Peripheral Blood Lymphocytes Research Articles

Afla Toxin B<SUB>1</SUB> in Fish Feeds and Experimental Acute Aflatoxicosis in an Air-Breathing Teleost

A general survey of three types of fish feeds for the incidence of toxigenic mold infestation revealed an alarming level of aflatoxigenic Aspergillus flavus contamination during storage. About 89% of A. flavus strains were toxigenic. The toxicity of crude toxin sample was assessed on Channa punctatuus. The experiment was conducted using pure crystalline aflatoxin B. Four different concentrations (10, 20, 30 and 40 mg/D) of pure toxin in 0.5 ml propylene glycol were administered intraperitoneally to four groups of fish along with a control. Observations were made on day 20, 40, 60 and 80. The treated fish showed loss of appetite, followed by listlessness and increased mucus over body surface. Hemorrhagic patches and tumors were prominent on liver. Histopathologically, liver was the most prominently affected organ leading to classical trabeular hepatoma. The excretory -kidney revealed some pre-neoplastic alteration arising from tubular epithelium. The head -kidney was atrophic, but proliferation of inter-renal lobules was prominent. The muscularis of the stomach showed severe proliferation. The intestine, pyloric caeca and gills were atrophic and necrosed. Formation of atypical glandular masses in mucosal region was marked in the intestine. The spleenic pulps were indistinguishable and showed a monotony of lymphocytes. The total and differential counts of leucocytes revealed a significant rise of lymphocytes in peripheral blood.

Newly identified common variable immunodeficiency in an elderly patient with rheumatoid arthritis

Common variable immunodeficiency is a group of heterogeneous diseases that are primary immunodeficiencies with a predominant lack of antibody synthesis. The most common manifestation is chronic infectious pathology of the respiratory tract, less often of the gastrointestinal tract, as well as septic arthritis. Non-infectious manifestations include autoimmune diseases, non-infectious diarrhea, cancer, benign lymphoid hyperplasia. In one patient, a combination of several syndromes is possible. Unlike most other primary immunodeficiencies that debut in early childhood, this pathology often manifests in the third or fourth decade of life, and sometimes later in apparently healthy people. Although the routine laboratory method for determining the concentration of the main classes of immunoglobulins makes it quite easy to identify patients with common variable immunodeficiency, often the diagnosis is made many years after the onset of the first clinical symptoms, when serious complications have already developed. The disease is characterized by a gradual onset, chronic progressive course and ineffectiveness of standard therapy. The main method of treatment is lifelong replacement therapy with intravenous immunoglobulins. The article considers a clinical case when an elderly patient with seronegative rheumatoid arthritis with a juvenile onset was diagnosed many years later. The results of immunological studies, including immunophenotyping of peripheral blood and bone marrow lymphocytes, are presented. Issues of differential diagnosis with secondary hypogammaglobulinemia and lymphoproliferative diseases are highlighted. It has been stated that until now, doctors of various specialties are not sufficiently familiar with the problem of primary immunodeficiencies, especially when it comes to adults, and non-infectious manifestations dominate in the clinical picture.

Systemic Inflammatory Status of Patients with Obstructive Sleep Apnea Syndrome and the Predictive Value of Heparin-Binding Protein.

Objective: To investigate the alteration of the systemic inflammatory status of patients with obstructive sleep apnea syndrome (OSAS) and the predictive value of heparin-binding protein (HBP) for OSAS. Methods: Patients with OSAS who were hospitalized in our hospital from 2020 January to 2022 December and diagnosed by polysomnography (PSG) (OSAS group, n = 79) were retrospectively studied and their relevant examination findings and demographic characteristics were recorded. Sex- and age-matched non-OSAS patients hospitalized at the same time were selected as the control group (control group, n = 28). Differences in peripheral blood neutrophil counts, lymphocyte counts, neutrophil-to-lymphocyte ratio (NLR), procalcitonin (PCT), and HBP were compared between the 2 groups. The predictive value of these markers for OSAS was analyzed using the receiver operating characteristic curve, and independent risk factors for OSAS were determined using logistic regression analysis. Peripheral blood was drawn from all patients in the fasting state of the morning. Results: The number of peripheral blood neutrophils, lymphocytes, and HBP was higher in the OSAS group than in the control group, and the differences were statistically significant (P all <.05), while the differences in NLR and PCT between the 2 groups were not statistically significant (P all >.05). Plasma HBP level had an area under the curve (AUC) of 0.79 (P < .0001) in determining OSAS, with a sensitivity of 80.49% and a specificity of 70.83% and the best cutoff value was >10.73 ng/ml. Combining body mass index (BMI), neutrophil, lymphocyte, and HBP improved the predictive value of OSAS with an AUC of 0.89 (P < .0001), a sensitivity of 80.49%, and a specificity of 87.50%. Logistic regression analysis showed that both BMI and HBP were independent risk factors for OSAS (PBMI < .001, PHBP < .01), while neutrophils and lymphocytes were not (P all >.05). Conclusions: This is the first study to objectively examine HBP in OSAS patients, and HBP is an independent risk factor that may serve as a diagnostic biomarker in OSAS. Patients with OSAS have an altered systemic inflammatory state and may be more prone to severe bacterial infections.

