Comparative study of TNF-α and IL-10 levels at different times of the course of COVID-19.

INTRODUCTION: SARS-CoV-2 infection (COVID-19) induces dysregulated production of pro- and anti-inflammatory cytokines, called the cytokine storm, leading to the development of severe pneumonia and ARDS. We aimed to examine the dynamic cytokine response on different days of the disease in adult COVID-19 patients. METHODOLOGY: Our study included 142 patients (with SARS-CoV-2 PCR-positive nasopharyngeal samples) with varying disease severity and admitted on different days of the disease. We examined the presence and mean levels of TNF-α and IL-10 and did a correlation and logistic regression analysis. RESULTS: TNF-α levels were high in all patients, with mean levels being the highest on day 5 of the disease. IL-10 was high only in a quarter of the patients. The levels of IL-10 were also the highest on day 5, which was significantly different from the mean levels on the other days of the disease. Average IL-10 levels were not any higher than the normal range on the other days. We found a significant positive correlation between the levels of TNF-α and IL-10 during the first week of the infection. In the second week, the positive correlation was no longer significant, and starting from day 10, there was even a slight negative correlation. IL-10 level increase showed prognostic significance for severe, but not the critical forms of the disease. CONCLUSIONS: The uncontrolled immune response to SARS-CoV-2 in the second week of the disease can be the result of dysregulated production of endogenous anti-inflammatory cytokines. This leads to a severe disease course and a possible unfavorable outcome.

Uncovering the Hidden Threat: A Case Report of Suspected Dengue Fever in Armenia.

Dengue is a viral infection transmitted by mosquitoes that causes fever, headache, joint pain, nausea, vomiting, and pain behind the eyes. In severe cases, it can progress to dengue hemorrhagic fever and dengue shock syndrome, which can be life-threatening. Armenia has not reported a single case of dengue to date and is non-endemic for this disease. However, it has been found that the vector of the disease, Aedes albopictus, is present in Armenia since 2016. The aim of this report is to present the imported case of suspected dengue hemorrhagic fever. A 23-year-old female who was admitted to the University Hospital experienced symptoms of general weakness, fever, joint pain, and chills after her return from Bali and had a three-day febrile period. A thorough examination revealed mosquito bites on her skin. On the fourth day of hospitalization, the patient's condition deteriorated. She started experiencing vaginal bleeding. On the same day, the patient noted a small petechiae rash sized 1-2 mm in diameter in the upper and lower extremities. The patient deteriorated, with progressive leukopenia and thrombocytopenia, and hypertransaminasemia. Screening tests for HIV and hepatitis A, B, C, and E were performed, and the results showed that the anti-hepatitis C antibody was positive, while the hepatitis C virus polymerase chain reaction was negative. The case was reported to the National Center for Disease Control and Prevention as an imported case of hemorrhagic fever. Unfortunately, no lab test was available there for confirmation of the diagnosis. The patient received IV infusion and symptomatic treatment. Her condition improved, and upon discharge, she was in a state of recovery. This case report highlights the importance of early diagnosis and appropriate treatment for hemorrhagic fevers, particularly dengue fever. The unavailability of diagnostic kits for dengue in Armenia highlights the need to invest in improving their availability. It also emphasizes the importance of maintaining dengue surveillance in non-endemic nations and carefully evaluating and monitoring febrile patients who have returned from dengue-endemic countries.

Molecular Epidemiology and Virulence of Non-Typhoidal Salmonella in Armenia.

In this work, we analysed human isolates of nontyphoidal Salmonella enterica subsp. enterica (NTS), which were collected from salmonellosis cases in Armenia from 1996 to 2019. This disease became a leading food-borne bacterial infection in the region, with the younger age groups especially affected. The isolates were characterised by serotyping, Enterobacterial Repetitive Intergenic Consensus (ERIC-PCR) typing, and whole genome sequencing (WGS). The main serotypes were S. Typhimurium, S. Enteritidis, and S. Arizonae. ERIC-PCR indicated a high degree of clonality among S. Typhimurium strains, which were also multidrug-resistant and produced extended spectrum beta-lactamases. During the study period, the frequency of S. Typhimurium and S. Arizonae isolations decreased, but with the increase in S. Enteritidis and other NTS. A total of 42 NTS isolates were subjected to WGS and explored for virulence-related traits and the corresponding genetic elements. Some virulence and genetic factors were shared by all NTS serotypes, while the main differences were attributed to the serotype-specific diversity of virulence genes, SPIs, virulence plasmids, and phages. The results indicated the variability and dynamics in the epidemiology of salmonellosis and a high virulence potential of human NTS isolates circulating in the region.

Kinetics of anti-nucleocapsid IgG response in COVID-19 immunocompetent convalescent patients.

