Diagnosis of thanatophoric dysplasia using clinical exome screening.

Bone dysplasias are a broad, heterogeneous group of diseases. Thanatophoric dysplasia is a rare bone dysplasia, but it is the most common lethal skeletal dysplasias. The major role in diagnostics plays a high-quality ultrasound examination in the 2nd trimester and the latest methods of genetic testing, including clinical exome testing. Knowing the correct diagnosis is crucial for the future of the fetus and the couple.

[A case of botulism in the Czech Republic and current possibilities for detecting the neurotoxin produced by Clostridium botulinum].

In the Czech Republic, botulism is a rare life-threatening disease. A total of 155 cases have been reported since 1960; according to the ISIN (formerly EPIDAT) database, there have been only three isolated cases since 2013, with the exception of a single occurrence of familial botulism in 2013. In our work, we present the occurrence of botulism after ingestion of pâté of untraceable origin by a couple who were hospitalized for botulotoxin food poisoning in July 2022. Their neurological symptoms were dominated by dysarthria. After administration of antibotulinum serum, their condition improved significantly. Patient samples were analyzed using affinity carriers and MALDI mass spectrometry, a modern highly sensitive technique for detecting the presence of botulinum neurotoxins. Unlike traditional detection by a difficult and costly biological experiment on mice, the above analysis does not require the killing of laboratory animals.

Autoimmunity, cancer and COVID-19 abnormally activate wound healing pathways: critical role of inflammation.

Recent evidence indicates that targeting IL-6 provides broad therapeutic approaches to several diseases. In patients with cancer, autoimmune diseases, severe respiratory infections [e.g. coronavirus disease 2019 (COVID-19)] and wound healing, IL-6 plays a critical role in modulating the systemic and local microenvironment. Elevated serum levels of IL-6 interfere with the systemic immune response and are associated with disease progression and prognosis. As already noted, monoclonal antibodies blocking either IL-6 or binding of IL-6 to receptors have been used/tested successfully in the treatment of rheumatoid arthritis, many cancer types, and COVID-19. Therefore, in the present review, we compare the impact of IL-6 and anti-IL-6 therapy to demonstrate common (pathological) features of the studied diseases such as formation of granulation tissue with the presence of myofibroblasts and deposition of new extracellular matrix. We also discuss abnormal activation of other wound-healing-related pathways that have been implicated in autoimmune disorders, cancer or COVID-19.

Monkeypox and Herpes Simplex Virus Type 2 Coinfection: Case Report of Perianal Lesions in HIV-Positive Patient.

ABSTRACT: We report a case of monkeypox and herpes simplex type 2 coinfection in an HIV-positive male patient who has sex with men. This case report describes a diagnostic approach for papular rash in the anal area of the male patient who has sex with men with a history of sexually transmitted disease. This is also the first documented case of monkeypox in the Czech Republic, which was confirmed after a retrospective review of swab samples from a previously hospitalized HIV-positive patient.

Decreased quality of care for Staphylococcus aureus bacteremia during the COVID-19 pandemic.

OBJECTIVES: Staphylococcus aureus bacteremia (SAB) is one of the most frequent bloodstream infections. High mortality of SAB can be significantly reduced by regular infectious disease (ID) consultations and appropriate clinical management. Because the pandemic of coronavirus disease 2019 (COVID-19) has had a negative impact on hospital ID service, it can be assumed that it has also led to decreased quality of care for SAB patients. METHODS: This study enrolled all (n = 68) patients with proven SAB who were hospitalized in Military University Hospital, Prague, in 2019 and 2020 and the quality of care indicators for SAB patients were compared. RESULTS: A total of 33 and 35 patients with SAB were hospitalized in our hospital in 2019 and 2020, respectively. The significant difference between the pandemic year 2020 and year 2019 was in ID consultations performed (74% vs. 100%; p = 0.002) and fulfilment of all quality of care indicators (66% vs. 93%; p = 0.012). Next, higher in-hospital mortality was observed in 2020 than in 2019 (6% vs. 23%; p = 0.085). There was no significant difference in the percentages of patients with performed echocardiographic examinations (66% vs. 83%; p = 0.156) and collected follow-up blood cultures (85% vs. 94%; p = 0.428). In addition, there was no difference between the two years in the adequate antibiotic therapy, sources, and bacterial origin of SAB. CONCLUSIONS: The quality of care of SAB patients significantly decreased during the COVID-19 pandemic in our institution.

