Raman spectroscopy of lymphocytes from patients with the Epstein-Barr virus infection.

In this study, Raman spectroscopy is applied to trace lymphocytes activation following contact with the Epstein-Barr virus (EBV) of the herpesvirus family. The biomarker of cell activation is found to be the 520 cm-1 band, indicating formation of immunoglobulins. The blood samples are obtained from patients diagnosed with infectious mononucleosis and treated at the University Hospital in Kraków. The lymphocytes' Raman spectra are collected using a mapping technique, exciting samples with a 514.5 nm line of Ar + laser. Measurements are performed on the 1st, 4th, 6th, 12th and 30th day of hospitalization, until the patient has recovered. The highest intensity of the immunoglobulin marker is observed on the 4th day of hospitalization, while the results of the blood count of patients show the greatest increase in the number of lymphocytes at the beginning of hospitalization. No activated lymphocytes were observed in the blood of healthy volunteers. Some information is provided by the evaluation of B-cell activation by estimating the activated areas in the cells, which are determined by the presence of the Ig marker. The 900 cm-1 band and band around 1450 cm-1 are also analyzed as markers of the presence of the latent membrane protein, LMP2A (and 2B), of the EBV viral protein. The anomalous degree of depolarization observed in B-cells in the course of EBV infection appears to be due to the influence of a virus protein, disrupting BCR signal transduction.

Course of Plasmodium infection studied using 2D-COS on human erythrocytes.

BACKGROUND: The threat of malaria is still present in the world. Recognizing the type of parasite is important in determining a treatment plan. The golden routine involves microscopic diagnostics of Giemsa-stained thin blood smears, however, alternative methods are also constantly being sought, in order to gain an additional insight into the course of the disease. Spectroscopic methods, e.g., Raman spectroscopy, are becoming increasingly popular, due to the non-destructive nature of these techniques. METHODS: The study included patients hospitalized for malaria caused by Plasmodium falciparum or Plasmodium vivax, in the Department of Infectious Diseases at the University Hospital in Krakow, Poland, as well as healthy volunteers. The aim of this study was to assess the possibility of using Raman spectroscopy and 2D correlation (2D-COS) spectroscopy in understanding the structural changes in erythrocytes depending on the type of attacking parasite. EPR spectroscopy and two-trace two-dimensional (2T2D) correlation was also used to examine the specificity of paramagnetic centres found in the infected human blood. RESULTS: Two-dimensional (2D) correlation spectroscopy facilitates the identification of the hidden relationship, allowing for the discrimination of Raman spectra obtained during the course of disease in human red blood cells, infected by P. falciparum or P. vivax. Synchronous cross-peaks indicate the processes taking place inside the erythrocyte during the export of the parasite protein towards the cell membrane. In contrast, moieties that generate asynchronous 2D cross-peaks are characteristic of the respective ligand-receptor domains. These changes observed during the course of the infection, have different dynamics for P. falciparum and P. vivax, as indicated by the asynchronous correlation cross-peaks. Two-trace two-dimensional (2T2D) spectroscopy, applied to EPR spectra of blood at the beginning of the infection, showed differences between P. falciparum and P. vivax. CONCLUSIONS: A unique feature of 2D-COS is the ability to discriminate the collected Raman and EPR spectra. The changes observed during the course of a malaria infection have different dynamics for P. falciparum and P. vivax, indicated by the reverse sequence of events. For each type of parasite, a specific recycling process for iron was observed in the infected blood.

Musculoskeletal manifestations of Lyme borreliosis - a review.

Lyme borreliosis (LB) is a zoonotic disease caused by Gram-negative bacteria Borrelia burgdorferi sensu lato. The majority of reported cases of LB originate in the northern hemisphere, mostly in the US and Europe. One of the typical manifestations of LB are musculoskeletal symptoms; they may appear in any of the three LB stages. The diagnosis is based on clinical manifestations and confirmed by serological tests. One course of antibiotic therapy is sufficient for LB to dissipate in most cases, although for some patients, the symptoms subside gradually even after completion of therapy. Patients who have been demonstrated to have specific antibodies but are symptomless should not be treated. In instances where the advised treatment proved to be unsuccessful, patients should be referred to rheumatologist for additional diagnostics. The goal of this review is to update physicians on current scientific knowledge on musculoskeletal manifestations of LB.

Mortality Following Clostridioides difficile Infection in Europe: A Retrospective Multicenter Case-Control Study.

