Identification and characterisation of functional Kir6.1-containing ATP-sensitive potassium channels in the cardiac ventricular sarcolemmal membrane.

BACKGROUND AND PURPOSE: The canonical Kir6.2/SUR2A ventricular KATP channel is highly ATP-sensitive and remains closed under normal physiological conditions. These channels activate only when prolonged metabolic compromise causes significant ATP depletion and then shortens the action potential to reduce contractile activity. Pharmacological activation of KATP channels is cardioprotective, but physiologically, it is difficult to understand how these channels protect the heart if they only open under extreme metabolic stress. The presence of a second KATP channel population could help explain this. Here, we characterise the biophysical and pharmacological behaviours of a constitutively active Kir6.1-containing KATP channel in ventricular cardiomyocytes. EXPERIMENTAL APPROACH: Patch-clamp recordings from rat ventricular myocytes in combination with well-defined pharmacological modulators was used to characterise these newly identified K+ channels. Action potential recording, calcium (Fluo-4) fluorescence measurements and video edge detection of contractile function were used to assess functional consequences of channel modulation. KEY RESULTS: Our data show a ventricular K+ conductance whose biophysical characteristics and response to pharmacological modulation were consistent with Kir6.1-containing channels. These Kir6.1-containing channels lack the ATP-sensitivity of the canonical channels and are constitutively active. CONCLUSION AND IMPLICATIONS: We conclude there are two functionally distinct populations of ventricular KATP channels: constitutively active Kir6.1-containing channels that play an important role in fine-tuning the action potential and Kir6.2/SUR2A channels that activate with prolonged ischaemia to impart late-stage protection against catastrophic ATP depletion. Further research is required to determine whether Kir6.1 is an overlooked target in Comprehensive in vitro Proarrhythmia Assay (CiPA) cardiac safety screens.

Robotic assisted surgery reduces ergonomic risk during minimally invasive colorectal resection: the VOLCANO randomised controlled trial.

PURPOSE: Minimally invasive surgery benefits patients but poor operating ergonomics causes musculoskeletal injuries in surgeons. This randomised controlled trial aims to assess whether robotic-assisted surgery with the open-console Versius® system can reduce surgeons' ergonomic risks during major colorectal resections. METHODS: Prospectively registered at ClinicalTrials.gov (NCT05262296) in March 2022. Adult patients requiring a minimally invasive colorectal resection were potentially eligible. Photographs taken at 2-min intervals were analysed using the objective Rapid Entire Body Assessment (REBA) posture analysis scale to calculate intraoperative surgeon ergonomic risk. Secondary outcomes included team communication (Oxford NOTECHS II), surgeon cognitive strain (modified NASA-TLX scale), and clinical outcomes. RESULTS: Sixty patients were randomised in a 2:1 ratio (40 robot, 20 laparoscopic). Mean age was 65yrs and 34 (57%) were male. Body Mass Index did not differ between the 2 groups (overall mean 29.0 ± 5) and there were equal proportions of left and right-colonic resections. REBA was significantly lower in the robotic arm (median robot REBA score 3 vs lap REBA 5 [p < 0.001]), equating to an injury risk category drop from "medium" to "low risk". There were no significant differences in team communication, operative duration, or patient outcomes. Surgeon cognitive strain was lower in robotic cases (mean robot 32.4 ± 10.3 vs lap 45.6 ± 14.3 [p < 0.001]). CONCLUSIONS: This trial demonstrates that robotic surgery with an open-console system reduces ergonomic risk scores and cognitive strain during colorectal resections, with no apparent detriment to team communication. This may therefore be a safe & feasible solution to the increasing problem of work-related musculoskeletal injuries in surgeons.

A Study to Assess Biochemical Profile of Tobacco Chewers with Chewing Habit Over a Period of 10 Years.

