Can the prophylactic administration of tranexamic acid reduce the blood loss after robotic-assisted radical prostatectomy? Robotic Assisted Radical Prostatectomy with tranEXamic acid (RARPEX): study protocol for a randomized controlled trial.

BACKGROUND: The prophylactic administration of tranexamic acid reduces blood loss during procedures at high risk of perioperative bleeding. Several studies in cardiac surgery and orthopedics confirmed this finding. The aim of this prospective, double-blind, randomized study is to evaluate the effect of tranexamic acid on peri-and postoperative blood loss and on the incidence and severity of complications. METHODS/DESIGN: Based on the results of our pilot study, we decided to conduct this prospective, double-blind, randomized trial to confirm the preliminary data. The primary endpoint is to analyze the effect of tranexamic acid on perioperative and postoperative blood loss (decrease in hemoglobin levels) in robotic-assisted radical prostatectomy. The additional endpoint is to analyze the effect of tranexamic acid on postoperative complications and confirm the safety of tranexamic acid in robotic-assisted radical prostatectomy. DISCUSSION: No study to date has tested the prophylactic administration of tranexamic acid at the beginning of robotic-assisted radical prostatectomy. This study is designed to answer the question of whether the administration of tranexamic acid might lower the blood loss after the procedure or increase the rate and severity of complications. TRIAL REGISTRATION: ClinicalTrials.gov NCT04319614. Registered on 25 March 2020.

Hand-assisted laparoscopic nephrectomy in morbidly obese patients.

INTRODUCTION: Prevalence of obesity is 30 % in the Czech Republic and is expected to increase further in the future. This disease complicates surgical procedures but also the postoperative period. The aim of our paper is to present the surgical technique called hand-assisted laparoscopic nephrectomy (HALS), used in surgical management of kidney cancer in morbid obese patients with BMI >40 kg/m2. METHODS: The basic cohort of seven patients with BMI >40 undergoing HALS nephrectomy was retrospectively evaluated. Demographic data were ana-lyzed (age, gender, body weight, height, BMI and comorbidities). The perioperative course (surgery time, blood loss, ICU time, hospital stay and early complications), tumor characteristics (histology, TNM classification, tumor size, removed kidney size) and postoperative follow-up were evaluated. RESULTS: The patient age was 3867 years; the cohort included 2 females and 5 males, the body weight was 117155 kg and the BMI was 40.3501 kg/m2. Surgery time was 7398 minutes, blood loss was 20450 ml, and hospital stay was 57 days; incisional hernia occurred in one patient. Kidney cancer was confirmed in all cases, 48-110 mm in diameter, and the largest removed specimen size was 210×140×130 mm. One patient died just 9 months after the surgery because of metastatic disease; the tumor-free period in the other patients currently varies between 1 and 5 years.  Conclusion: HALS nephrectomy seems to be a suitable and safe surgical technique in complicated patients like these morbid obese patients. HALS nephrectomy provides acceptable surgical and oncological results.

Colloid Osmotic Pressure Participates on the Post-transplant Lymphocele Pathogenesis.

The aim of the study was to evaluate the role of colloid osmotic pressure in post-transplant lymphocele pathogenesis. We have analyzed total plasmatic protein and albumin levels, and electrophoresis has been completed in blood samples before transplantation and in days 3 and 14 after transplantation in 50 patients with lymphocele (Lymphocele) and 198 patients without lymphocele (control), respectively. Colloid osmotic pressure (COP) was calculated according to the Hoefs formula. Statistically significant differences were confirmed in albumin levels (42.2 respectively 44.8 g/L) before transplantation (day 0); in total protein (52.5 resp. 55.5 g/L), in albumin (30.1 resp. 32.1 g/L), and COP (15.6 respectively 17.7 kPa) in day 3; and in total protein (52.8 resp. 58.9 g/L), in albumin (30.5 respectively 35.4 g/L), in COP (16.1 respectively 21.2 kPa) in day 14. A potentially critical albumin level was established in 44.1 g/L in the blood analyzed, but its sensitivity was only 62%. The main risk element for the lymphocele formation remains the surgeon's hand. We can proclaim the role of proteins and their COP in the post-transplant lymphocele formation as one of possible pathogenetic cofactors. It is responsible for the impaired mechanisms of the reabsorption the lymph back to the tissues. Better metabolic care could help to reduce incidence of this surgical complication.