We aimed to describe the clinical presentation, treatment, outcome and report on factors associated with mortality over a 90-day period in Clostridioides difficile infection (CDI). Descriptive, univariate, and multivariate regression analyses were performed on data collected in a retrospective case-control study conducted in nine hospitals from seven European countries. A total of 624 patients were included, of which 415 were deceased (cases) and 209 were still alive 90 days after a CDI diagnosis (controls). The most common antibiotics used previously in both groups were ß-lactams; previous exposure to fluoroquinolones was significantly (p = 0.0004) greater in deceased patients. Multivariate logistic regression showed that the factors independently related with death during CDI were older age, inadequate CDI therapy, cachexia, malignancy, Charlson Index, long-term care, elevated white blood cell count (WBC), C-reactive protein (CRP), bacteraemia, complications, and cognitive impairment. In addition, older age, higher levels of WBC, neutrophil, CRP or creatinine, the presence of malignancy, cognitive impairment, and complications were strongly correlated with shortening the time from CDI diagnosis to death. CDI prevention should be primarily focused on hospitalised elderly people receiving antibiotics. WBC, neutrophil count, CRP, creatinine, albumin and lactate levels should be tested in every hospitalised patient treated for CDI to assess the risk of a fatal outcome.

Decreased Expression of the High Mobility Group Box 1 (HMGB1) Gene in Peripheral Blood in Patients with Mild or Moderate Clostridioides difficile Infection.

Cytokines are mediators of inflammation induced in the course of Clostridioides difficile infection (CDI). High Mobility Group Box 1 (HMGB1) is a cytokine playing an important role in the pathogenesis of numerous inflammatory and autoimmune diseases. The aim of the study was to assess the HMGB1 gene expression in the course of CDI. We have performed a prospective case-control study- including 55 adult patients, among them 27 with CDI, who were hospitalized from October 2018 to February 2020 and 28 healthy volunteers. We assessed: a complete blood count with differential leukocyte count, blood creatinine, albumin, and C-reactive protein (CRP) levels. Then, the expression of the HMGB1 gene was evaluated using quantitative Real-Time PCR. Patients with CDI were found to have a significant increase in white blood cells (WBC), neutrophil count, and CRP levels, they also exhibited decreased levels of albumin compared with controls. The HMGB1 gene expression was significantly lower among patients with CDI compared with the control group and significantly, inversely correlated with CRP level in blood. In conclusion, we have observed a decreased expression of the HMGB1 gene in peripheral blood of patients with mild or moderate CDI, which hypothetically could reflect their diminished capability to fight the pathogen.

Clostridium difficile caused changes in fatty acids profile and resolvin D1 content in plasma of infected patients.

OBJECTIVES: Clostridium difficile infection (CDI) is an acute gastrointestinal infection caused by anaerobic, toxin-producing bacteria. During the course of CDI, there is a general inflammatory state. In order to gain a deeper understanding of the role of fatty acids (FAs) in the pathogenesis of acute infection we analyzed their plasma content in both patients with CDI and controls. METHODS: The study groups included 40 patients with CDI and 40 healthy volunteers. Plasma FA content was analyzed by gas chromatography, resolvin D1 (RvD1) level using ELISA assay, and we assessed the white blood cell (WBC) count, neutrophil count and C-reactive protein (CRP) level. RESULTS: Patients with CDI were characterized by significantly higher values of WBC, neutrophils, platelets and CRP compared with the control group. The saturated FA index was statistically higher and total n-3 FA was significantly decreased in the plasma of CDI patients as compared with the control group. RvD1 content was significantly higher in the control group as compared with patients with CDI. CONCLUSION: In patients with good outcomes, we probably observed the effective resolution of inflammation, as reflected in n-3 FA metabolism and their significant decrease in plasma. This may indicate the therapeutic role of n-3 FA in CDI infection.

Prevalence of Function-Dependent Temporomandibular Joint and Masticatory Muscle Pain, and Predictors of Temporomandibular Disorders among Patients with Lyme Disease.

