Study of family clusters of COVID-19 in the Ústí nad Labem Region with a focus on children - clinical picture, secondary transmission, antibody response.

OBJECTIVE: The article presents the results of a study of family clusters of COVID-19 whose aim was to describe the clinical course of the disease in children, to identify secondary transmission in households with infected children, and to find out the differences in the antibody response between children and adults. Data were collected from 3 March 2020 to 25 October 2020, that is at the time when the alpha or delta variants were not dominant. METHODS: A retrospective analysis of data on infected children in the Ústí nad Labem Region collected by the Regional Public Health Authority along with questionnaire data reported by their families. Serological tests were performed in study volunteers. RESULTS: The most common symptoms in children were headache, runny nose, and loss of smell and taste. The hospital admission rates were 0.8% in children and 2.4% in adults. The secondary transmission rate in families where the index case was a child reached 22.6%. It was higher when the index case was a child 11-15 years of age or asymptomatic. The secondary transmission rate in families where the index case was an adult equalled 77.1%. Children had higher IgG antibody levels than adults at post-infection months 4 and 5, but their seropositivity rate was comparable to that of adults. CONCLUSION: Children have a nonspecific clinical course of the disease, which is quite similar as other viroses, and rarely require hospital admission. Secondary transmission in families where the index case was a child is not uncommon and is also possible through contact with an asymptomatic child. After recovery, children have higher levels of antibodies at post-infection months 4 and 5.

The duration of SARS-CoV-2 shedding in patients recovering from COVID-19.

The knowledge of SARS-CoV-2 (severe acute respiratory syndrome coronavirus 2) shedding is highly relevant to the diagnosis, treatment and follow-up of patients with COVID-19 (coronavirus disease 2019). Deep nasopharyngeal swabs repeatedly collected from a cohort of one hundred patients with COVID-19 were tested for SARS-CoV-2 RNA using RT-PCR (real-time polymerase chain reaction). The median period of viral genome detectability was 15 days. Furthermore, the authors tested the hypothesis on the relationship between the severity of COVID-19 and the period in which the viral genome is detectable. They did not find any statistically significant difference in the duration of viral clearance between patients with asymptomatic to mild disease or severe disease.

Functionalized hydrogel microparticles prepared by microfluidics and their interaction with tumour marker carbonic anhydrase IX.

Microfluidics allows precise control of the synthesis of microparticles for specific applications, where size and morphology play an important role. In this work, we have introduced microfluidic chip design with dedicated extraction and gelation sections allowing to prepare hydrogel particles in the size range of a red blood cell. The influence of the extractive channel size, alginate concentration and type of storage media on the final size of the prepared alginate microparticles has been discussed. The second part of the work is dedicated to the surface modification of prepared particles using chitosan, pHPMA and the monoclonal antibody molecule, IgG M75. The specific interaction of the antibody molecule with an antigen domain of carbonic anhydrase IX, the transmembrane tumour protein associated with several types of cancer, is demonstrated by fluorescence imaging and compared to an isotypic antibody molecule.

Fluorescent Probes for Monitoring Cholesterol Trafficking in Cells.

Cellular cholesterol plays fundamental and diverse roles in many biological processes and affects the pathology of various diseases. Comprehensive and detailed understanding of the cellular functions and characteristics of cholesterol requires visualization of its subcellular distribution, which can be achieved by fluorescence microscopy. Many attempts have been made to develop fluorescent cholesterol reporters, but so far, none of them seems to be ideal for studying all aspects of cholesterol management. To meet the requirements for the right probe remains a great challenge, and progress in this field continues. The main objective of this review is to not only present the current state of the art, but also critically evaluate the applicability of individual probes and for what purpose they can be used to obtain relevant data. Hence, the data obtained with different probes might provide complementary information to build an integrated picture about the cellular cholesterol.

Synthesis and deposition of a Tröger's base polymer on the electrode surface for potentiometric detection of a neuroblastoma tumor marker metabolite.

The development of novel diagnostic tools is a primary goal in bioanalytical chemistry. Here we report the synthesis of Tröger's base functionalized with amino- and coumarin-units designed as a monomeric unit for the development of an electrochemical cancer sensor. The synthesized receptor was deposited onto a conducting support using electrochemical polymerization, characterized spectroscopically and tested potentiometrically towards metabolites used as tumor markers of neuroblastoma.

Striking antitumor activity of a methinium system with incorporated quinoxaline unit obtained by spontaneous cyclization.

