Low-Dose Gamma Radiation Fields Decrease Cell Viability, Damage DNA, and Increase the Expression of Hsp70 and p53 Proteins in Human Leukocytes

Ionizing radiations are tools in diagnosis and treatment of diseases. Leukopenia from exposure to ionizing radiation has been reported. Due to their radiosensitivity, leukocytes are a biological model to analyze cell damage. Therefore, cell viability, DNA damage, and Hsp70 and p53 expression in human leukocytes exposed to low-dose gamma radiation fields from a 137Cs source were evaluated. A decrease in cell viability, DNA damage and an increase in the expression of Hsp70 and p53 proportional to the radiation dose received was found, which was 0.2, 0.4, 0.6, 0.8 and 1.0 mGy.

胃癌源性外泌体对HLA-DR^(neg)leukocytes中Dicer1、PTEN基因的调控

目的通过观察胃癌源性的外泌体对HLA-DR^(^(neg))leukocytes中Dicer1、PTEN基因表达的影响,探讨肿瘤源性外泌体调控机体免疫抑制功能的机制。方法提取胃癌患者血清外泌体并鉴定,免疫磁珠法分选HLA-DR^(neg)leukocytes,吖啶橙染色后的外泌体与CFSE染色后的HLA-DR^(neg)leukocytes在37℃,5%CO_2条件下共培养12 h;采用QRT-PCR和Western blot方法分别检测共培养后胃癌患者与健康对照组中Dicer1、PTEN基因和蛋白的表达。结果成功分离胃癌患者外周血外泌体;同时观察到外泌体被HLA-DR^(neg)leukocytes有效摄取。QRT-PCR检测结果显示Dicer1 m RNA、PTEN m RNA的相对表达量分别是健康对照组的0.46±0.19、0.48±0.27倍(P<0.05);Western blot结果显示Dicer1、PTEN蛋白表达量是健康对照组的0.15±0.11、0.33±0.19倍(P<0.05)。结论胃癌来源的外泌体可以抑制HLA-DR^(neg)leukocytes中Dicer1、PTEN基因的表达,为今后进一步研究肿瘤来源的外泌体的免疫调控作用提供线索。

Leukocytes and oxidative stress： dilemma for sperm Function and male fertility

Spermatozoa are constantly exposed to the interphase between oxidation through high amounts of reactive oxygen species （ROS） and leukocytes, and reduction by means of scavengers and antioxidants. Considering the very special functions as being the only cells with such high polarization and exerting their functions outside the body, even in a different individual, the female genital tract, the membranes of these cells are chemically composed of an extraordinary high amount of polyunsaturated fatty acids. This in turn, renders them very susceptible to oxidative stress, which is defined as an imbalance between oxidation and reduction towards the oxidative status. As a result, ROS deriving from both leukocytes and the male germ cells themselves cause a process called ＇lipid peroxidation＇ and other damages to the sperm cell. On the other hand, a certain limited amount of ROS iS essential in order to trigger vital physiological reactions in cells, including capacitation or the acrosome reaction in sperm. The treatment of patients with antioxidants to compensate the oxidative status caused by oxidative stress is highly debated as uncontrolled antioxidative treatment might derail the system towards the reduced status, which is also unphysiological and can even induce cancer. This paradox is called the ＇antioxidant paradox＇. Therefore, a proper andrological diagnostic work-up, including the evaluation of ROS levels and the antioxidant capacity of the semen, has to he carried out beforehand, aimed at keeping the fine balance between oxidation and scavenging of vital amounts of ROS.

Gene expression of CCL8 and CXCL10 in peripheral blood leukocytes during early pregnancy in cows