The comprehension of a long-term humoral immune response against SARS-CoV-2 can shed light on the treatment and vaccination strategies of COVID-19 disease, improving the knowledge about this virus infection and/or re-infection. We assessed the IgG antibodies against SARS-CoV-2 nucleocapsid (N) protein (anti-SARS-CoV-2 (N) IgG) in 1441 COVID-19 convalescent patients within 15 months longitudinal study from middle-developed country. The main inclusion criteria was positive RT- PCR result on nasopharyngeal swab samples at least one month before antibody testing and absence of any induced or inherited immunodeficiency. 92.7% of convalescent patients' serum contained anti-SARS-CoV-2 (N) IgG and only 1.3% of patients had a delayed antibody response. In the majority of convalescent patients' the durability of antibodies lasted more than one year. The kinetics of anti-SARS-CoV-2 (N) IgG took a bell-shaped character-increased first 25-30 weeks, then started to decrease, but were still detectable for more than 15 months. We found that on the one hand anti-SARS-CoV-2 humoral response level correlates with disease severity, on the other, in particular, the level of peak antibodies correlates with age-older patients develop more robust humoral response regardless of sex, disease severity and BMI.

Assessment of pathomorphological characteristics of the oral mucosa in patients with HBV, HCV and HIV.

INTRODUCTION: Oral clinical manifestations in HBV HCV and HIV patients indicate a deterioration in general health status. The aim of the study was to assess pathomorphologic features of oral mucosa observed in patients with these diseases. METHODOLOGY: The study was conducted in N1 Dental Clinic of YSMU after M. Heratsi. The total number of patients taking part in the research was 120, including HBV (n = 40), HCV (n = 40) and HIV (n = 40). After biopsy and subsequent histological examination of the oral mucosa, statistical analysis was carried out using Excel 2013 and R software. RESULTS: Pathomorphological examination revealed inflammatory infiltrations in all samples collected from HBV, HCV and HIV patients. These changes included microcirculatory disorders in 98.3% of samples: fibrinous-like deposits lining the surface of erosions and ulcers on the oral mucosa (1.67%), fibrosis of the mucous membrane (70%), dystrophy of squamous epithelium (93.3%) and bone sequestration (3.3%). Comparative analysis of pathomorphological characteristics revealed distinct content of infiltrates: lymphoplasmacytic infiltration in patients with HBV and HCV, while HIV patients showed neutrophils infiltration and lack of plasmocytes. CONCLUSIONS: There are common abnormal morphological changes in the oral mucosa typical of all patients with HBV, HCV and HIV, as well as liver diseases specific to each of them. Inflammation in the patients with HIV indicated impairment of the humoral immune system. Understanding the distinct characteristic of inflammation in the oral cavity could be useful for early differential diagnosis and management of patients with HIV, HBV and HCV.

Extended-Spectrum ß-Lactamases in Human Isolates of Multidrug-Resistant Non-typhoidal Salmonella enterica.

A total of 291 non-duplicate isolates of non-typhoidal Salmonella (NTS) were collected from the fecal samples of patients with salmonellosis in Armenia and Georgia during 1996-2016. The isolates were tested for resistance to antimicrobials, including extended-spectrum ß-lactamases (ESBL). The high prevalence of multidrug-resistance (MDR) and ESBL-producer phenotypes was detected among Salmonella enterica subsp. enterica serovar Typhimurium (S. Typhimurium) isolates collected from patients in Armenia between 1996 and 2016. A total of 36 MDR NTS isolates were subjected to whole genome sequencing (WGS) to determine the genetic background of antimicrobial resistance (AMR) and mobile genetic elements. All ESBL-producing S. Typhimurium isolates belonged to the same sequence type (ST328). The ESBL-producer phenotype was associated with plasmid-encoded CTX-M-5 production. A range of other plasmids was associated with resistance to other antimicrobials, including the MDR phenotype.

Potential Involvement of Salmonella Infection in Autoimmunity.

In this work, we investigated the potential effects of nontyphoidal Salmonella infection on autoantibody (AA) formation. The titer and profiles of autoantibodies in the sera of patients with acute salmonellosis due to Salmonella enterica serovar Typhimurium (S. Typhimurium) or Salmonella enterica serovar Enteritidis (S. Enteritidis) infection, as well as in convalescent patients, were determined with indirect immunofluorescence. A significant increase of autoantibodies in acute diseases caused by both serotypes of Salmonella and during post infection by S. Enteritidis was detected. Antibody profile analysis by multivariate statistics revealed that this increase was non-specific and was not dependent on the infectious agent or disease stage. The results obtained suggest that nontyphoidal Salmonella infection contributes to the generation of autoantibodies and may play a role in autoimmune disease.

Epidemiology and economic losses of rotavirus infection associated with hospitalization of Armenian children.