Genomic Instability and Cytotoxicity Evaluation of Two Communities Exposed to Pesticides in the Mexicali Valley by the L-CBMN Assay.

The continuous biomonitoring of a population directly or indirectly exposed to pesticides could be an additional tool for decision makers to improve their health conditions. In this work, we performed biomonitoring on two groups of people from the Mexicali Valley who were continuously exposed to pesticides using the cytokinesis-block micronucleus cytome assay (L-CBMN) to evaluate cytotoxic and genotoxic damage in human peripheral blood lymphocytes. The study groups comprised 14 indigenous Cucapah with non-vegetarian habits (NV group) from Ejido el Mayor (32.12594°, -115.27265°) and 21 lacto-ovo vegetarian (LOV) persons from the Seventh-day Adventist Church of Ejido Vicente Guerrero (32.3961°, -115.14023°). The L-CBMN assay determines the nuclear division index (NDI), apoptosis, necrosis, micronuclei (MNs), nuclear buds (NBUDs), and nucleoplasmic bridges (NPBs). Our results show that, regardless of diet or daily habits, both the studied groups presented with cytogenotoxic damage compared with non-exposed pesticide individuals, without modifications to the nuclear division index. In the rest of the evaluated biomarkers, the NV group exhibited greater cytotoxic and genotoxic damage than the LOV group. Nevertheless, individuals practicing a lacto-ovo vegetarian diet (LOV) showed lower damage than those with non-vegetarian habits (NV), suggesting a better antioxidant response that helps decrease the genotoxic damage due to the enhanced intake of folates and antioxidants from a plant-based diet.

The Association Between Peripheral Blood Neutrophil-to-Lymphocyte Ratio and Cirrhosis Complications in Adults with Compensated Liver Cirrhosis in Ahvaz, Iran

Background: The neutrophil-to-lymphocyte ratio (NLR) is an inflammation biomarker in patients with liver cirrhosis. While increased NLR is linked to poor clinical outcomes and mortality in diseases like cancers, its association with cirrhosis complications has been sparsely examined. Objectives: This study aimed to evaluate the association between NLR and cirrhosis complications in patients with compensated liver cirrhosis. Methods: This retrospective cohort study was conducted on 256 patients with compensated liver cirrhosis visiting the Gastroenterology Clinic of Imam Khomeini Hospital, Ahvaz, Iran, in 2020. The peripheral blood lymphocyte and neutrophil counts were evaluated, and the NLR was calculated by dividing the absolute neutrophil count by the absolute lymphocyte count. Cirrhosis complications were assessed based on clinical and laboratory evaluation during 1 year of follow-up. Results: During the retrospective follow-up period, 59 patients (23.05%) experienced cirrhosis complications such as spontaneous bacterial peritonitis (n = 23), gastrointestinal bleeding (n = 22), and hepatic encephalopathy (n = 14). The baseline NLR, Model for End-Stage Liver Disease (MELD) score and Child-Pugh score were significantly higher in patients who later developed complications than in those who did not (P &lt; 0.0001). The NLR with an optimal cut-off of &gt; 1.95 had a sensitivity of 84.75% and specificity of 93.91% in predicting complications during the 1-year follow-up (AUC = 0.905, P &lt; 0.0001). Conclusions: Our results indicated that the NLR is a simple, non-invasive, and cost-effective marker for predicting short-term complications in patients with compensated liver cirrhosis.

