COVID-19 infection elicits textbook immune response.

Imunita po prodelané infekci SARS-CoV-2 byla dlouhodobe zpochybnována. Z jiných typu respiracních onemocnení vsak víme, ze bunky, které se vytvorí pri první infekci, dlouhodobe prezívají a pri opakované infekci je díky nim imunitní reakce rychlejsí a úcinnejsí. Popisuje se zvýsení hladin protilátek, jejich lepsí avidita a vznik nových variant. Jiz existující pametové B a T lymfocyty jsou pouzity jako "prototyp", který se dále vylepsuje. U reinfikovaných osob tak logicky klesá riziko vázných prubehu onemocnení. V predkládaném clánku jsou uvedeny výsledky dlouhodobého merení hladiny IgG protilátek proti proteinum S a N a IgA protilátek proti proteinu S u 4 osob, které prodelaly opakované infekce SARS-CoV-2 spojené s nárustem hladin protilátek a s méne závazným prubehem, nez mela primoinfekce. Tyto zkusenosti potvrzují i závery z nasí predchozí dlouhodobé studie imunity u senioru z roku 2020, v níz byla také pozorována reaktivace imunity u rekonvalescentu po pravdepodobné nové expozici SARS-CoV-2, ale bez prodelaného onemocnení. Prezentované výsledky potvrzují jiz dríve publikovaná fakta, tedy ze prodelání nemoci neposkytuje dlouhodobou ochranu pred reinfekcí, zvláste novou variantou viru, ale pokud jiz k reinfekci dojde, bývá její prubeh méne závazný nez pri primoinfekci.

Adherence and blood pressure control in patients with primary aldosteronism.

PURPOSE: The aim of our study was to evaluate the adherence to mineralocorticoid receptor (MR) antagonists and other antihypertensive therapy and blood pressure control in conservatively treated patients with primary aldosteronism (PA). MATERIALS AND METHODS: Conservatively treated subjects with previously confirmed PA (n-50, 64.5 ± 9 years of age, 24% women) were investigated via our outpatient hypertension clinic. All subjects underwent regular examinations in our clinic. In addition to basic laboratory and clinical parameters, 24 h ambulatory blood pressure monitoring (ABPM) (Spacelabs) was evaluated. Unplanned blood sampling for assessment of serum antihypertensive drug concentrations by the means of liquid chromatography-mass spectrometry was performed in all patients. In case of spironolactone, its active metabolite canrenone was also evaluated. Total non-compliance was then defined as the absence of all measured antihypertensive drugs. Partial non-compliance was calculated as the absence of serum levels of at least one, but not all antihypertensive drugs prescribed. RESULTS: Good blood pressure control was detected (mean 24 h systolic/diastolic BP 130 ± 12/77 ± 9 mmHg). The average number of antihypertensive drugs was 3.9 ± 1.5. All subjects were treated by MR antagonists. 44% of patients received spironolactone (average daily dose 45 ± 20 mg) and in the remaining 56% of subjects eplerenone was administered (average daily dose 80 ± 30 mg) due to spironolactone side effects. Assessment of antihypertensive drug concentrations revealed full adherence in 80% of all subjects, partial nonadherence was noted in the remaining 20% of subjects. MR antagonist levels were detected in almost all subjects (49 out of 50). CONCLUSIONS: Good blood pressure control and adherence to therapy were detected in conservatively treated patients with PA. Eplerenone had to be used quite often as male subjects did not tolerate dose escalation due to spironolactone side effects.

Diagnostics of infertile males in the 21st century - a traditional concept or a modern approach?

In developed countries, approximately 15% of couples suffer from infertility, i.e. they do not conceive within one year of a regular unprotected sexual intercourse. Since infertility is the only one diagnosis of a couple, and not of an individual, it is essential to examine the couple as the unit. Sperm analysis, i.e. native microscopic evaluation, has been used for decades as a golden standard for male fertile potential assessment. Sperm analysis, in its fundamental form, has been only morphological, and not functional evaluation of ejaculate, thus it might not give us reliable information about actual fertile potential of an individual male. On that account, new methods are being introduced to the clinical practice with a goal to improve diagnostics and subsequent treatment. The article presents these new methods, namely flow cytometry, and the impact of asymptomatic urogenital infections on fertility.

