Nitroblue Tetrazolium Reduction Test Research Articles

Pirarucu requires taurine to maximize growth and antioxidant status when fed diets high in plant-based ingredients

Taurine (Tau) is a free conditionally essential amino acid for most fish species that plays a significant nutritional role, mainly in carnivorous fish species. However, limited studies are still available regarding the role of Tau on pirarucu nutrition, an important Amazonian fish species. Therefore, this study used diets high in plant-based ingredients to investigate the effect of taurine (Tau) supplementation on juvenile pirarucu's growth, metabolism, and health (n = 115, Weight = 504.12 ± 19 g). Fish were fed five diets containing increasing levels of Tau: 0.95, 5.1, 9.6, 12.9, and 17.2 g/kg in a completely randomized design with three replicates per treatment. After a 60-day feeding trial, growth, hematology, blood chemistry, Tau content in muscle, carcass, and liver, antioxidant status, and immunological parameters were evaluated. The control group (0.95 g/kg Tau) showed the lowest growth. According to the segmented regression model, the recommended Tau level for maximizing the development of juvenile pirarucu was 4.49 g/kg of feed. Additionally, fish fed the lowest dietary Tau level showed the highest ROS (reactive oxygen species) concentration. Dietary Tau levels did not affect hematological parameters and blood chemistry (p > 0.05). Fish fed 9.6 g/kg of Tau showed increased catalase activity, while the control group had the worst result in NBT (nitroblue tetrazolium reduction test). Fish fed with the highest dietary Tau levels (12.9 and 17.2 g/kg) showed increased total serum immunoglobulin concentration. These results suggest that the optimal inclusion level of Tau in diets for juvenile pirarucu is 4.49 g/kg of feed and that this amino acid acts as a mitigator of oxidative stress.

Exploring the Relationship between Neutrophil Activation and Different States of Canine L. infantum Infection: Nitroblue Tetrazolium Test and IFN-γ.

This study aimed to investigate the role of neutrophils in canine leishmaniosis by assessing neutrophil activation and its relationship with different states of L. infantum infection and antibody and IFN-γ production. Dogs were categorized into five groups: healthy-seronegative (n = 25), healthy-seropositive (n = 21), LeishVet-stage I (n = 25), Leishvet-stage II (n = 41), and LeishVet-stage III-IV (n = 16). Results of the nitroblue tetrazolium reduction test (NBT) showed significantly higher neutrophil activation in stage I (median:17.17, range: [7.33-31.50]%) compared to in healthy-seronegative (4.10 [1.20-18.00]%), healthy-seropositive (7.65 [3.98-21.74]%), stage II (6.50 [1.50-28.70]%), and stage III-IV (7.50 [3.00-16.75]%) groups (p < 0.0001). Healthy-seropositive dogs also displayed higher values than all groups except stage I. Stages II and III-IV did not show significant differences compared to healthy-seronegative. Regarding IFN-γ, stage I dogs had higher concentrations (median:127.90, range: [0-3998.00] pg/mL) than healthy-seronegative (0 [0-109.50] pg/mL) (p = 0.0002), stage II (9.00 [0-5086.00] pg/mL) (p = 0.045), and stage III-IV (3.50 [80.00-548.80] pg/mL) (p = 0.02) dogs. Stage II dogs showed increased IFN-γ compared to healthy-seronegative dogs (p = 0.015), while stage III-IV dogs had no significant differences compared to healthy-seronegative dogs (p = 0.12). Healthy-seropositive dogs had elevated IFN-γ concentrations compared to healthy-seronegative dogs (p = 0.001) and dogs in stage III-IV (p = 0.03). In conclusion, neutrophil activation was higher in dogs with mild disease and healthy-seropositive dogs, and a relationship between neutrophil activation and the production of IFN-γ was found.

Effect of storage on the nitro blue tetrazolium reduction test in dog blood samples.