A novel pentamethinium salt was synthesized with an unforeseen expanded conjugated quinoxaline unit directly incorporated into a pentamethinium chain. The compound exhibited high fluorescence intensity, selective mitochondrial localization, high cytotoxicity, and selectivity toward malignant cell lines, and resulted in remarkable in vivo suppression of tumor growth in mice.

[Iatrogenic bile ducts injuries]. / Iatrogenní poranení zlucových cest.

AIM: To evaluate therapeutic options and outcomes of repair of iatrogenic bile duct injuries during cholecystectomy, which were solved in our institution over the past five years. The incidence of this injury is stated in the range of 0-0.4% for open cholecystectomy and 0-0.7% for laparoscopic cholecystectomy. METHODS: Authors present a group of ten patients who were operated on for iatrogenic bile duct injury incurred during cholecystectomy in 2005-2009. All patiens were refered from other hospitals. Three men and seven women aged 20-71 years. The bile duct injury occured twice during open procedure and during laparoscopic procedure in eight. Incomplete lesion was idenified in one case, complete lesions with tissue loss were found in nine patients. Right hepatic artery injuries were found in four patients with tissue loss injury. Nine patients required reconstruction of the biliary tract using hepaticojejunoanastomosis with Roux-Y loop. RESULTS: The bile leak occurred in two patients after reconstruction. In one patient was required early percutaneous transhepatic drainage. The early death occurred in a patient with a complicated course, where our reconstruction of the biliary tract was already in the field of advanced biliary peritonitis as a third operation during 7 days. All other patients are monitored postoperatively at regular intervals in our clinic. They carried out clinical examinations and monitoring of liver enzymes. In the long interval from reconstruction (6-12 months) anastomotic stenosis occurred in three patients. Postoperative radiological intervention in the form of dilation of anastomosis and prolonged transient transanastomotic drainage was necessary (the duration of drainage was 6-7 months). CONCLUSION: Iatrogenic bile duct injury is a serious condition threatening the patient's life from the progressive failure of liver function on the basis of secondary biliary cirrhosis. Due to the nature of lesions arising from laparoscopic cholecystectomy (loss tissue injuries, thermal damage to surrounding structures, the hepatic artery injuries) reconstructions are extremely difficult. For most patients reconstructive operations are the last possible surgical procedures in this area, except for liver transplantation. Hilar reconstructions have a higher probability of stenosis of the anastomosis. If they occur, there are repeated cholangitis, which pass into the secondary sclerosing cholangitis and cause secondary biliary cirrhosis, with all the consequences of disease (portal hypertension, bleeding esophageal varices). For these reasons, it is necessary for careful long-term postoperative monitoring of liver function and good interdisciplinary cooperation, especially with the intervention radiologist in management postoperatively evolving stenosis of anastomoses. It is necessary for the early identification and indication of radiological interventions in order to prevent damage to the liver parenchyma.

[Esophageal replacement using large intestine--experience with 109 cases]. / Náhrada jícnu tlustým strevem--zkusenosti se 109 prípady.

Use of large intestinal transplants in esophageal replacement is an uncommonly used alternative. The authors discuss its indications in benign, as well as in malignant disorders and, based on them, assign them into three groups. Based on historical data and their professional experience, they present reasons for the commonest uses of particular parts of the large intestine. They evaluate introperative, as well as long-term outcomes.

[Video-assisted esophageal resection for carcinoma--ten-year experience]. / Videoasistovaá resekce jícnu pro karcinom--desetileté zkusenosti.