The study was conducted to evaluate the systemic adverse effects of smokeless tobacco (SLT) on hematological as well as other biochemical parameters and find out if any correlation between them which may be worthy of creating awareness among the masses against its usage. The current observational study was carried out among 250 tobacco chewers, and complete hemogram, LFT profile, and electrolyte profile were studied. The mean values of hemoglobin (Hb) gram % was 12.74 ± 1.6, total leucocyte count (TLC/cu mm) is 6608.33 ± 1752.083, and platelet count lakh cell/cu mm is 2.55 ±0.806. The mean values of Na+ mmol/1 of the S. electrolyte profile was 132 ± 18.05 and K+ mmol/1 was 3.89 ± 0.538. The study deliberates imperative perception into smokeless tobacco-mediated effects on body systemic functions and reports a crucial part into SLT-mediated effects on biochemical profile and metabolism which can be revealed in promoting tobacco cessation.

Slowly activating voltage-gated potassium current potentiation by ML277 is a novel cardioprotective intervention.

Cardiovascular disease is thought to account for nearly a third of deaths worldwide, with ischemic heart disease, including acute coronary syndromes such as myocardial infarction, accounting for 1.7 million deaths per year. There is a clear need for interventions to impart cardioprotection against ischemia. Here, we show that the slowly activating voltage-gated potassium current (IKs) potentiator ML277 imparts cardioprotection against ischemia in cellular and whole-heart models by modulating the action potential duration. In three different metabolic inhibition and reperfusion models, an increased contractile recovery and cell survival was observed with ML277, indicative of protection. Finally, ML277 reduced infarct size in an ex vivo Langendorff coronary ligation model, including if only applied on reperfusion. In conclusion, potentiation of the IKs with ML277 imparted a cardioprotection that was equivalent to the protection reported previously by ischemic preconditioning. These data suggest that IKs potentiation may be therapeutically useful in acute coronary syndromes.

Mitigation of Cardiovascular Disease and Toxicity through NRF2 Signalling.

Cardiovascular toxicity and diseases are phenomena that have a vastly detrimental impact on morbidity and mortality. The pathophysiology driving the development of these conditions is multifactorial but commonly includes the perturbance of reactive oxygen species (ROS) signalling, iron homeostasis and mitochondrial bioenergetics. The transcription factor nuclear factor erythroid 2 (NFE2)-related factor 2 (NRF2), a master regulator of cytoprotective responses, drives the expression of genes that provide resistance to oxidative, electrophilic and xenobiotic stresses. Recent research has suggested that stimulation of the NRF2 signalling pathway can alleviate cardiotoxicity and hallmarks of cardiovascular disease progression. However, dysregulation of NRF2 dynamic responses can be severely impacted by ageing processes and off-target toxicity from clinical medicines including anthracycline chemotherapeutics, rendering cells of the cardiovascular system susceptible to toxicity and subsequent tissue dysfunction. This review addresses the current understanding of NRF2 mechanisms under homeostatic and cardiovascular pathophysiological conditions within the context of wider implications for this diverse transcription factor.

In vivo and in vitro validation of powdery mildew resistance in garden pea genotypes.

Powdery mildew is one of the serious diseases of garden pea which causes a large number of yield losses. Genetic resistance is quite effective, being cost-effective and environment friendly than fungicide applications. In the present studies an initial attempt has been made to identify resistant genotypes against powdery mildew disease developed from hybridization followed by validation of the disease. The experimental material comprised of 48 genotypes that includes 44 advanced breeding lines was evaluated for powdery mildew incidence in Randomized Complete Block Design with three replications at two locations under field conditions [Palampur (winter 2017-18 and 2018-19) and Kukumseri (summer 2018)] and in vitro at Palampur [detached leaf method and polyhouse conditions]. Ten lines viz., SP7, SN-1, SN-6-1, SN-7-1, SN-2, SN-5-2, SN-6-2, SN-10, SN-21 and SP-281 showed resistant reaction along with check Palam Sumool while 27 lines were identified as moderately resistant in comparison to susceptible check Azad P-1. Besides, six lines namely, SP-2, SP-5, SP-10, SP-24, SA-4 and SP-12-1 gave moderately susceptible reaction along with checks Pb-89 and Palam Priya. Only, SP-19 was categorized as susceptible. The high yielding lines SP-3, SP-6 and SP-22 showed moderately resistant reaction in both natural and artificial conditions. Validation of resistance using molecular markers revealed that neither the parental genotypes nor the progenies possess the er1 gene of JI1559. The er2 linked marker ScOPX-171700 was polymorphic between Palam Sumool and Palam Priya but the marker didn't show polymorphism between er2 harboring line (JI2480). These results suggested that the lines showing resistance under field conditions may have some other genes or alleles for resistance and further confirmation is needed by developing mapping populations with specific gene or gene combinations.