Background: The aim of the present study was to evaluate CCL8 and CXCL10 expression and its regulatory mechanism in peripheral blood leukocytes(PBLs) at the time of maternal recognition in cows. Blood samples were collected on 14, 15, 16, 17 and 18 d after artificial insemination(AI). Based on the day of return of estrus, cows were divided into three groups, pregnant(n = 5), early embryonic mortality(EEM; n = 5) and late embryonic mortality(LEM; n = 5). The gene expression levels in PBLs were assessed with quantitative real-time reverse transcription PCR.Results: The expression of CCL8 and CXCL10 mRNA in PBLs gradually increased from 14 to 18 d of pregnant cows and significant differences were observed on 18 d(P < 0.05), whereas no significant changes were observed both in EEM and LEM cows. Interferon-stimulated protein 15 k Da(ISG15), myxovirus-resistance gene(MX) 1 and MX2 mRNA expression in PBLs increased from 14 to 18 d which was significant on 18 d of pregnant cows as well as in LEM cows(P < 0.05), but no changes were observed in EEM cows. To determine whether the expression of CCL8 and CXCL10 in PBLs was regulated by pregnancy-related substances or not, expression level was assessed after exposure to interferon-τ(IFNT) and CCL16. Monocytes, granulocytes and lymphocytes were obtained using density-gradient centrifugation and flow cytometry. The addition of IFNT(100 ng/mL) and CCL16(100 ng/mL) to cultured PBLs increased the expression of CCL8 and CXCL10 mRNA(P < 0.05). The expression of ISG15, MX1 and MX2 mRNA in PBLs was also stimulated by IFNT and CCL16(P < 0.05).Conclusions: The expression of CCL8 and CXCL10 genes increased in PBLs during early pregnancy. Since IFNT stimulated CCL8 and CXCL10 expression in cultured PBLs, the increase of CCL8 and CXCL10 might be pregnancy-dependent events.The expression of both CCL8 and CXCL10 in PBLs was stimulated by CCL16 as wel as IFNT, suggesting a chemokine interaction that at least includes CCL8, CXCL10 and CCL16, and may play a role in regulating maternal recognition in cows.

Protective Effect of Distillate and Redistillate of Cow’s Urine in Human Polymorphonuclear Leukocytes Challenged With Established Genotoxic Chemicals

Objective From the ancient period cow’urine has been used as a medicine. In Veda, cow’urine was compared to the nectar. In Susrut, several medicinal properties of cow’ urine have been mentioned and are known to cause weight loss, reversal of certain cardiac and kidney problems, indigestion, stomach ache, edema, etc. However, the literature and scripture did not mention the antigenotoxic properties of cow’urine. Methods In the present investigation, the antigenotoxic/ antioxidant properties of cow’ urine distillate and redistillate were studied in vitro. The antioxidant status and volatile fatty acid levels were determined. Actinomycin-D (0.1ol/L) and hydrogen peroxide (150 mol/L) were used for inducing DNA strand break with 0.1% DMSO as negative control. Dose for the antigenotoxic effect of cow’ urine was chosen from the dose response study carried out earlier. Results Both actinomycin-D and H2O2 caused statistically significant DNA unwinding of 80% & 75% respectively (P<0.001) as revealed by fluorimetric analysis of DNA unwinding (FADU), and the damage could be protected with the redistilled cow urine distillate (1, 50 & 100 ) in simultaneous treatment with genotoxic chemicals. Conclusion The redistillate of cowurine was found to possess total antioxidant status of around 2.6 mmol, contributed mainly by volatile fatty acids (1500 mg/L) as revealed by the GC-MS studies. These fatty acids and other antioxidants might cause the observed protective effects.

A functional type I interferon pathway drives resistance to cornea herpes simplex virus type 1 infection by recruitment of leukocytes

Type I interferons are critical antiviral cytokines produced following herpes simplex virus type-1 （HSV-1） infection that act to inhibit viral spread. In the present study, we identify HSV-infected and adjacent uninfected corneal epithelial cells as the source of interferon-a. We also report mice deficient in the A1 chain of the type I IFN receptor （CDl18-/） are extremely sensitive to ocular infection with low doses （100 PFU） of HSV-1 as seen by significantly elevated viral titers in the cornea Compared to wild type （WT） controls. The enhanced susceptibil- ity correlated with a loss of CD4＋ and CD8＋ T cell recruitment and aberrant chemokine production in the cornea despite mounting an adaptive immune response in the draining mandibular lymph node of CDll8/ mice. Taken together, these results highlight the importance of IFN production in both the innate immune response as well as eliciting chemokine production required to facilitate adaptive immune cell trafficking.

Evaluation of autonomic nervous system by heart rate variability and differential count of leukocytes in athletes

Top Japanese sprinters were evaluated for their physical condition, autonomic function, blood chemistry, differential leukocyte count and blood lactate level before and after short, maximal exercise to explore methods of quantifying their conditioning level. Statistical analysis of data obtained before and 10 min after exercise were used to estimate the athletes’ autonomic capacity during recovery. Pre and post exercise variances in differential leukocyte count revealed strong correlations between neutrophil and sympathetic activity, and lymphocyte and parasympathetic activity. The results of the study demonstrated significant alterations in autonomic parameters and differential white blood cell count in response to maximal exercise.

