Sensitive SARS-CoV-2 detection, air travel Covid-19 testing, variant determination and fast direct PCR detection, using ddPCR and RT-qPCR methods.

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) monitoring in air traffic is important in the prevention of the virus spreading from abroad. The gold standard for SARS-CoV-2 detection is RT-qPCR; however, for early and low viral load detection, a much more sensitive method, such as droplet digital PCR (ddPCR), is required. Our first step was to developed both, ddPCR and RT-qPCR methods, for sensitive SARS-CoV-2 detection. Analysis of ten swab/saliva samples of five Covid-19 patients in different stages of disease showed positivity in 6/10 samples with RT-qPCR and 9/10 with ddPCR. We also used our RT-qPCR method for SARS-CoV-2 detection without the need of RNA extraction, obtaining results in 90-120 minutes. We analyzed 116 self-collected saliva samples from passengers and airport staff arriving from abroad. All samples were negative by RT-qPCR, while 1 was positive, using ddPCR. Lastly, we developed ddPCR assays for SARS-CoV-2 variants identification (alpha, beta, gamma, delta/kappa) that are more economically advantageous when compared to NGS. Our findings demonstrated that saliva samples can be stored at ambient temperature, as we did not observe any significant difference between a fresh sample and the same sample after 24 hours (p = 0.23), hence, saliva collection is the optimal route for sampling airplane passengers. Our results also showed that droplet digital PCR is a more suitable method for detecting virus from saliva, compared to RT-qPCR. Keywords: COVID-19; RT-PCR; ddPCR; SARS-CoV-2; nasopharyngeal swab; saliva.

Natural Taxanes: From Plant Composition to Human Pharmacology and Toxicity.

Biologically active taxanes, present in small- to medium-sized evergreen conifers of various Taxus species, are widely used for their antioxidant, antimicrobial and anti-inflammatory effects, but mostly for their antitumour effects used in the treatment of solid tumours of the breast, ovary, lung, bladder, prostate, oesophagus and melanoma. More of the substances found in Taxus plant extracts have medical potential. Therefore, at the beginning of this review, we describe the methods of isolation, identification and determination of taxanes in different plant parts. One of the most important taxanes is paclitaxel, for which we summarize the pharmacokinetic parameters of its different formulations. We also describe toxicological risks during clinical therapy such as hypersensitivity, neurotoxicity, gastrointestinal, cardiovascular, haematological, skin and renal toxicity and toxicity to the respiratory system. Since the effect of the drug-form PTX is enhanced by various Taxus spp. extracts, we summarize published clinical intoxications and all fatal poisonings for the Taxus baccata plant. This showed that, despite their significant use in anticancer treatment, attention should also be focused on the risk of fatal intoxication due to ingestion of extracts from these plants, which are commonly found in our surroundings.

Nutrition in Pediatric Intensive Care: A Narrative Review.

Nutrition support in pediatric intensive care is an integral part of a complex approach to treating critically ill children. Smaller energy reserves with higher metabolic demands (a higher basal metabolism rate) compared to adults makes children more vulnerable to starvation. The nutrition supportive therapy should be initiated immediately after intensive care admission and initial vital sign stabilization. In absence of contraindications (unresolving/decompensated shock, gut ischemia, critical gut stenosis, etc.), the preferred type of enteral nutrition is oral or via a gastric tube. In the acute phase of critical illness, due to gluconeogenesis and muscle breakdown with proteolysis, the need for high protein delivery should be emphasized. After patient condition stabilization, the acute phase with predominant catabolism converts to the anabolic phase and intensive rehabilitation, where high energy demands are the keystone of a positive outcome.

Perceptions of gender equity in departmental leadership, research opportunities, and clinical work attitudes: an international survey of 11 781 anaesthesiologists.

BACKGROUND: Women make up an increasing proportion of the physician workforce in anaesthesia, but they are consistently under-represented in leadership and governance. METHODS: We performed an internet-based survey to investigate career opportunities in leadership and research amongst anaesthesiologists. We also explored gender bias attributable to workplace attitudes and economic factors. The survey instrument was piloted, translated into seven languages, and uploaded to the SurveyMonkey® platform. We aimed to collect between 7800 and 13 700 responses from at least 100 countries. Participant consent and ethical approval were obtained. A quantitative analysis was done with χ2 and Cramer's V as a measure of strength of associations. We used an inductive approach and a thematic content analysis for qualitative data on current barriers to leadership and research. RESULTS: The 11 746 respondents, 51.3% women and 48.7% men, represented 148 countries; 35 respondents identified their gender as non-binary. Women were less driven to achieve leadership positions (P<0.001; Cramer's V: 0.11). Being a woman was reported as a disadvantage for leadership and research (P<0.001 for both; Cramer's V: 0.47 and 0.34, respectively). Women were also more likely to be mistreated in the workplace (odds ratio: 10.6; 95% confidence interval: 9.4-11.9; P<0.001), most commonly by surgeons. Several personal, departmental, institutional, and societal barriers in leadership and research were identified, and strategies to overcome them were suggested. Lower-income countries were associated with a significantly smaller gender gap (P<0.001). CONCLUSIONS: Whilst certain trends suggest improvements in the workplace, barriers to promotion of women in key leadership and research positions continue within anaesthesiology internationally.

Experiences With Expanded Criteria Donors: 10-Year Analysis of the Martin, Slovakia Transplant Center.