Acute phase of COVID-19 is associated with elevated plasmablasts in the blood.

OBJECTIVES: The study was aimed at the characterization of humoral immunity in acute SARS-CoV-2 infection. BACKGROUND: Humoral immunity plays a central role in the protection from infection due to SARS-CoV-2, causative agent of coronavirus diseases 2019 (COVID-19). PATIENTS AND METHODS: In 24 adult patients hospitalized with COVID-19, the functional subsets of circulating B-lymphocytes and SARS-CoV-2 specific IgA and IgG antibodies were analyzed using a flow cytometry and immunoassays, respectively. RESULTS: Circulating plasmablasts and memory B-lymphocytes were significantly elevated and regulatory B-lymphocytes significantly decreased in the patients in comparison with 11 age- and sex-matched SARS-CoV-2 seronegative healthy adults. Next, circulating plasmablasts correlated negatively with the levels of SARS-CoV-2 specific IgG antibodies, which were detectable in 9 out of 15 tested patients. In addition, SARS-CoV-2 specific IgA antibodies were detectable in 13 of 15 tested patients and did not demonstrate correlation with any B-lymphocyte subset. CONCLUSION: Severe course of COVID-19 is associated with significant changes of phenotypes of circulating B-lymphocytes and elevated circulating plasmablasts correlate with decreased SARS-CoV-2-specific IgG antibodies (Tab. 2, Fig. 3, Ref. 14).

Fatal Neutropenic Colitis and Clostridium Septicum Bacteremia in a Breast Cancer Patient.

A fatal case of 67-year-old female with metastatic breast cancer on chemotherapy complicated with febrile neutropenia, colitis and sepsis due to Clostridium septicum is presented. Important clinical symptoms, laboratory and radiology findings together with therapy and outcome of neuropenic colitis are also discussed.

Salmonella Paratyphi Infection: Use of Nanopore Sequencing as a Vivid Alternative for the Identification of Invading Bacteria.

In our study we present an overview of the use of Oxford Nanopore Technologies (ONT) sequencing technology on the background of Enteric fever. Unlike traditional methods (e.g., qPCR, serological tests), the nanopore sequencing technology enables virtually real-time data generation and highly accurate pathogen identification and characterization. Blood cultures were obtained from a 48-year-old female patient suffering from a high fever, headache and diarrhea. Nevertheless, both the initial serological tests and stool culture appeared to be negative. Therefore, the bacterial isolate from blood culture was used for nanopore sequencing (ONT). This technique in combination with subsequent bioinformatic analyses allowed for prompt identification of the disease-causative agent as Salmonella enterica subsp. enterica serovar Paratyphi A. The National Reference Laboratory for Salmonella (NIPH) independently reported this isolate also as serovar Paratyphi A on the basis of results of biochemical and agglutination tests. Therefore, our results are in concordance with certified standards. Furthermore, the data enabled us to assess some basic questions concerning the comparative genomics, i.e., to describe whether the isolated strain differs from the formerly published ones or not. Quite surprisingly, these results indicate that we have detected a novel and so far, unknown variety of this bacteria.

Clinical management of Staphylococcus aureus bacteremia.

Staphylococcus aureus (SAB) bacteremia is very serious and often fatal infection with the high incidence and lethality. Diagnosis of SAB must be followed by an appropriate diagnostic and therapeutic process. From the point of view of proper SAB management, it is essential to find the primary source of infection, which can be skin and soft tissue infections, catheter infections, infectious endocarditis, osteomyelitis, pneumonia or abscesses with hematogenous spread. After the SAB has been identified, it is crucial to determine the appropriate examination and treatment procedure in close collaboration with an infectious disease specialist, clinical microbiologist and clinical pharmacist.

Chronic Hepatitis C Virus Infection Modulates the Transcriptional Profiles of CD4+ T Cells.