Circulating NK and NKT cells in the diagnosis and treatment of immunological causes of female infertility - retrospective data analysis from the tertiary clinical center.

Immune dysregulation can cause embryo implantation failure, possibly due to mechanisms of innate non-adaptive immunity, including natural killer (NK) and natural killer T (NKT) cells. Retrospective analysis of relative counts and functional properties of NK and NKT peripheral blood cells in women with cellular immunopathology before and after immunomodulatory treatment was realized to evaluate these values in a proportion of clinical post-treatment pregnancies. For retrospective analysis, data were collected from 184 infertile women treated for abnormal functional properties and/or numbers of NK and NKT cells after stimulation with sperm and trophoblast antigens. Flow cytometric analyses of peripheral blood both before and during/after immunomodulatory treatment were performed to determine whether clinical pregnancy was achieved. Of 184 cases, immunomodulatory treatment contributed to clinical pregnancy in 109 women (59 %), all but 25 of whom required assisted reproduction techniques to become pregnant. Clinical pregnancy was associated with significantly lower numbers of circulating NK cells (p = 0.03) and significantly less activation of NK cells by trophoblasts (p < 0.001). Increased numbers of peripheral blood NK cells and their pathological activation by trophoblast antigens are immunological factors of infertility in women. However, when appropriately chosen, immunomodulatory treatment can make clinical pregnancy more likely.

COVID-19 antibody testing: examination of the antibody response to coronavirus in common practice.

Antibody identification is an important diagnostic tool that allows us to confirm a past SARS-CoV-2 infection. While antibody tests in the Czech Republic are yet to receive the attention they deserve, in the USA antibody testing has already become part of the diagnostic algorithms. The aim of this publication is to provide comprehensive information on the topic of antibody testing, the kinetics of antibody production, the types of tests and the issue of virus neutralization tests. Special attention is paid to the correct interpretation of test results and the issues surrounding reinfection.

What is the immunity of high school students to coronavirus?

Children and young people are at the least risk of coronavirus infection and the consequences of COVID-19. However, COVID-19 has dramatically affected their lives due to many months of school closures intended to prevent the spread of the infection. In the Czech Republic, secondary schools were closed from 14 October 2020 to 24 May 2021 with a short break before Christmas. However, the students were most often infected by their parents, who usually became infected at work. The aim of this study was to find out by means of antibody testing how many high school students in Havlíčkův Brod have undergone the infection. We used questionnaires to find out how many students were in quarantine. The group included 196 students aged 15-19. Examination of IgG antibodies against the S antigen (Abbott, Architect) was performed in June 2021. Antibodies were detected in 90 of 196 students (45.9%). Coronavirus infection was reported in 88 families. Antibodies were detected in 61 quarantined students (69.3%), while the remaining 27 students had neither antibodies, nor clinical signs, nor positive PCR test. Thus, these students are considered unsusceptible to the infection at the time of quarantine. Despite the interruption of teaching in schools and the associated reduction of contacts, a significant share of students met with the coronavirus. About half of the students in the sample are already largely resistant to the disease (have antibodies). Another part of the students was unsusceptible to the disease, because they did not fall sick despite having been in contact with an infected family member.

Antibodies from previous infection bring sufficient and long-term protection against COVID-19.

One of the basic manifestations of an adaptive anti-infective immunity is the production of specific antibodies. The presence of antibodies can be detected in serum and serves as one of the diagnostic tools used to confirm past infection. Very often it also serves as a confirmation that the body has acquired immunity to the disease. The appearance of COVID-19 has cast a shadow of doubt on these textbook examples of antibody behavior. Information that repeatedly questions antibody measurement and its significance has been circulating among professionals and the general public. The aim of our article is to summarize the current knowledge on the immunity acquired following SARS-CoV-2 infection and to present the results of antibody testing from four Czech laboratories which have been measuring these antibodies for over one year. Our data suggest that commonly available diagnostic methods reliably predict the results of a virus neutralization test, which is the gold standard of immunity detection. By acknowledging those with naturally acquired anti-infective immunity, in addition to vaccinated individuals, we will significantly increase the perceived level of collective immunity.

