Protein Kinase A Regulates Platelet Phosphodiesterase 3A through an A-Kinase Anchoring Protein Dependent Manner.

Platelet activation is critical for haemostasis, but if unregulated can lead to pathological thrombosis. Endogenous platelet inhibitory mechanisms are mediated by prostacyclin (PGI2)-stimulated cAMP signalling, which is regulated by phosphodiesterase 3A (PDE3A). However, spatiotemporal regulation of PDE3A activity in platelets is unknown. Here, we report that platelets possess multiple PDE3A isoforms with seemingly identical molecular weights (100 kDa). One isoform contained a unique N-terminal sequence that corresponded to PDE3A1 in nucleated cells but with negligible contribution to overall PDE3A activity. The predominant cytosolic PDE3A isoform did not possess the unique N-terminal sequence and accounted for >99% of basal PDE3A activity. PGI2 treatment induced a dose and time-dependent increase in PDE3A phosphorylation which was PKA-dependent and associated with an increase in phosphodiesterase enzymatic activity. The effects of PGI2 on PDE3A were modulated by A-kinase anchoring protein (AKAP) disruptor peptides, suggesting an AKAP-mediated PDE3A signalosome. We identified AKAP7, AKAP9, AKAP12, AKAP13, and moesin expressed in platelets but focussed on AKAP7 as a potential PDE3A binding partner. Using a combination of immunoprecipitation, proximity ligation techniques, and activity assays, we identified a novel PDE3A/PKA RII/AKAP7 signalosome in platelets that integrates propagation and termination of cAMP signalling through coupling of PKA and PDE3A.

Rethinking the 'one-stop' neck lump clinic: a novel pathway beyond coronavirus disease 2019.

OBJECTIVES: UK guidelines advocate 'one-stop' neck lump assessment for cancer referrals. This paper reports the pilot of a novel pre-clinic ultrasound pathway, presents outcomes, and discusses strengths and limitations in the context of the coronavirus disease 2019 pandemic. METHODS: Two-week-wait cancer referral patients with a neck lump were allocated a pre-clinic ultrasound scan followed by a clinic appointment. Demographic, patient journey and outcome data were collected and analysed. RESULTS: Ninety-nine patients underwent ultrasound assessment with or without biopsy on average 8 days following referral. Patients were followed up on average 14.1 days (range, 2-26 days) after initial referral. At the first clinic appointment, 45 patients were discharged, 10 were scheduled for surgery, 12 were diagnosed with cancer, 6 were referred to another specialty and cancer was excluded in 19 patients. Retrospectively, four ultrasounds were performed unnecessarily. CONCLUSION: Pre-clinic ultrasound scanning is an alternative to the one-stop neck lump pathway. This study demonstrates fewer clinic visits, faster diagnosis and a low proportion of unnecessary scans, whilst minimising face-to-face consultations and aerosol-generating procedures.

Molecular Research Comparing the Probabilities of Burkholderia Cepacia Bacterium Diagnosis Procedures.

Burkholderia cepacia is found as part of the B. cepacia complex (Bcc), a collection of highly pathogenic organisms. The Bcc is present almost everywhere in nature; however, it is most prevalent in damp settings, plant roots, and soils. Moreover, Bcc is a major source of morbidity and death in patients due to its high intrinsic antibiotic resistance. The present study aims to isolate and identify gram-negative aerobic bacteria from clinical samples derived from a variety of pathological diseases and investigate the bacterium's virulence factors and genes. The current study included 250 specimens collected from patients suffering from diabetic foot ulcers, urine, burn, wound, sputum, and discharge from the eyes. The samples were collected from both sexes with the age range of 1-75 years. The recorded data showed that males had a higher frequency of infection (79.2%) than females (52%). The results revealed that 7.6% of infected females were between 1-15 years old, while 22% of infected males were aged between 31-45 years. In addition, 26.8% of infected patients (both males and females) were aged between 31-45 years.

Colorectal cancer in patients with SARS-CoV-2: a systematic review and meta-analysis.