The aim was to determine the occurrence of temporomandibular disorders (TMDs) in patients with Lyme disease (LD), and to estimate the contribution of factors that may identify TMD among LD patients. In seventy-six (N = 76) adult patients with LD (mean age 57.6 ± 14.6 years) and 54 healthy non-Lyme volunteers with a mean age of 56.4 ± 13.5 years, possible function (i.e., non-pain) diagnoses were established using the Research Diagnostic Criteria of Temporomandibular Disorders (RDC/TMD). Pain diagnoses were established by means of the function-dependent dynamic and static tests. The two groups did not significantly differ in the frequency of disc displacements diagnoses and function-dependent pain diagnoses. LD showed a significantly higher frequency (p < 0.001) of osteoarthrosis than the control group. For the prediction of pain diagnoses in LD patients, the single regression analyses pointed out an association with age, sleep bruxism (SB), and awake bruxism (AB). Two predictors (i.e., SB (p = 0.002) and AB (p = 0.017)) were statistically significant in the final multiple variable model. The frequency of TMD in patients with LD based on function-dependent tests was not significantly different from that in the control group. This investigation suggests that the contribution of bruxism to the differentiation between patients with Lyme and TMD is high.

Analysis of cases of Lyme arthritis in patients hospitalized in Infectious Diseases Department, University Hospital in Cracow.

Lyme disease is an emerging problem in Poland. Analysis has been undertaken of the medical documentation of 86 patients hospitalized in the Infectious Diseases Department, University Hospital in Cracow in 2013-2016, suspected of Lyme arthritis. It has especially considered medical history including potential exposure to the infection, detailed characteristics of the symptoms, diagnostic challenges and results of the treatment. Only some patients had a history of erythema migrans and not all of them recalled tick-bite. The majority of the patients had affected large joints, especially knee joints, and polyarthritis was rarely observed. Symptoms were resolved completely or partially after antibiotic treatment in most patients. The diagnosis of Lyme arthritis in areas endemic for Lyme disease is still a diagnostic challenge in patients with other rheumatic diseases, including osteoarthritis.

Clostridium difficile infection: review.

Clostridium difficile (C. difficile) is a Gram-positive, spore-forming, anaerobic bacillus, which is widely distributed in the intestinal tract of humans and animals and in the environment. In the last decade, the frequency and severity of C. difficile infection has been increasing worldwide to become one of the most common hospital-acquired infections. Transmission of this pathogen occurs by the fecal-oral route and the most important risk factors include antibiotic therapy, old age, and hospital or nursing home stay. The clinical picture is diverse and ranges from asymptomatic carrier status, through various degrees of diarrhea, to the most severe, life threatening colitis resulting with death. Diagnosis is based on direct detection of C. difficile toxins in feces, most commonly with the use of EIA assay, but no single test is suitable as a stand-alone test confirming CDI. Antibiotics of choice are vancomycin, fidaxomicin, and metronidazole, though metronidazole is considered as inferior. The goal of this review is to update physicians on current scientific knowledge of C. difficile infection, focusing also on fecal microbiota transplantation which is a promising therapy.

Differences between palpation and static/dynamic tests to diagnose painful temporomandibular disorders in patients with Lyme disease.

OBJECTIVES: The aim was to determine the frequency of Research Diagnostic Criteria for Temporomandibular Disorders (RDC/TMD)-based pain diagnoses and dynamic/static tests-based pain diagnoses, and to assess the agreement of palpation tests with static/dynamic tests. MATERIALS AND METHODS: Eighty-six (N = 86) adult patients with Lyme disease (mean age 57.0 ± 14.3 years; male/female ratio was 42/44) were examined according to techniques described in the RDC/TMD. Additionally, dynamic/static tests were performed. For RDC/TMD-based pain diagnoses and dynamic/static tests-based pain diagnoses, descriptive frequencies were calculated. Differences between the frequency of palpation-based diagnoses and of dynamic/static-based diagnoses as well as the agreement between pain diagnoses established with the two diagnostic approaches were assessed. RESULTS: RDC/TMD-based pain diagnoses were made in 61 patients for myofascial pain and in 11 patients for arthralgia and/or osteoarthritis. Based on dynamic/static tests, mainly myogenous pain was diagnosed in 6 patients, and a mainly arthrogenous pain in 5. The agreement of palpation tests with static/dynamic tests in Lyme disease population was poor. CONCLUSION: A high prevalence of TMD symptoms was found in patients with Lyme disease. The results suggest that using palpation tests alone could overestimate primary TMDs when comorbid conditions are present. CLINICAL RELEVANCE: Dynamic/static tests should be used as part of the routine TMD assessment. In case of Lyme disease as the actual cause of the facial pain, while the dentist might be suspecting TMD when dynamic/static TMD tests are negative, referral to an appropriate specialist for the diagnosis and treatment of Lyme disease needs to be made.

Meningoencephalitis caused by Lactobacillus plantarum - case report.