Implementation of robotic hernia surgery using the Versius® system.

This case series aims to demonstrate that hernia surgery is safe and feasible using the Versius® robotic system from CMR Surgical, and to describe the surgical techniques used. It is the first series published using this novel system. Forty-one consecutive hernia repair cases were completed using Versius®, including inguinal and ventral hernias. Data were collected prospectively on a number of pre-, peri-, and postoperative outcomes. Techniques are described for robotic transabdominal preperitoneal repair of inguinal hernia, and intraperitoneal onlay mesh repair of ventral hernia. Thirty-two inguinal and nine ventral hernia repairs were performed over a 12-month period. The population were 88% male with a mean body mass index of 27.4 ± 3.5. There were no conversions to open surgery. Median length of stay was 0 days. Six patients (15%) experienced urinary retention, and there were 2 further minor complications with no major complications, readmissions or reoperations. Use of the Versius® system for robotic hernia surgery is safe, with comparable results to existing robotic systems. Implementation is possible with minimal changes to established surgical techniques.

Morpho-molecular genetic diversity and population structure analysis in garden pea (Pisum sativum L.) genotypes using simple sequence repeat markers.

Garden pea (Pisum sativum L.) is a self-pollinated plant species which played an important role for the foundation of modern genetics. Genetic diversity among 56 garden pea genotypes was assessed using 12 morphological descriptors, 19 quantitative traits and 8 simple sequence repeat (SSR) markers. Eight morphological descriptors were found polymorphic, and highest Shannon diversity index was recorded for pod curvature (1.18). Mahalanobis D2 illustrating genetic divergence arranged 56 genotypes into six clusters, with the highest inter-cluster distance between clusters IV and VI (18.09). The average values of Na (number of alleles), Ne (effective number of alleles), I (Shannon's Information index), PIC (polymorphism information content), Ho (observed heterozygosity) and He (expected heterozygosity) were 3.13, 1.85, 0.71, 0.36, 0.002 and 0.41, respectively. Pair wise genetic distance among all pairs of the genotypes varied from 0.33 to 1.00 with an average of 0.76. Based on genetic distance, the genotypes were classified into two main clusters (A and B) by cluster analysis, whereas structure analysis divided the genotypes into four sub-populations. The SSR makers indicated that present of genetic variability among the studied genotypes. When, we compared the groups formed by agro-morphological and molecular data, no genotypes were observed, indicating that both stages of characterization are crucial for a better understanding of the genetic variability. Hybridization between genetically diverse genotypes can be exploited to expend the genetic variability and introduce new traits in the pea breeding program.

Selective protein kinase C inhibition switches time-dependent glucose cardiotoxicity to cardioprotection.

Hyperglycaemia at the time of myocardial infarction has an adverse effect on prognosis irrespective of a prior diagnosis of diabetes, suggesting glucose is the damaging factor. In ex vivo models of ischaemia, we demonstrated that deleterious effects of acutely elevated glucose are PKCα/ß-dependent, and providing PKCα/ß are inhibited, elevated glucose confers cardioprotection. Short pre-treatments with high glucose were used to investigate time-dependent glucose cardiotoxicity, with PKCα/ß inhibition investigated as a potential mechanism to reverse the toxicity. Freshly isolated non-diabetic rat cardiomyocytes were exposed to elevated glucose to investigate the time-dependence toxic effects. High glucose challenge for >7.5 min was cardiotoxic, proarrhythmic and lead to contractile failure, whilst cardiomyocytes exposed to metabolic inhibition following 5-min high glucose, displayed a time-dependent protection lasting ∼15 min. This protection was further enhanced with PKCα/ß inhibition. Cardioprotection was measured as a delay in contractile failure and KATP channel activation, improved contractile and Ca2+ transient recovery and increased cell survival. Finally, the effects of pre-ischaemic treatment with high glucose in a whole-heart coronary ligation protocol, where protection was evident with PKCα/ß inhibition. Selective PKCα/ß inhibition enhances protection suggesting glycaemic control with PKC inhibition as a potential cardioprotective therapeutics in myocardial infarction and elective cardiac surgery.