[Tissue cyanoacrylate glue in partial nephrectomy - our own experience and assessment]. / Tkánové kyanoakrylátové lepidlo u resekce ledviny - zhodnocení vlastních zkuseností.

INTRODUCTION: The effect of cyanoacrylate glues was repeatedly evaluated in resections of liver, lungs and others organs, but minimally in kidney resection. The aim of our study was to evaluate the use of cyanoacrylate tissue glue in open and laparoscopic partial nephrectomy. METHOD: We evaluated a cohort of 32 patients who underwent partial nephrectomy with treatment of the resected area using cyanoacrylate glue. Laparoscopic resection was done in 23 patients, while 9 patients underwent an open procedure. Surgical results, complications and oncologic results were assessed. RESULTS: The median follow-up duration was 46 months. The mean time of open partial nephrectomy was 104 minutes and that of laparoscopic resection was 154 minutes; blood losses reached 250 ml and 184 ml, respectively. No serious complications occurred, and all patients survived the 3-year follow-up without any tumor recurrence. CONCLUSION: Hemostasis using the cyanoacrylate glue Glubran II was effective and safe. On the other hand, it increased the costs of the surgery and was not necessary.Key worlds: partial nephrectomy laparoscopic resection of kidney cyanoacrylate tissue glue.

A new method for immediate derivatization of hydroxyl groups by fluoroalkyl chloroformates and its application for the determination of sterols and tocopherols in human serum and amniotic fluid by gas chromatography-mass spectrometry.

A new method has been described for efficient derivatization of secondary alicyclic hydroxyl groups in steroids by alkyl chloroformates (RCFs). Cholesterol, an essential human sterol and a steroid precursor in eukaryotic cells, was used as a model for treatment with various RCFs in an aqueous and non-aqueous environment. While the cholesterol hydroxyl group did not react completely with any of the tested RCFs reagents in the former case, trifluoroethyl chloroformate (TFECF) or heptafluorobutyl chloroformate (HFBCF) fully converts cholesterol and related metabolites into the corresponding mixed carbonates under anhydrous conditions in seconds. The acylation reaction was combined with liquid-liquid microextraction (LLME) between isooctane and acetonitrile phase. The sample preparation requires just a stepwise addition of 50µl isooctane with 5µl of a pyridine catalyst, 100µl acetonitrile and 100µl isooctane with dissolved 5µl of the fluoroalkyl chloroformate reagent to a dried sample. The protocol developed in this study was successfully tested for GC-MS analysis of 12 important model steroids and four main tocopherols. Each analyte provided a single peak with excellent GC separation properties, well defined EI spectra containing diagnostic fragment ions suitable for their identification and quantitation. The new method was further validated for the determination of six diagnostic non-cholesterol sterols and four main tocopherols in human serum and in amniotic fluid. Satisfactory data were obtained in terms of calibration, quantitation limits (for sterols and tocopherols, 0.05 and 0.15µg/ml, respectively), within-run precision (0.9-19.5%) and between-run precision (0.2-19.0%), accuracy (82-115%) and recovery (90-110%). The validated method was successfully applied to GC-MS analysis of the analytes in woman sera and amniotic fluids and the results are well-comparable with those reported by other authors. The presented work demonstrates for the first time capability of the RCFs to derivatize alicyclic hydroxyls in steroids and tocopherols metabolites for GC-MS with excellent reaction rates, highest reaction yields, minor reagent consumption and easy conjunction with LLME methods.

[Biochemic evidence of the presence of urine in a drain following surgery]. / Biochemický prukaz prítomnosti moci v drénu po chirurgickém výkonu.

OBJECTIVE: Evaluation of the laboratory investigations of the postoperative wound drains liquids containing urine and establishing the borderline levels for confirmation of the urine presence. PATIENTS GROUP AND THE METHODS: Biochemical analysis of the drain liquids, urine and blood serum from the 34 patients with confirmed urine leak out of the urinary tract. The urea, creatinine, natrium, potassium and chloride levels were measured in all samples. RESULTS: The mean natrium, potassium, chlorides, urea and creatinine mean levels were 93.8 mmol/l, 15.9 mmol/l, 77.0 mmol/l, 73.3.