Specific strains of Lactobacillus spp. are widely used as probiotic agents but it has been repeatedly reported that may have a pathogenic potential. We present the report on a case of meningoencephalitis caused by Lactobacillus plantarum in a 63-year-old man with newly diagnosed metastatic planoepitheliale lung cancer. The patient was hospitalised due to newly diagnosed cancer and during the course of hospitalisation developed symptoms of neuroinfection. On the basis of the symptoms and results of the conducted tests the patient was diagnosed with bacterial meningoencephalitis. In microbiological tests of the blood and cerebrospinal fluid L. plantarum was cultured. During the course of antibiotic therapy the patient's condition improved. Lactobacilli are now recognised as a causative agent of infection, most notably bacteraemia. To our knowledge, this is the fourth documented case of Lactobacillus-associated neuroinfection, and only the second in an adult. Lactobacilli cause mostly opportunistic infections in immunocompromised individuals.

The level of fecal calprotectin significantly correlates with Clostridium difficile infection severity.

INTRODUCTION: Fecal calprotectin (FC) rises significantly in intestinal inflammation accompanied by neutrophil activation - such as Clostridium difficile infection (CDI). The aim of the study was to evaluate the benefit of FC testing in assessing the severity of CDI. MATERIALS AND METHODS: The study group included 76 patients with CDI hospitalized in the Jagiellonian University Hospital in Krakow from July 2017 till January 2018. FC levels were measured using an EIA (Enzyme Immunoassay). Demographic, clinical information and blood tests were recorded using standardized data collection forms. The selection of patients into non-severe and severe groups was carried out in accordance with the ESCMID criteria (European Society of Clinical Microbiology and Infectious Diseases) and some modi cations to those criteria were proposed. RESULTS: the studied population included 76 patients (39 men and 37 women) with CDI aged from 24 to 98 years (mean: 72). Median calprotectin level was 739 (Q25-Q75: 612-799 µg/g), characteristic of patients with colitis. A statistically significant difference in FC concentration in patients with severe vs non-severe CDI was observed (severe - 770 vs non-severe - 659 µg/g, p = 0.009). FC directly correlated with platelets level; however, no correlation between FC level and the blood parameters prognostic for CDI (leukocyte, neutrophil count, albumin, creatinine levels) was found. CONCLUSION: FC level is an indication of ongoing intestinal inflammation in CDI patients. FC level significantly correlated with CDI severity, which demonstrates that FC could serve as a predictive marker for assessing CDI severity.

Fatty acids and selected endocannabinoids content in cerebrospinal fluids from patients with neuroinfections.

Neuroinfections are a significant medical problem and can have serious health consequences for patients. Their outcome, if not fatal, can be associated with permanent residual deficits. Cerebrospinal fluid (CSF) examination is commonly used for meningitis confirmation. Fatty acids (FA) are precursors of lipid mediators with pharmacological activity. They actively modulate inflammation as well as contribute to its resolution. Therefore the aim of this study was to determine the FA and selected endocannabinoids (ECB) content in the CSF obtained from patients with bacterial (BM) and viral meningitis (VM) using chromatographic techniques. A significantly lower level of saturated FA was found in patients with BM and VM as compared to controls. There was a significantly higher concentration of long-chain monounsaturated FA and polyunsaturated n-6 FA in the CSF obtained from patients with neuroinfection. Moreover, a significant reduction of n-3 FA in CSF obtained from patients with BM and VM was demonstrated. The highest amount of ECB was detected in the CSF of patients with VM: eicosapentaenoyl ethanolamide (1.65 pg/mL), docosahexaenoyl ethanolamide (655.5 pg/mL) and nervonoyl ethanolamide (3.09 ng/mL). Results indicate the participation of long-chain monounsaturated and polyunsaturated FA and their derivatives in the inflammatory process and likely in the process of resolution of inflammation during neuroinfection. It seems that the determination of the FA and ECB profile in CSF may be a valuable biomarker of health and may allow the development of new pharmacological strategies, therapeutic goals and fatty acids supplementation necessary in the fight against inflammation of the central nervous system.

Clostridium difficile infection in young people - 2 case reports.