A genetic study of a Brazilian cohort of patients with X-linked hypophosphatemia reveals no correlation between genotype and phenotype.

X-linked hypophosphatemia (XLH) is the most common inherited form of rickets, and it is caused by pathogenic inactivating variants of the phosphate-regulating endopeptidase homolog X-linked (PHEX) gene. The main purpose of this study is to identify the presence of a genotype-phenotype correlation in a cohort of XLH patients. This is a retrospective study including patients diagnosed with hypophosphatemic rickets, confirmed by clinical, radiological, and laboratory findings. Medical records were reviewed for phenotypic analyses. Genomic DNA was extracted from the peripheral blood lymphocytes, and PHEX sequencing was performed by exomic NGS sequencing. The Wilcoxon rank-sum test and the two-tailed Fisher's exact test were employed for the statistical analyses of this study. A total of 41 patients were included in this study, and 63.41% (26/41) of the patients were female. The mutation analyses identified 29.27% missense variants and 29.72% nonsense variants, most of them were considered deleterious (66.41%). Six novel deleterious variants in the PHEX gene were detected in seven patients. The median concentrations of pretreatment serum calcium, phosphorus, and parathyroid hormone (PTH) were not significantly different among patients with different genotypes. An orthopedic surgery due to bone deformity was required in 57.69%. Our analysis did not identify any specific genotype as a predictor. No significant genotype-phenotype correlation was found, suggesting that the recognition of subjacent pathogenic mutation in the PHEX gene may have limited prognostic value. Despite this finding, genetic testing may be useful for identifying affected individuals early and providing appropriate treatment.

Genotoxicity of a synthetic plant growth regulator, Forchlorfenuron (CPPU), on human lymphocytes using chromosome aberration assay

Forchlorfenuron (FCF, also known as CPPU), which belongs to the group of phenylurea cytokinins, is one of the most widely used synthetic plant growth regulators (PGRs) worldwide. Although FCF plays a crucial role in cellular growth and differentiation by promoting cell division in plants, it disrupts higher-order septin assembly in other eukaryotic organisms, including humans. Despite its widespread use, no study has been found investigating the genotoxic effects of this synthetic PGR on humans. Hence, this investigation was designed to examine the potential cyto-genotoxicity of a commercial formulation of FCF on human peripheral blood lymphocytes (PBLs) using chromosome aberrations (CAs) and mitotic index (MI) endpoints. The whole blood cultures were treated with 0.25, 0.50, 1.00, and 2.00 µg/ml concentrations of a commercial form of FCF. According to the results, FCF significantly enhanced the percentage of cells containing structural CAs at the concentrations of 1.00 and 2.00 µg/ml for both treatment times (24 and 48 h), in comparison to the negative control (P&lt;0.05). Besides, in cultures exposed to FCF concentrations of 0.50, 1.00, and 2.00 µg/ml, the total CA/cell ratio was significantly higher (P&lt;0.05). In addition, FCF was found to have cytotoxic activity on human PBLs at all treatments (except for the lowest concentration at 24 h) by significantly reducing the MI compared to the negative control (P&lt;0.05). The findings of this investigation indicate the first time that a commercial formulation of FCF (0.50-2.00 µg/ml) may have genotoxic and cytotoxic potential on human lymphocytes.

Effects of low-dose ionizing radiation on genomic instability in interventional radiology workers

Interventional radiologists are chronically exposed to low-dose ionizing radiation (IR), which may represent a health risk. The aim of the present study was to evaluate genomic instability by analyzing chromosomal aberrations, micronuclei, and 53BP1 DNA repair foci in peripheral blood lymphocytes of radiologists. Based on the IAEA guidelines on biodosimetry using dicentrics, the average protracted whole-body dose in radiologists were estimated. Since preleukemic fusion genes (PFG) are the primary events leading to leukemia, we also studied their presence by RT-qPCR and FISH. No significant difference in 53BP1 foci and incidence of PFG (MLL-AF4, MLL-AF9, AML1-ETO, BCR-ABL p190) was found in cells of interventional radiologists in comparison to controls. However, our results showed an increased frequency of micronuclei and various types of chromosomal aberrations including dicentrics in interventional radiologists. The average protracted whole body estimated dose was defined at 452.63 mGy. We also found a significantly higher amplification of the MLL gene segment and increased RNA expression in cells of interventional radiologists in comparison to controls. In conclusion, our results showed that long-term low-dose IR induces genomic instability in interventional radiologists.