The nitro blue tetrazolium (NBT) reduction test (NBTT) has been used for measuring the metabolic activity of phagocytes of mammals. Activated neutrophils transform NBT into formazan in the cytoplasm. The NBTT can detect the activation of neutrophils in peripheral blood and is used to assess neutrophil function in dogs. However, the NBTT is not used frequently in the clinical setting, as samples should be processed after blood collection. The aim of this study was to evaluate the effect of storage on NBTT in dog blood samples. Residual EDTA blood samples from 22 dogs were included of different ages, breeds, and sex. The buffy coat layer was separated from the blood and incubated with 0.1% NBT. The NBTT was performed at 0, 24, 48, and 72 h after the collection of blood. Blood samples were stored at 4°C until the tests were performed. Blood smears were evaluated by light microscopy, and the NBT reduction rate was reported, which represents the percentage of activated neutrophils. The NBT reduction rate was calculated after counting 300 neutrophils in each slide. The means of NBTT in dog blood samples at 0, 24, 48, and 72 h were 8.3%, 8.5%, 8.7%, and 7.8%, respectively. No significant differences were observed between time points. This study showed that the NBTT can be performed up to 72 h after the collection of canine blood if correctly refrigerated at 4°C. This finding supports the performance of the NBTT in the clinical setting.

Oncolytic Newcastle disease virus effects on immune response - a new issue in cancer treatment.

Millions of people are diagnosed with cancer each year, and fighting it puts a heavy financial burden on communities and governments. Numerous advances have been made in the field of cancer; one of the newest methods is using oncolytic viruses. This study aimed to evaluate the effect of oncolytic Newcastle disease virus wild-type strains (NDV-WTS) on the immune system. Forty mice were divided into four groups (10 animals in each group). The control group received phosphate buffered saline, and experimental group 1 (NDV-WTS 1), experimental group 2 (NDV-WTS 2), and experimental group 3 (NDV-WTS 3) received 10-1, 10-2, and 10-3 titers of Newcastle virus on 0, 14th, and 28th days. On the 31st day, 100 µL of Newcastle virus was injected into the left footpads of animals. After 48 hours, delayed-type hypersensitivity (DTH) reactions were measured. On the 33rd day, peritoneal macrophages were isolated. Then proliferation of the cells was measured by the methyl-thiazolyl-tetrazolium (MTT) test. Neutral red uptake and respiratory burst of peritoneal macrophages were also assessed. Data were analyzed using statistical software SPSS, version 19. The results of the DTH test showed that footpad swelling in control, NDV-WTS 1, NDV-WTS 2, and NDV-WTS 3 groups were 23.5%, 23.5%, 23.6% and 23.6%. No significant differences were seen between the groups in this regard (P &gt; 0.05). A negative nitroblue tetrazolium (NBT) reduction test as an indicator of macrophage's respiratory burst, showed no significant difference between the groups (P &gt; 0.05). The neutral red uptake assay and MTT test showed no significant differences between the groups (P &gt; 0.05). The results of this study showed that NDV-WTS in doses of 10-1, 10-2, and 10-3 have no adverse effects on healthy normal cells.

MEGAMOX MEDIATES ECO-FRIENDLY SYNTHESIS OF SELENIUM NANOPARTICLES AS IMMUNOMODULATORY, ANTIOXIDANT AND ANTIMICROBIAL AGENTS

The research herein includes methodology for eco-friendly preparation of selenium nanoparticles (SeNPs) using the broad-spectrum antibiotic, Megamox (Mega.). Characterization of SeNPs was done using UV-Visible spectroscopy, FTIR, XRD, DLS and TEM imaging. Additionally, the immunomodulatory, antioxidant and antimicrobial activities of both SeNPs and Mega. were checked against different strains of bacteria and fungi. TEM and DLS images showed that SeNPs are polydisperse spheres with mean diameter of 22.4 nm. FTIR analysis indicated that the hydroxyl and nitrogen moieties in Mega. were effective for reduction plus binding manner. According to the results of the nitro blue tetrazolium reduction test, both SeNPs and Mega. presented high intracellular killing activity, which confirmed their immunostimulatory effect. Antioxidant activity of SeNPs and Mega. were 90 and 82 %, respectively. SeNPs presented great activity facing multi-drug resistant bacteria and TB. SeNPs are considered promising cost-effective and eco-friendly anti-bacterial and anti-fungal agents that can represent a new potential nano-platform in both medical and infectious diseases control.