BACKGROUND: Chronic hepatitis C (CHC) is associated with altered cell-mediated immune response. OBJECTIVE: The aim of the study was to characterize functional alterations in CD4+ T cell subsets and myeloid-derived suppressor cells (MDSCs) during chronic hepatitis C virus (HCV) infection. Methodology. The expression levels of the lineage-defining transcriptional factors (TFs) T-bet, Gata3, Rorγt, and Foxp3 in circulating CD4+ T cells and percentages of MDSCs in peripheral blood were evaluated in 33 patients with CHC, 31 persons, who had spontaneously cleared the HCV infection, and 30 healthy subjects. Analysis. The CD4+ T cells TFs T-bet (T-box expressed in T cells), Foxp3 (Forkhead box P3 transcription factor), Gata3 (Gata-binding protein 3), and Rorγt (retinoic-acid-related orphan receptor gamma) and activation of CD8+ T cells, as well as percentages of MDSCs, were measured by multicolor flow cytometry after intracellular and surface staining of peripheral blood mononuclear cells with fluorescent monoclonal antibodies. RESULT: The patients with CHC had significantly lower percentages of CD4+ T cells expressing Rorγt and Gata3 and higher percentages of Foxp3-expressing CD4+ T cells than healthy controls and persons who spontaneously cleared HCV infection. The ratios of T-bet+/Gata3+ and Foxp3+/Rorγt+ CD4+ T cells were the highest in the patients with CHC. In the patients with CHC, the percentages of Gata3+ and Rorγt+ CD4+ T cells and the percentages of T-bet+ CD4+ T cells and CD38+/HLA-DR+ CD8+ T cells demonstrated significant positive correlations. In addition, the percentage of CD38+/HLA-DR+ CD8+ T cells correlated negatively with the percentage of MDSCs. CONCLUSION: Chronic HCV infection is associated with downregulation of TFs Gata3 and Rorγt polarizing CD4+ T cells into Th2 and Th17 phenotypes together with upregulation of Foxp3 responsible for induction of regulatory T cells suppressing immune response.

Production and use of convalescent plasma in COVID-19 treatment, taking into account the experience in the Central Millitary Hospital Prague.

One of the available treatment alternatives for COVID-19 is the administration of convalescent plasma (CP), blood plasma obtained from people who have undergone the disease. Administration of anti-SARS-CoV-2 antibodies in plasma is a method of passive specific immunization with an expected therapeutic response. CP can also be used for production a specific immunoglobulin. Experience from previous epidemic infections, caused by the coronaviruses SARS-CoV-1 and MERS-CoV, shows that CP contains neutralizing antibodies against the virus, which are probably the main source of its therapeutic potential. However, other immune mechanisms cannot be ruled out, such as antibody-induced cellular cytotoxicity and/or phagocytosis. The use of CP for the treatment of COVID-19 spread during the first half of year 2020 in many countries worldwide and relatively common is also in the Czech Republic, where, at the end of August 2020, about 100 patients were treated with CP. The production and use of CP is governed by the national multidisciplinary guidelines from April 2020 and the recommended therapeutic dose are 2 TU RP (400-450 mL), resp. 4-6 mL/kg. CP is indicated mainly in severe cases of COVID-19, which require oxygen support, ideally within 2-3 days after diagnosis, but our and foreign experience shows a beneficial effect of CP even in moderately severe cases that do not need oxygen treatment.

A case of severe course of COVID-19 treated with experimental therapy.

70-year-old high-risk patient with severe course of COVID-19 hospitalized for progressive dyspnea due to extensive bilateral pneumonia caused by SARS-CoV-2. The patient was treated with hydroxychloroquine and azithromycin from day one of hospital stay. Because of progression on the therapy, the convalescent plasma was administered on day three of hospitalization. The patient subsequently improved and was discharged home on day eleven of the hospital stay. Risk factors of severe course of the infection, complications and potential therapies of COVID-19 are discussed.

Frequent Recurrences of Genital Herpes Are Associated with Enhanced Systemic HSV-Specific T Cell Response.

OBJECTIVES: Genital herpes simplex virus (HSV) infection is controlled by HSV-specific T cells in the genital tract, and the role of systemic T cell responses is not fully understood. Thus, we analysed T cell responses in patients with recurrent genital herpes (GH). METHODS: T cell responses to HSV-1 and HSV-2 native antigens and the expression of HLA-DR and CD38 molecules on circulating CD8+ T cells were analysed in adults with high frequency of GH recurrences (19 patients) and low frequency of GH recurrences (7 patients) and 12 HSV-2 seronegative healthy controls. The study utilized the interferon-γ Elispot assay for measurement of spot-forming cells (SFC) after ex vivo stimulation with HSV antigens and flow cytometry for analysis of the expression of activation markers in unstimulated T cells. RESULTS: The patients with high frequency of GH recurrences (mean number of recurrences of 13.3 per year) had significantly enhanced HSV-specific T cell responses than the HSV-2 seronegative healthy controls. Moreover, a trend of higher numbers of SFC was observed in these patients when compared with those with low frequency of GH recurrences (mean number of recurrences of 3.3 per year). Additionally, no differences in CD38 and HLA-DR expression on circulating CD8+ T cells were found among the study groups. CONCLUSIONS: Frequency of GH recurrences positively correlates with high numbers of systemic HSV-specific T cells.