Influence of Body Mass Index and Duration of Disease on Chromosome Damage in Lymphocytes of Patients with Diabetes.

It is well-established that patients with diabetes mellitus (DM) have a higher incidence of several types of cancer. The precise mechanisms of this association are still unknown, but obesity and chronic inflammation-induced reactive oxygen species (ROS) are thought to be the main risk factors. ROS may produce different DNA damage, which could eventually lead to cancer. The main objective of this study was to evaluate the relation of chromosome aberrations (CA) with disease status, demographics, and clinical parameters in 33 subjects with type 1 DM (T1DM), 22 subjects with type 2 DM (T2DM), and 21 controls. CAs were analyzed in cultured peripheral blood lymphocytes and subdivided into chromatid (CTA)- and chromosome (CSA)-type aberrations. Compared with controls, higher levels of CTAs and CSAs were observed in T1DM (p = 0.0053 and p = 0.0203, respectively) and T2DM (p = 0.0133 and p = 0.00002, respectively). While there was no difference in CTAs between T1DM and T2DM, CSAs were higher in T2DM (p = 0.0173). A significant positive association between CTAs and disease duration (rs = 0.2938, p = 0.0099) and between CSAs and disease duration (rs = 0.4306, p = 0.0001), age (rs = 0.3932, p = 0.0004), and body mass index (BMI) (rs = 0.3502, p = 0.0019) was revealed. After multiple regression analysis, duration of disease remained significant for CTA, CSA, and CAs (p = 0.0042, p = 0.00003, and p = 0.00002, respectively). For CSA, BMI and the use of statins were the other important confounding variables (p = 0.0105 and p = 0.0763). Thus, this study demonstrated that both T1DM and T2DM patients had a higher number of all types of aberrations than controls, which increases with the prolonged disease duration. Higher BMI was associated with a higher frequency of CSA. The use of statins might be beneficial for reducing chromosome damage, but further investigations are needed to confirm this association.

Efficacy of a synthetic peptide Chlamydia pecorum major outer membrane protein vaccine in a wild koala (Phascolarctos cinereus) population

Chlamydiosis is a significant disease affecting Eastern Australian koala (Phascolarctos cinereus) populations, impacting individual animal welfare and fecundity and therefore influencing population dynamics. The aim of this study was to investigate the effect of a synthetic peptide vaccine based on 4 components of the Chlamydia pecorum major outer membrane protein (MOMP), over an 18-month period in a koala population severely impacted by chlamydiosis. Wild koalas were recruited into a vaccination or a placebo treatment group on a random allocation, then followed through a period of 18 months, with recapture at 6 monthly intervals. Vaccination did not alter clinical disease expression or chlamydial shedding from the ocular or urogenital sites. Vaccination did not stimulate a significant plasma anti-MOMP IgG response, when compared to the placebo group. There was no significant effect of vaccination on IFN-γ and IL-17A mRNA expression of peripheral blood lymphocytes when stimulated with rMOMP. We have demonstrated that a synthetic peptide vaccination against chlamydiosis is not an effective management tool in a koala population with a high prevalence of C. pecorum infection and related disease. The lack of antigenic response found in this study suggests that further research utilising a larger, full-length antigen is an avenue worth investigation if we are to consider vaccination as a part of a management strategy in diseased koala populations.

Correction of immune status from hospitalized COVID-19-patients with immunotropic drug added to the basic treatment