Иммунорегулирующий потенциал омега-3 полиненасыщенных жирных кислот: использование для профилактики частой инфекционной заболеваемости у детей

В последние годы все больше исследований направлено на изучение иммуномодулирующей роли длинноцепочечных полиненасыщенных жирных кислот (ДЦПНЖК), практическое применение которых потенциально способно предотвращать развитие острых инфекционных респираторных заболеваний и аллергических состояний. Из множества ДЦПНЖК наибольший интерес вызывают омега-3-кислоты. В статье приведен обзор некоторых исследований, а также данные собственного исследования о влиянии диетической добавки, содержащей комплекс омега-3-кислот («Омегамi Розумна дитина»), на снижение заболеваемости ОРЗ детей с рекуррентными формами острой респираторной инфекционной патологии. Иммунотропное действие диетической добавки к рациону питания «Омегамi Розумна дитина» у детей характеризовалось усилением функционирования врожденного и адаптивного иммунитета, а иммунорегулирующая направленность проявилась в ограничении провоспалительной настроенности иммунной системы у детей, в виде снижения НСТ-теста, количества CD25 и уменьшения сывороточного содержания ФНО-α при отсутствии сопутствующей иммуносупрессии противомикробного защитного потенциала.

Study of in-vitro anti-inflammatory and immunomodulatory effect of Ayurvedic plants – Murva

Background and Aims: Toxic chemicals, pathogens and damaged cells trigger the inflammatory process and cause disease in the kidney, heart, brain, liver, lung and reproductive system. The present study evaluates the anti-inflammatory and immunomodulatory effect of Ayurvedic plants under the Murva category. Methods: Marsdenia tenacissima is an acceptable source of Murva. Sansevieria roxburghiana is considered as Murva in west Bengal. Methanol extracts of both plants were used to evaluate anti-inflammatory and immunomodulatory effects by certain in vitro methods. The in vitro anti-inflammatory effects of methanol extract Marsdenia tenacissima (MEMT) and methanol extract of Sansevieria roxburghiana (MESR) were assessed by heat and hypotonic induced haemolysis on the red blood cells of rats. Nitro blue tetrazolium (NBT) assay and inhibition of TNF-α release in DAL cell lines were conducted to assess the immunomodulatory potential of extracts. Results: MEMT, MESR and a combination of MEMT&amp;MESR showed significant (p&lt;0.001) inhibition of haemolysis on heat and hypotonic induced methods compared with the standard Diclofenac Sodium. In NBT reduction test, MEMT showed a more significant (p&lt;0.001) result than MESR. Similarly, the inhibition of TNF-α release was also significantly enhanced by 400 µg/ mL of MEMT. Conclusion: The present research results revealed that Murva is a safe Ayurveda drug for the treatment of cancer and cardiac disease with the protection of systemic immunity and set free from an inflammatory condition.

Non-specific factors of children protection with streptococcal tonsillitis and infected with herpes virus type 6

The aim of the work was to study the characteristics of the response of non-specific protective factors in tonsillitis of streptococcal etiology in children infected and uninfected with HHV-6 infection at different periods of the disease. Materials and research methods. In 78 children aged 3–15 years, inflammation markers and phagocytic activity of peripheral blood neutrophils (PAN) were determined using the nitroblue tetrazolium reduction test. The studies were carried out in the acute period and in the period of convalescence. Mathematical and statistical processing of the data obtained was carried out using Microsoft Excel 2003 and Statistica 6.0 programs. The significance of differences between the mean values was determined using the Student's test (t). Results. It was found that infection of children with HHV-6 leads to a decrease in the functional activity of neutrophils in the onset of streptococcal tonsillitis compared with uninfected patients (p&lt;0.05). Also, the acute period of streptococcal tonsillitis in patients with co-infection is accompanied by a violation of the functional reserve of the oxygen-dependent mechanism of bactericidal neutrophils: minimal opportunities were found in tonsillitis on the background of HHV-6 infection (p&lt;0.05). By the period of convalescence, a decrease in the indicators of the functional activity of neutrophils, increased in the acute period, was recorded, but in children infected with HHV-6, a complete recovery of indicators to the physiological norm did not occur. One of the features of streptococcal tonsillitis in children with HHV-6 infection is an increase in serum gamma globulin level (p˂0.05), lack of significant dynamics of decrease in the ASL-O indicator. Conclusions. The presence of VHL-6t infection in a child with streptococcal tonsillitis adversely affects the state of nonspecific protective factors, in particular PAN. The obtained data must be used to optimize the management of patients at different periods of the disease, including at the stages of dispensary observation

