Nitroblue Tetrazolium Reduction Research Articles

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&MESR showed significant (p<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<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.

Comparative Study of Antioxidant and Pro-Oxidant Properties of Homoleptic and Heteroleptic Copper Complexes with Amino Acids, Dipeptides and 1,10-Phenanthroline: The Quest for Antitumor Compounds.

In a search for new antitumoral agents, a series of homoleptic copper(II) complexes with amino acids and dipeptides, as well as heteroleptic complexes containing both dipeptides and 1,10-phenanthroline, were studied. Furthermore, a single-crystal structure containing alanyl-leucinato ([Cu3(AlaLeu)3(H2O)3(CO3)]·PF6·H2O), which is the first homotrinuclear carbonato-bridged copper(II) complex with a dipeptide moiety, is presented. To assess possible antitumor action mechanisms, we focused on the comparative analysis of pro- and antioxidant behaviors. Pro-oxidant activity, in which the reactive oxygen species (ROS) formed by the reaction of the complexes with H2O2 produce oxidative damage to 2-deoxy-d-ribose, was evaluated using the TBARS method. Additionally, the antioxidant action was quantified through the superoxide dismutase (SOD)-like activity, using a protocol based on the inhibitory effect of SOD on the reduction of nitrobluetetrazolium (NBT) by the superoxide anion generated by the xanthine/xanthine oxidase system. Our findings show that Cu–amino acid complexes are strong ROS producers and moderate SOD mimics. Conversely, Cu–dipeptide–phen complexes are good SOD mimics but poor ROS producers. The activity of Cu–dipeptide complexes was strongly dependent on the dipeptide. A DFT computational analysis revealed that complexes with high SOD-like activity tend to display a large dipole moment and condensed-to-copper charge, softness and LUMO contribution. Moreover, good ROS producers have higher global hardness and copper electrophilicity, lower copper softness and flexible and freely accessible coordination polyhedra.

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<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<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

Статеві й вікові особливості змін імунологічних показників у хворих на рак щитоподібної залози і хворих на токсичний зоб після радіойодотерапії

Мета: провести порівняльний аналіз цитотоксичної активності NK-клітин і метаболічної активності нейтрофілів (НФ) до і після радіойодотерапії (РЙТ) у хворих на диференційований рак щитоподібної залози (ДРЩЗ) залежно від статі, віку, наявності віддалених метастазів і у хворих на токсичний зоб (ТЗ) залежно від статі. Матеріали та методи. Визначили імунологічні показники в трьох групах тиреоїдектомованих хворих із ДРЩЗ. У групу 1 ввійшли 48 осіб (30 жінок, 18 чоловіків) молодого віку (від 11 до 39 років) без віддалених метастазів ДРЩЗ. Групу 2 становили 40 хворих (23 жінки, 17 чоловіків) молодого віку (від 16 до 40 років) із віддаленими метастазами ДРЩЗ. Групу 3 становили 55 хворих (30 жінок, 25 чоловіків) старшого віку (від 41 до 75 років) із віддаленими метастазами ДРЩЗ. Середня кількість курсів РЙТ у жінок групи 1 становила 1,10 ± 0,04, у чоловіків — 1,50 ± 0,24, у жінок групи 2 — 5,4 ± 0,7, у чоловіків — 6,3 ± 0,8. У хворих старшого віку (група 3) середня кількість курсів РЙТ у жінок становила 5,4 ± 0,3, у чоловіків — 5,5 ± 0,7. Визначали імунологічні показники в 30 хворих (22 жінки, 8 чоловіків) із ТЗ віком від 23 до 67 років. Хворі на ТЗ отримали курс РЙТ. Імунологічне дослідження проводили напередодні й у ранній термін (5 чи 6 діб) після РЙТ. Цитотоксичну активність NК-клітин проти індикаторних клітин-мішеней визначали, використовуючи спектрофотометрію. Метаболічну активність НФ досліджували в стандартному тесті відновлення нітросинього тетразолію. Результати. У ранній термін після РЙТ активність NK-клітин знижується в чоловіків молодого віку групи 1. У чоловіків молодого віку групи 2 активність NК-клітин після РЙТ знижується більшою мірою, ніж у групі 1. Активність NК-клітин у чоловіків старшого віку (група 3) і до, і після РЙТ істотно нижча, ніж у жінок, і значно нижча, ніж у чоловіків молодого віку. Після РЙТ метаболічна активність НФ у хворих на ДРЩЗ чоловіків і жінок збільшується в групі 1 і в чоловіків групи 2. У жінок групи 1 цей показник значно більший, ніж у чоловіків. У чоловіків і жінок групи 2 метаболічна активність НФ після РЙТ істотно менша, ніж у хворих групи 1. У хворих на ТЗ осіб чоловічої статі активність NK-клітин напередодні РЙТ нижча, а метаболічна активність НФ значно вища, ніж у жінок. Після РЙТ збільшення активності NК-клітин у хворих на ТЗ більшою мірою відбувається в осіб жіночої статі, а метаболічної активності НФ — в осіб чоловічої статі. Висновки. Порівняльний аналіз результатів дослідження раннього впливу 131І на цитотоксичну активність NK-клітин і метаболічну активність НФ виявив особливості змін цих показників у хворих на ДРЩЗ залежно від віку, статі, наявності віддалених метастазів, у хворих на ТЗ — залежно від статі.