COVID-19-free workplace: Measuring the level of antibodies against coronavirus as a basis for testing strategy in companies.

The Czech Republic is one of the countries most affected by the coronavirus pandemic - approximately 16% of the population had a positive PCR test, 2-3 times more people underwent infection without undergoing this examination. It is particularly useful for employers to know how many employees have already contracted the infection and for how many people are still at risk of the coronavirus infection. For this purpose, it is appropriate to examine IgG antibodies. However, the testing strategy is different at present - antigen testing is mandatory to look for infectious individuals, regardless of human immunity. The aim of the pilot study was to determine the number of immune individuals after infection at three clinics of GENNET s.r.o. At the same time, unvaccinated individuals who had not had COVID-19 or had undergone it more than three months ago were tested with antigen tests. The cohort included 297 subjects, of whom 182 were not vaccinated (61.3 %) and 115 subjects (38.7 %) were after the vaccination. Of the unvaccinated, 71 people had in the past a positive PCR test (39 %), another 18 people had positive IgG antibodies without infection (9.9 %) and 38 people (20.9 %) had negative IgG antibodies. So far, 55 persons (30.2 %) have not been examined. If we add people vaccinated and people with antibodies, then 74.3 % of employees of the GENNET Archa clinic, 68 % of employees from the GENNET Kostelní clinic and 58.1 % from the GENNET Liberec clinic were immune to infection. 153 individuals on average (60 of whom had antibodies) were tested for the antigen test in four rounds. The infection was detected in two people. Both belonged to the group without tested antibodies. No person with antibodies was tested positive for antigen. People who have antibodies after vaccination or infection are substantially less prone to infection and have a low risk of continuing to spread the virus. By examining antibodies, employers will gain a better overview of the situation in the workplace. Based on our study, we recommend including antibody testing into antiepidemic measures and limit antigen testing to seronegative individuals.

Current possibilities of diagnostics and treatment of immunological causes of female infertility.

The immune system plays an important role in many processes of human reproduction. During pregnancy, mother's body has to accept the semialogenic fetus, therefore the role of immune processes has a high importance. Tolerance of the fetus by the mother's immune system is ensured by a complex of immune mechanisms, the knowledge of which brings us to the new insights into human reproduction processes and in seeking of new ways to modulate immunity in repeated embryo implantation failures, miscarriages, premature births, preeclampsia, and other fertility disorders and pregnancy complications. The review article is a summary of current possibilities of immunological laboratory diagnostics in reproductive immunology, presents indications for these tests and their interpretation, and mentions possible methods of therapeutic immune intervention.

Anti-SARS-CoV-2 antibody testing in IBD healthcare professionals: are we currently able to provide COVID-free IBD clinics?

Coronavirus disease 2019 (COVID-19), caused by Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2), has spread worldwide triggering a pandemic during the year 2020. The proportion of persons infected with SARS-CoV-2 whose infection remained subclinical is not known. However, such information is important to determine whether the control measures currently employed are sufficient to halt the spread of the virus. Current study has examined the seroprevalence of anti-SARS-CoV-2 antibodies in a population of 92 healthcare professionals working with patients with inflammatory bowel disease (IBD). The enzyme-linked immunosorbent assay (ELISA) test system for SARS-CoV-2 IgG from EUROIMMUN Medizinische Labordiagnostika AG (Germany) was used. Very low herd antibody-mediated immunity was proven, less than 2%, although we have been faced with the COVID-19 pandemic for several months. Anti-SARS-CoV-2 IgG antibody testing is currently unable to provide sufficient information about our anti-infectious immunity.

Age is not a disease: Evolution of protective antibodies against SARS-CoV-2 in seniors from the Břevnice nursing home.

