Cell-free DNA in plasma as an essential immune system regulator.

The cell-free DNA (cfDNA) is always present in plasma, and it is biomarker of growing interest in prenatal diagnostics as well as in oncology and transplantology for therapy efficiency monitoring. But does this cfDNA have a physiological role? Here we show that cfDNA presence and clearance in plasma of healthy individuals plays an indispensable role in immune system regulation. We exposed THP1 cells to healthy individuals' plasma with (NP) and without (TP) cfDNA. In cells treated with NP, we found elevated expression of genes whose products maintain immune system homeostasis. Exposure of cells to TP triggered an innate immune response (IIR), documented particularly by elevated expression of pro-inflammatory interleukin 8. The results of mass spectrometry showed a higher abundance of proteins associated with IIR activation due to the regulation of complement cascade in cells cultivated with TP. These expression profiles provide evidence that the presence of cfDNA and its clearance in plasma of healthy individuals regulate fundamental mechanisms of the inflammation process and tissue homeostasis. The detailed understanding how neutrophil extracellular traps and their naturally occurring degradation products affect the performance of immune system is of crucial interest for future medical applications.

[Hemicorporectomy as the Highest Grade of En Bloc Sacrectomy]. / Hemikorporektomie jako nejvyssí stupen en bloc resekce sakra.

Hemicorporectomy or translumbar amputation is an extensive surgical procedure consisting in removing the lower portion of the body. Thakur et al. found a total of 71 hemicorporectomies described in literature before 2017. In the form of a case study we present the case of our patient with terminal pelvic osteomyelitis, in whom hemicorporectomy was subsequently performed, namely from the spine surgery perspective. The man, 19 years old, was exposed to high-voltage electricity and fell down from a height of 4 meters. He suffered an instable comminuted fracture of T10 (AO A3.3.) with paraplegia (Frankel A) and multiple third-degree burns affecting 25% of his total body surface area. Subsequently, the patient underwent a total of 16 surgical procedures performed by medical experts in various specialties (orthopaedic surgery, general surgery, plastic surgery, urology, vascular surgery), but in spite of that the extensive pelvic osteomyelitis has not been successfully managed. At first, urine and stool diversion were performed. After 3 weeks, i.e. 18 months after the injury, the removal of the lower portion of the body was scheduled. The hemicorporectomy was divided into 4 stages. The surgery started by posterior transecting the spine at L4-L5 segment with nerve root and dural sac ligation and treating the bleeding venous plexus in the spinal canal. After turning the patient to the supine position, the second stage of the operation followed, consisting in transecting large vessels and harvesting a musculocutaneous flap from the right thigh. During the third stage of the surgery the separation of the L4-L5 motion segment was completed by the transaction of the anterior longitudinal ligament and m. psoas major, subsequently followed by the amputation of the lower portion of the body. During the last stage of the surgery, the wound was closed by musculocutaneous flap from the fight thigh with preserved a. femoralis. The patient was discharged to home in a generally good condition 127 days after the amputation of the lower portion of the body. Now, 1 year after the surgery, the patient enjoys good physical as well as mental health. Hemicorporectomy is an extensive surgical technique, which can despite multiple complications be offered to patients with otherwise unmanageable condition. Terminal pelvic osteomyelitis is currently the most frequent diagnostic indication and the resulting condition makes possible a long-term survival of the patient in a satisfactory condition. The spinal surgeon is an irreplaceable member of the multidisciplinary team performing the surgical procedure, the primary treatment of the spinal column considerably limits blood losses. Key words: hemicorporectomy, en bloc sacrectomy, terminal pelvic osteomyelitis, sacral tumors.

Enhanced recovery after surgery and mini-invasive approaches in rectal cancer surgery - short-term outcomes.