OBJECTIVES: Kidneys from expanded criteria donors with diagnosis of brain death have become a part of the organ transplant program, which have thus increased the number of transplants. MATERIALS AND METHODS: In this retrospective analysis, we identified the expanded criteria donors in a group of 156 kidney donors at our center. Basic parameters of the donors before kidney recovery were collected. Graft function, graft survival, and patient survival at 1, 3, and 5 years posttransplant were compared in expanded criteria versus standard criteria donors. RESULTS: Expanded criteria donors were significantly older than standard criteria donors (P < .001), had higher body mass index (P = .006), and had more frequent arterial hypertension (P < .001) and diabetes mellitus (P = .004) in their histories. When we considered the estimated glomerular filtration rate, graft function in the first 6 months after transplant was significantly worse in kidneys from expanded criteria donors (P = .011). In addition, recipients of grafts from expanded criteria donors had significantly worse survival in the first year posttransplant (P = .023); however, no differences in graft survival were observed. CONCLUSIONS: From the long-term aspect, graft function and graft and patient survival in cases of kidneys from expanded criteria donors were comparable to results with kidneys from standard criteria donors. Expanded use of organs available for transplant is important due to the constantly increasing demands versus limited offers of organs.

Donor Body Mass Index as a Risk Factor for Delayed Onset of Graft Function.

Delayed graft function continues to pose a significant challenge to clinicians in the context of kidney transplantation. The objective of this retrospective, 5-year analysis is to identify the parameters of beating heart donors and those of recipients that affect the delayed development of graft function. The monitored group was composed of 152 beating heart donors and 179 recipients. Delayed graft function was identified in 32 (17%) patients. The predictor for development of delayed graft function was the body mass index of the donor (odds ratio: 1.1473; 95% confidence interval [CI]: 1.0017-1.3140; P = .0472), and the independent risk factors were donor body mass index 30 to 34.9 kg/m2 (hazard ratio [HR]: 6.0215; 95% CI: 1.4188-25.556; P = .0149), donor body mass index ≥35 kg/m2 (HR: 13.5484; 95% CI: 1.4575-125.938; P = .0220), and abuse of alcohol in the donor's history (HR: 1.779; 95% CI: 1.0679-2.964; P = .0270).

Complications of Liver Resection in Geriatric Patients

Abstract: Introduction and aims. Liver resection is the treatment of choice for many primary and secondary liver diseases. Most studies in the elderly have reported resection of primary and secondary liver tumors, especially hepatocellular carcinoma and colorectal metastatic cancer. However, over the last two decades, hepatectomy has become safe and is now performed in the older population, implying a paradigm shift in the approach to these patients. Material and methods. We retrospectively evaluated the risk factors for postoperative complications in patients over 65 years of age in comparison with those under 65 years of age after liver resection (n = 360). The set comprised 127 patients older than 65 years (35%) and 233 patients younger than 65 years (65%). Results. In patients younger than 65 years, there was a significantly higher incidence of benign liver tumors (P = 0.0073); in those older than 65 years, there was a significantly higher incidence of metastasis of colorectal carcinoma to the liver (0.0058). In patients older tan 65 years, there were significantly more postoperative cardiovascular complications (P = 0.0028). Applying multivariate analysis, we did not identify any independent risk factors for postoperative complications. The 12-month survival was not significantly different (younger versus older patients), and the 5-year survival was significantly worse in older patients (P = 0.0454). Conclusion. In the case of liver resection, age should not be a contraindication. An individualized approach to the patient and multidisciplinary postoperative care are the important issues.

Complications of liver resection in geriatric patients.

Introduction and aims. Liver resection is the treatment of choice for many primary and secondary liver diseases. Most studies in the elderly have reported resection of primary and secondary liver tumors, especially hepatocellular carcinoma and colorectal metastatic cancer. However, over the last two decades, hepatectomy has become safe and is now performed in the older population, implying a paradigm shift in the approach to these patients. MATERIAL AND METHODS: We retrospectively evaluated the risk factors for postoperative complications in patients over 65 years of age in comparison with those under 65 years of age after liver resection (n = 360). The set comprised 127 patients older than 65 years (35%) and 233 patients younger than 65 years (65%). RESULTS: In patients younger than 65 years, there was a significantly higher incidence of benign liver tumors (P = 0.0073); in those older than 65 years, there was a significantly higher incidence of metastasis of colorectal carcinoma to the liver (0.0058). In patients older than 65 years, there were significantly more postoperative cardiovascular complications (P = 0.0028). Applying multivariate analysis, we did not identify any independent risk factors for postoperative complications. The 12-month survival was not significantly different (younger versus older patients), and the 5-year survival was significantly worse in older patients (P = 0.0454). CONCLUSION: In the case of liver resection, age should not be a contraindication. An individualized approach to the patient and multidisciplinary postoperative care are the important issues.

[Rare diagnostics of infective endocarditis after kidney transplantation]. / Raritná diagnostika infekcnej endokarditídy po transplantácii oblicky.

INTRODUCTION: Infective endocarditis in a patient after kidney transplantation is a serious infective complication which increases the risk of loss of the graft and also the mortality of patients. The most important predisposing factor is the immunosuppressive therapy - mainly induction immunosuppression.Material and case description: 250 patients underwent kidney transplantation throughout the period of 12 years in the Transplant Center Martin. This set of patients included 5 patients (2 %) after heart valve replacement. We present the case of a patient after kidney transplantation with development of endocarditis of the bioprosthesis of the aortic valve one month after successful kidney transplantation. Diagnostics of endocarditis by standard procedures (examination by transthoracic echocardiogram, transesophageal echocardiography, hemocultures) was unsuccessful. We rarely diagnosed endocarditis only by PET-CT examination with a consequent change of the antibiotic treatment and successful managing of this post-transplant complication. CONCLUSION: Endocarditis after kidney transplantation is a serious complication which significantly worsens the mortality of patients. The risk of development of infective endocarditis after transplantation is also increased by induction, mainly by antithymocyte globulin. Diagnostics only by PET-CT examination is rare; however, in this case it fundamentally changed the approach to the patient and led to a successful treatment.