Calprotectin and Calgranulin C as Biomarkers of Pancreatic Tumors: Baseline Levels and Level Changes after Surgery.

Pancreatic tumors and their surgical resection are associated with significant morbidity and mortality, and the biomarkers currently used for these conditions have limited sensitivity and specificity. Because calprotectin and calgranulin C serum levels have been demonstrated to be potential biomarkers of certain cancers and complications of major surgery, the levels of both proteins were tested in the current study in patients with benign and malignant pancreatic tumors that were surgically removed. The baseline serum levels and kinetics of calprotectin and calgranulin C during the 7-day postoperative period were evaluated with immunoassays in 98 adult patients who underwent pancreatic surgery. The baseline serum levels of calprotectin and calgranulin C in patients with malignant (n = 84) and benign tumors (n = 14) were significantly higher (p < 0.01) when compared to those in the healthy controls (n = 26). The serum levels of both proteins were also significantly (p < 0.05) higher in patients with benign tumors than in those with malignant tumors. After surgery, the serum levels of calprotectin and calgranulin C were significantly (p < 0.01) higher than their baseline values, and this elevation persisted throughout the seven days of the follow-up period. Interestingly, starting on day 1 of the postoperative period, the serum levels of both proteins were significantly (p < 0.05) higher in the 37 patients who developed postoperative pancreatic fistulas (POPFs) than in the patients who had uneventful recoveries (n = 61). Moreover, the serum levels of calprotectin and calgranulin C demonstrated a significant predictive value for the development of POPF; the predictive values of these two proteins were better than those of the serum level of C-reactive protein and the white blood cell count. Taken together, the results of this study suggest that calprotectin and calgranulin C serum levels are potential biomarkers for pancreatic tumors, surgical injury to the pancreatic tissue and the development of POPFs.

Selected Biomarkers Correlate with the Origin and Severity of Sepsis.

The microbial etiology and source of sepsis influence the inflammatory response. Therefore, the plasma levels of cytokines (IL-6, IL-8, and IL-10), chemokines (CCL2/MCP-1, MIP-1ß), heparin-binding protein (HBP), soluble CD14 (sCD14), and cortisol were analyzed in blood from septic patients obtained during the first 96 hours of intensive care unit hospitalization. The etiology was established in 56 out of a total of 62 patients enrolled in the study. Plasma concentrations of MCP-1, sCD14, IL-6, and IL-10 were significantly higher in patients with community-acquired pneumonia (CAP; n = 10) and infective endocarditis (IE; n = 11) compared to those with bacterial meningitis (BM; n = 18). Next, cortisol levels were higher in IE patients than in those with BM and CAP, and at one time point, cortisol was also higher in patients with gram-negative sepsis when compared to those with gram-positive infections. Furthermore, cortisol and MCP-1 levels correlated positively with the daily measured SOFA score. In addition, HBP levels were significantly higher in patients with IE than in those with BM. Our findings suggest that MCP-1, sCD14, IL-6, IL-10, cortisol, and HBP are modulated by the source of sepsis and that elevated MCP-1 and cortisol plasma levels are associated with sepsis-induced organ dysfunction.

[A Case of a patient with AIDS and pneumocystis pneumonia complicated by cytomegalovirus pneumonitis].

We report a case of a 39-year-old male admitted for respiratory failure. On admission, the patient was diagnosed with advanced HIV infection and Pneumocystis jirovecii pneumonia (PJP). The patient's condition improved following specific PJP therapy but then deteriorated. The patient was subsequently diagnosed with cytomegalovirus pneumonitis and treated with ganciclovir. The severe course of both opportunistic infections required long-term care at an intensive care unit. Despite complications, the patient was discharged after 108 inpatient days in a stable clinical condition. The case demonstrates a rare coincidence of PJP and cytomegalovirus pneumonitis while also emphasizing the importance of correct diagnosis, treatment and interdisciplinary care which, despite poor prognosis, may lead to successful cure of serious simultaneous opportunistic infections in AIDS.

[Outpatient parenteral antibiotic therapy (OPAT)]. / Ambulantní parenterální antibiotická terapie (OPAT).