INTRODUCTION: The aim of this study was to evaluate short-term outcomes of patients undergoing mini-invasive rectal resection within an ERAS (enhanced recovery after surgery) protocol. METHODS: A prospectively managed database of patients undergoing rectal operations performed at our department between January 2015 and April 2020 was retrospectively analyzed. An ERAS protocol was implemented into clinical practice at our department in April 2016 and mini-invasive rectal procedures in May 2016. The ERAS group consisted of all patients who underwent mini-invasive rectal resections or amputations within the ERAS protocol. The control group consisted of patients who underwent open procedures and received standard perioperative care. The extracted data included basic patient characteristics, surgical data, postoperative recovery parameters, 30-day morbidity, length of postoperative stay and 30-day rehospitalization. RESULTS: A total of 110 patients were included in the study: 67 patients in the ERAS group and 43 in the control group. Within the ERAS group 47 patients underwent robotic procedures and 20 had laparoscopic procedures. Patients in the ERAS group had significantly better clinical and laboratory recovery parameters except for postoperative nausea and vomiting. A significantly lower incidence of paralytic ileus (20.9% vs. 3%) and a shorter length of postoperative stay (13 days vs. 9 days) was found in the ERAS group. The rehospitalization rate and 30-day morbidity were not different between the ERAS and control group. CONCLUSIONS: Implementation of the ERAS protocol in combination with mini-invasive approaches leads to better short-term postoperative outcomes after rectal surgery.

Serum hepcidin is increased in ANCA-associated vasculitis and correlates with activity markers.

Hepcidin is a key regulator of iron metabolism and plays an important role in many pathologies. It is increased by iron administration and by inflammation, while erythropoiesis downregulates its expression. It decreases iron availability and thus contributes to anemia of chronic diseases. The aim of the study was to measure hepcidin as a marker and pathogenetic factor in ANCA-associated vasculitis (AAV). Hepcidin plasma concentration was measured by the immunological method in 59 patients with AAV and compared to patients with non-vasculitic etiology of chronic kidney disease, patients on hemodialysis (HD), with systemic lupus erythematodes (SLE) and to healthy controls and blood donors, and was correlated with the parameters of iron metabolism, inflammation, activity of the process and kidney function. Hepcidin concentration was increased in patients with AAV, SLE and HD and correlated positively with C-reactive protein, serum ferritin and creatinine, and negatively with hemoglobin and serum transferrin. In active form of AAV it correlated with the clinical scoring system (BVAS). Hepcidin can thus be considered as a pathogenetic factor of anemia in AAV and can be used for evaluation of inflammation in AAV and as an additional marker in active forms of the disease.

Proteomic approach for identification of IgA nephropathy-related biomarkers in urine.

Proteinuria is often used as a surrogate marker in monitoring and predicting outcome in patients with chronic kidney diseases, but it is non-specific. IgAN belongs to the most common primary glomerulonephritis worldwide with serious prognosis. The main aim of this work was to assess differences in urine proteins in patients with IgA nephropathy and to identify abnormal proteins as potential biomarkers of IgA nephropathy or the renal disease. In our pilot project, we selected 20 patients and compared them with 20 healthy volunteers. Protein quantification was performed using iTRAQ (isobaric tag for relative and absolute quantitation) labeling method. The peptides were separated by the isoelectric focusing method (IEF) and nano-LC with C18 column and identified by mass spectrometry using MALDI-TOF/TOF MS. Proteins´ lists obtained from IEF-LC-MS-MS/MS analysis were combined and contained 201 proteins. It was found out that 113 proteins were common in both experiments. 30 urinary proteins were significantly up- or down-regulated in patients with IgA nephropathy. We characterized potential biomarkers such as alpha-1-antitrypsin, apolipoprotein A-I, CD44 antigen or kininogen. Potential biomarkers of IgAN should be validated in further studies.

Iron Overload Causes Alterations of E-Cadherin in the Liver.

Iron overload causes tissue damage in the liver, but its initial effects at the molecular and cellular level are not well understood. Epithelial cadherin (E-cad) is a major adhesion protein in adherens junctions and is associated with several signal transduction pathways. Dysfunction of E-cad causes instability of adherens junctions, which leads to cell invasion, cell migration, and carcinogenesis. We found in liver samples from iron-overloaded mice that the apparent molecular mass of E-cad was reduced from 125 to 115 kDa in sodium dodecyl sulphate polyacrylamide gel electrophoresis under reducing conditions and immunoblotting, and that the cellular expression of E-cad was decreased in immunohistochemistry. The mRNA level of E-cad, however, did not change significantly, suggesting that the alterations are posttranslational. Interestingly, incubation of control liver extracts with Fe2+ alone also produced the same mobility shift. Neither an oxidant nor an antioxidant influenced this shift in vitro, suggesting that reactive oxygen species, which are generated by iron and known to cause damage to macromolecules, are not involved. Treatment of the 115 kDa E-cad with deferoxamine, an iron chelator, thus removing Fe2+, shifted the molecular mass back to 125 kDa, demonstrating that the shift is reversible. The observation also implies that the alteration that causes the mobility shift is not due to transcriptional control, deglycosylation, and proteolysis. This reversible mobility shift of E-cad has not been previously known. The alteration of E-cad that causes the mobility shift might be an initial step to liver diseases by iron overload.

