagining the future in a case of psychogenic amnesia

We report a case of psychogenic amnesia and examine the relationships between autobiographical memory impairment, the self, and ability to imagine the future. Case study JH, a 60 year old male, experienced a 6 year period of pervasive psychogenic amnesia covering all life events from childhood to the age of 53. JH was tested during his amnesic period and again following hypnotherapy and the recovery of his memories. JH’s amnesia corresponded with deficits in self-knowledge and imagining the future. Results are discussed with reference to models of self and memory and processes involving remembering and imagining

Signal transduction events induced by extracellular guanosine 5′triphosphate in excitable cells

A better understanding of the physiological effects of guanosine-based purines should help clarify the complex subject of purinergic signalling. We studied the effect of extracellular guanosine 5′triphosphate (GTP) on the differentiation of two excitable cell lines that both have specific binding sites for GTP: PC12 rat pheochromocytoma cells and C2C12 mouse skeletal muscle cells. PC12 cells can be differentiated into fully functional sympathetic-like neurons with 50′00 ng ml−1 of nerve growth factor, whereas serum starvation causes C2C12 cells to differentiate into myotubes showing functional excitation–contraction coupling, with the expression of myosin heavy chain proteins. Our results show that GTP enhances the differentiation of both of these excitable cell lines. The early events in guanosine-based purine signal transduction appear to involve an increase in intracellular Ca2+ levels and membrane hyperpolarization. We further investigated the early activation of extracellular-regulated kinases and phosphoinositide 3-kinase in GTP-stimulated PC12 and C2C12 cells, respectively. We found that GTP promotes the activation of both kinases. Together, our results suggest that, even if there are some differences in the signalling pathways, GTP-induced differentiation in both cell lines is dependent on an increase in intracellular Ca2+

Your space or mine? : Mapping self in time

Peer reviewedPublisher PD

In-situ formation of solidified hydrogen thin-membrane targets using a pulse tube cryocooler

An account is given of the Central Laser Facility's work to produce a cryogenic hydrogen targetry system using a pulse tube cryocooler. Due to the increasing demand for low Z thin laser targets, CLF (in collaboration with TUD) have been developing a system which allows the production of solid hydrogen membranes by engineering a design which can achieve this remotely; enabling the gas injection, condensation and solidification of hydrogen without compromising the vacuum of the target chamber. A dynamic sealing mechanism was integrated which allows targets to be grown and then remotely exposed to open vacuum for laser interaction. Further research was conducted on the survivability of the cryogenic targets which concluded that a warm gas effect causes temperature spiking when exposing the solidified hydrogen to the outer vacuum. This effect was shown to be mitigated by improving the pumping capacity of the environment and reducing the minimum temperature obtainable on the target mount. This was achieved by developing a two-stage radiation shield encased with superinsulating blanketing; reducing the base temperature from 14 0.5 K to 7.2 0.2 K about the coldhead as well as improving temperature control stability following the installation of a high-performance temperature controller and sensor apparatus. The system was delivered experimentally and in July 2014 the first laser shots were taken upon hydrogen targets in the Vulcan TAP facility.</p