INTRODUCTION: Rotavirus (RV) is the leading cause of severe diarrhea-associated morbidity and mortality among children worldwide. Limited data exist on the epidemiology and burden of rotavirus gastroenteritis in Armenia. The purpose of this study is to estimate the economic losses and describe the epidemiological characteristics of rotavirus infections in hospitalized children in Armenia. METHODOLOGY: A retrospective chart review was performed of all children (aged < 5 years) with Rotavirus infection admitted to the "Nork" Republican Infection Clinical Hospital in Yerevan, the capital of Armenia, between January and July 2014. Criteria of inclusion were age under 5 years old and presence of RV antigen in stool by enzyme linked immunosorbent assay. RESULTS: The total number of patients was 126; average age was 28.7 ± 13.3 months; 54.8% were male. The highest number of cases (31.8%) was observed in April. Most of the patients (71.4%) were hospitalized in the first three days, demonstrating an acute onset of the disease. In total 19% of the patients had received RV vaccine and, despite this, were infected with RV. Based on bacteriological examination of stool, 18.3% of patients had RV infection combined with pathogenic or conditional pathogenic microflora. All patients spent 817 days in total in the hospital. Economic losses associated with hospitalization were 16340000 AMD (≈ 33346 USD). CONCLUSION: Rotavirus carries significant morbidity and economic losses. Comprehensive estimates of the disease characteristics and introduction of a national immunization program against RV initiated in 2012 may decrease this burden. Further studies to evaluate the feasibility and cost-effectiveness of such a program are warranted.

A retrospective chart review study to describe selected zoonotic and arboviral etiologies in hospitalized febrile patients in the Republic of Armenia.

BACKGROUND: Scant information is available on the infectious causes of febrile illnesses in Armenia. The goal of this study was to describe the most common causes, with a focus on zoonotic and arboviral infections and related epidemiological and clinical patterns for hospitalized patients with febrile illnesses of infectious origin admitted to Nork Infectious Diseases Clinical Hospital, the referral center for infectious diseases in the capital city, Yerevan. METHOD: A chart review study was conducted in 2014. Data were abstracted from medical charts of adults (≥18 years) with a fever (≥38 °C), who were hospitalized (for ≥24 h) in 2010-2012. RESULTS: Of the 600 patients whose charts were analyzed, 76 % were from Yerevan and 51 % were male; the mean age (± standard deviation) was 35.5 (±16) years. Livestock exposure was recorded in 5 % of charts. Consumption of undercooked meat and unpasteurized dairy products were reported in 11 and 8 % of charts, respectively. Intestinal infections (51 %) were the most frequently reported final medical diagnoses, followed by diseases of the respiratory system (11 %), infectious mononucleosis (9.5 %), chickenpox (8.3 %), brucellosis (8.3 %), viral hepatitis (3.2 %), and erysipelas (1.5 %). Reviewed medical charts included two cases of fever of unknown origin (FUO), two cutaneous anthrax cases, two leptospirosis cases, three imported malaria cases, one case of rickettsiosis, and one case of rabies. Engagement in agricultural activities, exposure to animals, consumption of raw or unpasteurized milk, and male gender were significantly associated with brucellosis. CONCLUSION: Our analysis indicated that brucellosis was the most frequently reported zoonotic disease among hospitalized febrile patients. Overall, these study results suggest that zoonotic and arboviral infections were not common etiologies among febrile adult patients admitted to the Nork Infectious Diseases Clinical Hospital in Armenia.

Systemic Concentrations of Short Chain Fatty Acids Are Elevated in Salmonellosis and Exacerbation of Familial Mediterranean Fever.

Gut microbiota-produced short chain fatty acids (SCFAs) play an important role in the normal human metabolism and physiology. Although the gradients of SCFAs from the large intestine, where they are largely produced, to the peripheral blood as well as the main routes of SCFA metabolism by different organs are known well for the healthy state, there is a paucity of information regarding how these are affected in disease. In particular, how the inflammation caused by infection or autoinflammatory disease affect the concentration of SCFAs in the peripheral venous blood. In this work, we revealed that diseases caused either by infectious agents (two Salmonella enterica serovars, S. Enteritidis, and S. Typhimurium) or by the exacerbation of an autoinflammatory disease, familial Mediterranean fever (FMF), both result in a significantly elevated systemic concentration of SCFAs. In the case of salmonellosis the concentration of SCFAs in peripheral blood was significantly and consistently higher, from 5- to 20-fold, compared to control. In the case of FMF, however, a significant increase of SCFAs in the peripheral venous blood was detected only in the acute phase of the disease, with a lesser impact in remission. It seems counterintuitive that the dysbiotic conditions, with a reduced number of gut microorganisms, produce such an effect. This phenomenon, however, must be appraised within the context of how the inflammatory diseases affect the normal physiology. We discuss a number of factors that may contribute to the "leak" and persistence of gut-produced SCFAs into the systemic circulation in infectious and autoinflammatory diseases.