[Latarjet procedure for the treatment of inveterated shoulder dislocations]. / Pouzití operace podle Latarjeta pri inveterované luxaci ramene.

PURPOSE OF THE STUDY: Transfer of the tip of the coracoid process to the anterior margin of the glenoid is a procedure indicated in shoulder instability due to bone lesion. However, it is also used in inveterated shoulder dislocations refractory to conservative treatment that require open reduction of the joint. The bone block thus created allows for more effective and safer rehabilitation than does a temporary rigid Ki-wire fixation of the humeral head. The aim of this retrospective study was to evaluate the group of patients treated for irreducible inveterated shoulder dislocations by the Latarjet procedure. MATERIAL: Between 2005 and 2009, 16 patients with inveterated anterior shoulder dislocations were operated on. The group comprised 11 men and five women with an average age of 58 years. In all patients the duration of shoulder dislocation was longer than 3 days and reduction that would keep the humeral head in the glenoid socket was not possible even under general anaesthesia. The reasons for the late treatment included a delayed visit to the doctor's because the injury was not considered serious, the dislocation being missed in the presence of other more serious conditions, and even an overlooking by the attending physician. The longest time between the dislocation and its treatment was 23 days. Alcohol abuse was evident in some of the patients. METHODS: In the patients with irreducible inveterated shoulder dislocations who were referred to us, closed reduction was attempted in the first place.When this failed or when spontaneous dislocation subsequently occurred, open revision surgery was carried out. After coagula and synovia were removed from the glenoid socket, the head still had a tendency to dislocate spontaneously. Subsequently, the Latarjet procedure ;was performed this involved osteotomy of the coracoid process, retaining the attachment of the short head of the biceps brachii, and its transposition, through a split in the subscapularis ten- don, to the anterior lower margin of the glenoid. The process was fixed with two traction screws to create a firm bone barrier preventing dislocation. RESULTS: The functional outcome was assessed at 6 months after surgery, ;using the Constant score its average value was 60 points (35 to 85). From the evaluation of individual cases it appeared that the longer the time of shoulder dislocation, the worse the functional outcome of treatment. In comparison with the patients treated by open reduction with a temporary rigid Ki-wire fixation, the Latarjet procedure was clearly beneficial. DISCUSSION: Before the Latarjet procedure has been adopted, inveterated dislocations were treated by open reduction and a temporary fixation of the head in the socket by means of Ki-wires for three weeks. The functional outcomes following inveterated dislocation and rigid fixation were pitiable. Although the shoulder was articulated, its motion was restricted and painful. The procedure described here allows the patient to start early rehabilitation while maintaining shoulder stability. CONCLUSIONS: Inveterated irreducible shoulder dislocation is a rare diagnosis which is usually associated with the patient's personality disorder or is missed in patients with a disorder of consciousness. The treatment is difficult, but with the use of the procedure described here it is possible, with some limitation, to restore the shoulder function and its range of motion.

Downregulation of myc promoter-binding protein 1 (MBP-1) in growth-arrested malignant B cells.

Normal human B lymphocytes are sensitive to the growth-inhibitory action of TGF-beta1 whereas malignant B lymphoma cells are mostly resistant to TGF-beta1 effects. We have shown in our previous work that, TGF-beta1 treatment resulted in significant growth inhibition of the DoHH2 cell line. In the present study we showed that TGF-beta1-induced growth arrest was associated with notable downregulation of the myc-binding protein-1 (MBP-1). Moreover, our results indicated that c-Myc overexpression in TGF-beta1-arrested malignant B cells is mediated by binding of MBP-1, as a transcription repressor, to the (+118/+153) element of the promoter region of the myc gene.