BACKGROUND: Patients with colorectal cancer (CRC) are more likely to develop severe course of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection and experience increased risk of mortality compared to SARS-CoV-2 patients without CRC. OBJECTIVES: To estimate the prevalence of SARS-CoV-2 infection in CRC patients and analyse the demographic parameters, clinical characteristics and treatment outcomes in CRC patients with COVID-19 illness. METHODS: For this systematic review and meta-analysis, we searched Proquest, Medline, Embase, Pubmed, CINAHL, Wiley online library, Scopus and Nature for studies on the incidence of SARS-CoV-2 infection in CRC patients, published from December 1, 2019 to December 31, 2021, with English language restriction. Effect sizes of prevalence were pooled with 95% confidence intervals (CIs). Sub-group analyses were performed to minimize heterogeneity. Binary logistic regression model was used to explore the effect of various demographic and clinical characteristics on patient's final treatment outcome (survival or death). RESULTS: Of the 472 papers that were identified, 69 articles were included in the systematic review and meta-analysis (41 cohort, 16 case-report, 9 case-series, 2 cross-sectional, and 1 case-control studies). Studies involving 3362 CRC patients with confirmed SARS-CoV-2 (all patients were adults) were analyzed. The overall pooled proportions of CRC patients who had laboratory-confirmed community-acquired and hospital-acquired SARS-CoV-2 infections were 8.1% (95% CI 6.1 to 10.1, n = 1308, 24 studies, I2 98%, p = 0.66), and 1.5% (95% CI 1.1 to 1.9, n = 472, 27 studies, I2 94%, p < 0.01). The median patient age ranged from 51.6 years to 80 years across studies. The majority of the patients were male (n = 2243, 66.7%) and belonged to White (Caucasian) (n = 262, 7.8%), Hispanic (n = 156, 4.6%) and Asian (n = 153, 4.4%) ethnicity. The main source of SARS-CoV-2 infection in CRC patients was community-acquired (n = 2882, 85.7%; p = 0.014). Most of those SARS-CoV-2 patients had stage III CRC (n = 725, 21.6%; p = 0.036) and were treated mainly with surgical resections (n = 304, 9%) and chemotherapies (n = 187, 5.6%), p = 0.008. The odd ratios of death were significantly high in patients with old age (≥ 60 years) (OR 1.96, 95% CI 0.94-0.96; p < 0.001), male gender (OR 1.44, 95% CI 0.41-0.47; p < 0.001) CRC stage III (OR 1.54, 95% CI 0.02-1.05; p = 0.041), CRC stage IV (OR 1.69, 95% CI 0.17-1.2; p = 0.009), recent active treatment with chemotherapies (OR 1.35, 95% CI 0.5-0.66; p = 0.023) or surgical resections (OR 1.4, 95% CI 0.8-0.73; p = 0.016) and admission to ICU (OR 1.88, 95% CI 0.85-1.12; p < 0.001) compared to those who survived. CONCLUSION: SARS-CoV-2 infection in CRC patient is not uncommon and results in a mortality rate of 26.2%. Key determinants that lead to increased mortality in CRC patients infected with COVID-19 include older age (≥ 60 years old); male gender; Asian and Hispanic ethnicity; if SARS-CoV-2 was acquired from hospital source; advanced CRC (stage III and IV); if patient received chemotherapies or surgical treatment; and if patient was admitted to ICU, ventilated or experienced ARDS.

Inhibition of Arginine Methylation Impairs Platelet Function.

Protein arginine methyltransferases (PRMTs) catalyze the transfer of methyl groups to arginine residues in proteins. PRMT inhibitors are novel, promising drugs against cancer that are currently in clinical trials, which include oral administration of the drugs. However, off-target activities of systemically available PRMT inhibitors have not yet been investigated. In this work, we study the relevance of arginine methylation in platelets and investigate the effect of PRMT inhibitors on platelet function and on the expression of relevant platelet receptors. We show that (1) key platelet proteins are modified by arginine methylation; (2) incubation of human platelets with PRMT inhibitors for 4 h results in impaired capacity of platelets to aggregate in response to thrombin and collagen, with IC50 values in the µM range; and (3) treatment with PRMT inhibitors leads to decreased membrane expression and reduced activation of the critical platelet integrin αIIbß3. Our contribution opens new avenues for research on arginine methylation in platelets, including the repurposing of arginine methylation inhibitors as novel antiplatelet drugs. We also recommend that current and future clinical trials with PRMT inhibitors consider any adverse effects associated with platelet inhibition of these emerging anticancer drugs.

The use of computed tomography as a first-line imaging modality in patients with primary hyperparathyroidism.