INTRODUCTION: Esophageal procedures belong to the most complicated gastrointestinal procedures. Therefore, esophageal carcinoma patients have been and still are concentrated into specialized centres, equipped with all diagnostic and therapeutic means. At the Ist Surgical Clinic of the LF UP (Medical Faculty of the Palacky University) in Olomouc, esophageal surgery has a long lasting tradition. In the management of malignant esophageal disorders, the clinic prefers the use subtotal esophageal resection, attaching the esophagus transplant to a cervical esophageal stud, to the use of more saving resection with the anastomosis site in the mediastinum. In order to substitute for the resected esophagus, the authors normally use a tubulized gastric flap. In cases, where the stomach cannot be used, the authors use transverse colon to replace the resected esophagus. Classical esophageal procedures have nearly entirely been replaced by video-assisted procedures. Most esophageal resections are performed using video-assisted laparoscopic transhiatal approach. In the management of esophageal tumors which are located at the level of tracheal bifurcation, or below it, the authors use right- sided thoracoscopic approach. MATERIAL AND METHODS: During the past 10 years, 178 patients have undergone subtotal esophageal resection for esophageal carcinoma. In 81 patients (45.5%), the esophagus was resected using classical approach. 35 subjects underwent transhiatal "blind"extirpation according to Orringer, in 46 subjects right-sided thoracotomy was used (subjects with tumors located in the middle third of the thoracic esophagus and higher). Video-assisted approach was used in 97 patients, which included 88 subjects with laparoscopic transhiatal modification of the procedure and 9 subjects, who underwent video-thoracoscopy. OUTCOMES: The mean procedure duration was 242 minutes. The authors recorded the following complications: pneumothorax in 29 patients (16%) and n. laryngeus recurrens palsy in 16 subjects (9%). 13 patients (7%) developed a fistule in anastomosis, which was managed by drainage of the cervical wound. Pulmonary complications were recorded in 55 patients (31%). The mean duration of hospitalization was 12 days, intraoperative death rate was 4.5%. Only 2 out of 8 deaths occurring during the early postoperative period, were surgery-related. In the both cases, mediastinitis developed, resulting from a transponate necrosis in one of the cases and from a bronchial fistule in the other subject. Five subjects exited because of ARDS, which included one case of myocardial infarction. CONCLUSION: When esophageal carcinoma is managed at a clinic equipped with the latest modern diagnostic and therapeutical means, the procedure of esophageal resection is a fairly safe procedure with low death and morbidity rates.

Serological response to tick-borne encephalitis (TBE) vaccination in the elderly--results from an observational study.

BACKGROUND: In the elderly, the incidence of tick-borne encephalitis (TBE) is higher than in younger individuals. At the same time, the responsiveness of the elderly to vaccination may be decreased. Product information for TBE vaccines therefore recommends determining antibody response after two of the three doses required for primary immunization in persons over 60. OBJECTIVE: To determine the proportion of persons aged 60 years or older with insufficient antibody response after administration of two doses of TBE vaccine. METHODS: A retrospective analysis of TBE-specific antibody levels as determined by Immunozym ELISA in serum samples of 185 vaccinees > 60 years of age after the first two of the three vaccinations required for primary immunisation. Patients received either FSME-IMMUN Adult or Encepur Adults at one vaccination centre in the Czech Republic. In a subset of 80 vaccinees, two additional ELISAs and a neutralization test (NT) were performed. RESULTS: Overall, 33 of the 185 individuals (18%) included in the Immunozym analysis had antibody concentrations below putative protective levels after two doses. Geometric mean concentrations and seropositivity rates were higher in individuals vaccinated with FSME-IMMUN than in those vaccinated with Encepur. While the differences in mean antibody levels and/or seropositivity rates in ELISAs for the Neudörfl strain (used in FSME-IMMUN) were significant, no significant differences were obtained for Enzygnost, which uses the Karlsruhe strain (used in Encepur). CONCLUSIONS: After two doses of TBE vaccine, one in five persons aged 60 years or older may be without protective antibodies, confirming the importance of testing the antibody response after the second vaccination and administering an additional dose in the event of insufficient response.

[Analytical methods at the boundary between biochemistry and immunology]. / Vysetrovací metody na pomezí biochemie a imunologie.

The article presents assays shared by both disciplines; i.e the specialization 801 (Clinical Biochemistry) and 813 (Laboratory of Allergology and Immunology), and it covers over 60 tests. It is intended as reference information for general practitioners, as well as for laboratory workers at the beginning of their studies. Emphasis is laid on some facts which GPs ordering laboratory determinations should be familiar with; the pre-analytical stage (specimen collection, their transport and storage), the analytical reliability of particular methods, their accessibility, frequency and total costs. Analytical techniques are only mentioned marginally as their understanding is not essential for GPs. Assuming the readers' fundamental experience of diagnostic applications of the methods discussed; we only comment some obsolete or rare tests. The shared assays cover selected analyses of proteins (incl. some enzymes and tumormarkers), ions, osmolality and quantitative microscopic urinalysis.

Optimization of the thickness of a conducting polymer, polyaniline, deposited on the surface of poly(vinyl chloride) membranes: a new way to improve their potentiometric response.