Влияние пептида GHK-D-Ala на механизмы врожденного иммунитета и процессы перекисного окисления липидов в условиях инфицированной раны

The objective is to study the effects of Gly-His-Lys-D-Ala (GHK-D-Ala) peptide on mechanisms of innate immunity and lipid peroxidation processes in infected wound. Materials and methods. The experiments were carried out on 80 Wistar male rats. The infected wound was modeled by applying a full-layer wound on the back and performing primary wound surgery with primary suturing 24 hours later. GHK-D-Ala peptide was injected at a dose of 0.5 µg/kg into the wound area every day for 30 days. The phagocytic activity of blood neutrophils was studied in spontaneous and stimulated nitroblue tetrazolium reduction test (NBT-test). The activity of lipid peroxidation (LPO) processes in rats’ blood serum was assessed by the content of malondialdehyde (MDA) and acylhydroperoxides (AHP). Results. GHK-D-Ala administration had a positive effect on both phagocytosis mechanisms and the level of LPO products in comparison with the control group. There was a significant increase in NST-positive neutrophils in the spontaneous test against peptide administration on days 3, 7, 10, and 30 (p˂0.001) and in the stimulated NST test on days 3, 7, and 10 (p &lt; 0.001). The functional neutrophil reserve had been significantly increased by day 10 (p˂0.05) and had been significantly lower by day 30 (p &lt; 0.05). Phagocytic index and phagocytic number were significantly higher on days 3 and 10 (p &lt; 0.01). The phagocyte activity index significantly increased on days 3 and 10 (p &lt; 0.05-0.001).A significant decrease in the MDA concentration was demonstrated on days 3, 7, 10, and 30 (p&lt;0,001). There was a significant decrease in the content of AGP on days 3, 7, 10 and 30 (p &lt; 0.001). Conclusion. The corrective effect of Gly-His-Lys-D-Ala peptide on the phagocytic activity of granulocytes and the processes of lipid peroxidation in infected skin wounds was established.

Effect of different doses of the tissue biostimulant on some indicators of the immune status of heifers at the age of 6 months

The article presents the results, the purpose of which was to study the effect of the tissue biostimulant obtained from the slaughterhouse tankage of antler deers on some indicators of the immune status of heifers at the age of 6 months. Experimental studies were carried out in 2020 on the basis of Prigorodnoye Uchkhoz JSC of the Industrial District of Barnaul, the Altai Territory. For the experiment, we selected one-month-old heifers with an average live weight of 51.0 kg. Throughout the experiment, the animals of the experimental groups were given 1 injection in 1, 2, 3, 4, 5 and 6 months of raising with an interval of 30 days. The study preparation was administered subcutaneously into the lower third of the neck. Animals in the control group were administered with saline from 1 to 5 months of raising at a dose of 3 ml/head, at 6 months. - 6 ml/head, to analogs of the I experimental group from 1 to 5 months - 2 ml/head. At 6 months. - 4 ml/head, in II - from 1 to 5 months. - 3 ml/head, at 6 months - 6 ml/head, in III - from 1 to 5 months. - 4 ml/head at 6 months - 8 ml/head. According to the results of the experiment, it was found that the heifers of experimental group II had the best immune status, in which the largest increase of the indicators of spontaneous Nitro Blue Tetrazolium Reduction Test (NBT-test) was noted - by 12.5% (p≤0.01), of stimulated NBT-test - by 10.0% (p ≤0.05), of phagocytic index - by 5.8% (p≤0.05) in comparison with the control.