BACKGROUND: The success of minimally invasive parathyroidectomy (MIP) relies on accurate localization of the abnormal parathyroid glands. Concordant findings on ultrasound (US) and 99mTc-scintigraphy (sestamibi) are currently considered the 'gold standard'. Computed tomography (CT) has also recently been used in preoperative planning. We sought to assess the accuracy of CT for localization of abnormal parathyroid glands in such patients. METHODS: An audit of 75 patients with primary hyperparathyroidism (PHPT) who underwent neck US and CT between 2017 and 2019 at our center as their first-line imaging. RESULTS: All 75 patients underwent US and CT and 54 (72.0%) also had sestamibi. CT alone identified a potential target in all patients, of which the location was correct in 63 (84.0%). The overall combined sensitivity of US and CT was 88% (95% CI 78-94) and was higher than the combined sensitivity of US and sestamibi (65% [95% CI 53-76]; p < 0.001). Twenty-one patients (28.0%) had an ectopic gland, and the sensitivity of US and CT was 86% (95% CI 64-96) versus US and sestamibi (57% [95% CI 34-77]; p = 0.016). For adenomas < 1.0 g (n = 36; 48%), the accuracy of CT was 81% (95% CI 64-91) compared with 62% (95% CI 44-77) for US and sestamibi (p = 0.04). The correct preoperative diagnosis of multiglandular disease (n = 9; 12%) seemed to be the most difficult, with similar accuracy for US and sestamibi (40% [95% CI 14-73]) and US and CT (50% [95% CI 20-80]) (p > 0.99). CONCLUSION: The combination of US and CT was able to correctly identify the location of the abnormal parathyroid in 88% of patients and, in comparison with US and sestamibi, had better diagnostic accuracy, especially for smaller and ectopic adenomas. This finding suggests that US and CT could be considered as a first-line imaging modality in patients with PHPT considered for MIP.

Coronin 1 Is Required for Integrin ß2 Translocation in Platelets.

Remodeling of the actin cytoskeleton is one of the critical events that allows platelets to undergo morphological and functional changes in response to receptor-mediated signaling cascades. Coronins are a family of evolutionarily conserved proteins implicated in the regulation of the actin cytoskeleton, represented by the abundant coronins 1, 2, and 3 and the less abundant coronin 7 in platelets, but their functions in these cells are poorly understood. A recent report revealed impaired agonist-induced actin polymerization and cofilin phosphoregulation and altered thrombus formation in vivo as salient phenotypes in the absence of an overt hemostasis defect in vivo in a knockout mouse model of coronin 1. Here we show that the absence of coronin 1 is associated with impaired translocation of integrin ß2 to the platelet surface upon stimulation with thrombin while morphological and functional alterations, including defects in Arp2/3 complex localization and cAMP-dependent signaling, are absent. Our results suggest a large extent of functional overlap among coronins 1, 2, and 3 in platelets, while aspects like integrin ß2 translocation are specifically or predominantly dependent on coronin 1.

A Transoral Excision of a Deep Parotid Lobe Lesion Using Ultrasound-Guided Wire Localization: A Multi-disciplinary Team Approach.

PURPOSE: Removing deep parotid lobe lesions often requires a mandibulotomy procedure which carries with it a risk of significant morbidity. A transoral approach may help mitigate against complications but is technically demanding due to limited access in an area with a close relation to the internal carotid artery. METHODS: We describe a multi-disciplinary approach with the use of intraoral ultrasound-guided wire localization of a low-grade acinic cell carcinoma located in the deep lobe of the right parotid gland. RESULTS: Our multi-disciplinary approach facilitated the complete removal of this deep lobe parotid lesion via a transoral approach with minimal post-operative sequelae or complications. CONCLUSION: In selected cases, with appropriate expertise, this dynamic approach can potentially be used even for malignant disease to limit post-operative morbidity when managing small deep parotid lobe lesions.

Alterations in Platelet Alpha-Granule Secretion and Adhesion on Collagen under Flow in Mice Lacking the Atypical Rho GTPase RhoBTB3.