Clostridium difficile infection (CDI) remains one of the most important healthcare-associated infections of the last two decades. The pathogen is a Gram-positive, toxin-producing, anaerobic, rod-shaped and sporeforming bacterium; it is ubiquitous in the human environment. Transmission occurs by the fecal-oral route. The consequence of the action of bacterial toxins is initially a local inflammatory reaction, which then goes into systemic inflammation. Clinical presentation is varied; some patients are asymptomatic, in symptomatic form the main symptom is diarrhea of varying severity, which is sometimes accompanied by acute abdominal pain, nausea, vomiting and high fever. Risk factors of CDI include prior antibiotic use, increasing age and recent hospitalisation. Extremely rarely does CDI occur in immunocompetent patients under 30 years of age, even if previously treated with an antibiotic. Here presented are two untypical cases of CDI development in the lower age group, in the presence of additional risk factors of comorbid gastrointestinal tract infections. Both developed infections are following recent infection - Salmonella enterididis in the first case and Salmonella typhi in the second case. Therefore, the article also contains basic principles for the diagnosis and treatment of Salmonella spp.

Analysis of risk factors and outcomes of Clostridium difficile infection.

INTRODUCTION: Clostridium difficile (C. difficile) is a Gram-positive, anaerobic rod-shaped bacteria, widely spread in the human environment. In the last decade, the frequency and severity of Clostridium difficile infection (CDI) have been increasing, making this particular disease one of the most significant nosocomial infections. The aim of our study was an analysis of CDI risk factors, its course and consequences. MATERIALS AND METHODS: Medical documentation of the patients treated for CDI in the University Hospital in Cracow and St Anne's Hospital in Miechów has been analysed. The analysis focused on epidemiological data, blood parameters, comorbidities, recurrence rate, and complication rate (deaths included). As part of risk factors analysis, antibiotic use or hospitalisation in a period of 3 months before the episode of infection was considered relevant. Blood tests have been performed using routinely employed, standard methods. RESULTS: We evaluated data of 168 people infected with C. difficile, out of which there were 102 women (61%) and 66 men (39%). The median age of the patients was 74 years for the entire population with 76 years for women and 71 years for male patients. One hundred thirteen people (67%) had been previously hospitalised, and 5 person was a pensioner of a nursing home. 99 people (59%) were treated with antibiotics within 3 months before the first episode of infection. An average length of the hospital stay because of CDI was 11 days. One hundred thirty persons (77%) experienced only 1 episode whereas 38 people (23%) had more than 1 episode of infection. The person with the largest number of recurrences had 9 of them. CONCLUSIONS: The development of CDI is an increasing problem in a group of hospitalised persons, particularly of an old age. The general use of beta-lactam antibiotics is the cause of a larger number of infections with C. difficile. Vast majority of patients have had at least one typical risk factor of CDI.

The presence of IL-8 +781 T/C polymorphism is associated with the parameters of severe Clostridium difficile infection.

PURPOSE: There is large variation in the clinical manifestations of Clostridium difficile infection (CDI). We also still can not predict which patients are more susceptible to reinfection with CDI. The aim of our study was to evaluate the effect of gene single nucleotide polymorphisms (SNP) of proinflammatory cytokines, specifically IL-1ß, IL-8 on the development, clinical course and recurrence of CDI. METHODS: We performed a prospective study of adults (130 people ≥ 18 years) including 65 patients with CDI treated in tertiary hospital and 65 healthy persons. The following 3 variants were analyzed for the occurrence of gene polymorphisms in patients with CDI versus the control group: IL-1ß +3953 A/G (rs1143634), IL-1ß -31 A/G (rs1143627), and IL-8 +781 T/C (rs2227306). Then, we assessed the correlation between these genetic polymorphisms and biochemical parameters important in CDI course, CDI severity as well as CDI recurrence. RESULTS AND CONCLUSIONS: The presence of genetic polymorphisms of IL-1ß +3953 A/G, -31 A/G and IL-8 +781 T/C did not have an effect on the development or recurrence of CDI. The presence of IL-8 +781 T/C polymorphism is associated with the severe CDI.

[Diagnostic difficulties in amebic liver abscess].

Amebiasis is a widespread parasitic infection caused by the human-specific protozoan Entamoeba histolytica (E.- histolytica). Tropical and subtropical regions with poor socioeconomic and sanitary conditions belong to endemic areas. The highest rates of E. histolytica infection are observed in India, Mexico, Africa, some parts ofCentral and South America. Up to 90% of infections remain asymptomatic, about 10% of patients develop amebic colitis. About 10% of symptomatic individuals may present with an extraintestinal manifestation, mostly amebic liver abscess (ALA). Clinical symptoms of ALA appear within 5 months after an exposition to E. histolytica cysts. Anamnesis revealing a travel to endemic area plays a crucial role in a diagnostic process, which is further supported by an physical examination, radiological findings, serology and parasitology test. The following article presents the difficulties which may occur when the ALA is suspected in a patient traveling from endemic areas.