Antimicrobial and immunomodulatory potential of nanoscale hierarchical one-dimensional zinc oxide and silicon carbide materials

This work reports the comparison of antimicrobial and immunomodulatory activities between nanoscale hierarchical ZnO nanorods (NRs) and SiC nanowires (NWs). The fabricated one-dimensional (1D) materials possess various nanoscale sizes, morphologies, geometries, and crystal directions. A developed hexamethylenetetramine-assisted hydrothermal regime was followed for preparing wurtzite–ZnO NRs with 40 nm average width, 1 μm length, and [0001] growth orientation. The antibacterial activity of NRs was compared with that of SiC NWs, which had 50–80 nm width and [111] direction. The antibacterial activity of the hierarchical NRs and NWs was tested using different microorganisms including Gram-positive and Gram-negative bacteria and fungi strains. The microbial inactivation of the hierarchical 1D nanomaterials was evaluated by inhibition zone, minimal inhibition concentration (MIC), cell viability assays, and nitroblue tetrazolium reduction test. Cell apoptosis was elucidated by confocal laser scanning microscopy. The scanning electron microscope results showed that [0001]-ZnO NRs led to higher loss in bacterial viability than [111]-SiC NWs. Also, ZnO NRs exhibited lower MIC values than SiC NWs with 3.9, 7.9, 31.25, and 7.91 μg/mL for P. aeruginosa, A. baumannii, S. aureus ATCC 29213, S. typhi ATCC 6539, respectively. This is caused by the higher surface area, smaller width and length, hexagonal wurtzite structure, and highly exposed [0001] polar surface grown along the c-axis of ZnO NRs.

Tetrandrine Induces HL-60 Cell Differentiation

To investigate the differentiation of acute promyelocytic leukemia (APL) cells induced by Tetrandrine targeting MUC1. HL-60 cells at logarithmic growth phase were choosen as object of reaserch and were divided into four groups: control, ATRA, Tetrandrine and ATRA+Tetrandrine group. Cell morphologic changes in each group were observed by microscopy. The change of cell differentiation ability was analyzed by nitro-blue tetrazolium (NBT) reduction test. The expression of cell surface differentiation antigen CD11b was measured by flow cytometry. The expression of MUC1 protein was detected by Western blot. The differentiation of HL-60 cell could be induced by Tetrandrine. The NBT reduction-positive rate of ATRA+Tetrandrine group was significantly higher than that in ATRA group and Tetrandrine group (P<0.05). The percentage of CD11b positive cells in ATRA+Tetrandrine group(43.62％±1.38％) was significantly higher than that in Tetrandrine group(15.25％±2.11％), ATRA group (28.84％±7.53％) and control group(8.16％±1.01％) (P<0.05). The content of MUC1 protein in Tetrandrine group was significantly lower than that in control group and ATRA group (P<0.05). Tetrandrine and ATRA can synergize to promote the differentiation and maturation of HL-60 cells, and the mechanism may be related with MUC1 expression.

The effect of acute somatic pain on the killing activity of neutrophils in newborn rats

The immune system is subject to all sorts of influences. Pain is one of them, accompanying an organism’s existence. It is essential to be aware of and account for age-related characteristics of the innate immunity in order to adequately assess their dynamics in ontogenesis. The literature is scarce on the changes to the killing activity of neutrophils occurring in newborns in response to acute pain. The aim of this study was to detect potential changes to the phagocytic activity of neutrophils in response to an algogenic stimulus in newborn rats. The experiments were carried out in 3-5-day-old rats. Two groups were formed: the control group and the main group, in which acute pain was modelled. Blood samples were collected 2, 30–60 and 120–180 minutes after exposure to the algogenic stimulus. The microbicidal activity of neutrophils was measured using a spectrophotometric modification of the spontaneous/stimulated nitroblue tetrazolium (NBT) reduction test. The results were compared using the Mann-Whitney U test. In the first hour following pain modeling, the stimulated NBT reduction test demonstrated an increase in the measured parameters from 71.5 to 87.4 a.u. (р &lt; 0.001); the spontaneous NBT reduction test showed an increase from 50.7 to 58.6 a.u. (p &lt; 0.01) 30 to 60 min after exposure. The most pronounced change of the microbicidal activity coefficient was observed 2 min after pain modeling, increasing from 1.40 to 1.72 a.u (р &lt; 0.001). By the end of the experiment, the measured parameters approximated their initial values. During the analysis, we accounted for the fact that the neutrophil response to the algogenic stimulus was unfolding in the setting of microbial colonization occurring in newborns.

