Vitrified turkey ovarian tissue cultured and assessed through gene expression patterns: A potential screening tool.

Biobanking of turkey ovarian tissue has the potential to play a crucial part in preserving female genetics. To date, ovarian tissue has only been vitrified using a standard protocol, with immediate analyses after warming, therefore, long-term cryoinjury is unknown. Long-term cryoinjury was investigated here by in-ovo culturing, fresh (non-vitrified), a purposefully suboptimal poor vitrification (PV), and the standard vitrified (StV) protocol. Assessments were performed via cellular morphological changes and mRNA gene expression differences, immediately (day 0) or after 2, 4, or 6 days of in-ovo culturing. On day 0, the mRNA levels of heat-shock protein A2 (HSPA2) were lowest in the fresh tissue, and increased 5-fold in the StV treatment, and 18-fold in the PV treatment. Whereas, by day 6, growth determining factor 9 (GDF9) mRNA levels within the fresh tissue were over 3-fold and 21-fold higher than StV and PV treatments, respectively. After 6 days of in-ovo culture the follicle density was highest in the fresh ovarian tissue (4701 ± 950 #/mm3), followed by the StV (1601 ± 300 #/mm3), with PV having the lowest density (172 ± 145 #/mm3). This shows that although the density of follicles was higher in StV versus PV, a considerable number (∼65 %) were lost compared to the fresh treatment. Additionally, the HSPA2 expression could be an early screening tool, whereas GDF9 expression could be a late screening tool, used to assess turkey ovarian tissue vitrification protocols. We conclude that the StV protocol should be further optimized to try and improve follicle numbers post-warming.

Fractured Sealing Rings in a Fenestrated Anaconda Stent-Graft.

An 81 year-old man presented with an asymptomatic juxtrarenal abdominal aortic aneurysm and was subsequently treated with a fenestrated endovascular Anaconda stent-graft. Surveillance imaging within the first postoperative year demonstrated a lower proximal sealing ring fracture. In the second postoperative surveillance year, the upper proximal sealing ring was also fractured with extension of the wire into the right paravertebral space. Despite these sealing ring fractures, there were no endoleak nor visceral stent complications and the patient continued on standard surveillance protocols. There are an increasing number of reports of fractured proximal sealing rings with the fenestrated Anaconda platform. Those analysing the surveillance scans of patients treated with this device should stay vigilant for the development of this complication.

Short-Term Treatment with Rho-Associated Kinase Inhibitor Preserves Keratinocyte Stem Cell Characteristics In Vitro.

Primary keratinocytes including keratinocyte stem cells (KSCs) can be cultured as epidermal sheets in vitro and are attractive for cell and gene therapies for genetic skin disorders. However, the initial slow growth of freshly isolated keratinocytes hinders clinical applications. Rho-associated kinase inhibitor (ROCKi) has been used to overcome this obstacle, but its influence on the characteristics of KSC and its safety for clinical application remains unknown. In this study, primary keratinocytes were treated with ROCKi Y-27632 for six days (short-term). Significant increases in colony formation and cell proliferation during the six-day ROCKi treatment were observed and confirmed by related protein markers and single-cell transcriptomic analysis. In addition, short-term ROCKi-treated cells maintained their differentiation ability as examined by 3D-organotypic culture. However, these changes could be reversed and became indistinguishable between treated and untreated cells once ROCKi treatment was withdrawn. Further, the short-term ROCKi treatment did not reduce the number of KSCs. In addition, AKT and ERK pathways were rapidly activated upon ROCKi treatment. In conclusion, short-term ROCKi treatment can transiently and reversibly accelerate initial primary keratinocyte expansion while preserving the holoclone-forming cell population (KSCs), providing a safe avenue for clinical applications.

The effects of ischaemic conditioning on lung ischaemia-reperfusion injury.

Ischaemia-reperfusion injury (IRI) encompasses the deleterious effects on cellular function and survival that result from the restoration of organ perfusion. Despite their unique tolerance to ischaemia and hypoxia, afforded by their dual (pulmonary and bronchial) circulation as well as direct oxygen diffusion from the airways, lungs are particularly susceptible to IRI (LIRI). LIRI may be observed in a variety of clinical settings, including lung transplantation, lung resections, cardiopulmonary bypass during cardiac surgery, aortic cross-clamping for abdominal aortic aneurysm repair, as well as tourniquet application for orthopaedic operations. It is a diagnosis of exclusion, manifesting clinically as acute lung injury (ALI) or acute respiratory distress syndrome (ARDS). Ischaemic conditioning (IC) signifies the original paradigm of treating IRI. It entails the application of short, non-lethal ischemia and reperfusion manoeuvres to an organ, tissue, or arterial territory, which activates mechanisms that reduce IRI. Interestingly, there is accumulating experimental and preliminary clinical evidence that IC may ameliorate LIRI in various pathophysiological contexts. Considering the detrimental effects of LIRI, ranging from ALI following lung resections to primary graft dysfunction (PGD) after lung transplantation, the association of these entities with adverse outcomes, as well as the paucity of protective or therapeutic interventions, IC holds promise as a safe and effective strategy to protect the lung. This article aims to provide a narrative review of the existing experimental and clinical evidence regarding the effects of IC on LIRI and prompt further investigation to refine its clinical application.