Cellular immunity plays an important role in the control of SARS-CoV-2. Lymphopenia and a decrease in the functional activity of cells may be among the main reasons for deterioration of clinical outcomes of the disease. Usage of the bacterial therapeutic vaccine Immunovac-VP-4 during the inflammation phase may be promising for immunomodulation of the cellular immunity. The aim of our study was to evaluate the dynamics of lymphocyte subpopulations in hospitalized patients with COVID-19 upon combining the basic therapy with immunotropic drug based on the antigens from opportunistic pathogens. The study included 45 patients (18-70 years old) admitted with a confirmed diagnosis of moderate/severe infection caused by the COVID-19 virus. In addition to basic therapy, 33 persons of this group received Immunovac-VP-4 by a combined nasal-oral method. Subpopulation activity of peripheral blood lymphocytes in patients over time (at baseline, on the 14th and 30th day after hospitalization) was studied by flow cytometry by means of FC-500 Cytomics (Beckman Coulter, USA) using monoclonal antibodies (mAb) (Immunotech, France). In the group receiving only standard therapy, an increased number of T lymphocytes was detected on day 14 (79.9 (75.5-81.6), p = 0.00252), on day 30 from the start of treatment (78.4 (74.25-79. 2), p = 0.03662), and a decrease in B lymphocytes on day 14 (10.6 (7.78-11.63), p = 0.03236), on day 30 (7.85 (6.25-11.1), p = 0.01352) relative to baseline parameters upon admission. We revealed more pronounced changes in the parameters of cellular immunity relative to the initial parameters, i.e., an increased proportion of T lymphocytes on the 14th day (80.1 (73.8-84.2), p = 0.00018), and 30th day from starting the treatment (80.2 (76-81.9)), T helpers at 14 days after treatment (50.2 (43-57), p = 0.00694), cytotoxic T cells by 30th day of therapy (26.35 (24-29.4), p = 0.0114), decrease in B lymphocytes on day 14 (13.1 (8.2-16.9), p = 0 00158), on the 30th day from the start of treatment (8.2 (7.6-9.7), p &lt;0.00001), and a transient decrease in NK cells on the 14th day (3.7 (2,1-6.3), p = 0.00308), with their recovery on the 30th day of observation to 8.6 (6-12.5) in the Immunovac-VP-4 group. Modulation of cellular immunity may be important for the virus clearance.

In Vivo and In Vitro Efficacy of Trastuzumab Deruxtecan in Uterine Serous Carcinoma.

Uterine serous carcinoma (USC) is a rare, biologically aggressive variant of endometrial cancer with a high recurrence rate and poor prognosis. HER2 overexpression (3+ positivity) by IHC and/or FISH ERBB2 gene amplification is detected in approximately one-third of patients with USC. Clinical trials incorporating trastuzumab with standard chemotherapy have recently demonstrated improved progression-free and overall survival in advanced-stage or recurrent USC that overexpresses HER2. However, a large number of patients with USC eventually developed resistance to trastuzumab. Trastuzumab deruxtecan (T-DXd) is a novel HER2-directed antibody-drug conjugate with a topoisomerase I inhibitor payload recently approved by the Food and Drug Administration (FDA) for multiple tumor indications. Here, we investigated the in vitro and in vivo efficacy of T-DXd in primary USC cell lines and xenografts with different HER2 expression. T-DXd-induced cell growth suppression in HER2-overexpressing cell lines in vitro, increased early and late apoptosis as assessed by annexin and propidium iodide staining, and, similarly to trastuzumab, T-DXd-induced significant antibody-dependent cellular cytotoxicity in the presence of peripheral blood lymphocytes. While negligible activity was detected against USC cell lines with low HER2 expression, T-DXd demonstrated significant bystander killing against USC tumors with low/negligible HER2 when such cells were admixed with HER2 3+ tumor cells in vitro. T-DXd showed tumor growth suppression in in vivo USC PDX models that overexpress HER2 at 3+ levels, prolonging survival when compared with controls, with minimal toxicity. Future clinical trials are warranted in patients with USC failing trastuzumab treatment.

Simulating Space Conditions Evokes Different DNA Damage Responses in Immature and Mature Cells of the Human Hematopoietic System.

The impact of space radiation and microgravity on DNA damage responses has been discussed controversially, largely due to the variety of model systems engaged. Here, we performed side-by-side analyses of human hematopoietic stem/progenitor cells (HSPC) and peripheral blood lymphocytes (PBL) cultivated in a 2D clinostat to simulate microgravity before, during and after photon and particle irradiation. We demonstrate that simulated microgravity (SMG) accelerates the early phase of non-homologous end joining (NHEJ)-mediated repair of simple, X-ray-induced DNA double-strand breaks (DSBs) in PBL, while repair kinetics in HSPC remained unaltered. Repair acceleration was lost with increasing LET of ion exposures, which increases the complexity of DSBs, precluding NHEJ and requiring end resection for successful repair. Such cell-type specific effect of SMG on DSB repair was dependent on the NF-кB pathway pre-activated in PBL but not HSPC. Already under unperturbed growth conditions HSPC and PBL suffered from SMG-induced replication stress associated with accumulation of single-stranded DNA and DSBs, respectively. We conclude that in PBL, SMG-induced DSBs promote repair of radiation-induced damage in an adaptive-like response. HSPC feature SMG-induced single-stranded DNA and FANCD2 foci, i.e., markers of persistent replication stress and senescence that may contribute to a premature decline of the immune system in space.