Evaluation of immunotropic activity of gold nanocolloid in chickens

Gold nanoparticles (AuNPs) are one of the most examined nanomaterials, but information about their immunogenic potential is still insufficient. Understanding interaction of AuNPs with immune system is essential in designing their safety and possibilities of biomedical applications. An experiment was conducted to determine immunotropic activity of gold nanocolloid (AuNPs) administered orally to chickens depending on dose and duration time. 162 birds were assigned to 9 experimental groups of 18 birds each. The control group (C) did not receive AuNPs. Groups: T10.5, T11.0, T11.5, T12.0, received nano-gold in a rate of 0.5 mg/kg body weight/d, 1.0 mg/kg body weight/d, 1.5 mg/kg body weight/d and 2.0 mg/kg body weight/d in 8–14, 22–28 and 36–42 days of the life. The birds in groups T20.5, T21.0, T21.5, T22.0, received nano-gold in the same doses, but only in 8–10, 22–24 and 36–38 days of life. Phagocytic activity of leukocytes was determined in vitro using Staphylococcus aureus 209P strain, their respiratory burst activity was quantified by nitroblue tetrazolium reduction test. Serum lysozyme content was determined by the turbidimetric method. The Wintrobe method was used to determine the erythrocyte sedimentation rate. Ceruloplasmin in the blood plasma was estimated by the p-phenylenediamine colorimetric method. The level of chicken immunoglobulins: IgA, IgM and IgY and interleukin IL-6 in the blood were determined using ELISA tests. The lowest dose of AuNPs, independently on duration time had no effect on immune parameters of chickens. In all other groups receiving nano-gold for a shorter period (T2), there was an increase in the respiratory burst activity of leukocytes and a drop in lysozyme activity in blood. The higher doses (1.5 and 2.0 mg/kg body weight/d) of the nano-gold administered for the longer time period had a pro-inflammatory effect, as indicated by an increase in the level of interleukin 6 and ceruloplasmin activity as well as the erythrocyte sedimentation rate. They also contributed to an elevation of class IgA and IgY contents in blood. The results of the study revealed that the influence of nano-gold on immune response of chickens were dependent both on dose and duration time. Long lasting administration of higher doses of AuNPs contributed to adverse effect in form of inflammation response. To avoid the development of inflammatory reaction, administered dose of nano-gold should not exceed 1.0 mg/kg body weight/d.

Chronic Granulomatous Disease: An Unreported Mutation

Chronic Granulomatous Disease (CGD) is caused by defects in the phagocyte NADPH oxidase and occurs in approximately 1:200,000 births worldwide. It presents with early onset of severe recurrent bacterial and fungal infections. This is a case of a 9-year old male with severe, recurrent bacterial infections since 3 weeks of age. Initial Nitroblue tetrazolium (NBT) reduction tests were normal but a DNA analysis revealed a previously unreported homozygous mutation in CYBB, p.S418Y. Dihydrorhodamine (DHR) test showed poor neutrophil oxidation consistent with X-linked CGD. Definitive microbiologic diagnosis is essential for directing therapy for recurrent bacterial and fungal infections. Treatment of infections should be aggressive. Lifelong bacterial and fungal prophylaxis is necessary for prolonged survival. We report a case of confirmed CGD with the previously unreported mutation.

Cases of Impaired Oxidative Burst in HIV-Exposed Uninfected Infants' Neutrophils-A Pilot Study.

An increased risk of serious bacterial infections in HIV-exposed uninfected (HEU) infants has been demonstrated. Although neutrophils are essential for the protection of infants against bacterial infections, no study has investigated their profile in HEU infants to date. In this study, we assessed the function of neutrophils in HEU infants using the nitroblue tetrazolium reduction test. Among 25 HEU infants, 9 (36%) showed a reduced ability of their neutrophils to produce reactive oxygen species upon stimulation with bacteria. No alteration of total neutrophil counts was noted in the blood of HEU infants indicating that the alteration observed in the 36% of HEU infants may only be functional. Conclusively, impaired neutrophil function could be a factor of vulnerability in HEU infants.