Cyclosporin A Prevents Ovarian Graft Rejection, and Permits Normal Germ Cell Maturation Within the First 5 Weeks Post-transplantation, in the Domestic Turkey (Meleagris gallopavo).

Biobanked ovaries collected from recently hatched poults can only be revived through transplantation, using a recipient bird. The main hurdle in transplantation is preventing graft rejection, which appears as lymphocytic infiltration upon histologic evaluation of the graft. In this study, the condition of the transplants [immunological compatibility (auto- vs. allotransplants), donor age, time in holding media, and temperature of holding media] and treatment of recipient poults with varying immunosuppressants [mycophenolate mofetil (MFM), cyclophosphamide (CY), and cyclosporin A (CsA)] were studied to determine which factors could reduce lymphocytic infiltration, during the first 35 days post-transplantation. Lymphocytic infiltration was determined via cytoplasmic CD3 (T cell) and nuclear PAX5 (B cell) expression. There was no significant difference in the percent of cytoplasmic CD3 or nuclear PAX5 immunostained area between the unoperated group and the autotransplants, by 6 days post-transplantation. However, the allotransplants had more (P < 0.05) positive cytoplasmic and nuclear immunostained areas compared to autotransplants, irrespective of donor age, time in holding media or temperature of the media. By 14 days post-transplantation, the CsA 25 and 50 mg/kg/day treatment groups had less (P < 0.05) CD3 and PAX5 positive areas in their allotransplants, compared to the unsuppressed group. At 35 days post-transplantation, the CsA 25 mg/kg/day allotransplant group also had less (P < 0.05) CD3 and PAX5 positive areas compared to the unsuppressed group. The CsA 25 mg/kg/day transplants also had a similar ovarian follicular size compared to the unoperated group, although they contained fewer (P < 0.05) follicles based on follicular density. Donor age, duration in holding media, temperature of media, and treatment of recipients with MFM or CY had no effect on reducing lymphocytic infiltration. However, immunological compatibility was associated with decreased lymphocytic infiltration, as autotransplants had little lymphocytic infiltration. Treatment of recipients with CsA at 25 mg/kg/day was also associated with reduced lymphocytic infiltration and allowed transplants to develop normally during the first 35 days post transplantation.

Three world wars: Fiscal-monetary consequences.

Directed by a consolidated government budget constraint, we compare US monetary­fiscal responses to World Wars I and II and the War on COVID-19.

Virtual Interactive Surgical Skills Classroom: A Parallel-group, Non-inferiority, Adjudicator-blinded, Randomised Controlled Trial (VIRTUAL).

OBJECTIVE: This study evaluated the efficacy of virtual classroom training (VCT) in comparison to face-to-face training (FFT) and non-interactive computer-based learning (CBL) for basic surgical skills training. DESIGN: This was a parallel-group, non-inferiority, prospective randomised controlled trial with three intervention groups conducted in 2021. There were three intervention groups with allocation ratio 1:1:1. Outcome adjudicators were blinded to intervention assignment. Interventions consisted of 90-minute training sessions. VCT was delivered via the BARCO weConnect platform, FFT was provided in-person by expert instructors and CBL was carried out by participants independently. The primary outcome was post-intervention Objective Structured Assessment of Technical Skills score, adjudicated by two experts and adjusted for baseline proficiency. The assessed task was to place three interrupted sutures with hand-tied knots. SETTING: This multicentre study recruited from five medical schools in London. PARTICIPANTS: Inclusion criteria were medical student status and access to a personal computer and smartphone. One hundred fifty-nine eligible individuals applied online. Seventy-two participants were randomly selected and stratified by subjective and objective suturing experience prior to permuted block randomization. RESULTS: Twenty-four participants were allocated to each intervention, all were analysed per-protocol. The sample was 65.3% female with mean age 21.3 (SD 2.1). VCT was non-inferior to FFT (adjusted difference 0.44, 95% CI: -0.54 to 1.75, delta 0.675), VCT was superior to CBL (adjusted difference 1.69, 95% CI: 0.41-2.96) and FFT was superior to CBL (adjusted difference 1.25, 95% CI: 0.20-2.29). The costs per-attendee associated with VCT, FFT and CBL were £22.15, £39.69 and £16.33 respectively. Instructor hours used per student for VCT and FFT were 0.25 and 0.75, respectively. CONCLUSIONS: VCT provides greater accessibility and resource efficiency compared to FFT, with similar educational benefit. VCT has the potential to improve global availability and accessibility of surgical skills training.

