Circulating Cell-Free DNA Extraction from Liquid Biopsy for Cancer Research.

As the number of cancer patients globally increases, a need for reliable biomarkers including circulating tumour DNA from liquid biopsy for diagnosis, prognosis and monitoring of the disease is rising. Currently, mainly tissue samples from biopsy are used, but there are certain limitations: firstly, it is an invasive technique, and secondly, in some cases it is almost impossible to obtain an acceptable tissue sample. This could be changed by using circulating cell-free DNA from liquid biopsy, which also gives the possibility of repeated examination. Here, we focus on the options of isolating circulating cell-free DNA from plasma samples using two isolation techniques: precision manual QIAamp Circulating Nucleic Acid Kit and automatic MagNA Pure Compact (MPC) using Nucleic Acid Isolation Kit I. Manual extraction gave significantly better yields of circulating tumour DNA (P < 0.05). This DNA also had less contaminants (organic compounds or proteins). DNA obtained by both tested methods of isolation is suitable for subsequent molecular genetic methods.

The effect of surgery on the levels of matrix metalloproteinases in patients with inguinal hernia.

Matrix metalloproteinases (MMPs) are associated with the alteration of extracellular matrix. The purpose of this study was to investigate how the levels of matrix metalloproteinases and their inhibitors - TIMPs are influenced by the presence of inguinal hernia as well as by its surgical treatment. The studied group consisted of 25 patients with inguinal hernia and 21 healthy controls for comparison. Two blood samples - before and after the treatment were collected from patients. Serum concentrations of MMPs and TIMPs were analysed by multiplex immunoassays. There was a difference in circulating levels of MMPs in patients before the surgery compared to healthy controls - the concentrations of MMP-2 and MMP-9 were significantly lower (p=0.026, p=0.018, respectively). After the surgery, the levels of MMPs, especially MMP-2 (p<0.0001), were significantly decreased in patients compared to the preoperative values, apart from MMP-9. On the contrary, MMP-9 showed significant increase after the surgery (p<0.0001). Circulation levels of TIMP-2 in patients were significantly decreased in comparison with controls (p=0.004), whereas levels of TIMP-1 were similar to controls. Both tested metalloproteinase inhibitors showed a significant decrease in detected levels (TIMP-1 p=0.0004; TIMP-2 p<0.0001) after the procedure compared to the preoperative values. The levels of MMPs, especially MMP-2 and MMP-9, and their inhibitors TIMP-1 and TIMP-2 are involved by the presence of inguinal hernia as well as are influenced by the surgery.

Rectal cancer - current treatment strategy and the tumor regression grade evaluation after neoadjuvant therapy in patients who underwent surgery at the I. Department of Surgery, General University Hospital in Prague between 2012 and 2016.

INTRODUCTION: The article contains a summary of the issues of staging and therapy with an emphasis on the neoadjuvant treatment and associated tumor regression grade with the analysis of our own group of patients. METHODS: Retrospective analysis of patients with rectal cancer who underwent a surgery at the 1st Department of Surgery - Thoratic, Abdominal and Injury Surgery; First Faculty of Medicine, Charles University in Prague and General University Hospital in Prague, Czech Republic, focusing on those who underwent neoadjuvant chemoradiotherapy and their pathologists evaluated tumor regression grade after the resection. RESULTS: The group consists of 161 patients operated on between 2012 and 2016. 47 patients underwent neoadjuvant oncological treatment with further evaluation of the tumor regression grade by a pathologist, a scoring system according to Ryan was used. A complete pathological response was elicited in 10.4% of patients, no response in 35.4% of patients, and partial tumor regression in 54.2%. CONCLUSION: Although there is a difference in our results compared to foreign publications, the proportion of patients remains comparable. Studies evaluating the advantages versus disadvantages of neoadjuvant therapy will certainly follow, and the question of the suitability of surgical treatment as the only curative solution is partially raised.

Intra-abdominal candidiasis in surgical intensive care unit - epidemiology characteristics and trends.