Honey bee venom combined with 1,25-dihydroxyvitamin D3as a highly efficient inducer of differentiation in human acute myeloid leukemia cells.

Most cancer cells exhibit a defect in their capacity to mature into nonreplicating adult cells and existing in a highly proliferating state. Differentiation therapy by agents such as 1,25-dihydroxyvitamin D3(1,25-(OH)2 VD3) represents a useful approach for the treatment of cancer including acute myeloid leukemia. Human myeloid leukemia cell lines are induced to terminal differentiation into monocyte lineage by 1,25-(OH)2 VD3. However, usage of these findings in the clinical trials is limited by calcemic effects of 1,25-(OH)2 VD3. Attempts to overcome this problem have focused on a combination of low concentrations 1,25-(OH)2 VD3 with other compounds to induce differentiation of HL-60 cells. In this study, the effect of honey bee venom (BV) and 1,25-(OH)2 VD3, individually and in combination, on proliferation and differentiation of human myeloid leukemia HL-60 cells were assayed. In this in vitro study, toxic and nontoxic concentrations of BV and 1,25-(OH)2 VD3 were tested using Trypan blue stained cell counting and (3[4, 5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide) assay. In addition, differentiation of cells was assayed using a Wright-Giemsa staining and nitroblue tetrazolium reduction test. Data were analyzed by a one-way analysis of the variance test using SPSS software. Our findings showed that both the BV and 1,25-(OH)2 VD3, in a dose and time-dependent manner, caused cell death at high concentrations and inhibited cell proliferation at lower concentrations. About 5 nM of 1,25-(OH)2 VD3 induced differentiation of HL-60 cells to monocytes after 72 h. 2.5 μg/ml of BV suppressed proliferation of HL-60 cells but had not any effects on their differentiation, whereas in combination with 5 nM of 1,25-(OH)2 VD3, it enhanced antiproliferative and differentiation potency of 1,25-(OH)2 VD3. These results indicate that BV potentiates the 1,25-(OH)2 VD3-induced HL-60 cell differentiation into monocytes.

Клініко-діагностичні та лікувально-профілактичні особливості синдрому асептичного менінгіту у хворих з кістозними менінгіомами головного мозку

Objective – to investigate the frequency of aseptic meningitis syndrome (AMS), to describe the peculiarities of diagnostics, clinical signs of AMS and its differential diagnostic criteria, as well as features of immune parameters for patients with cystic meningiomas with normal postoperation recovery and with AMS progression. Materials and methods. To describe the symptoms and determine the frequency of AMS there had been analyzed the medical records of 54 patients with cystic meningiomas of various localization and histological structure, being operated at Extracerebral Tumor Department of the SI Romodanov Neurosurgery Institute NAMS of Ukraine during 2012-2016. Immunity was estimated for 18 patients with brain tumors of different histostructures and postoperative AMS (meningiomas – in 13, gliomas of low degree anaplasia – in 2, gliomas of high degree anaplasia – in 3) in comparison with normal postoperative recovery for 7 patients with tumors of the brain without occuring of AMS (meningiomas – in 3, glioblastoma – in 1, metastasis of cancer – in 1). Studies were conducted in dynamics (weekly): at the 1st-7th, 8th-14th and 15th day or further after the surgery. The phenotype of peripheral blood lymphocytes (CD3+, CD4+, CD8+, CD4+/CD8+) was determined using monoclonal antibodies (IMK Plus, Becton Dickinson, USA) by means of flow cytometry (FASCalibur E2085), IgA, IgM and IgG, serum immunoglobulin level – by means of simple radial immunodiffusion in gel (Mancini method). The functional activity of neutrophils was estimated by the phagocytosis test according to phagocytic activity and phagocytic number, as well as by nitro blue tetrazolium reduction test. Ten patients with stable compression of the roots of lumbar spine were considered as the control group. Results. AMS occurred for 8 (14.81 %) patients. The clinical picture of AMS developed quickly on the 3rd-7th day after the operation. In the clinical blood count, leukocytosis was noted without significant fluctuations, high ESR. The analysis of the peripheral blood immunity parameters of patients with AMS showed no signs of infectious or autoimmune inflammation. The changes were indolent, atypical and short-term. The rates returned to normal ones within 3 weeks. In our opinion, the following laboratory indicators can show the risk of AMS developing in the preoperative period: a decrease in the number of lymphocytes and eosinophils, an increase in the number of monocytes of the peripheral blood. It is possible to assume, that a greater risk of AMS developing arises in special areas of localization (convextic-associated with the liquoronal pathways, the ventricles of the cerebrum and the posterior cranial fossa) and tumors of individual histological variants (anaplastic cystic meningiomas and glial tumors with cystic component, with necrosis and sarcolysis of the tumor itself). Conclusions. Eradication of cystic meningioma of the brain, the prevention of cystic fluid entering the cerebrospinal fluid, the rational use of glucocorticoids within 5-10 days after the operation can significantly reduce the frequency of ASM occurrence. When evidence of meningitis appear, it is mandatory to differentiate between it and bacterial meningitis and to extract cystic components from the cerebrospinal fluid and reduce the inflammatory response of the cerebral meninges by means of glucocorticoids.