The Radical-Scavenging Activity of a Purified and Sequenced Peptide from Lactic Acid Fermentation of Thunnus albacares By-Products.

Yellowfin tuna by-products (Thunnus albacares) were processed to produce radical-scavenging peptides from hydrolysis by lactic acid fermentation (LAF) with Lactobacillus plantarum, papaya fruit (Carica papaya), and molasses as a carbon source for 72 h. A 15-kDa peptide was purified; after de novo sequencing, it was determined that fragments are rich in hydrophobic and neutral amino acids. The results suggest this effect is mainly to the hydrophobicity of the amino acids in their sequence. Further work is on progress to assess the ability of peptides to provide stability in lipids or in other types of samples sensitive to the action of free radicals.

The Origins and Vulnerabilities of Two Transmissible Cancers in Tasmanian Devils.

Transmissible cancers are clonal lineages that spread through populations via contagious cancer cells. Although rare in nature, two facial tumor clones affect Tasmanian devils. Here we perform comparative genetic and functional characterization of these lineages. The two cancers have similar patterns of mutation and show no evidence of exposure to exogenous mutagens or viruses. Genes encoding PDGF receptors have copy number gains and are present on extrachromosomal double minutes. Drug screening indicates causative roles for receptor tyrosine kinases and sensitivity to inhibitors of DNA repair. Y chromosome loss from a male clone infecting a female host suggests immunoediting. These results imply that Tasmanian devils may have inherent susceptibility to transmissible cancers and present a suite of therapeutic compounds for use in conservation.

Brief history of US debt limits before 1939.

Between 1776 and 1920, the US Congress designed more than 200 distinct securities and stated the maximum amount of each that the Treasury could sell. Between 1917 and 1939, Congress gradually delegated all decisions about designing US debt instruments to the Treasury. In 1939, Congress began imposing a limit on the par value of total federal debt outstanding. By summing Congressional borrowing authorizations outstanding each year for each bond, we construct a time series of implied federal debt limits before 1939.

The impact of glycaemic variability on the surgical patient.

PURPOSE OF REVIEW: Diabetes is the most prevalent long-term metabolic condition and its incidence continues to increase unabated. Patients with diabetes are overrepresented in the surgical population. It has been well recognized that poor perioperative diabetes control is associated with poor surgical outcomes. The outcomes are worst for those people who were not recognized as having hyperglycaemia. RECENT FINDINGS: Recent work has shown that preoperative recognition of diabetes and good communication between the clinical teams at all stages of the patient pathway help to minimize the potential for errors, and improve glycaemic control. The stages of the patient journey start in primary care and end when the patient goes home. The early involvement of the diabetes specialist team is important if the glycated haemoglobin is more than 8.5%, and advice sought if the preoperative assessment team is not familiar with the drug regimens. To date the glycaemic targets for the perioperative period have remained uncertain, but recently a consensus is being reached to ensure glucose levels remain between 108 and180 mg/dl (6.0 and 10.0 mmol/l). There have been a number of ways to achieve these - primarily by manipulating the patients' usual diabetes medications, to also allow day of surgery admission. SUMMARY: glycaemic control remains an important consideration in the surgical patient.

Anti-semaphorin 4D immunotherapy ameliorates neuropathology and some cognitive impairment in the YAC128 mouse model of Huntington disease.