Comparative Evaluation of Suture Versus Sutureless Surgery in Mandibular Third Molar Impactions.

Purpose: The purpose of this study is to compare the suture versus sutureless surgery in impacted mandibular third molar and to evaluate the morbidity and complications associated with each technique. Materials and Methods: A total of 50 patients with asymptomatic impacted mandibular third molars were randomly divided into two groups of 25 patients each. Radiographs were taken to assess the angulation and degree of eruption in the third molar. A small modified Szmyd, V-shaped flap was raised in all cases, and teeth were extracted. In Group I-Suture group (suture was used to close the flap), and in Group II-Sutureless group (no suture used to close the flap). The post-operative pain, swelling, trismus, haemorrhage, periodontal pocket, and alveolar osteitis were evaluated at 24 h, 48 h, 5th days, 7th days, and 2 weeks after surgery. The statistical analysis was done using the Chi-square "t" test and Independent Samples "t" test. Observations and Results: Pain, swelling, and trismus were found to be significantly reduced especially in the immediate post-operative period in the sutureless group as compared to the suture group (p < 0.001). There were no incidences of intra-operative and post-operative haemorrhage in any case. Follow-up of all the patients showed that there was no difference in periodontal sequelae and alveolar osteitis. Conclusions: Sutureless surgery with small flap was found to be less invasive, time-saving, and also a cost-effective method. This technique significantly reduced the early crucial phase of patient discomfort and demonstrates good results.

Initiation and feasibility of a multi-specialty minimally invasive surgical programme using a novel robotic system: A case series.

INTRODUCTION: There are a number of small case series examining new robotic surgical systems, but this is the first large case series assessing the feasibility of the Versius® system from CMR Surgical (1 Evolution Business Park, Cambridge, UK) in a multi-specialty setting. MATERIALS AND METHODS: All patients undergoing Versius®-assisted surgery in a previously robot-naïve centre were consented for collection of data on demographics, pre-, intra-, and postoperative outcomes. Data collection was performed prospectively from the start of the robotic surgical programme. RESULTS: 160 operations were performed over a 19-month period, including 68 colorectal, 60 gynaecology, and 32 general surgery cases. The conversion rate to open surgery was 4.4% for colorectal, and 0% for gynaecology and general surgery. Median length of stay was 6 days for colorectal, 1 day for gynaecology, and 0 days for general surgery. Other outcomes were comparable to existing literature for robotic assisted surgery. CONCLUSION: The Versius® system is safe and feasible for use in a multi-specialty minimally invasive surgery programme, including colorectal, general surgical & gynaecological cases, and operative volume can be safely and easily scaled up in a district general hospital setting without prior robotic surgical experience.

Characterization of a high throughput human stem cell cardiomyocyte assay to predict drug-induced changes in clinical electrocardiogram parameters.

Human induced pluripotent stem cell derived cardiomyocytes (hIPSC-CM's) play an increasingly important role in the safety profiling of candidate drugs. For such models to have utility a clear understanding of clinical translation is required. In the present study we examined the ability of our hIPSC-CM model to predict the clinically observed effects of a diverse set of compounds on several electrocardiogram endpoints, including changes in QT and QRS intervals. To achieve this, compounds were profiled in a novel high throughput voltage-sensitive dye platform. Measurements were taken acutely (30 min) and chronically (24 h) to ensure that responses from compounds with slow onset kinetics or that affected surface ion channel expression would be captured. In addition, to avoid issues associated with changes in free drug levels due to protein binding, assays were run in serum free conditions. Changes in hIPSC-CM threshold APD90 values correlated with compound plasma exposures that produced a +10 ms change in clinical QTc (Pearson r2 = 0.80). In addition, randomForest modeling showed high predictivity in defining TdP risk (AUROC value = 0.938). Risk associated with QRS prolongation correlated with an increase in action potential rise-time (AUROC value = 0.982). The in-depth understanding of the clinical translatability of our hIPSC-CM model positions this assay to play a key role in defining cardiac risk early in drug development. Moreover, the ability to perform longer term studies enables the detection of compounds that may not be highlighted by more acute assay formats, such as inhibitors of hERG trafficking.