In the nursing home in Břevnice, 21 out of 23 seniors were infected with the SARS-CoV-2 virus in the spring of 2020. Three of them died from the infection. This study monitors the antibodies formation in group of seniors who have overcome the infection. Between April and November (1-8 months after the infection), the antibodies were measured repeatedly within the period of several weeks. The virus neutralizing tests was used as well as the ECLIA assay by Roche (total immunoglobulins), CMIA assay by Abbott (IgG) and ELISA assay by Euroimmun (IgG and IgA). Six months from the infection, neutralizing antibodies were detected in 18 out of 20 seniors. High levels of neutralizing antibodies correlated with higher levels of immunoglobulins and were a good predictor of an increase of IgG in the autumn during the second wave of the epidemic in the Czech Republic. During the autumn wave, neither any of the clients, nor any staff contracted the virus, although the prevalence of PCR positivity in the Vysočina region reached around 5 %. The antigen tests also came out all negative. This study shows that in the senior population, the production of protective antibodies follows a normal pattern, and the seniors are probably immune to a repeated infection for at least 8 months from the first infection. Based on these results, we would like to open the discussion on the testing for seroprevalence in nursing homes, possible changes to the epidemiologic regime in relation to the risk of infection, and about vaccination schemes in these centers.

Testing for COVID-19: a few points to remember.

Diagnostic approaches to COVID-19 include clinical history, PCR tests for the presence of SARS-CoV-2 virus and detection of antibodies. By combining these three approaches, the seroprevalence of anti-SARS-CoV-2 antibodies can be examined in healthcare teams. The aim of the study was to examine the seroprevalence of anti-SARS-CoV-2 antibodies in a population of healthcare professionals 6 - 8 weeks after the first COVID-19 case was detected in the Czech Republic. A total of 269 subjects were enrolled in the study (187 women, 82 men) with a median age of 45.9 years (21 - 71 years). We used a questionnaire to ascertain travel history and clinical signs of any respiratory tract infection. Blood samples were collected, and IgG levels were analysed in all samples. The level of IgA antibodies was analysed in those positive for IgG. PCR testing was performed in cases testing positive for presence of antibodies. The enzyme-linked immunosorbent assay (ELISA) test system for SARS-CoV-2 from Euroimmun (Germany) was used to analyse immunoglobulin levels. 17 % of the tested cohort reported symptoms compatible with COVID-19 and 35.8 % reported history of international travel. There were 5 subjects positive IgG cases (of 269; 1.85 %), and one IgA positive and IgG borderline positive subject (0.37 %). There was only one PCR positive subject. Anti SARS-CoV-2 antibodies were thus detected in 2.22% of participating health professionals. This article shows the pitfalls of the testing methods and highlights the necessity of using a correct testing algorithm, considering the character of the tested population and the expected low prevalence.

Exploratory drilling: how to set up, carry out, and evaluate a seroprevalence study.

Seroprevalence studies represent a very important tool to find out what fraction of population has already met with the new type of coronavirus (e.g. SARS-CoV-2). Without these data, it is almost impossible for the state authorities to manage the epidemic and adopt rational measures. This article brings the results of a medium-sized seroprevalence study which was carried out in the spring of 2020 in South Bohemia. In the Strakonice and Písek regions, the ELISA method was used to test the prevalence of IgA and IgG antibodies in 2011 subjects, volunteers from general public and selected professions working in areas with a higher exposure to the infection. The study showed that already in May 2020, 2.9% of inhabitants of the Strakonice region and 1.9% of inhabitants of the Písek region had antibodies against the coronavirus. These numbers imply that for each PCR positive person, there were at least fifty others who had probably already undergone the infection. The article points out three types of problems that might occur in such a study. First, the study must be planned correctly, and possible outcomes must be pre-assessed. Second, an appropriate test must be selected with known parameters. This enables us to correctly estimate the share of false positive and false negative results. Third, the data must be evaluated in a reasonable way and correct inference must be performed. We offer a set of recommendations how to manage these issues and how to solve problems that inevitably arise in such a large-scale testing.