Effect of Human Umbilical Cord-derived Mesenchymal Stem Cells on Proliferation and Differentiation of Leukemia Cells

To investigate the effect of human umbilical cord-derived mesenchymal stem cells(HUC-MSC) on the proliferation and differentiation of NB4 treated with all-trans retinoid acid (ATRA) and its underlining mechanisms . Human umbilical cord mesenchymal stem cells were isolated from umbilical cord of newborns. Co-culture system was established by HUC-MSC and NB4 in vitro. The experiment was divided into 4 groups: NB4 group (NB4 cells alone) , NM group (NB4 cells co-cultured with HUC-MSC) , NA group (NB4 cells treated with ATRA) , NMA group (NB4 cells co-cultured with HUC-MSC and treated with ATRA) . NB4 cells were counted by a microscopy, NB4 proliferation was monitored by CCK-8 assay, NB4 differentiation was assessed by Wright ' s staining and nitroblue tetrazolium reduction test. IL-6 levels in the culture supernatant of different groups were tested by ELISA kit. Quantitative PCR was used to detect the transcription level of CDKN1A, CCND1 and Survivin. NB4 and HUC-MSC in the co-culturing systems were in good condition with a slight repression of NB4 proliferation by HUC-MSC. HUC-MSC could collaborate with ATRA to induce significant NB4 differentiation. Consistent with this finding, IL-6 expression levels of co-cultured groups were remarkably higher than that in any other groups or the group of HUC-MSC alone. The quantitative PCR analysis showed that the levels of CDKN1A and CCND1 mRNA expression were increased or decreased respectively in the co-cultured groups. HUC-MSC co-culture can reduce proliferation but promote the differentiation of NB4 cells, suggesting that this effect may be closely related with the secretion of IL-6 which can affect the expression of some factors in vitro.

Human Umbilical Cord-derived Mesenchymal Stem Cells Secrete Interleukin-6 to Influence Differentiation of Leukemic Cells

To investigate the effect of human umbilical cord-derived mesenchymal stem cells (UC-MSC) on the differentiation of leukemic cells. The co-culture system of UC-MSC with acute promyelocytic leukemic cell line NB4 cells was constructed in vitro,and the differentiation status of the leukemic cells was assessed by cell morphology,nitroblue tetrazolium reduction test,and cell surface differentiation marker CD11b. UC-MSC induced the granulocytic differentiation of NB4 cells. When UC-MSC and a small dose of all-trans retinoic acid were applied together,the differentiation-inducing effect was enhanced in an additive manner. Interleukin (IL)-6Ra neutralization attenuated differentiation and exogenous IL-6-induced differentiation of leukemic cells. UC-MSC can promotd granulocytic differentiation of acute promyelocytic leukemia cells by way of IL-6 and presented additive effect when combined with a small dose of all-trans retinoic acid.