[Five-year follow up in patients after surgery for clear cell renal carcinoma]. / Petileté sledování nemocných po operaci svetlobunecného karcinomu ledviny.

OBJECTIVE: Evaluation of the five-year follow-up of patients treated for clear-cell renal carcinoma in dependency on its characteristic and extension. PATIENTS' COHORT AND METHODS: Retrospective analysis of five-year follow-up of 269 patients treated for clear-cell renal carcinoma in dependency on T, N, M and clinical tumor stage; presence of lymphatic and distant metastases, cellular grade and tumor necrosis. Tumor-specific surviving is presented on Kaplan-Meier curves. RESULTS: Five-year patients surviving with tumor T1, T2 and T3 were 91.8%, 70.7% and 34.6% respectively. Surviving patients without lymphatic metastases was 85%; with metastases was 6.9%. Surviving patients without distant metastases was 84.1%; with metastases was 0%. Surviving of patients with clinical tumor stage I, II, III and IV were 94.6%, 83.8%, 48.7% and 0% respectively. Surviving of patients with tumor cellular grade G1, G2, G3 and G4 were 87.5%, 95.5%, 64.2% and 27.6% respectively. Surviving patients without tumor necrosis was 92.4%; with metastases was 31.9%. CONCLUSION: Tumor size and the presence of metastases are very significant factors to patient survival. Histological characteristics as a nuclear grade and a tumor necrosis have been important prognostic factors for the future of the patient. Early diagnosis and aggressive surgical treatment are the most important for the positive course of the disease.

Gas chromatographic analysis of amino acid enantiomers in Carbetocin peptide hydrolysates after fast derivatization with pentafluoropropyl chloroformate.

A novel sample preparation protocol for gas chromatographic (GC) analysis of amino acid enantiomers in peptides was developed. It comprises traditional acid hydrolysis, a novel treatment of the analytes with a fluoroalkyl chloroformate and GC/FID separation of enantiomers on a chiral capillary column. The major improvements consist in that the derivatization step proceeds in organic-aqueous media within seconds and the amino acid derivatives are volatile enough to suit the temperature range of the chiral Chirasil-Val capillary column. The approach was found beneficial for chiral analysis of pharmaceutically important Carbetocin peptide.

[Dacryolithiasis--analysis of 15 cases]. / Dakryolitiáza--rozbor 15 prípadu.

Dacryoliths are found in 15-20% patients subject to dacryocystorhinostomy. Dacryoliths are formed by proteins or calcium and phosphate salts. In the development or dacryolithiasis participates chronic inflammation of the sac, it is found more frequently in women where some part is played by the structure of the lacrimal duct, hormonal influence and the use of cosmetic preparations. A typical sign of the disease in the variability of complaints and results of clinical examinations. Assessment of the diagnosis before surgery can be facilitated by dacryocystography. The method of choice in dacryolithiasis is dacryocystorhinostomy. The authors present their own group of 16 dacryolithis which they observed 10x in women and 5x men. The concrements were examined by the method of infrared spectroscopy and gas chromatography. All were formed by proteins; calcium and phosphate salts were not found In all patients after dacryocystorhinostomy the complaints disappeared.

Brain maldevelopment and neurobehavioural deviations in adult rats treated neonatally with indomethacin.