Huntington disease (HD) is an inherited, fatal neurodegenerative disease with no disease-modifying therapy currently available. In addition to characteristic motor deficits and atrophy of the caudate nucleus, signature hallmarks of HD include behavioral abnormalities, immune activation, and cortical and white matter loss. The identification and validation of novel therapeutic targets that contribute to these degenerative cellular processes may lead to new interventions that slow or even halt the course of this insidious disease. Semaphorin 4D (SEMA4D) is a transmembrane signaling molecule that modulates a variety of processes central to neuroinflammation and neurodegeneration including glial cell activation, neuronal growth cone collapse and apoptosis of neural precursors, as well as inhibition of oligodendrocyte migration, differentiation and process formation. Therefore, inhibition of SEMA4D signaling could reduce CNS inflammation, increase neuronal outgrowth and enhance oligodendrocyte maturation, which may be of therapeutic benefit in the treatment of several neurodegenerative diseases, including HD. To that end, we evaluated the preclinical therapeutic efficacy of an anti-SEMA4D monoclonal antibody, which prevents the interaction between SEMA4D and its receptors, in the YAC128 transgenic HD mouse model. Anti-SEMA4D treatment ameliorated neuropathological signatures, including striatal atrophy, cortical atrophy, and corpus callosum atrophy and prevented testicular degeneration in YAC128 mice. In parallel, a subset of behavioral symptoms was improved in anti-SEMA4D treated YAC128 mice, including reduced anxiety-like behavior and rescue of cognitive deficits. There was, however, no discernible effect on motor deficits. The preservation of brain gray and white matter and improvement in behavioral measures in YAC128 mice treated with anti-SEMA4D suggest that this approach could represent a viable therapeutic strategy for the treatment of HD. Importantly, this work provides in vivo demonstration that inhibition of pathways initiated by SEMA4D constitutes a novel approach to moderation of neurodegeneration.

Patient evaluation of early discharge after hip arthroplasty: development of a measure and comparison of three centres with differing durations of stay.

OBJECTIVE: We compared patients' evaluation of care between a surgical unit with a rapid discharge policy and two comparison units to test the hypothesis that the centre with rapid discharge has outcomes that are not inferior to those of the comparison sites. DESIGN: Cross-sectional cohort study. SUBJECTS: Consecutive consenting patients undergoing primary hip arthroplasty during 12 months in: a unit that had reduced postoperative stay to median three days; a specialised orthopaedic surgery treatment centre with median stay of five days; a traditional unit with median stay of six days (N = 316, 125, 119, respectively). METHODS: Six weeks postoperatively, patients completed a specially developed questionnaire measuring their evaluation of care and recovery, together with measures of function and quality of life for validation purposes. RESULTS: Factor analysis of questionnaire responses identified two independent components of patients' evaluation: problems in staff care and problems in physical recovery. Neither component was impaired in the unit with rapid discharge: similar proportions of patients reported recovery problems in each site (odds radios (ORs) for the two comparators versus unit with rapid discharge: 0.96, 1.18); and more patients reported care problems in the two comparator sites (ORs 2.97, 2.16). CONCLUSION: Duration of stay after primary hip arthroplasty can be reduced to three days without intensive pre- or postoperative care, without detriment to patient evaluation.

Different responses of selected hormones to three types of exercise in young men.

Exercise is a potent stimulus for release of growth hormone (GH), cortisol, testosterone and prolactin, and prolonged exercise inhibits insulin secretion. These responses seem to be specific to the type of exercise but this has been poorly characterised primarily because they have not been compared during exercise performed by the same individuals. We investigated hormone responses to resistance, sprint and endurance exercise in young men using a repeated measures design in which each subject served as their own control. Eight healthy non-obese young adults (18-25 years) were studied on four occasions in random order: 30-s cycle ergometer sprint (Sprint), 30-min resistance exercise bout (Resistance), 30-min cycle at 70 % VO(2max) (Endurance), and seated rest in the laboratory (Rest). Cortisol, GH, testosterone, prolactin, insulin and glucose concentrations were measured for 60 min after the four different interventions. Endurance and sprint exercise significantly increased GH, cortisol, prolactin and testosterone. Sprint exercise also increased insulin concentrations, whereas this decreased in response to endurance exercise. Resistance exercise significantly increased only testosterone and glucose. Sprint exercise elicited the largest response per unit of work, but the smallest response relative to mean work rate in all hormones. In conclusion, the nature and magnitude of the hormone response were influenced by exercise type, perhaps reflecting the roles of these hormones in regulating metabolism during and after resistance, sprint and endurance exercise.

Effects of anaesthesia on the inflammatory response to injury.

PURPOSE OF REVIEW: The systemic inflammatory response to injury is essential for wound healing and recovery in concert with other endocrinological, metabolic and immunological changes. However, recent studies suggest that a hyperinflammatory state is associated with adverse perioperative outcomes. Therefore interventions that modulate the inflammatory response, surgical, anaesthetic and pharmacological, may enhance recovery with fewer complications. RECENT FINDINGS: Basic research on wound biology has shown the importance of genetic variability in determining the initial inflammatory response. Clinically, studies of cardiac surgery predominate in which genetic polymorphisms have been shown to result in a hyperinflammatory state. SUMMARY: The use of an interleukin-1 receptor antagonist to control wound pain and limit local inflammation is under consideration. The role of glucocorticoids in obtunding the inflammatory response to injury with improved outcome requires confirmation with better-quality trials. Systemic lidocaine is anti-inflammatory but is effective only in abdominal surgery. NSAIDs are neglected, despite their widespread clinical use and merit detailed investigation.