The correlation between human seminal plasma sialoproteins and ejaculate parameters.

Sialic acids are negatively charged carbohydrates that are components of saccharide chains covalently linked to macromolecules. Sialylated glycoproteins are important for most biological processes, including reproduction, where they are associated with spermatogenesis, sperm motility, immune responses, and fertilization. Changes in the glycoprotein profile or sialylation in glycoproteins are likely to affect the quality of ejaculate. The aim of this study was to determine differences in the degree of sialylation between normozoospermic ejaculates and ejaculates with a pathological spermiogram using two lectins, Sambucus nigra (SNA) and Maackia amurensis (MAL II/MAA) recognizing α-2,6 or α-2,3 linkage of Sia to galactosyl residues. Our results show a close relationship between seminal plasma (SP) sialoproteins and the presence of anti-sperm antibodies in the ejaculate, apoptotic spermatozoa, and ejaculate quality. Using mass spectrometry, we identified SP sialoproteins such as, semenogelins, glycodelin, prolactin-inducible protein, lactotransferrin, and clusterin that are associated with spermatozoa and contribute to the modulation of the immune response and sperm apoptosis. Our findings suggest a correlation between the degree of SP glycoprotein sialylation and the existence of possible pathological states of spermatozoa and reproductive organs. Glycoproteins sialylation represents a potential parameter reflecting the overall quality of ejaculate and could potentially be utilised in diagnostics.

Cardiovascular Complications in Pheochromocytoma and Paraganglioma: Does Phenotype Matter?

BACKGROUND: Adrenaline-producing tumors are mostly characterized by a sudden release of catecholamines with episodic symptoms. Noradrenergic ones are usually less symptomatic and characterized by a continuous overproduction of catecholamines that are released into the bloodstream. Their effects on the cardiovascular system can thus be different. The aim of this study was to determine the prevalence of cardiovascular complications by catecholamine phenotype. METHODS: We retrospectively analyzed data on the prevalence of cardiovascular events in 341 consecutive patients with pheochromocytoma and paraganglioma treated from 1995 to 2023. Biochemical catecholamine phenotype was determined based on plasma or urinary catecholamines and metanephrines. RESULTS: According to the phenotype, 153 patients had noradrenergic pheochromocytoma and paraganglioma and 188 had adrenergic pheochromocytoma and paraganglioma. In the whole sample, the incidence of serious cardiovascular complications was 28% (95 patients), with no difference between the phenotypes or sexes. The noradrenergic phenotype had significantly more atherosclerotic complications (composite end point of type 1 myocardial infarction and symptomatic peripheral artery disease; odds ratio, 3.58 [95% CI, 1.59-8.83]; P=0.003), while the adrenergic phenotype more often had type 2 myocardial infarction and takotsubo-like cardiomyopathy (OR, 0.24 [95% CI, 0.09-0.57]; P=0.002). These changes remained even after adjustment for conventional risk factors of atherosclerosis. CONCLUSIONS: We found a 28% incidence of cardiovascular complications in a consecutive group of patients with pheochromocytoma and paraganglioma. Patients presenting with a noradrenergic phenotype have a higher incidence of atherosclerotic complications, while the adrenergic phenotype is associated with a higher incidence of acute myocardial damage due to takotsubo-like cardiomyopathy.

Subtype Identification of Surgically Curable Primary Aldosteronism During Treatment With Mineralocorticoid Receptor Blockade.

