Imaging Immune Cells Using Fc Domain Probes in Mouse Cancer Xenograft Models

Tracking immune responses is complex due to the mixture of cell types, variability in cell populations, and the dynamic environment. Tissue biopsies and blood analysis can identify infiltrating and circulating immune cells; however, due to the dynamic nature of the immune response, these are prone to sampling errors. Non-invasive targeted molecular imaging provides a method to monitor immune response, which has advantages of providing whole-body images, being non-invasive, and allowing longitudinal monitoring. Three non-specific Fc-containing proteins were labeled with near-infrared dye IRDye800CW and used as imaging probes to assess tumor-infiltrating immune cells in FaDu and A-431 xenograft models. We showed that Fc domains localize to tumors and are visible by fluorescent imaging. This tumor localization appears to be based on binding tumor-associated immune cells and some xenografts showed higher fluorescent signals than others. The Fc domain alone bound to different human immune cell types. The Fc domain can be a valuable research tool to study innate immune response.

Nafamostat reduces systemic inflammation in TLR7-mediated virus-like illness

Background The serine protease inhibitor nafamostat has been proposed as a treatment for COVID-19, by inhibiting TMPRSS2-mediated viral cell entry. Nafamostat has been shown to have other, immunomodulatory effects, which may be beneficial for treatment, however animal models of ssRNA virus infection are lacking. In this study, we examined the potential of the dual TLR7/8 agonist R848 to mimic the host response to an ssRNA virus infection and the associated behavioural response. In addition, we evaluated the anti-inflammatory effects of nafamostat in this model. Methods CD-1 mice received an intraperitoneal injection of R848 (200 μg, prepared in DMSO, diluted 1:10 in saline) or diluted DMSO alone, and an intravenous injection of either nafamostat (100 μL, 3 mg/kg in 5% dextrose) or 5% dextrose alone. Sickness behaviour was determined by temperature, food intake, sucrose preference test, open field and forced swim test. Blood and fresh liver, lung and brain were collected 6 h post-challenge to measure markers of peripheral and central inflammation by blood analysis, immunohistochemistry and qPCR. Results R848 induced a robust inflammatory response, as evidenced by increased expression of TNF, IFN-γ, CXCL1 and CXCL10 in the liver, lung and brain, as well as a sickness behaviour phenotype. Exogenous administration of nafamostat suppressed the hepatic inflammatory response, significantly reducing TNF and IFN-γ expression, but had no effect on lung or brain cytokine production. R848 administration depleted circulating leukocytes, which was restored by nafamostat treatment. Conclusions Our data indicate that R848 administration provides a useful model of ssRNA virus infection, which induces inflammation in the periphery and CNS, and virus infection-like illness. In turn, we show that nafamostat has a systemic anti-inflammatory effect in the presence of the TLR7/8 agonist. Therefore, the results indicate that nafamostat has anti-inflammatory actions, beyond its ability to inhibit TMPRSS2, that might potentiate its anti-viral actions in pathologies such as COVID-19.

The lineage of coronavirus SARS-CoV-2 of Russian origin: Genetic characteristics and correlations with clinical parameters and severity of coronavirus infection

The identification of new SARS-CoV-2 and human protein and gene targets, which may be markers of the severity and outcome of the disease, are extremely important during the COVID-19 pandemic. The goal of this study was to carry out genetic analysis of SARS-CoV-2 RNA samples to elucidate correlations of genetic parameters (SNPs) with clinical data and severity of COVID-19 infection.Material and Methods. The study included viral RNA samples isolated from 56 patients with COVID-19 infection who received treatment at the City Hospital No. 40 of St. Petersburg from 04/18/2020 to 04/18/2021. Patients underwent physical examination with the assessments of hemodynamic and respiratory parameters, clinical risk according to National Early Warning Score (NEWS), computed tomography (CT) of the chest, and laboratory studies including clinical blood analysis, assessment of ferritin, C-reactive protein (CRP), interleukin-6 (IL-6), lactate dehydrogenase (LDH), D-dimer, creatinine, and glucose levels. All patients tested positive for SARS-CoV-2 RNA by polymerase chain reaction (PCR). Single nucleotide polymorphisms (SNPs) in viral RNA were identified through the creation of cDNA libraries by targeted sequencing (MiSeq Illumina). Bioinformatic analysis of viral samples was performed using the viralrecon v2 pipeline with the further annotation via Pangolin and Nextlade. Sampled genomes were visualized using the Integrative Genomics Viewer (IGV) software. Statistical data processing (descriptive statistics and graphical analysis of data relationships from diff erent tables) was performed using a GraphPad device on the Prism 8.01 platform.Results. A comparative analysis of SNP frequencies in the virus genome in samples from deceased and discharged patients was carried out. The SNPs associated with risk of death (OR > 1), neutral SNPs (OR = 1), and protective SNPs (OR < 1) were identifi ed. Patient samples were infected with 14 lines of SARS-CoV-2, fi ve of which (B.1.1.129, B.1.1.407, B.1.1.373, B.1.1.397, and B.1.1.152) were of Russian origin. The SNPs in the samples infected with the strains of non-Russian origin were associated with an increased risk of mortality (OR = 2.267, 95% confi dence interval 0.1594-8.653) compared to the SNPs in the samples obtained from the group of patients infected with the strains of Russian origin. Positive correlations were identifi ed between the average SNP number, nonsynonymous SNPs, and S-protein SNPs with the degree of respiratory failure, total NEWS score, CT-based form of disease, duration of treatment with mechanical ventilation, disease outcome, levels of LDH, glucose, D-dimer, and ferritin, and RNA amount in the PCR test. S-protein SNPs negatively correlated with the leukocyte and neutrophil counts.