OBJECTIVES: Intra-abdominal candidiasis (IAC) is an invasive fungal infection representing the most common type of invasive Candida infection in surgical intensive care units (ICUs). Recently, decreased antifungal susceptibility and progressive shift in the aetiology of invasive candidiasis has been observed worldwide. We explored IAC epidemiology in surgical ICU. MATERIAL AND METHODS: We retrospectively reviewed the records of 64 patients with IAC admitted at our surgical ICU over a 4-year period (2013-2016). IAC incidence, microbiological results, antifungal therapy, and mortality were analysed. RESULTS: The cumulative IAC incidence was 18.4 cases per 1000 admissions (2013: 12.6; 2014: 17.7; 2015: 16.8; 2016: 24.5), including hospital-acquired IAC incidence (2013: 9.8; 2014: 13.3; 2015 10.1; 2016: 13.3) and community-acquired IAC incidence (2013: 2.8; 2014: 4.4; 2015: 6.7; 2016: 11.2). Candida albicans represented the most common species (n = 35, 50.0%) followed by Candida glabrata (n = 15, 21.4%), Candida tropicalis (n = 6, 8.6%) and other yeasts (each < 5.0%). Incidence rate of C. albicans (2013: 7(78%); 2014: 10(59%); 2015: 6(35%); 2016: 12(44%)) and incidence rate of C. non-albicans (2013: 2(22%); 2014: 7(41%); 2015: 9(53%); 2016: 14(52%)) were different in trend. All fungal isolates were susceptible to echinocandins, amphotericin B and voriconazole. Regarding fluconazole susceptibility, C. krusei (n = 3) was resistant and C. glabrata (n = 9) was susceptible-dose dependent (SDD). The ratio of SDD C. glabrata isolates to all isolated C. glabrata strains was 9/15 (60%) (2013: 0/2; 2014: 0/2; 2015: 1/3; 2016: 8/8). Decreased fluconazole susceptibility for C. glabrata isolates was reported in both community-acquired IAC (n = 3) and hospital-acquired IAC (n = 6). Overall 30-day mortality rate was 25.0% (16/64). CONCLUSIONS: We have revealed slowly raising of overall IAC incidence, more increasing trend in incidence of community-acquired IAC compared to rather steady incidence of hospital-acquired IAC. During period 2013-2016 we have observed a significant shift in the aetiology of IAC towards an increased proportion of non-albicans Candida species, particularly C. glabrata. Acquired decreased fluconazole susceptibility was related to C. glabrata isolates exclusively. Emergence of decreased antifungal susceptibility has been preceded by increase of non-albicans Candida isolates.

PIPAC - overview of the method and first use in the Czech Republic.

PIPAC is a new technique for intra-abdominal administration of aerosol chemotherapy in a gaseous environment (capnoperitoneum). It can be indicated for peritoneal spread of various origins, most commonly ovarian cancer, stomach cancer and colorectal cancer. Due to its mini-invasiveness, the application can be repeated. The article provides a brief overview of current views of PIPAC and describes the first experience with PIPAC in the Czech Republic.

Splanchnic vein closures.

Closures in the splanchnic venous system (SVS) represent a broad medical problem. Anatomically, individual or even multiple sections of SVS may be affected at the same time. Main sections of SVS include the venous liver outflow system, the portal vein, and the upper mesenteric vein and its basin. Thrombosis is clearly the predominant cause of closure. The closures can present as acute, subacute, chronic occult or chronic manifest. The main pathological and anatomical units are the Budd-Chiari syndrome (BCS), extrahepatic portal vein obstruction (EHPVO) and mesenteric vein thrombosis (MVT). Advanced laboratory, imaging and intervention methods substantially modify the approach to prevention, diagnosis and treatment; surgical approach also plays a role. The problem of SVS closures is interdisciplinary.

Surgical treatment of alveolar echinococcosis: a single centre experience and systematic review of the literature.

INTRODUCTION: Alveolar echinococcosis (AE) is a zoonosis caused by Echinococcus multilocularis. AE is primarily localised in the liver. Echinococcus multilocularis imitates tumour-like behaviour. It can metastasise through blood or lymphatic system to distant organs. Echinococcosis often remains asymptomatic due to its long incubation period and indistinct symptoms. Clinical symptoms are determined by the parasites location. Diagnosis of echinococcosis is based on medical history, clinical symptoms, laboratory tests, serology results, imaging methods and final histology findings. Surgical removal of the cyst with a safety margin, followed by chemotherapy is the therapeutic method of choice. CASE REPORT: We present a case report of alveolar echinococcosis in a thirty-year-old female patient in whom we surgically removed multiple liver foci of alveolar echinococcosis. The disease recurred after two years and required another surgical intervention. CONCLUSIONS: Alveolar echinococcosis is a disease with a high potential for a complete cure provided that it is diagnosed early and that the recommended therapeutic procedures are strictly adhered to.

Inverted Meckel´s diverticulum: a rare cause of ileo-ileal intussusceptions and lower gastrointestinal bleeding.