Oral cytarabine ocfosfate pharmacokinetics and assessment of leukocyte biomarkers in normal dogs.

Cytosine arabinoside (Ara-C) is a nucleoside analog prodrug utilized for immunomodulatory effects mediated by its active metabolite Ara-CTP. Optimal dosing protocols for immunomodulation in dogs have not been defined. Cytarabine ocfosfate (CO) is a lipophilic prodrug of Ara-C that can be administered PO and provides prolonged serum concentrations of Ara-C. Provide pharmacokinetic data for orally administered CO and determine accumulation and functional consequences of Ara-CTP within peripheral blood leukocytes. Three healthy female hound dogs and 1 healthy male Beagle. Prospective study. Dogs received 200 mg/m2 of CO PO q24h for 7 doses. Serum and cerebrospinal fluid (CSF) CO and Ara-C concentrations were measured by liquid chromatography-tandem mass spectroscopy (LC-MS/MS). Complete blood counts, flow cytometry, and leukocyte activation assays were done up to 21 days. Incorporation of Ara-CTP within leukocyte DNA was determined by LC-MS/MS. Maximum serum concentration (Cmax ) for Ara-C was 456.1-724.0 ng/mL (1.88-2.98 μM) and terminal half-life was 23.3 to 29.4 hours. Cerebrospinal fluid: serum Ara-C ratios ranged from 0.54 to 1.2. Peripheral blood lymphocyte concentrations remained within the reference range, but proliferation rates poststimulation were decreased at 6 days. Incorporation of Ara-CTP was not saturated and remained >25% of peak concentration at 13 days. Oral CO may produce prolonged serum Ara-C half-lives at concentrations sufficient to induce functional changes in peripheral leukocytes and is associated with prolonged retention of DNA-incorporated Ara-CTP. Application of functional and active metabolite assessment is feasible and may provide more relevant data to determine optimal dosing regimens for Ara-C-based treatments.

Особенности использования лимфоцитарного теста для биологической дозиметрии в ранние сроки после облучения

When eliminating the consequences of large-scale radiation accidents, primary triage of victims is of key importance during the early phase of medical evacuation. Information about lymphocyte counts (blood test) per unit of peripheral blood volume can be used for this purpose. The study was aimed to validate the method of using a lymphocyte test for prediction of acute radiation injury severity in the first days after the exposure associated with the radiation mass casualty incident, given peripheral blood was tested once. We performed correlation analysis of the data of laboratory studies focused on quantifying lymphocytes in peripheral blood of victims during the first days following the Chernobyl disaster and other radiation accidents on the territory of the countries of the former USSR (115 individuals), including radiation accidents with gamma neutron radiation (20 individuals). It was found that with the lymphocyte concentration of 0.2–1.0 × 109/L on day 2 after exposure, the absolute error of estimated dose was ±1.5 Gy in case of gamma exposure and ±1.3 Gy in case of exposure to gamma neutron radiation. When the lymphocyte concentration exceeds 1.0 × 109/L, mild acute radiation syndrome (ARS) is predicted, given the average dose is below 2.0 Gy; when the lymphocyte concentration is less than 0.2 × 109/L&lt; the estimated average dose exceeds 4.0 Gy, which corresponds to severe or extremely severe ARS. Thanks to the lymphocyte test accessibility and simplicity, this biological dosimetry method can occupy a worthy position in the diagnosis of radiation injury associated with large-scale accidents, since the results of cytogenetic tests are not available within first days after the accident.

Identification, expression pattern and functional characterization of IFN-γ involved in activating JAK-STAT pathway in Sebastes schlegeli.