Platelet-Rich Fibrin as an Aid to Soft- and Hard-Tissue Healing.

INTRODUCTION: Contemporary published data present confounding results on use of PRF in soft- and hard-tissue healing in the oral cavity, and many authors have suggested for further studies to reach the definitive conclusion. AIM: Our main objective therefore was to evaluate soft-tissue healing and osseous regeneration (by using VIXWIN PRO software) in extraction sites of mandibular third molars with substantial sample size to understand the effect of PRF in bony defects. METHODOLOGY: Sixty patients had their bilaterally impacted third molars (120 sites) extracted in the split mouth study, following which platelet-rich fibrin was placed in one of the sockets. Patients were followed up clinically and radiographically, and pain score, presence of infection, exudation of graft and VIXWIN PRO software were used to evaluate healing of soft tissue and bone. RESULT AND CONCLUSION: Our study advocates the use of PRF for enhanced soft- and hard-tissue healing. Though the osseous regeneration could be differentiated in both the groups at second month interval only, pain scores were better with PRF at most instances. Subsequent phase to the research should include histopathological investigations for ancillary support.

Comparative Analysis of the Anesthetic Efficacy of 0.5% Ropivacaine Versus 2% Lignocaine Hydrochloride with Adrenaline (1:80,000) for Inferior Alveolar Nerve Block in Surgical Removal of Impacted Mandibular Third Molars.

OBJECTIVE: To compare the onset, duration and depth of anesthesia, postoperative pain, duration of analgesia and adverse reaction between 0.5% ropivacaine and 2% Lignocaine hydrochloride with adrenaline (1:80,000) in 40 patients having bilaterally impacted mandibular third molars. PATIENTS AND METHODS: A prospective, randomized, single blind study was carried out among 40 patients requiring surgical removal of bilaterally impacted mandibular third molars having similar "difficulty index." The onset of action, duration and depth of anesthesia, duration of analgesia, postoperative pain and adverse reactions of 0.5% ropivacaine and 2% lignocaine hydrochloride with 1:80,000 adrenaline were evaluated. All patients were infiltrated intradermally with 0.5 ml of 0.5% ropivacaine as test dose to rule out any allergic reaction. The surgical extractions of the impacted third molars were done using the standard surgical procedure. RESULTS: 0.5% Ropivacaine had higher depth of anesthesia, longer duration of action and postoperative analgesic effect than 2% Lignocaine hydrochloride with 1:80,000 adrenaline with no adverse effects. CONCLUSION: 0.5% ropivacaine is safe, efficacious, clinically acceptable and equally potent local anesthetic agent when compared to 2% lignocaine with 1:80,000 adrenaline in oral and maxillofacial surgery for longer duration of surgeries.

Correction to: Evaluation of the Variations in the Levels of Perioperative Inflammatory Markers After Open Reduction and Internal Fixation of Maxillofacial Fractures.

[This corrects the article DOI: 10.1007/s12663-018-1174-4.].

Drugs in phase I and II clinical development for the prevention of stroke in patients with atrial fibrillation.