Studies on the flow and distribution of leukocytes in mesentery microcirculation of rats

AIM To study the effect of leukocyteendothelium interaction (LEI) on the flow anddistribution of leukocytes in microcirculationunder physiological condition.METHODS A microcirculation image multipleparameter computer analysis system (MIMPCAS)was used to study the flow and distribution ofleukocytes in mesentery microcirculation of ratsIn vivo.RESULTS The difference of visible leukocyteflux (VLF) was as high as 131 times in thearterioles and venules with similar diameter andblood velocity. The visible leukocytes rolledalong the blood vessel wall as a" jerky"movement. The frequency distribution of thevisible leukocyte velocity (VLV) showed a "twopeak" Curve. The low peak value was at 10 pm/ s-- 15 pm/ s while the high peak fell between25 pm/ s-- 30 pm/ s. With the increase of diameterof venules, VLF increased while the VLVremained at the same level. With the increase ofRBC velocity, VLV trends to elevate and VLF tofal I down.CONCLUSION The results herein might providea basic theory for the study on the mechanism ofLEI under physiological condition and novelmethods for the prevention and treatment of highLEI in many pathological

Serum Resistin Level and Polymorphonuclear Leukocytes Dysfunctions in Children on Regular Hemodialysis

Resistin is a secretory adipocytoine, which is expressed mainly in humans by inflammatory cells especially macrophages. Resistin serum levels are elevated in end-stage renal diseases of people having an increased risk of infections as a result of impaired polymorphonuclear leukocytes (PMNLs) functions. Objectives: To evaluate neutrophil functions (phagocytosis and oxidative burst) in children with end-stage renal disease (ESRD) on regular hemodialysis and to shed light on the contribution of resistin on neutrophil functions. Patients and Methods: The study included 40 children with ESRD on regular hemodialysis. Their ages ranged from 6 to 12 years, and they were selected from children attending the pediatric hemodialysis unit of AL-Zahraa Hospital, Al-Azher University during the period from October 2012 to December 2013. Another group of 40 apparently healthy children with matched age and sex with the patient group served as a control. Serum resistin, phagocytic index and nitro blue tetrazolium test (NBT%) were assessed in both groups. Results: There was a statistically more significant increase in resistin serum levels in cases than in controls;it was (3.25 ± 0.86 ng/ml) and (0.25 ± 0.16 ng/ml) respectively

A TWO-PARAMETER MODEL FOR THREE TYPES OF NUCLEPORE FILTRATION OF LEUKOCYTES

Micropore filtration of leukocytes is one of the main methods for evaluating leukocytes' deformability. Here a biofluid mechanical model was proposed for three cases: (i) filtration under gravity; (ii) constant pressure filtration; (iii) constant flow rate filtration. In previous models, constant blood cell resistance was assumed. In this paper, when evaluating the filter resistance to leukocytes, not only the effect of the change in the driving pressure but also the difference in the deformability of individual leukocytes are taken into consideration. Moreover, based on Moessmer's experimental results, a probability distribution function for the transit time of polymorphonuclear cells (PMNs) through the filter is assumed. Finally, numerical curves are obtained for the above three types of filtration and the comparison between the theoretical and experimental results of PMNs for case (i) turns out to be satisfactory. The two parameters in the model, A and B represent leukocytes' deformability under low and high pressure respectively.

Transfusion of Ipscs-Derived Leukocytes to Kill Cancer

Here we propose that the rejuvenation of leukocytes with iPSC technology in vitro and transfusion of cancer cellresistant white blood cells back to human body provide a prospective therapy for cancer patients.

Assessment of malathion and its effects on leukocytes in human blood samples

In the present paper,we report a reproducible,cost effective,fast response method for detection of malathion and its effects on leukocytes in different human blood groups.Spectroscopic methods（UV-Vis spectrometry）and Fourier transform infrared coupled with solid phase extraction were applied for analyzing malathion content in human blood plasma.The spiking levels of malathion in the range of 0.1-1.7μg/mL were extracted from blood plasma samples using SPE.The present active functional groups（C=0;P-O-C;-OH;P=S）were also characterized.The recovery rate of malathion was 80%±4.5%.The calculated correlation coefficient was 0.9799,indicating the linearity of the results.The limit of detection（LOD）and limit of quantification（LOQ）were（0.1-1.7）μg/mL and（0.3-1.5）μg/mL,respectively.Malathion〈1.0μg/mL showed no significant change while higher levels of malathion exposure（1.5μg/mL and 3.0｜ig/mL）reduced the number of white blood cells.In conclusion,the spectroscopic results may be useful to understand the mechanism of other pesticides such as methyl parathion and parathion.