IFN-γ (interferon gamma) is a critical cytokine in the immune system involved both directly and indirectly in antiviral activity, stimulation of bactericidal activity, antigen presentation and activation of macrophages via the Janus kinase/signal transducer and activator of transcription (JAK-STAT) pathway. The IFN-γ function is best described in cell defense against intracellular pathogens in mammals, but IFN-γ cytokine-induced metabolic change and its role in anti-infection remain unknown in teleost fish. In this study, a novel IFN-γ (SsIFN-γ) was identified from black rockfish (Sebastes schlegeli) by rapid amplification of cDNA ends (RACE). The open reading frame (ORF) of SsIFN-γ encoded a putative protein of 215 amino acids and shares 60.2%-93.5% overall sequence identities with other teleost IFN-γ. SsIFN-γ was distributed ubiquitously in all the detected tissues and immune cells, which was highly expressed in the spleen, gills, head kidney by quantitative real-time PCR. The mRNA expression of SsIFN-γ was significantly upregulated in the spleen, head kidney, head kidney (HK) macrophages and peripheral blood lymphocytes (PBLs) during pathogen infection. Meanwhile, the recombinant protein (rSsIFN-γ) exhibited an immunomodulatory function to enhance respiratory burst activity and nitric oxide response of HK macrophages. Furthermore, rSsIFN-γ could effectively upregulate the expression of macrophage proinflammatory cytokine, the expression of JAK-STAT signaling pathway related genes and interferon-related downstream genes in the head kidney and spleen. Luciferase assays showed ISRE and GAS activity were obviously enhanced after rSsIFN-γ treatment. These results indicated that SsIFN-γ possessed apparent immunoregulatory properties and played a role in fighting pathogen infection which will be helpful to further understanding of the immunologic mechanism of teleosts IFN-γ in innate immunity.

Psychophysiological Assessment of Individual Personality in Two Patients with Severe Local Radiation Injuries (LRI) of Hand and Acute Radiation Sickness (ARS) I Degree (30 Years of Follow-up)

Psychophysiological examination was carried out on two patients, former border guards, who suffered on October 05, 1982 as a result of accidental contact with a source of gamma radiation (137Cs) while on duty while serving on the Iranian-Azerbaijani border in the ranks of the Soviet army. They suffered ARS of the 1st degree with extremely uneven gamma irradiation in the form of local radiation injuries of the hands of varying severity. The assessment of the aver -age dose for the whole body according to the frequency of dicentrics in cultures of peripheral blood lymphocytes in patient No. 1 and patient No. 2 is the same and is 0,26 Gy. Psychophysiological research was carried out in dynamics in 2001-2013 using the automated software and methodological complex «Expert». According to the MMPI data, both victims have a hyperthymic type of psychophysiological adaptation with varying degrees of severity. Patient No. 2 is a chronically hyperthymic personality with a significant increase in the profile on the 9th scale and with its gradual increase in dynamics, which reflects the persistence of hyperthymic personality traits and causes an overstrain of mental adaptation, with an increase in the dynamics of the tendency to explain their difficulties and problems by somatic distress (1Hs). Patient No. 1 has a hyperthymic personality type with a moderate increase in profile dynamics on the ninth scale. A personality trait is indicated by a high, growing in dynamics, indicator on the K scale, which determines behavior depending on social approval and concern for one’s social status. The appearance in dynamics (2013) of a peak on scales 1 (preoccupation with the state of health) and 4 (emotional tension in direct behavior) determines the insufficient ability to internalize the social norm, i.e. insufficient ability to perceive this norm as part of one’s own settings (a combination of profile peaks on scales 4 and 9). The presence of profile peaks on the 7th scale and the scales of the neurotic triad reflect the tendency to various options for the implementation of antisocial attitudes in a socially acceptable way, the desire to appear in a favorable light (scale L). Sufficiently high intelligence and demonstrative behavior determine good adaptation to the environment with self-confidence, high social adaptability, and resilience in overcoming difficulties.