INTRODUCTION: Atrial fibrillation is the most frequently diagnosed cardiac arrhythmia globally and is associated with ischemic stroke and heart failure. Patients with atrial fibrillation are typically prescribed long-term anticoagulants in the form of either vitamin K antagonists or non-vitamin K antagonist oral anticoagulants; however, both carry a potential risk of adverse bleeding. AREAS COVERED: This paper sheds light on emerging anticoagulant agents which target clotting factors XI and XII, or their activated forms - XIa and XIIa, respectively, within the intrinsic coagulation pathway. The authors examined data available on PubMed, Scopus, and the clinical trials registry of the United States National Library of Medicine (www.clinicaltrials.gov). EXPERT OPINION: Therapies targeting factors XI or XII can yield anticoagulant efficacy with the potential to reduce adverse bleeding. Advantages for targeting factor XI or XII include a wider therapeutic window and reduced bleeding. Long-term follow-up studies and a greater understanding of the safety and efficacy are required. Atrial fibrillation is a chronic disease and therefore the development of oral formulations is key.

Evaluation of the Variations in the Levels of Perioperative Inflammatory Markers After Open Reduction and Internal Fixation of Maxillofacial Fractures.

INTRODUCTION: Postoperative infectious complications are commonly encountered in open reduction and internal fixation (ORIF) of maxillofacial fractures. An early diagnosis of infectious processes is the key in preventing morbidity/mortality which could be in the form of loss of hardware and sepsis. To prevent these, various markers of inflammation have been studied in different disciplines of surgery but are found scarce in maxillofacial practice. MATERIAL AND METHOD: The present study was designed to evaluate the perioperative variations in the levels of inflammatory markers. We analysed temperature, TLC, DLC, ALT, AST and CRP in 50 patients of ORIF. Their values were recorded preoperatively as well as at 24 h, 48 h, third day and seventh day postoperatively. The correlation of inflammatory markers with the type of anaesthesia and length of surgery were also analysed. RESULTS: The ranges of various markers in the perioperative phase were: temperature (97.6 ºF-99.2 ºF), TLC (5100/mm3-18200/mm3), neutrophils (51-91%), AST (12-86 IU/L), ALT (12-96 IU/L) and CRP (1.2-150 mg/L). Mean values of all the inflammatory markers achieved their peak values within 24 h postoperatively. These values showed a decline thereafter, with the day 3 and day 7 values being even lower than their preoperative values. This fall in the values was highly significant (p < 0.001) except ALT where the fall was significant (p < 0.05). The data obtained could be used as a reference range by the surgeons for monitoring the recovery of the patient. It could also help in timely interception and expeditious management of an infectious episode in the postoperative phase.

Microbial and Enzymatic Degradation of Synthetic Plastics.

Synthetic plastics are pivotal in our current lifestyle and therefore, its accumulation is a major concern for environment and human health. Petroleum-derived (petro-)polymers such as polyethylene (PE), polyethylene terephthalate (PET), polyurethane (PU), polystyrene (PS), polypropylene (PP), and polyvinyl chloride (PVC) are extremely recalcitrant to natural biodegradation pathways. Some microorganisms with the ability to degrade petro-polymers under in vitro conditions have been isolated and characterized. In some cases, the enzymes expressed by these microbes have been cloned and sequenced. The rate of polymer biodegradation depends on several factors including chemical structures, molecular weights, and degrees of crystallinity. Polymers are large molecules having both regular crystals (crystalline region) and irregular groups (amorphous region), where the latter provides polymers with flexibility. Highly crystalline polymers like polyethylene (95%), are rigid with a low capacity to resist impacts. PET-based plastics possess a high degree of crystallinity (30-50%), which is one of the principal reasons for their low rate of microbial degradation, which is projected to take more than 50 years for complete degraded in the natural environment, and hundreds of years if discarded into the oceans, due to their lower temperature and oxygen availability. The enzymatic degradation occurs in two stages: adsorption of enzymes on the polymer surface, followed by hydro-peroxidation/hydrolysis of the bonds. The sources of plastic-degrading enzymes can be found in microorganisms from various environments as well as digestive intestine of some invertebrates. Microbial and enzymatic degradation of waste petro-plastics is a promising strategy for depolymerization of waste petro-plastics into polymer monomers for recycling, or to covert waste plastics into higher value bioproducts, such as biodegradable polymers via mineralization. The objective of this review is to outline the advances made in the microbial degradation of synthetic plastics and, overview the enzymes involved in biodegradation.