Intussusception due to an inverted Meckel's diverticulum in adults is rare. We present a case report of a 28-year-old man with Meckel's diverticulum as a cause of ileo-ileal intussusceptions and lower gastrointestinal bleeding. The patient was admitted to the hospital with incomplete small bowel obstruction, abdominal pain and massive rectal bleeding. Surprisingly, pre-operative abdominal USG revealed inhomogeneous target-like mass in the right lower abdominal quadrant as intussusception with an intraluminal polypoid lesion. The patient underwent urgent laparotomy which confirmed ileo-ileal intussusception. The involved segment of small intestine (70 cm) was resected. The surgical specimen contained an elongated polypoid lesion of 8×2.5×2 cm within the ileal lumen. Histopathological examination ascertained an inverted Meckel's diverticulum. In the discussion, we deal with diagnosis and treatment issues of Meckel's diverticulum. Key words: inverted Meckel's diverticulum - intussusceptions - gastrointestinal bleeding - acute abdomen.

[Hamartoma of the abdominal cavity and retroperitoneum- a review and a case report]. / Hamartomy dutiny brisní a retroperitonea - souhrnné sdelení doplnené kazuistikou.

METHODS: We have reviewed recent as well as older literature with the intention of compiling a summary report on hamartomas of the abdominal cavity and retroperitoneum. INTRODUCTION: Hamartoma of the abdominal cavity and retroperitoneum is a rare condition which has received relatively little attention. RESULTS: The most commonly affected organs are the liver, spleen, digestive tract, pancreas and kidneys. By its nature, hamartoma is a benign lesion with a good prognosis. However, neoplastic transformation may occur and hamartomas need to be regarded as potentially malignant. Patients are very often asymptomatic. Signs and symptoms depend on the organ involved and are caused, to different degrees, by obstruction, surrounding tissue compression, rupture or tumor bleeding. In imaging diagnostics, it is usually not possible to safely distinguish between a hamartoma and a malignant lesion. Therefore, the patient is preferably indicated for surgery (or endoscopic removal) for the purpose of histological verification and definitive treatment. CONCLUSION: Hamartomas of the abdominal cavity and retroperitoneum are relatively rare, benign and often asymptomatic diseases which, however, may manifest as compression, obstruction, perforation, bleeding or even malignant transformation. The recent case report of splenic hamartoma presented at the end of this review proves that surgical management is often the only rational option to obtain definitive confirmation of the diagnosis.Key words: splenic hamartoma hamartoma of the liver hamartoma of the pancreas hamartomatous polyps of the colon multiple hamartoma syndrome.

[Management of acute postoperative pain following thoracotomy - state of the art]. / Lécba akutní pooperacní bolesti po torakotomii - soucasné moznosti.

UNLABELLED: Postoperative pain management is an important part of complex perioperative care in patients undergoing thoracotomy, irrespective of the procedure type. Adequate pain relief leads to early mobilisation, improves respiratory functions and decreases global stress response. Thus, good perioperative pain management significantly reduces postoperative complications. Currently, numerous analgesic methods are available for the management of acute postthoracotomy pain including patient- or nurse-controlled systemic administration of analgesics, infiltration with local anaesthetics, intrapleural or intercostal nerve blockades and neuroaxial blocks (paravertebral, intrathecal, epidural). The aim of this review is to analyze the currently used methods in postthoracotomy pain management, their benefits in the light of current guidelines, and potential risks. KEY WORDS: thoracotomy - chest surgery analgesia.

[Current status regarding surgical treatment of pancreatic cancer in the Czech Republic]. / Aktuální stav chirurgické lécby karcinomu pankreatu v Ceské republice.

INTRODUCTION: The aim is to map the current situation in the surgical treatment of pancreatic cancer in the Czech Republic. This information has been obtained from surgical treatment providers using a simple questionnaire and by identifying the so called high volume centres. The information has been collected in the interest of organizing and planning research projects in the field of pancreatic cancer treatment. METHOD: We addressed centres known to provide surgical treatment of pancreatic cancer. A simple questionnaire formulated one question about the total number of pancreatic resections, also separately for the diagnoses PDAC - C25, in the last two years (2014 and 2015). Other questions focused on the use of diagnostic methods, neoadjuvant therapy, preoperative assessment of risks, the possibility of rapid intraoperative histopathology examination, Leeds protocol, monitoring of morbidity and mortality including long-term results, and the method of postoperative follow-up and treatment. ÚZIS (Institute of Health Information and Statistics of the Czech Republic) was addressed with a request to analyze the frequency of reported total numbers for DPE, LPE, TPE and to do the same with respect to diagnosis C 25 for the last two years, available for the entire Czech Republic (2013, 2014). RESULTS: Altogether 19 institutions were identified by the preceding audit, which reported more than 10 pancreatic resections annually; these institutions were addressed with the questionnaire. Sixteen institutions responded to the questions, 13 of them completely. CONCLUSION: The majority of potentially radical surgeries for PDAC in the Czech Republic are carried out at 6 institutions. All of the institutions that participated in the survey collect data about morbidity and mortality and monitor their results. KEY WORDS: pancreas cancer outcomes surgery.