Repeated depositions of polyaniline (PANI) have been used to control the thickness of the polymeric film deposited on poly(vinyl chloride) (PVC) membrane surface. The oxidation of aniline was carried out in a dispersion mode, i.e. in the presence of poly(N-vinylpyrrolidone) (PVP). Two kinds of PVC were used for this purpose: a non-plasticized PVC for the study of PANI deposition and PVC, plasticized with nitrophenyl octyl ether (NPOE), as a prototype of a liquid membrane electrode. The results of UV-visible and FTIR spectroscopies and electron microscopy showed that (1) the film thickness increased by about equal increments of approximately 40 nm after each polymerization, and (2) the interface with PVC was constituted by PANI film and adhering PANI-PVP colloidal particles. The various thicknesses of the deposited PANI films affected the potentiometric response of the NPOE/PVC membrane with and without an anion-exchanger. The potentiometric anionic response was observed with a minimal thickness of PANI film on the blank NPOE/PVC membrane. Sensitivity of the PANI film to pH occurred only with a blank NPOE/PVC membrane coated with a thick polymeric film, while it was strongly suppressed by the presence of a lipophilic anion-exchanger, tridodecylmethylammonium chloride (TDDMACl), in the membrane, regardless of the thickness of the polymer film. The thickness of the PANI film did not affect the anionic selectivity pattern of TDDMACl-based membranes to any great extent, but its presence improved and stabilized their potentiometric characteristics (sensitivity, linear-response range).

p38 MAPK plays an essential role in apoptosis induced by photoactivation of a novel ethylene glycol porphyrin derivative.

In this study, we provide evidence that photostimulation of various cancer cells preloaded with a new photosensitizing compound, tetrakis-meso-(4-ethyleneglycol-2,3,5,6-tetrafluorophenyl) porphyrin (PORF-TEG), results in rapid activation of the cell death machinery. PORF-TEG, although primarily localized in lysosomes, induces mitochondria-driven apoptosis. The induction of apoptosis is accompanied by immediate and sustained activation of p38 mitogen-activated protein kinase (MAPK) and transient activation of c-Jun N-terminal kinase (JNK). Conversely, the inhibition of p38 by PD 169316 or SB202190 and by the p38alpha dominant-negative mutant as well as the deletion of the p38alpha gene (MEFs-KO) protected cells from apoptosis, whereas inhibition of JNK did not. Activation of the p38 signaling pathway occurs upstream of caspase activation. In addition, preincubation of cells with scavengers of reactive oxygen species attenuated p38 and caspase activation and increased cell survival, thus connecting reactive oxygen species formation with the activation of the p38 pathway. Later events included degradation of Bcl-2, activation of tBid, and cleavage of Bad and Mcl-1. The data suggest a key role for p38 MAPK in PORF-TEG-photoinduced apoptosis.

[Analysis of regressive changes in the lung carcinoma lymphatic metastases following induction chemotherapy]. / Analýza regresivních zmen v lymfatických metastázách plicního karcinomu po indukcní chemoterapii.

BACKGROUND: Prognosis of higher stages of non-small cell lung cancer (NSCLC) is very poor. Only 13% of patients in stage IIIA survive 5 years after the diagnosis determination. The purpose of neoadjuvant (inductive) therapy is to eliminate haematogenous and lymphogenous metastases and to cause a cytoreduction in the primary tumor before the resection. Especially detection of lymphogenous metastases is the main factor in the indication of the pre-operative chemotherapy, but this therapy is frequently indicated according to CT without any bioptic verification. The authors suppose such a discrepancy in radiological and histo-pathological staging to influence in a positive sense rather optimistic results of therapeutic protocols, which include the pre-operative chemotherapy. METHODS: A biopsy of the lymph nodes was performed as a part of the staging. If the metastasis was proved, the lymph node was labeled and the patient was treated by 3 cycles of the pre-operative chemotherapy. The 57 labeled lymph nodes were removed during the subsequent lung resection and lymphadenectomy. An analysis of regressive alterations in the lymphatic metastases of lung cancer after an inductive chemotherapy and comparison with the pre-operative bioptic findings have quantificated the chance of the inductive therapy to eliminate lymphatic metastases. RESULTS: The clinical down-staging was stated in 21 cases (36.8%), but the viable malignant cells rested in the majority 50 (87.7%) of the taken labeled lymph nodes 57 (100%) even after the neoadjuvant therapy. On the other hand, the satisfactory tumor regression changes was proven in 49 (86%) lymphatic metastases. CONCLUSIN: The results of the study may modify an indicating judgment in the therapy of locally advanced stages of lung cancer because it has brought a new view to the results of neoadjuvant therapy.

Temporary aorto-aortal bypass used during extended pneumonectomy for lung cancer.