BACKGROUND: Current guidelines and consensus documents recommend withdrawal of mineralocorticoid receptor antagonists (MRAs) before primary aldosteronism (PA) subtyping by adrenal vein sampling (AVS), but this practice can cause severe hypokalemia and uncontrolled high blood pressure. Our aim was to investigate if unilateral PA can be identified by AVS during MRA treatment. METHODS: We compared the rate of unilateral PA identification between patients with and without MRA treatment in large data sets of patients submitted to AVS while off renin-angiotensin system blockers and ß-blockers. In sensitivity analyses, the between-group differences of lateralization index values after propensity score matching and the rate of unilateral PA identification in subgroups with undetectable (≤2 mUI/L), suppressed (<8.2 mUI/L), and unsuppressed (≥8.2 mUI/L) direct renin concentration levels were also evaluated. RESULTS: Plasma aldosterone concentration, direct renin concentration, and blood pressure values were similar in non-MRA-treated (n=779) and MRA-treated (n=61) patients with PA, but the latter required more antihypertensive agents (P=0.001) and showed a higher rate of adrenal nodules (82% versus 67%; P=0.022) and adrenalectomy (72% versus 54%; P=0.01). However, they exhibited no significant differences in commonly used AVS indices and the area under the receiving operating characteristic curve of lateralization index, both under unstimulated conditions and postcosyntropin. Several sensitivity analyses confirmed these results in propensity score matching adjusted models and in patients with undetectable, or suppressed or unsuppressed renin levels. CONCLUSIONS: At doses that controlled blood pressure and potassium levels, MRAs did not preclude the identification of unilateral PA at AVS. REGISTRATION: URL: https://www.clinicaltrials.gov; Unique identifier: NCT01234220.

Unilaterally Selective Adrenal Vein Sampling for Identification of Surgically Curable Primary Aldosteronism.

BACKGROUND: Adrenal venous sampling is recommended for the identification of unilateral surgically curable primary aldosteronism but is often clinically useless, owing to failed bilateral adrenal vein cannulation. OBJECTIVES: To investigate if only unilaterally selective adrenal vein sampling studies can allow the identification of the responsible adrenal. METHODS: Among 1625 patients consecutively submitted to adrenal vein sampling in tertiary referral centers, we selected those with selective adrenal vein sampling results in at least one side; we used surgically cured unilateral primary aldosteronism as gold reference. The accuracy of different values of the relative aldosterone secretion index (RASI), which estimates the amount of aldosterone produced in each adrenal gland corrected for catheterization selectivity, was examined. RESULTS: We found prominent differences in RASI values distribution between patients with and without unilateral primary aldosteronism. The diagnostic accuracy of RASI values estimated by the area under receiver operating characteristic curves was 0.714 and 0.855, respectively, in the responsible and the contralateral side; RASI values >2.55 and ≤0.96 on the former and the latter side furnished the highest accuracy for detection of surgically cured unilateral primary aldosteronism. Moreover, in the patients without unilateral primary aldosteronism, only 20% and 16% had RASI values ≤0.96 and >2.55. CONCLUSIONS: With the strength of a large real-life data set and use of the gold reference entailing an unambiguous diagnosis of unilateral primary aldosteronism, these results indicate the feasibility of identifying unilateral primary aldosteronism using unilaterally selective adrenal vein sampling results. REGISTRATION: URL: https://www. CLINICALTRIALS: gov; Unique identifier: NCT01234220.

Tumour volume and radiotherapy prolongation in locally advanced head and neck cancer patients treated with radical IMRT.

AIMS: The relationship of tumour volume, radiotherapy treatment time and other prognostic factors affecting prognosis was evaluated. METHODS: 184 patients with locally advanced head and neck cancer were treated with radical intensity modulated radiotherapy (IMRT) and compared retrospectively. RESULTS: In the multivariate analysis the overall survival was dependent on gross tumour volume (GTV), clinical stage (CS), radiotherapy treatment time (RTT) and p16 status. Local control was influenced by GTV, overall RTT and age. DFS was significantly affected by GTV, CS, RTT, p16 status and concomitant chemotherapy (CHT). CONCLUSIONS: The tumour volume and the radiotherapy treatment time were the most significant prognostic factors with the best outcomes in patients with GTV ≤ 55 cc and RTT ≤ 48 days (mean LC 8.1, DFS 7.1 and OS 6.4 years) and worst outcomes with GTV > 55 cc and RTT >48 days (mean LC 4.4, mean DFS 3.2 and mean OS 2.6 years).