The consequences of early discharge after hip arthroplasty for patient outcomes and health care costs: comparison of three centres with differing durations of stay.

OBJECTIVE: To compare outcomes from hip arthroplasty between a surgical unit with a rapid discharge policy and two comparison units to test the hypothesis that the centre with rapid discharge has outcomes that are not inferior to the comparison sites. DESIGN: Prospective cohort study. SUBJECTS: Consecutive consenting patients receiving primary hip arthroplasty during 12 months beginning July 2006 in three UK National Health Service surgical units. One has shortened postoperative stay to median three days; one was a new treatment centre with median stay of five days; the third was a traditional unit with median stay of six days (N = 316, 119, 87, respectively). METHODS: Patients were assessed preoperatively and six weeks postoperatively. The primary indicator of function was the Oxford Hip Score. Additional secondary measures included further self-report indicators of function and quality of life and health service costs. RESULTS: Patient outcome in the unit with rapid discharge was not impaired by comparison with the other sites on any measure: Oxford Hip Score decreased from 49 to 27 in the short-stay unit, from 40 to 30 in the treatment centre and from 43 to 32 in the traditional unit. Cost of arthroplasty was least in the short-stay unit, although there was potential for cost savings in each. CONCLUSION: Short postoperative stay after hip arthroplasty can be achieved without intensive patient preparation or post-discharge care and without compromising short-term patient outcome or increasing health care costs. Longer term follow-up is needed.

Early discharge following hip arthroplasty: patients' acceptance masks doubts and concerns.

OBJECTIVE: To describe patients' experience of accelerated discharge after hip arthroplasty in order to test the acceptability to patients of economically driven shortening of post-operative stay. METHODS: Patients (n = 35) who had received primary total hip replacement up to 12 weeks previously were recruited from two UK orthopaedic units, one of which has pioneered short post-operative stay (3-4 days), and another one of which retains a traditional regimen of discharge after 6-7 days. Patients were interviewed about their experience of care, focusing particularly on their views related to length of stay and with particular attention to patients' well-known tendency to mask critical views of their care. Transcripts were analysed thematically to identify the ways that patients evaluated their care and whether these differed between sites. RESULTS: Patients were primarily concerned with how attentive and informative hospital staff had been and did not refer to length of stay spontaneously. When prompted about this, they did not question their discharge time, although those in the more traditional unit could not countenance more rapid discharge. Patients in the unit with accelerated discharge described concerns about the consequences of early discharge for them or their family, particularly managing pain and mobility problems at home and needing more support. CONCLUSIONS: Patients' traditional beliefs about the necessity of prolonged convalescence are not a barrier to early discharge after hip arthroplasty. Nevertheless, some patients' acceptance of early discharge masks doubts and concerns. More intensive post-operative management may be needed if clinical care is not to suffer.

Growth hormone responses to 3 different exercise bouts in 18- to 25- and 40- to 50-year-old men.

Exercise is a potent stimulus for growth hormone (GH) release, although aging appears to attenuate this response. The aim of this study was to investigate GH responses to different exercise stimuli in young and early middle-aged men. Eight men aged 18-25 y and 8 men aged 40-50 y completed 3 trials, at least 7 days apart, in a random order: 30 s cycle-ergometer sprint (sprint), 30 min resistance exercise bout (resistance), 30 min cycle at 70% maximal oxygen consumption (endurance). Blood samples were taken pre-, during, and post-exercise, and area under the GH vs. time curve was calculated for a total of 120 min. Mean blood lactate concentrations and percentage heart rate maximum at which the participants were working were not different between groups in any of the trials. In both groups, blood lactate concentrations were significantly lower in the endurance trial than in the sprint and resistance trials. There were no significant differences in resting GH concentration between groups or trials. GH AUC was significantly greater in the young group than the early middle-aged group, in both sprint (531 (+/-347) vs. 81 (+/-54) microg.L-1 per 120 min, p = 0.003) and endurance trials (842 (+/-616) vs. 177 (+/-137) microg.L-1 per 120 min, p = 0.010). Endurance exercise elicits a greater GH response than sprint and resistance exercise; however, aging per se, factors associated with aging, or an inability to achieve a sufficient absolute exercise intensity results in a smaller GH response to an exercise stimulus in early middle-aged men.