Typical Rho GTPases, such as Rac1, Cdc42, and RhoA, act as molecular switches regulating various aspects of platelet cytoskeleton reorganization. The loss of these enzymes results in reduced platelet functionality. Atypical Rho GTPases of the RhoBTB subfamily are characterized by divergent domain architecture. One family member, RhoBTB3, is expressed in platelets, but its function is unclear. In the present study we examined the role of RhoBTB3 in platelet function using a knockout mouse model. We found the platelet count, size, numbers of both alpha and dense granules, and surface receptor profile in these mice were comparable to wild-type mice. Deletion of Rhobtb3 had no effect on aggregation and dense granule secretion in response to a range of agonists including thrombin, collagen, and adenosine diphosphate (ADP). By contrast, alpha-granule secretion increased in mice lacking RhoBTB3 in response to thrombin, collagen related peptide (CRP) and U46619/ADP. Integrin activation and spreading on fibrinogen and collagen under static conditions were also unimpaired; however, we observed reduced platelet accrual on collagen under flow conditions. These defects did not translate into alterations in tail bleeding time. We conclude that genetic deletion of Rhobtb3 leads to subtle alterations in alpha-granule secretion and adhesion to collagen without significant effects on hemostasis in vivo.

Mini Dental Implants in the Management of The Atrophic Maxilla and Mandible: A New Implant Design and Preliminary Results.

Although the edentulous population in the UK is falling, those that are rendered edentulous are becoming edentate later in life and with significantly resorbed ridges. This creates a challenge because the management of such patients and their ability to adapt to new dentures is impaired later in life. Despite widespread endorsement of two implants to retain lower complete dentures, the inability to comply has resulted in elderly patients with compromised ability to function and unable to eat a healthy diet. Mini dental implants may offer an ideal solution for the elderly edentulous population who may not be keen on invasive surgery for the placement of conventional dental implants. Further work is required to show the longevity of these restorations, however, existing research and clinical experience show that they potentially offer a simple solution to this group of patients. This paper presents the development of a new design of mini implant, based on clinical problems encountered during a pilot randomised controlled trial. The design of the new implant specifically aims to overcome problems in managing severely atrophic ridges. A preliminary survival study shows survival rates to be equivalent to other mini dental implants and highly satisfactory in the short to medium term.

Bringing SASI back: Single session selective arterial secretin injection and transarterial embolization of intrahepatic pancreatic neuroendocrine metastasis in a MEN-1 patient.

SASI (selective arterial secretin injection) is a form of ASVS (arterial stimulation and venous sampling) used to localize pancreatic gastrinomas. This report aims to review the protocol for SASI and demonstrate its utility in localizing functional and nonfunctional gastrinomas. Even if a patient has a pancreatic mass and a laboratory profile fitting a specific endocrine syndrome, these may or may not be associated as has been previously demonstrated with adrenal vein sampling. We present a case where a patient underwent simultaneous SASI and bland embolization of a hepatic metastasis to facilitate partial pancreatectomy for Zollinger-Ellison syndrome.

Percutaneous transgastrostomic interventional radiology-operated endoscopy facilitates foreign body removal using rigid endobronchial forceps.

Retained foreign bodies within the stomach and proximal small bowel may be problematic in patients with prior cerebrovascular injury or head, neck and esophageal malignancy, given the increased vulnerability of this patient population to complications from aspiration and increased difficulty of esophagogastroduodenoscopy in cases of tumoral obstruction. This article presents an alternative method for foreign body retrieval through an existing gastrostomy tract, which offers the benefits of fast procedure times, reduction in radiation dose and fluoroscopy time, and allows for safer retrieval of foreign bodies by using direct visualization. This technique may be performed entirely by interventional radiologists.

A qualitative analysis among regular waterpipe tobacco smokers in London universities.

BACKGROUND: Waterpipe smoking is popular in the West despite the significant harm associated with its use. Little is known about the habits and practices of British smokers. DESIGN: A total of 32 regular waterpipe smokers attended focus groups, where trained facilitators explored knowledge, belief and attitudes to waterpipes. Transcripts were thematically analysed and grouped into seven broad categories. RESULTS: Waterpipes played a clearly defined social role, accentuated by reduced harm perception and greater social acceptance than cigarettes. Knowledge about waterpipes was lacking, partly due to the lack of media attention. Addiction was described as a 'social addiction', although this may mask true nicotine addiction. Waterpipe smokers were less interested in quitting due to intermittent smoking patterns, reinforcing the belief of reduced exposure to harm. Legislative issues were explored, including the need to legislate on waterpipes on a par with cigarettes. CONCLUSION: Waterpipe smokers exhibited specific habits and attitudes not traditionally seen in cigarette smokers. It is important to encourage education to dispel the myths surrounding reduced harm perception and addiction to tackle its strong social acceptance. Legislation should play a more active role in prevention and education, and more emphasis should be placed on gathering nationwide epidemiological data to gauge the potential for escalation.