Treatment of locally advanced pancreatic cancer by percutaneous and intraoperative irreversible electroporation: general hospital cancer center experience.

The aim of this study was to evaluate the safety of irreversible electroporation (IRE) and the outcome of patients undergoing IRE of locally advanced pancreatic cancer (PC). Twenty-one patients with unresectable PC underwent open (n=19) or percutaneous (n=2) IRE of the tumor using the Nanoknife system with two electrodes that were repositioned several times to affect the whole mass. The size of the tumor was 39±10mm with a range from 21 to 65mm. Five patients underwent neoadjuvant chemotherapy and seven patients were treated with chemotherapy after IRE. Complications occurred in five patients, which resulted in prolongation of the average hospital stay from 10 to 34 days. There was no mortality in the first postoperative month. Median survival after IRE was 10.2 months compared to 9.3 months in a matched cohort (hazard ratio = .54, p = .053). The quality of life was declining slowly. 81% of time after IRE the Karnofsky performance status was ≥70 and sharp decline occurred approximately 8 weeks before death.In conclusion, IRE is a safe palliative treatment option for a percentage of patients with locally advanced pancreatic carcinoma. The patients treated with open IRE lived a decent life until 8 weeks before their death. We believe that IRE of pancreatic carcinoma can be regarded as an option, if imaging or explorative laparotomy show that R0 resection in not possible.

[Irreversible electroporation in the treatment of locally advanced pancreatic cancer]. / Ireverzibilní elektroporatizace v lécbe lokálne pokrocilého karcinomu pankreatu.

INTRODUCTION: Irreversible electroporation (IRE) is a quite novel method of tissue ablation. Its mechanism of action that does not use thermal energy is the most important feature of the method. Current experience with IRE in animal studies and in clinical practice are summarized in the paper. In particular, the paper is focused on using IRE in locally advanced pancreatic carcinoma. METHOD: The basic principle of IRE is that it causes micropores in the phospholipid membrane of cells. This leads to an impairment of cellular homeostasis and programmed cell death - apoptosis. Because of absence of protein denaturation this method spares tubular structures like vessels and ducts. This is the key feature that allows to use IRE in the pancreas where common thermic ablative procedures cannot be used for difficult anatomic circumstances and resulting injury of surrounding structures. PRE-CLINICAL AND CLINICAL STUDIES: The ability to spare vascular structures and ducts was confirmed in many animal studies. Subsequently, IRE was safely utilized also in human liver, pancreas, lung and kidneys. IRE in the treatment of advanced pancreatic cancer: Most experience with IRE ablation has been gathered for locally advanced pancreatic carcinoma where clinical studies published in the recent 5 years have provided encouraging results. CONCLUSION: Irreversible electroporation is a safe method used to decrease tumour mass in pancreatic cancer. Further studies are needed to determine its therapeutic efficiency.

[Basic principles of diagnosis and treatment of secondary peritonitis - recommendations of experts with the support of SIS]. / Základní principy diagnostiky a lécby sekundární peritonitidy - doporucení odborníku s podporou SIS.

Secondary peritonitis is the most common cause of inflammatory acute abdomen treated at general surgery departments. Only early and correct diagnosis may improve the prognosis of these patients. The authors compiled an interdisciplinary review of the basic principles of diagnosis and treatment of secondary peritonitis, which reflects current findings supported by evidence-based medicine. The work is based on published international literature but also shares opinions and experiences of the selected specialists. The presented work in its extent is not meant to substitute an in-depth study of the issue, but to allow a basic and quick review of the topic.

[The development of surgical treatment in liver and biliary tract diseases]. / Vývoj operativy onemocnení jater a zlucových cest.