The risk of neurodevelopmental toxicity was studied in indomethacin (INDO), an inhibitor of prostaglandin synthesis, which is used in at-risk neonates to prevent the consequences of brain intraventricular haemorrhage or to accelerate the closure of patent ductus arteriosus. Model experiments were carried out in rats of the Wistar strain and Konárovice breed. The drug dose (2 mg/kg, s.c.) was applied to rat pups either once or twice in the following way: (1) on postnatal day 4 (PD:4) or postnatal days 4 and 5 (PD:4-5), i.e. model of brain ontogenic developmental stage in human fetus/preterm neonate of 7-month-gestational age; (2) on postnatal day 9 (PD:9) or postnatal days 9-10 (PD:9-10), i.e. model of brain ontogenic stage in full-term human newborn. The rats were followed up during development (body weight, maturation) until adulthood (age 3-9 months) using tests of behaviour (open field, social memory), nociception (tail flick, plantar test), reproduction and brain neurobiological analysis. The results were evaluated by comparison of litter-mates: treated vs control. No differences between INDO and controls were found in developmental landmarks, adult social memory or reproduction. The pattern of behavioural and neuroendocrine deviations in adult animals was dependent on the ontogenic stage exposed to drug insult. INDO rats of the groups PD:4 and PD:4-5 revealed depression of open field motor activity and emotional reactivity, and higher pituitary weight with lower TSH content. On the other hand, deviations in adult INDO groups PD:9 and PD:9-10 were characterized by pain hypersensitivity, lower pituitary weight with unchanged TSH content and deficit of monoamine transmission in the hypothalamus.

Neuro-immuno-teratogenicity of drugs used in neonatal pharmacotherapy in relation to the ontogenic stage at the time of their administration.

The risk of functional teratogenicity of two drugs used in neonatal pharmacotherapy was studied: indomethacin (INDO) and dexamethasone (DEX). Model experiments were carried out in Wistar strain rats, breed Konárovice, which received single subcutaneous drug injection (INDO 2 mg/kg, DEX 1 mg/kg) on postnatal day 4 (PD:4; model of human fetus/preterm newborn of 6-7-month-gestational age) or on postnatal day 9 (PD:9; model of full-term human neonate). The rats were followed up during development (body weight, maturation) till late adulthood (age 6-8 months) using tests of cognition, immune reactivity and biochemical brain analysis. The results evaluated by comparing treated and control litter-mates indicated that the functional teratogenic risk was significantly higher in DEX than in INDO. DEX-rats revealed disorganization of developmental processes: retardation of body growth, but acceleration of sensory development (pinna and eye opening), retarded male sexual maturation. Adult DEX-rats (age 6 months) of both series (PD:4, PD:9) had deficit of short-term memory (social recognition test). Disturbances of immune reactivity (decrease of humoral and rise of cell-mediated immune response) appeared both in adult INDO and DEX-rats (age 7 months), but only in the PD:9 series i.e. when the drugs were administered at a higher stage of the ontogenic development simulating neonatal period in humans. This finding may be warning from the clinical point of view for the neonatological practice.

Chloroformates in gas chromatography as general purpose derivatizing agents.

Chloroformates with simplest alkyls, i.e. methyl, ethyl or isobutyl, already known as favourable reagents for treating amino groups in gas chromatography for years, were revealed randomly as exceptionally rapid esterification agents. Unlike the rather poor results achieved with chloroformate-mediated ester formation in organic chemistry, the pyridine-catalyzed esterification of carboxylic acids appeared to proceed at the analytical microscale smoothly. Along with the catalyzer, an alcohol should also be present in the medium, accompanied by acetonitrile or water, according to the character of the compounds treated. Reaction conditions were optimized for various classes of carboxylic acids and a uniquely rapid derivatization of amino acids in aqueous ethanol was shown to be possible. Most of the analytes, e.g. acidic metabolites in physiological fluids, could be treated directly in the aqueous matrix. A simultaneous analysis of, e.g., amino and fatty acids or amines and their acidic catabolytes was proven to be possible. Along with the low-molecular-mass reagents, still some others, i.e. the hexyl, menthyl or pentafluorobenzyl ones, found their application fields. Results of optimized reaction conditions and a wide range of applications of chloroformate-mediated derivatization in various disciplines have been summarized in this review.

Simple approach for analysis of plasma oxo-, hydroxy- and dicarboxylic acids.

Following deproteinization of plasma with organic solvents the supernatant was shaken with hexane and cation-exchange resin in an Eppendorf tube to remove fatty and amino acids and the medium was subjected to direct treatment with ethyl chloroformate under catalytic influence of pyridine. A subsequent extraction of the immediately formed ethyl esters with a drop of chloroform enabled us to subject the sample to gas chromatographic (GC) analysis. Since ketocarboxylic acids do not require a preliminary oximation the total time of sample workup and analysis takes only several minutes.