The power of words: sources of anxiety in patients undergoing local anaesthetic plastic surgery.

INTRODUCTION: With local or regional anaesthesia being employed for more as well as more complex surgical procedures, an increasing number of patients remain fully conscious during their operation. This is generally perceived as being advantageous to the patient as less time is spent in hospital and the side effects of general anaesthesia are avoided. However, there is no direct measure of the patient experience during 'awake surgery', in particular of which aspects of the process may be distressing. METHODS: Seventy patients undergoing day case plastic surgery under local anaesthesia were asked to complete a short questionnaire immediately following their operation. This was designed to identify specific factors likely to either increase or reduce anxiety during surgery. The questionnaire was initially validated on a pilot group of ten patients. RESULTS: Unsurprisingly, painful stimuli such as injections were identified as potential stressors. More interestingly, the data highlighted that some commonly used surgical terms such as 'knife' and 'scalpel' provoke considerable anxiety in the conscious patient. This varied according to age and sex with younger and female patients being most vulnerable. Other events identified as potential stressors, such as casual conversations and movements among theatre staff, were actually shown to be non-stressful and, in some cases, stress relieving. CONCLUSIONS: Technical jargon used by surgical staff can elevate anxiety levels among patients who are awake for their operation. Careful consideration of the words we use may reduce this, particularly in female patients.

Hospital outcome of coronary artery bypass grafting and coronary endarterectomy.

To compare hospital mortality and postoperative complications in patients with severe coronary artery disease undergoing coronary artery bypass grafting with or without coronary endarterectomy, 100 consecutive patients were divided into 2 groups of 50 patients each. Group 1 had coronary endarterectomy, and group 2 had no coronary endarterectomy. There were 87 men and 13 women, with a mean age of 54.79 +/-7.8 years; 48% had stable angina. The right coronary artery was endarterectomized in 22 patients, the left anterior descending artery in 21, and the left circumflex in 7. There were no significant differences in outcomes. There was 1 hospital death in each group. Perioperative myocardial infarction occurred in 2 patients in group 1 and 1 in group 2. Endarterectomy is a suitable option to achieve complete revascularization in patients with refractory angina and severe diffuse disease.

Inspired nitric oxide and modulation of oxidative stress during cardiac surgery.

Evidence in the literature is contradictory regarding the precise role of nitric oxide (NO) in modulating systemic inflammatory response induced by cardiopulmonary bypass (CPB). We studied the impact of inspired NO gas on physiological function and markers of inflammation-oxidative stress for subjects (n=15, age 62+/-4.5 and 12/3 M/F) scheduled for coronary artery bypass graft (CABG) operation. Outcomes from subjects that received 5 ppm and 20 ppm of inspired NO (n=5/group) were compared to those not given NO gas. Breath-to-breath measurement commenced at the start of intubation and continued up to 4h later. Indices of cardiovascular function, alveolar-capillary gas exchange and haematological parameters were not significantly different in outcomes for the inspired NO groups as compared with control. We observed a reduction in mean systemic arterial in all subjects at 30 min and 4h after bypass when compared with pre bypass values. Markers of systemic inflammatory response and oxidative stress increased during CPB particularly at 4h and 24h after the initiation of bypass. In contrast, we observed a reduction in expired NO, at 24h after surgery in the groups given inspired NO. In addition, there was also a significant reduction in oxidative stress markers in blood at 24h after surgery for the groups given inspired NO as compared with the control group. In contrast, cytokines response remained similar in all the three groups at all time points. The results suggested that inspired NO gas has an antioxidant property that reduces the levels of cell death, and is not associated with significantly worse-off physiological outcomes.

Coronary heart disease service framework for revascularization in Pakistan.

Many centers across the country have used collaborative techniques to identify problems and come up with innovative solutions. Excellent improvements have been made in every aspect of the patient's journey through the cardiac surgery services, such as decreased length of stay, reduced clinical variation and costs, and improved outcome. We looked at how the cardiac surgical team at our center is helping to improve services for patients undergoing coronary revascularization. Improvements are not just focused on waiting lists or operating rooms but reflect the wider experience of patients and their families.