Effect of Antioxidants on Endothelial Cell Reactive Oxygen Species (ROD Generation and Adhesion of Leukocytes to Endothelial Cells

Objective To investigatewhether antioxidants inhibit adhesion of leukocytes to endothelium and furthermore, whether all antioxidants regulate NF-KB activation through a redox sensitive mechanism. Methods The effect of the antioxidative substances pyrrolidin dithiocarbamat (PDTC), dichloroisocumarin (DCI), chrysin and probucol on the endothelial leukocyte adhesion were examined under near physiological flow conditions. The antioxidative activity of antioxidants was measured in a DCF fluorescence assay with flow cytometry. The activation of NF-kB in endothelial cells was investigated in a gel shift assay. Results PDTC and probucol did not show an inhibitory effect to the formation of intracellular H2O2 in TNFa activated human vascular endothelial cells (HUVEC) . Chrysin showed a moderate effect. DCI showed a strong antioxidative effect. In contrast, PDTC and chrysin inhibited the adhesion of HL 60 cells to TNFa-stimulated HUVEC. DCI and probucol did not have influence on the adhesion within the area of the examined shear stresses. Only PDTC inhibited the TNFa-induced activation of NF-KB in endothelial cells. Conclusion The inhibition of the endothelial leukocyte adhesion by antioxidative substances is not to be explained by its antioxidative characteristics only. The inhibitory effect of PDTC on NF-KB activation was probably not related to its antioxidative properties. Endothelial cell Antioxidants NF-kappa-B

Effects of glucocorticoids on leukocytes:Genomic and non-genomic mechanisms

Glucocorticoids(GCs)have been widely used as immunosuppressants and antiinflammatory agents to treat a variety of autoimmune and inflammatory diseases,and they fully exert their anti-inflammatory and immune-regulating effects in the body.The effect of GCs on white blood cells is an important part of their action.GCs can cause changes in peripheral blood white blood cell counts by regulating the proliferation,differentiation,and apoptosis of white blood cells.Although the total number of white blood cells,neutrophil counts,lymphocytes,and eosinophils increases,the counts of basic granulocytes and macrophages decreases.In addition,GCs can regulate the activation and secretion of white blood cells,inhibit the secretion of a variety of pro-inflammatory cytokines,the expression of chemokines,and promote the production of anti-inflammatory cytokines.For patients on GC therapy,the effects of GCs on leukocytes were similar to the changes in peripheral blood caused by bacterial infections.Thus,we suggest that clinicians should be more cautious in assessing the presence of infection in children with long-term use of GCs and avoid overuse of antibiotics in the presence of elevated leukocytes.GCs work through genomic and non-genomic mechanisms in the human body,which are mediated by GC receptors.In recent years,studies have not fully clarified the mechanism of GCs,and further research on these mechanisms will help to develop new therapeutic strategies.

Human sperm quality and lipid content after migration into normal ovulatory human cervical mucus containing low numbers of leukocytes

The aim of this study was to investigate whether a relationship exists between the presence of low numbers of leukocytes in normal ovulatory cervical mucus and sperm quality and lipid content after migration. The percentages of live, motile and morphologically normal spermatozoa, movement parameters assessed by computer-aided sperm analysis （CASA）, and ionophore-induced acrosome reaction measured by flow cytometry were determined before and after migration. High-performance liquid chromatography with ultraviolet detection was used to measure the sperm lipid content, including the various diacyl subspecies. The number of leukocytes found in solubilized mucus samples was counted using a haemocytometric method. Overall, the presence of leukocytes in the cervical mucus samples did not significantly influence sperm motility and morphology, sperm kinematic parameters, or the sperm content in sphingomyelin or cholesterol. In contrast, after migration, the decrease in various sperm diacyls and the level of induced acrosome reaction was significantly less pronounced in mucus samples containing ≥10^4 leukocytes than in mucus samples with no or rare leukocytes whereas the level of induced acrosome reaction was higher. The present data suggest that the low level of leukocytes found in normal ovulatory cervical mucus could influence the process of sperm lipid remodelling/capacitation.