INTRODUCTION: Surgical liver and biliary tract diseases remain to be among the most serious and the incidence of illnesses requiring surgical treatment has been increasing. It is therefore essential to consider the new possibilities of oncological and miniinvasive care, including ablation treatment, and to evaluate their importance and integration in the treatment system. MATERIAL AND METHODS: The article presents a retrospective analysis of three consecutive sets of patients from 1991-2011. Our monitoring focused primarily on bile duct lesions and on primary and secondary tumours of the liver. Assessments included the extent and method of the operation, neoadjuvant and adjuvant care, complications, lethality, and survival rate. RESULTS: Our study confirmed that the number of surgically treated secondary tumours (colorectal carcinoma metastases) rose by 9% as a result of downstaging (i.e., the combination of chemotherapy and oncological treatment or RFA) and the use of embolization via the portal vein and sequential resections. With no fundamental changes recorded in hepatocellular carcinoma treatment, resection or transplantation still appears to be the only curative treatment method. One-year survival is minimal without these procedures. We recorded a slight increase in the incidence of cholangiocarcinoma from 6 to 10.3%. The results of extrahepatic and hepatic form resections range from 0 to 90%, depending on the stage of the disease. Biliary duct lesions are treated more frequently at the time of laparoscopic surgery; they are diagnosed later or when stenosis has already developed, and hepaticojejunoanastomosis is the most widely applied procedure in treating the lesions. The authors indicate the ablation methods in hepatic and bile duct tumour treatment only when resection treatment is not possible. CONCLUSION: Surgical resection treatment in malignant diseases of the liver and biliary tract remains the primary treatment method with a high curative potential. Neoadjuvant combined treatment increases the eligibility of patients for such surgical procedures. In biliary tract lesions, we also regard the surgical approach as dominant. The overview of results for individual periods implies the necessity of interdisciplinary care in order to achieve better results in surgical management.

[Idiopathic inflammatory bowel disease - advancements in surgical treatment]. / Idiopatické strevní zánety - vývoj chirurgické lécby.

INTRODUCTION: Treatment of idiopathic inflammatory bowel disease is constantly developing. Biological therapy has become a standard part of conservative treatment, and gene and cell therapy of these diseases is in preclinical phase. Surgical therapy also offers some progress in the treatment, such as the increasingly preferred laparoscopic approach offering the numerous benefits of minimally invasive surgery or a tendency to perform stapled anastomosis. MATERIAL AND METHODS: A retrospective analysis of patients with a diagnosis of idiopathic inflammatory bowel operated on at the First Department of Surgery, General University Hospital in the years 2007-2011 was performed. RESULTS: Within this period, 179 patients diagnosed with Crohns disease were operated on. 30 patients underwent acute operation and 149 patients were indicated for elective surgery. In the same period, 40 patients with ulcerative colitis were indicated for surgery, of whom 22 patients for acute surgery and 18 for elective surgery. CONCLUSION: Multidisciplinary approach in the treatment of patients with inflammatory bowel disease is crucial and patients should be treated in specialized centres. New possibilities of conservative treatment and progress in surgical therapy mutually correlate, and thus the choice of a correct therapeutic procedure requires specific cooperation between the surgeon and the gastroenterologist.

[Protein S100b in differential diagnosis of brain concussion and superficial scalp injury in inebriated patients]. / Protein S-100b v diferenciální diagnostice komoce mozkové a povrchního poranení hlavy u pacientu v ebriete

INTRODUCTION: Concussion cannot be differentiated from superficial scalp injury, especially in inebriated or uncooperative patients. This can have serious medical or forensic consequences. The aim of the study was to determine whether serum concentrations of S-100b in mild traumatic brain injury (MTBI) patients are significantly higher than those in patients with superficial scalp injury with scalp wound and alcohol intoxication. MATERIAL AND METHODS: A total of 50 patients with head injury, 25 with mild concussion without scalp wound and alcohol intoxication, 25 superficial scalp injury patients with scalp wound and clinical signs of inebriety. Neurological status and cranial CT scan were evaluated 60-120 minutes after injury in all the 50 patients to exclude focal cerebral injury or skull fracture. RESULTS: The serum levels of S-100b were significantly increased in patients with concussion (median 0.36 ± 0.15 µg/l ) in comparison with the group of patients with scalp injury and alcohol intoxication (median 0.09 ± 0.002 µg/l). All 50 patients had a normal cranial CT finding and neurological status. In all superficial scalp injury patients alcohol intoxication was confirmed (0.96 - 3.11s). CONCLUSION: We proved significantly higher values of S-100b in patients with brain concussion. Diagnostically decisive value of S-100b concentration in the serum was set at 146 µg/l and higher (94% sensitivity and 100% specificity). Alcohol intoxication (up to 3.11 s) and scalp wound seem to have had no crucial impact on serum S-100b level.