Simultaneous profile analysis of plasma amino and organic acids by capillary gas chromatography.

A simultaneous GC analysis of more than 20 amino and nearly 30 non-amino organic acids abundant in plasma is for the first time possible. Isolation of the analytes from the plasma matrix is not necessary, keto acids do not require a preliminary oximation. An instantaneous derivatization of the acids with ethyl chloroformate takes place directly in the medium after deproteinization. Less than 30 min are required to prepare a plasma sample for the GC analysis.

[Perinatal pharmacotherapy and the risk of functional teratogenic defects]. / Perinatální farmakoterapie a riziko funkcne teratogenních defektu.

Perinatal period, which is characterized by intensive histogenesis and cytodifferentiation of the already shaped organs, is a highly vulnerable phase for fetal/neonatal brain and immune system-organs with high similarity in receptor equipment. Even fine deviations in the programmed developmental processes induced by drugs initiate disorders in the formation of neural network, cytoarchitectonics and receptor-transmitter communication systems. This pathology is not evident at birth, but forms the basis for various functional defects of neuro-psycho-immunocompetence which become apparent gradually during further maturation or even in adulthood. Clinical recognition of such functional teratogenic action of drugs is hampered by the long time interval (upto decades) between the drug administration and its consequences, making the identification of causal relations very difficult. Consequently, experimental research is necessary under the precondition of adequate animal models with sufficient validity for the extrapolation on human level. The authors suggest the principles of such approach using drug application in neonatal rats with lifelong follow-up of behaviour, immune reactivity and brain biochemical analysis. The evaluation of functional teratogenic risk in three drugs used in the treatment of risk pregnancies and risk neonates (dexamethazone, fenoterol, diazepam) is presented.

Separation of selenium analogues of sulphur-containing amino acids by high-performance liquid chromatography and high-resolution gas chromatography.

The historically conditioned adaptation of living organisms to chemically corresponding elements is influenced in nature by anthropogenic activities in many regions, the selenium-sulphur pair being one example of such a case. The separation of selenomethionine, selenoethionine and selenocystine was studied by HPLC and high-resolution GC. Ion-exchange chromatography followed by temperature-programmed GC gives the possibility of the analytical separation of trace amounts of selenomethionine in a complex mixture of common amino acids. Diastereoisomers of selenocystine were identified by HPLC in the AccQ-Tag mode.

Organic acid profiling by direct treatment of deproteinized plasma with ethyl chloroformate.

Carboxylic acids in plasma or serum can be conveniently determined by capillary gas chromatography (GC) following treatment with ethyl chloroformate (ECF). The mixed organic solvent used for plasma deproteinization is also suitable as medium for the subsequent reaction step. Thus, isolation of the compounds of interest is not necessary. Before treatment of the supernatant with ECF, the neutral lipids and amino acids are removed easily by hexane extraction and cation-exchange chromatography. Ketocarboxylic acids do not require a preliminary oximation. Capillary columns with a length of 15 m and a polar silicone phase proved to be ideal for the separation of mixtures of derivatized keto-, hydroxy-, mono- and dicarboxylic acids. The run time is less than 30 min.

Blood pressure changes induced by chronic insulin treatment in Wistar rats.

Hyperinsulinaemia may play a causal role in the development of hypertension in obese hypertensives. However, experimental evidence supporting this statement is inappropriate. The main purpose of this study was to evaluate the chronic effects of insulin administration on blood pressure, total-body glucose metabolism and urinary catecholamine excretion. After 10 weeks of insulin injection blood pressure was substantially increased in insulin-treated animals compared to those treated with saline (125 +/- 2 vs 108 +/- 2 mm Hg, p < 0.001). There were no differences in glycaemia, plasma triglyceride levels and free fatty acid levels between these two groups. Plasma level of corticosterone was increased in both insulin-treated and saline-treated rats as compared to untreated animals suggesting that the level of stress was similar in both injected groups. The urinary excretion of norepinephrine and dopamine was increased in the insulin-injected group by about 120% and 310%, respectively. Our data clearly indicate that long-term insulin administration increased blood pressure but the underlying mechanisms remain to be elucidated.