Spatial-spectral identication of abnormal leukocytes based on microscopic hyperspectral imaging technology

Screening and diagnosing of abnormal Leukocytes are crucial for the diagnosis of immune diseases and Acute Lymphoblastic Leukemia(ALL).As the deterioration of abnormal leukocytes is mainly due to the changes in the chromatin distribution,which signicantly affects the absorption and reflection of light,the spectral feature is proved to be important for leukocytes classication and identication.This paper proposes an accurate identication method for healthy and abnormal leukocytes based on microscopic hyperspectral imaging(HSI)technology which combines the spectral information.The segmentation of nucleus and cytoplasm is obtained by the morphological watershed algorithm.Then,the spectral features are extracted and combined with the spatial features.Based on this,the support vector machine(SVM)is applied for classication ofve types of leukocytes and abnormal leukocytes.Compared with different classication methods,the proposed method utilizes spectral features which highlight the differences between healthy leukocytes and abnormal leukocytes,improving the accuracy in the classication and identication of leukocytes.This paper only selects one subtype of ALL for test,and the proposed method can be applied for detection of other leukemia in the future.

Investigation of the immune effects of Scutellaria baicalensis on blood leukocytes and selected organs of the chicken＇s lymphatic system

Background： The health of chickens and the welfare of poultry industry are central to the efforts of addressing global food security. Therefore, it is essential to study chicken immunology to maintain and improve its health and to find novel and sustainable solutions. This paper presents a study on investigation of the effect of Scutellaria baicalensis root（SBR） on the immune response of broiler chicken, especially on lymphocytes and heterophils reactivity, regarding their contribution to the development of immunity of the chickens.Methods： The 121-day-old Hubbard Hi-Y male broiler hybrids were randomly assigned to four treatment groups,three SBR supplemented groups（0.5, 1.0, and 1.5% of SBR） and one control group. Each treatment was replicated five times with six birds per replicate pen in a battery brooder. Blood was collected after 3-（rd） and 6-（th）wk of the experiment, and hemoglobin and hematocrit values were determined, as well as total leukocyte count and differential count were performed. Nitroblue tetrazolium test and phagocytosis assay as nonspecific immune parameters and humoral immune responses to the antigenic challenge by sheep red blood cells were performed.Moreover, the ability of peripheral blood lymphocytes to form radial segmentation（RS） of their nuclei was analyzed.Body weight and relative weight of spleen, liver, and bursa of Fabricius were recorded.Results： Results showed that mean heterophile/lymphocyte ratio increased in the SBR groups compared to the control group and the blood of the chickens showed lymphocytic depletion. The results also demonstrated that the relative weight of bursa of Fabricius and spleen in groups fed with SBR significantly decreased compared to the control group. This study also showed that the addition of SBR significantly inhibited the formation of RS of nuclei compared to some cytotoxic substances.Conclusion： We found that SBR supplementation should be carefully evaluated when given to poultry. The excess intake of SBR supplementation may cause immunologic inhibition and may negatively affect the development of immune organs. SBR has inhibited the formation of radial segmentation nuclei showing antimetastatic properties and also the phagocytosis of chicken heterophils.

ANTAGONISTIC EFFECTS OF GRANULOCYTE DERIVED INHIBITORS AND CYTOKINES ON APOPTOSIS IN POLYMORPHONUCLEAR LEUKOCYTES

An extract (G-INH) made from mature human granulocytes freshly isolated from normai blood causes human neutrophils to undergo apoptosis in vitro as shown by morphologic changes and by the typical ladder pattern of small DNA fragments noted on agarose gel electrophoresis of isolated DNA. Apoptosis occurs in from 20% to 30% of neutrophils over 24 hours of culture in vitro and the addition of G-INH to the medium causes a dose-related increase in the incidence of apoptosis. Heating G-INH at 60t for 30 minutes does not destroy its capacity to induce apoptosis but GM-CSF, G-CSF, and to a lesser extent IL-1β, antagonize this action. IL-3 does not diminish G-INH induced apoptosis of neutrophils. Substances, released from, mature neutrophils may participate in regulating the survival of other neutrophils, particularly in sites where the cells are in close proximity as in the marrow. Self destruction of post-mitotic neutrophils in marrow may thus represent an-other level at which regulation of cell production

Changes of glucocorticoid receptors in peripheral leukocytes in patients with bronchial asthma

The maximal binding capacity(R0) and dissociation constant(Kd) of glucocorticoid receptors(GR) in peripheral leucocytes and the cortisol(F) level in plasma were estimated in 17 patients with bronchial asthma.The R0 of GR was 12908+2501 sites cell which wa