Optical cytosensors for detection of circulating tumour cells

Blood analysis is an established approach to monitor various diseases, ranging from heart defects and diabetes to cancer. Among various tumor markers in the blood, circulating tumor cells (CTCs) have...

Detection of Anti-Echinococcus granulosus Antibodies in Humans: An Update from Pakistan

Human cystic echinococcosis (CE) is a zoonotic disease caused by the larval stage of Echinococcus granulosus sensu lato that causes economic losses by affecting livestock and also poses a public health threat worldwide. The present study is the first retrospective report on the seroprevalence of anti-E. granulosus antibodies in humans in Pakistan. The study used data from 93 blood analysis reports of patients suspected of having CE from different medical centers in Lahore, Pakistan. Out of 93 sera samples, 20 (21.5%) were seropositive, and higher seropositivity (17.2%) was recorded with the indirect hemagglutination test (IHA) than with enzyme-linked immunosorbent assay (ELISA). The findings indicated that age, gender, and year had no significant relationship with the seropositivity of CE. The current study provides directions towards the management of the disease in the near future in Pakistan.

Features of influenza’s course in combination with other viruses in children

The objective. To evaluate the frequency and laboratory characteristics of pneumonia in children when influenza is combined with other respiratory viruses.Materials and methods. We examined 72 children aged 1 month to 17 years who were hospitalized in a specialized hospital in 2017–2019 with influenza type A (72 %) or B (28 %). The main group included 36 patients who had a laboratory-confirmed combination of influenza with other respiratory viruses, and the comparison group included 36 cases in which influenza was the only pathogen detected.Results. The analysis of the frequency of complications showed that in the main group, pneumonia developed significantly more often – in 22 % (8 children), and in the comparison group – in 6 % (2 children, p < 0.05).Conclusions. 1) The combination of influenza with other respiratory viruses is a risk factor for the development of pneumonia in children. 2) There are no significant changes in the indicators of clinical blood analysis in pneumonia developing against the background of influenza, including in combination with other respiratory viruses.

The Relationship between Routine Blood Parameters and the Prognosis of COVID-19 Patients in the Emergency Department

The aim of this study is to investigate the routine blood parameters of COVID-19 patients at the time of admission to the emergency department and their relationship with the severity of the disease and prognosis. A total of 500 patients, who were diagnosed with severe COVID-19 and hospitalized in the intensive care unit between 01.04.2020 and 01.02.2021 in the emergency department of a pandemic hospital, were retrospectively analyzed. Demographic, clinical, and laboratory data of the patients were obtained from the hospital registry system. Neutrophil-to-lymphocyte ratio (NLR), monocyte-to-lymphocyte ratio (MLR), and platelet-to-lymphocyte ratio (PLR) were calculated using neutrophil, lymphocyte, monocyte, and platelet counts. These patients were divided into two groups: survivors and deceased. All parameters obtained from routine blood analysis were statistically compared between these two groups. While 280 out of 500 patients survived, 220 died. Of all patients, the mean age was 67 years and 51.8% were males. There was a significant difference between the two groups in terms of age, gender, length of hospital stay, need for mechanical ventilation, white blood cell, neutrophil, lymphocyte, monocyte, eosinophil, platelet counts, CRP, ferritin, procalcitonin values, NLR, MLR, and PLR ( p < 0.001 for all). While NLR alone and MLR + NEU and NLR + PLR + MLR combinations had the highest AUC values (0.930, 0.947, and 0.939, respectively), MLR and PLR alone showed the lowest AUC values (0.875 and 0.797, respectively). The sensitivity, specificity, positive predictive values (PPVs), and negative predictive values (NPVs) in the prediction of death according to the cutoff values of the parameters have been determined. A significant correlation was determined between age, NLR, MLR, and PLR and duration of hospital stay ( p < 0.001 for all). Routine blood parameters and NLR, MLR, and PLR can assist emergency physicians to identify the severity and early prognosis of COVID-19 patients.