[Arthroscopic stabilization procedure for multidirectional shoulder instability]. / Artroskopická stabilizace ramene pri multidirekcionální nestabilite.

PURPOSE OF THE STUDY: The authors describe an arthroscopic procedure for multidirectional shoulder instability and its results in a group of their patients. They present a new argument to the widely accepted view that multidirectional instability of the shoulder is not an indication to surgical treatment. MATERIAL: A total of nine shoulders in seven patients were indicated to surgical intervention because of multidirectional shoulder instability. The average age of the patients was 19 years. Two patients underwent a bilateral procedure. Based on the preoperative clinical findings, two groups were distinguished: the patients with loose shoulders who, with overhead use of the arm, suffered from recurrent subluxations dorsally or caudally, and those who were able to voluntarily dislocate the glenohumeral joint as well as to reduce it spontaneously (habitual posterior dislocation). The patients were evaluated at one year follow-up using the Rowe and Zarins criteria. Patients with unidirectional instability of the shoulder were not included in the study. METHODS: All patients diagnosed with multiple recurrent posterior dislocations of the shoulder were indicated for surgery. After clinical examination, arthroscopy was carried out in a lateral recumbent position and, according to the findings, first capsulorrhaphy of the posterior capsule of the joint and reattachment of the labrum were performed, and then a similar intervention was carried out on the anterior part of the joint. For the operation, only two incisions were made, one for insertion of the arthroscope, the other for the instruments, and they were used interchangeably. Both absorbable and non-absorbable anchors were used and the capsule was fixed with a sliding knot. Subsequently, the shoulder was immobilized in a Desault bandage for 6 weeks, then rehabilitation was carried out. Full activity including sports was allowed beginning with the third month after surgery. RESULTS: Shoulder stability was achieved in all patients. After the procedure, no voluntary dislocation of the joint in the posterior direction or subluxations of the shoulder with overhead use of the arm were recorded. All patients, who had had serious problems pre-operatively, were satisfied with the results and would undergo the arthroscopic procedure again. Therefore, the therapy can be regarded as very effective. DISCUSSION: Our results of operative treatment contradict the generally accepted view that multidirectional shoulder instability is not indicated for surgical treatment, and the only therapy recommended to the patients is shoulder muscle strengthening, with avoidance of excessive motion range. This approach has persisted since the time when open surgery permitted either an anterior or a posterior intervention, but not both at the same stage. Today, arthroscopic reconstruction of the whole capsule is possible, with only a minimal interference with the shoulder muscles. Therefore, its benefits for the patients are undisputable. CONCLUSIONS: Arthroscopic stabilization of the shoulder for the treatment of multidirectional instability is a demanding procedure; however, if performed technically well, it is very efficient and provides good outcomes for patients with this diagnosis.

Plasma catecholamines: follow-up on 10-year study in health and cardiovascular disease.

The predictive value of blood pressure (BP), heart rate (HR), and catecholamines in terms of any subsequent development of cardiovascular disease was investigated. Systolic (S) and diastolic (D) BP, HR, epinephrine (E) and norepinephrine (NE) were measured three times a year in 1980, 1984, and 1989 on 20 clinically healthy subjects, 18 patients with 'essential hypertension', and 22 patients with angina pectoris. Of the 22 patients in the latter group, 15 died during a 2-year follow-up (1990-1991). Each individual data series was analyzed by single cosinor to assess the circannual variation. Results were summarized by population-mean cosinor for each group. Parameter tests were used to compare the circannual rhythm characteristics among the different patient groups. A circannual rhythm was invariably demonstrated on a group basis (P < 0.05). Differences in MESOR and/or circannual amplitude were found among the different groups. In particular, patients with angina pectoris who will die within the 2-year follow-up differ in terms of their E and NE from all other patient groups, a difference already detected at the beginning of the study, more than 10 years before they die. A similar separation is not achieved in terms of BP or HR.

[Fracture of the neck of the talus with pantalar dislocation (case report)]. / Zlomenina krcku hlezenní kosti s pantalární luxací.