Involvement of the aortic arch, or the descending aorta, is not uncommon in left-sided lung tumours. The direct invasion of the aortic wall is generally considered a contraindication in lung resection. However, there are a limited number of reports of full thickness resections of the aorta during lung cancer surgery. They may be accomplished as a partial resection using a patch, or as a local tubular resection with reconstruction by a graft. In order to prevent ischaemia of the spinal cord, a cardiopulmonary bypass is usually recommended. The authors present a case report of a full thickness tubular resection of the descending aorta during pneumonectomy for centrally located lung carcinoma. The direct invasion to the descending aorta was only confirmed intraoperatively. After tumour dissection, two aortal cannulas were inserted into the aorta: the first one into the aortic arch over the left subclavian artery, the other one into the descending aorta over the diaphragm. They were then bypassed without a pump. The descending aorta was cross-clamped and replaced by a Dacron graft. There were no early complications : the patient has already survived two years after surgery with no recurrence.

Potentiometric response and mechanism of anionic recognition of heterocalixarene-based ion selective electrodes.

The ion selective electrode (ISE)-based potentiometric approach is shown to be an effective means of characterizing the anion recognition sites in the molecular receptor calix[2]pyridino[2]pyrrole (CPP). In particular, potentiometric pH-measurements involving the use of experimental PVC-membranes based on CPP revealed the existence of both mono- and diprotonated forms of the receptor under readily accessible conditions. Based on these analyses, apparent surface protonation constants for this heterocalixarene were found to lie between 8.5-8.9 (pK(B1)) and 3.3-3.8 (pK(B2)). CPP was found to interact with targeted anionic analytes based on both coulombic and hydrogen bond interactions, as inferred from varying the kinds of ionic sites present within the membrane phase. Potentiometric selectivity studies revealed that CPP preferred "Y-shaped" anions (e.g. acetate, lactate, benzoate) over spherical anions (e.g. fluoride and chloride), fluoride over chloride within the set of spherical anions, and the ortho-isomer over the corresponding meta- and para-isomers in the case of hydroxybenzoate (salicylate and congeners). In the context of this study, the advantages of potentiometric determinations of acetylsalicylic acid using optimized PVC-membranes based on CPP relative to more conventional PVC-membrane ISEs based on traditional anion exchanger were also demonstrated.

Functionalization of PVC membrane with ss oligonucleotides for a potentiometric biosensor.

A novel application of a single stranded (ss) oligonucleotide as an active component of polymeric membrane in an ion-selective electrode (ISE) is described. The original oligonucleotides, oligo(dA)(15), modified by cholesterol, triphenylmethyl and hexadecyl derivatives, were immobilized into poly(vinyl chloride) (PVC) membrane using extraction protocol. In parallel, the adsorption protocol was used to immobilize unmodified oligo(dA)(15) on the PVC membrane based on tridodecylmethyammonium chloride (TDDMA(+)Cl(-)). Immobilization of ss oligonucleotide probe through spacer was more effective for the potentiometric detection of the hybridization between complementary oligonucleotides. It was found that cholesterol-oligo(dA)(15) modified membranes were sensitive toward complementary oligo(dT)(15) in the concentration range 2-80 nM at pH 7. An explanation for the detection mechanism is proposed.

[Duplication of the terminal ileum--a case review]. / Duplikatura terminálního ilea--kazuistika.

Duplications of the gastorintestinal tract are fairly rare developmental anomalies. They may occur throughout the gastrointestinal tract, however, most commonly, they occur in the small intestine. They are usually diagnosed in early childhood, however, they may be asymptomatic first and cause symptoms as late as in adulthood. Symptoms of,,urgent abdominal conditions", ranging from symptoms of intestinal obstruction, perforation to symptoms of bleeding from the GIT, prevail. A rare finding of a carcinoma in the duplication has been recorded. The etiopathogenesis has not been fully understood. The treatment is surgical. The authors present a case-review of the terminal ileum duplication in a child with symptoms of intestinal obstruction, resp. invagination.

Quo vadis porphyrin chemistry?

This review summarizes recent developments in the area of porphyrin chemistry in the direction of biological applications. Novel synthetic methodologies are reviewed for porphyrin synthesis, porphyrin analog synthesis, stable porphyrinogens -- calixpyrroles, expanded porphyrins. Unique biological properties of those compounds are desribed with focus on photodynamic therapy (PDT) and molecular recognition properties. Special attentions given to metalloporphyrins with potential to affect heme degradation and CO formation.

[The technique of securing sentinel lymph nodes in tumors of the distal oesophagus and the stomach ]. / Technika získání sentinelových uzlin u tumoru v oblasti distálního jícnu a zaludku.

The authors discuss the technique of identification and biopsy of sentinel lymph nodes in tumors of the upper gastrointestinal tract. They describe difficulties and problems of the technique, in which the identified sentinel lymph node is obtained mainly employing endoscopical and minimally invasive techniques.