A Review on Systematic Investigation of Leucocytes Identification and Classification Techniques for Microscopic Blood Smear

Healthcare services are an important part of human beings and healthcare services are changing with new and innovative technologies. In recent day’s healthcare sector performing very crucial role in metamorphose of traditional health services to e-health technologies. This proposal provides an error-free and improved technology-based blood analysis service for the identification of leucocytes in blood samples of humans. Leucocytes play a vital and important character in human immune systems. This system helps to protect the body from suffering from leukemia. Leukemia, a blood cancer, nowadays is commonly found in all age persons. Leukemia is a type of disease and image processing techniques and algorithms can play a crucial role in disease diagnostic methodology. Identification of leukocytes in blood smear provides important information to pathologist as well as doctors to analyze and predicts different types of diseases, such as cancer. However, this analysis is critical and major complexities which results in errors and also takes a lot of time for analysis. Most of the time, the laboratory practitioners and doctors are interested only in leucocytes in blood smear. Medical image processing techniques strongly supports in their critical diagnosis and better results.

Unusual cause of low abdominal pain

A 53-year-old woman, G2P2, was referred to the outpatient clinic because of chronic abdominal pain. Abdominal examination revealed tenderness in the right lower quadrant without peritoneal signs and blood analysis was normal. To exclude slow transit constipation a pellet study was performed: plain abdominal radio-graphy showed the presence of all ingested pellets and an abnormal, slightly lateralized and angulated position of the intrauterine device, a levonorgestrel intrauterine system called Mirena® which was placed five years earlier (figure 1). Subsequently an abdominal computed tomography was executed.

Experience of patients with herpetic ganglioneuritis observation in the neurological hospital

Objective — to evaluate the clinical, laboratory and functional parameters of patients with herpetic trigeminal ganglioneuritis in the neurological department. Methods and subjects. The case anamnesis of 43 patients (26 (60.5 %) female and 17 (39.5 %) male) with a diagnosis of herpetic trigeminal ganglioneuritis were retrospectively analyzed: clinical presentation, data on general blood and urine tests, biochemical blood tests, electrocardiography, ultrasound examination of the main arteries of the brain, organs of the abdominal cavity and small pelvis, rheoencephalography. Results. Out of 43 hospitalized patients, 31 (72.1 %) had lesion of the first branch (64.5 % of women and 35.5 % of men), 9 (20.9 %) had lesion of the second branch, and 3 (7.0 %) ) — III branch of the trigeminal nerve. Patients older than 46 years predominated (86.0 %). In general, right‑sided lesion was observed in 22 cases (51.2 %), left‑sided — in 21 (48.8 %) cases. In women, right‑sided lesions prevailed (16 patients — 61.5 %), while in men — left‑sided lesions (11 patients — 64.7 %). The main complaint, in addition to rashes, was local pain, which was mainly of a burning character (24 patients — 55.8 %) with simultaneous itching (15 patients — 34.9 %). Severe pain was observed in 23 (53.5 %), moderate in 14 (32.6 %) and mild in 6 cases (13.9 %). Hyperesthesia accompanied pain in 28 (65.1 %) cases, while only 6 (14.0 %) patients demonstrated hypoesthesia. In the case of involvement of the I branch, herpetic kerato‑uveitis was registered in 9 (29.0 %) cases. Edema of the periocular region was observed in 17 patients (54.8 %). In the case of elderly patients, intellectual and mind functions decrease (45 %), positive subcortical reflexes (46.5 %), and moderate coordination disorders (34.9 %) were observed. Among the general and functional analyses carried out, attention was drawn to an increase ESR (39.5 %), as well as metabolic changes in the myocardium on the ECG (65.1 %). In the case of older age group, comorbidities were dominated by bychronic cerebrovascular accident (45 %), arterial hypertension (40.0 %), type 2 diabetes mellitus (25.0 %), and osteochondrosis (20.0 %). Conclusions. In the case of herpetic lesions, clinical observations confirm the predominant lesion of the I branch of the trigeminal nerve, more marked predominance of elderly women with right‑sided symptoms in the pathological process was observed. Beside the background of typical rashes, severe baking local pain, accompanied by itching and hyperesthesia, dominates in the clinical picture. Despite sometimes delayed hospitalization due to attempts at outpatient or self‑treatment, a general blood analysis often reveals markers of inflammation. The complexity and variability of complaints, as well as examination results, can be mainly explained by the elderly age of patients.