Traditionally, parenteral (i. e. intravenous) antimicrobial therapy has been used in inpatients with various bacterial infections. In recent decades there has been growing experience with outpatient parenteral antimicrobial therapy, mainly in the USA and western Europe. This article provides basic information on OPAT, based on available literature and the author´s experience on running OPAT service in the UK.

[New definitions for sepsis and septic shock - Sepsis-3].

The article discusses new definitions for sepsis and septic shock called Sepsis-3. The definitions are put in the historical and factual context of the 1992 definition and their extended 2003 version. Also mentioned are potential impacts on clinical practice, with it-being clear that the new definition shifts the sepsis issues more to intensive care as it emphasizes organ failure. In prehospital care, emergency departments and general wards of hospitals where patients are triaged, a new scoring system, the so-called quick SOFA, may be used. In this approach, stress is placed on impaired consciousness, a drop in systolic pressure and tachypnea but its role in more precise identification is yet to be verified in common clinical practice.

Endemic hepatitis E in the Czech Republic.

BACKGROUND: An increasing incidence of endemic hepatitis E (HE) has been reported in developed countries. Thus, an evaluation of the clinical characteristics of the disease and the utility of the current diagnostic methods is warranted. METHODS: Fifty-one adult acute patients with HE hospitalized in a single center between the years 2009 and 2012 were evaluated. Serological and molecular techniques (detection of hepatitis E virus [HEV] RNA from stool and serum samples by quantitative reverse transcription polymerase chain reaction) with sequencing and phylogenetic analysis were used for diagnosis, and the clinical, laboratory, and epidemiological parameters of the patients were evaluated. RESULTS: Forty-nine (96.1%) patients had acute endemic HE and 2 (3.9%) had an imported infection. In the cohort of patients with acute symptomatic HE (n = 47), men outnumbered women (40:7), the patients were in older middle age (mean, 60.57 years), and they had elevated median values of total bilirubin (6.67 mg/dL), alanine aminotransferase (2288.82 U/L), aspartate aminotransferase (1251.76 U/L), gamma-glutamyl transferase (360.53 U/L), and alkaline phosphatase (197.06 U/L). Serology was positive in 50 (98%) of the patients, and 1 case was diagnosed by polymerase chain reaction only. HEV RNA was detected in at least 1 specimen from 84.3% of the patients, and 28 of 29 tested isolates belonged to genotype 3. The eating of meat, innards, other home-prepared pork products, or the tasting of raw meat before cooking were the most frequently reported data (reported by 25 patients [49.0%]). CONCLUSIONS: Large numbers of the endemic cases of HE were caused by HEV genotype 3, and the clinical characteristics of endemic HE were demonstrated.

Multiparametric analysis of the specific immune response against SARS-CoV-2.

BACKGROUND: SARS-CoV-2, which causes COVID-19, has killed more than 7 million people worldwide. Understanding the development of postinfectious and postvaccination immune responses is necessary for effective treatment and the introduction of appropriate antipandemic measures. OBJECTIVES: We analysed humoral and cell-mediated anti-SARS-CoV-2 immune responses to spike (S), nucleocapsid (N), membrane (M), and open reading frame (O) proteins in individuals collected up to 1.5 years after COVID-19 onset and evaluated immune memory. METHODS: Peripheral blood mononuclear cells and serum were collected from patients after COVID-19. Sampling was performed in two rounds: 3-6 months after infection and after another year. Most of the patients were vaccinated between samplings. SARS-CoV-2-seronegative donors served as controls. ELISpot assays were used to detect SARS-CoV-2-specific T and B cells using peptide pools (S, NMO) or recombinant proteins (rS, rN), respectively. A CEF peptide pool consisting of selected viral epitopes was applied to assess the antiviral T-cell response. SARS-CoV-2-specific antibodies were detected via ELISA and a surrogate virus neutralisation assay. RESULTS: We confirmed that SARS-CoV-2 infection induces the establishment of long-term memory IgG+ B cells and memory T cells. We also found that vaccination enhanced the levels of anti-S memory B and T cells. Multivariate comparison also revealed the benefit of repeated vaccination. Interestingly, the T-cell response to CEF was lower in patients than in controls. CONCLUSION: This study supports the importance of repeated vaccination for enhancing immunity and suggests a possible long-term perturbation of the overall antiviral immune response caused by SARS-CoV-2 infection.