The author describes the treatment of a fracture of the neck of the talus with pantalar dislocation. In this injury, healing prognosis depends on the preservation of blood supply to the talus, because there is high risk of necrosis development in the body of the talus. He reports a good functional outcome of conservative therapy by closed reduction followed by immobilization for 3 months and avoidance of using the leg for 6 months. Three years after the injury the patient was without any necrotic signs in the body of the talus, and returned fully to his sports activities at the same level as before the injury.

[Perioperative hypothermia and its sequelae]. / Perioperacní hypotermie a její dusledky.

UNLABELLED: Perioperative hypothermia is associated with the development of haemocoagulation, cardiovascular and metabolic disorders leading to an increased morbidity and mortality. The objective of the investigation was to assess the extent of hypothermia and its clinical and laboratory consequences. A group of 30 patients subjected to elective radical laparotomy on account of colorectal carcinoma was divided into to equivalent groups. To the first group heated infusions were administered, to the second group not heated ones. In all patients the central and peripheral temperature, rate of postoperative normalization of the temperature, postoperative thermal comfort, consumption of analgesics and biochemical and haematological parameters were monitored. RESULTS: In patients with non-heated infusions more marked and longer perioperative hypothermia was recorded with a significant alteration of the number of leucocytes and thrombocytes. In the other investigated indicators there was no significant difference between the two groups. Hypothermia did not cause serious complications in any of the patients. CONCLUSION: Although no serious clinical complications induced by hypothermia were recorded, the authors recommend an active approach and provisions for the perioperative maintenance of body temperature as a standard of contemporary perioperative care.

[A rare case of necrotizing fasciitis and subsequent sepsis with multiorgan failure caused by Vibrio vulnificus]. / Vzácný prípad nekrotizující fasciitidy a následné sepse s multiorgánovým selháním zpusobený agens Vibrio vulnificus.

The authors report a rare case of necrotizing fasciitis, sepsis caused by a strain of Vibrio vulnificus rare in Czech Republic.

[Vascular changes in essential hypertension after treatment with enalapril]. / Cévní zmeny u esenciální hypertenze po lécbe enalaprilem.

A group of patients with essential hypertension and healthy subjects from the same work place were investigated for a period of four weeks and were subjected, in addition to a basic clinical medical examination, to an ergometric examination ECHO cardiography, occlusion plethysmography, and concurrently serum levels of adrenaline (A), noradrenaline (NA), renin, cyclic adenosine monophosphate (cAMP) beta receptors before and after four weeks treatment with enalapril were assessed. The results are consistent with available data from the literature where administration of inhibitors of the angiotensin converting enzyme (ACE) caused a reduction of the peripheral vascular resistance and increased blood flow assessed by means of occlusion plethysmography. Similar results were recorded after captopril administration for a one year period, as reported by Agabiti-Rose (1). Statistically significant differences were detected between hypertonics given placebo and those given enalapril for four weeks, as regards blood flow at rest and peripheral vascular resistance at rest. There was also a significant difference when hypertonic and normotonic subjects were compared. The results are presented in tables and figures.

[Relation between echocardiographic indicators and beta 2-adrenergic receptors in essential hypertension during treatment with enalapril]. / Vztahy mezi echokardiografickými ukazateli a beta 2-adrenergními receptory u esenciální hypertenze v prubehu lécby enalaprilem.

A group of 22 men with essential hypertension stage I and II according to WHO, mean age 29.1 +/- 1.4 years, was divided at random into two sub-groups. Eleven patients were treated for four weeks with enalapril (E), 10 mg/day, 11 patients were given placebo (P), also for a period of four weeks. The patients were subjected to echocardiographic examination before and after four-week therapy with enalapril or placebo. In patients treated with E a significant reduction of the left ventricular wall and the interventricular septum was observed. There was also a highly significant decline of the weight of the left ventricle and the index of left ventricular mass. Enalapril led to a decline of the specific binding capacity and affinity of beta 2-adrenergic lymphocyte receptors. In the group given P no differences were found during the examination repeated after four weeks. By means of multidimensional linear regression estimates a close relationship was revealed between the difference in the weight of the left ventricle, the difference of the index of left ventricular mass and the difference of specific binding capacity and affinity of beta 2-adrenergic lymphocyte receptors.