Влияние внутриклеточной регуляции метаболизма на популяционный состав лимфоцитов периферической крови человека

Metabolic activity has a significant impact on the differentiation, proliferation and functioning of T cells. Different lymphocyte subpopulations use, to varying degrees, glycolysis and mitochondrial metabolism, whose main regulators are hypoxia-inducible factor 1-alpha (HIF-1α) and sirtuin 3 (SIRT3), respectively. The purpose of this paper was to study changes in the population composition of peripheral blood lymphocytes in humans depending on the level of the intracellular metabolic regulators SIRT3 and HIF-1α. Materials and methods. 227 residents of the city of Arkhangelsk and the Arkhangelsk Region were examined (mean age 42 ± 11 years). Absolute lymphocyte count was determined using the Sysmex XS 500i haematology analyser, while CD3+, CD4+, CD8+, CD10+, CD25+ and CD95+ phenotypes content, by indirect immunoperoxidase reaction. Intracellular adenosine triphosphate (ATP) concentration was measured using the luciferase bioluminescence method. HIF-1α and SIRT3 concentrations were measured in lymphocyte lysate using enzyme immunoassay. To divide the total sample into groups according to SIRT3 and HIF-1α content, k-means clustering was utilized. Results. Changes in SIRT3 and HIF-1α intracellular concentrations correlated with ATP concentration. It was found that in the group with high HIF-1α content, the proportion of CD4+, CD8+, CD10+ and CD25+ lymphocytes was greater than in the group with high SIRT3 concentration, which had a greater proportion of CD95+ lymphocytes. Thus, the content of intracellular metabolic regulators that regulate ATP production pathways in the cell, i.e. oxidative phosphorylation (SIRT3) and glycolysis (HIF-1α), affects the population composition of lymphocytes and is therefore important for assessing the immune response.

No increase in translocated chromosomal aberrations, an indicator of ionizing radiation exposure, in childhood thyroid cancer in Fukushima Prefecture

To investigate the effects of radiation exposure due to the Fukushima nuclear power plant accident, following the disaster Fukushima Prefecture launched thyroid ultrasound examinations of residents who were generally younger than 18 years at the time of the earthquake. As the rate of pediatric thyroid cancer was higher than expected, we conducted biological dose assessment based on the frequency of translocated chromosome (Tr) aberrations using peripheral blood lymphocytes. Tr formation frequency was compared among the thyroid cancer (n = 38, median age 18 years, age range 12–26 years), thyroid-related disease (n = 30, median age 21 years, age range 15–28 years), and healthy controls (n = 31, median age 22 years, age range 20–23 years) groups. Tr aberration frequency was initially significantly higher in the thyroid cancer than in the other two groups; however, differences among the groups disappeared after adjusting for history of CT scan, as 92%, 67%, and 28% of those in the thyroid cancer, thyroid-related disease, and control groups, respectively, had undergone CT previously. Therefore, the significant difference in the initial number of Tr formations is presumably due to radiation exposure from CT. Accordingly, the effects of medical exposure on the chromosomes of children and adolescents should be noted.

In vitro Radio Protective Effects of Metformin on Human Lymphocytes Irradiated with Gamma Rays

Radio protective effects of metformin and its ability to alter the spontaneous and induced genotoxic and cytotoxic levels effects on human peripheral blood lymphocytes were investigated in this study. Metformin, a hypoglycemic oral drug, is a biguanide derived from Galega officinalis that is widely utilized in controlling type 2 diabetes mellitus. Whole blood samples from 10 healthy donors (5 males and 5 females) were exposed to two doses of gamma-rays (1 and 2 Gray). Lymphocytes in cultures were treated with metformin (10 and 50µM) before gamma-irradiation. Cytokinesis-block micronucleus test was used to evaluate the protective effects of metformin on radiation induced genomic damage, cytostasis and cytotoxicity, via scoring micronuclei and nucleoplasmic bridges in once divided binucleated cells as well as counting nuclear division index. The results of the current study revealed that the increase in micronuclei and nucleoplasmic bridges rate is associated with the decrease of nuclear division index in human lymphocytes exposed to gamma radiation in a dose dependent manner. Metformin effectively decreased the rate of spontaneous micronuclei and nucleoplasmic bridges paralleled with the control as well as increased nuclear division index. Moreover, treatment of whole blood samples with metformin (10 and 50 µM), 2 h preceding to irradiation, remarkably reduced rate of micronuclei and nucleoplasmic bridges accompanied with an increase in nuclear division index rate. The results introduced metformin to be an effective radio protector against DNA damage induced by gamma radiation in human lymphocytes and that it can be used to develop radio protective materials for protection cells of cancer patients from the genomic damage prompted via radiotherapy.