Intestinal Infections Among Febrile Hospitalized Patients in the Republic of Armenia: A Retrospective Chart Review.

In the past, several enteric outbreaks in 1996, 1998, 1999, and 2003 caused by Salmonella typhi, a Gram-negative bacterium, have occurred in Armenia. This study describes the demographic, epidemiological, and clinical characteristics of febrile hospitalized patients with intestinal infections in Armenia. Using a chart review study design, medical data from adult patients who were hospitalized at the Nork hospital during 2010-2012 were reviewed. A total of 600 medical charts were reviewed. Of these, 51 % were diagnosed with intestinal infections. Among these patients, 59 % had an intestinal infection of known etiology, with three main pathogens identified: Salmonella sp. (32 %), Shigella sp. (32 %), and Staphylococcus aureus (18 %). After controlling for the calendar year, age in years, and gender, patients detected with Salmonella sp. were more likely to reported the presence of a family member with similar signs or symptoms [odds ratio (OR) 9.0; 95 % CI 2.4-33.7] and the lack of a water tap at home (OR 3.9; 95 % CI 1.7-9.5). Evidence indicates that Salmonella sp., Shigella sp., and S. aureus as the most common etiologies reported among febrile hospitalized patients. A high percentage of patients had intestinal infections of unknown etiology; thus, improvement in laboratory capacity (enabling more advanced tests, such as polymerase chain reaction) would increase the identification of the enteropathogens causing disease in Armenia.

Differential induction of total IgE by two Salmonella enterica serotypes.

The main goal of this study was to establish how the inflammation caused by infection with two different Salmonella enterica serotypes, S. Typhimurium and S. Enteritidis, may lead to the predisposition to allergy as measured by total IgE level in the blood. Infection by S. Typhimurium did not affect the systemic IgE concentration while in S. Enteritidis-infected patients there was a significant 3.5-fold increase. This effect was especially profound in patients >4 years old, with up to the 8-fold increase above the norm. The degree of dysbiosis in these two infections measured with the comparative counts of cultivated bacteria showed an inverse relationship with the IgE concentration. Earlier we reported the elevated level of IL-17 in patients infected by S. Enteritidis. In the current study a significant correlation was found between the concentrations of IL-17 and IgE suggesting a possible role played by this cytokine in triggering the production of IgE in response to S. Enteritidis infection.

Management of familial Mediterranean fever by colchicine does not normalize the altered profile of microbial long chain fatty acids in the human metabolome.

In our previous works we established that in an autoinflammatory condition, familial Mediterranean fever (FMF), the gut microbial diversity is specifically restructured, which also results in the altered profiles of microbial long chain fatty acids (LCFAs) present in the systemic metabolome. The mainstream management of the disease is based on oral administration of colchicine to suppress clinical signs and extend remission periods and our aim was to determine whether this therapy normalizes the microbial LCFA profiles in the metabolome as well. Unexpectedly, the treatment does not normalize these profiles. Moreover, it results in the formation of new distinct microbial LCFA clusters, which are well separated from the corresponding values in healthy controls and FMF patients without the therapy. We hypothesize that the therapy alters the proinflammatory network specific for the disease, with the concomitant changes in gut microbiota and the corresponding microbial LCFAs in the metabolome.

Profiles of Microbial Fatty Acids in the Human Metabolome are Disease-Specific.

The human gastrointestinal tract is inhabited by a diverse and dense symbiotic microbiota, the composition of which is the result of host-microbe co-evolution and co-adaptation. This tight integration creates intense cross-talk and signaling between the host and microbiota at the cellular and metabolic levels. In many genetic or infectious diseases the balance between host and microbiota may be compromised resulting in erroneous communication. Consequently, the composition of the human metabolome, which includes the gut metabolome, may be different in health and disease states in terms of microbial products and metabolites entering systemic circulation. To test this hypothesis, we measured the level of hydroxy, branched, cyclopropyl and unsaturated fatty acids, aldehydes, and phenyl derivatives in blood of patients with a hereditary autoinflammatory disorder, familial Mediterranean fever (FMF), and in patients with peptic ulceration (PU) resulting from Helicobacter pylori infection. Discriminant function analysis of a data matrix consisting of 94 cases as statistical units (37 FMF patients, 14 PU patients, and 43 healthy controls) and the concentration of 35 microbial products in the blood as statistical variables revealed a high accuracy of the proposed model (all cases were correctly classified). This suggests that the profile of microbial products and metabolites in the human metabolome is specific for a given disease and may potentially serve as a biomarker for disease.