Experimental study on disorders in the activity of macrophages in the acute period of different severities of cerebral stroke

BACKGROUND: The development of immunosuppression in the acute phase of cerebral stroke may lead to infectious and inflammatory complications. However, data on the functional activity of macrophages are insufficient to understand the pathogenesis of stroke.
 AIM: This study aimed to investigate the functional activity of macrophages in the acute period of experimental cerebral stroke (ECS) of different severities.
 MATERIALS AND METHODS: In an experimental study on 45 Wistar rats, the functional and spontaneous metabolic activity of macrophages in the spleen and abdominal cavity and the induced metabolic activity of peritoneal macrophages were evaluated through the reduction of nitro blue tetrazolium via spectrophotometry. The spontaneous and induced adhesive activity and the inhibition of the modified adhesive activity of peritoneal macrophages were examined via crystal violet staining by using a colorimetric method. ECS was modeled by the rotational movements of mandrel-knife inserted into the region of capsula interna (c.i.) on the left by guiding a needlecannula with the subsequent damage of vessels and the formation of mild (local incision of vessels in the region of c.i.), moderate (additional introduction of autologous blood in the area of c.i.), and severe (cutting of vessels from c.i. to pia mater with the subsequent introduction of autologous blood into the damaged area) hemorrhage. Data were evaluated in 72 h after different severities of ECS were modeled.
 RESULTS: In 72 h after ECS with different severities was modeled, the functional and spontaneous metabolic activity of splenic macrophages significantly decreased compared with those in the control group. The functional, spontaneous, and induced metabolic activity of peritoneal macrophages significantly reduced compared with those in the control group. The adhesive activity of peritoneal macrophages significantly increased compared with that of the control group. These disorders exacerbated in animal models with a more severe degree of stroke.
 CONCLUSION: In the acute period of ECS, the functional activity of macrophages decreased, whereas their adhesive activity increased. These phenomena enhanced in more severe models of stroke.

The Polarity of Fullerene C60 Derivatives for Enhanced Photodynamic Inactivation.

In this article, four novel fulleropyrrolidines derivatives were synthesized to study how the effect of polarity and positive charge distribution can influence the efficacy of photodynamic inactivation treatments to kill bacteria. The design of the photosensitizers was based on DFT calculations that allowed us to estimate the dipolar moment of the molecules. Neutral compounds bearing N-methyl bis-acetoxy-ethyl (1) and bis-hydroxyethyl (2) amine were the starting material to obtain the dicationic analogs N,N-dimethyl bis-methoxyethyl (3), and bis-acetoxy-ethyl) (4) methylammonio. As expected from fullerene C60 derivatives, compounds 1-4 absorb in the UV region, with a peak at 430 nm, a broader range of absorption up to 710 nm, and exhibit weak fluorescence emission in toluene and reverse micelles. In the biomimetic AOT micellar system, the highest singlet oxygen photosensitization was found for compounds 1, followed by 3, 2, and 4. Whereas 4 was the most effective reducing nitro blue tetrazolium in the presence of β-NADH. The influence of type I and type II mechanism on the photodynamic activity of compounds 3 and 4 was further examined in the presence of L-tryptophan and two reactive oxygen species scavengers. In vitro experiments indicated that the compounds with the highest dipolar moments, 3 (37.19 D) and 4 (38.46 D), inactivated methicillin-resistant Staphylococcus aureus and Escherichia coli bacteria using an energy dose <2.4 J cm-2 . No inactivation was observed for the neutral analogs with the lowest dipolar moments. These findings help to optimize sensitizer structures to improve photodynamic inactivation.

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.

Blood and cell infiltrate neutrophilic leucocytes As inflammation markers in chronic endometritis: A prospective non-randomised controlled trial

Background. Inflammation declares itself with the presence of cellular tissue infiltrate, which composition reflects the inflammation type. Chronic inflammation is predominated by mononuclear cell infiltration with a certain amount of neutrophils, which role and significance are not fully understood to date.Objectives. Assessment of the infiltrated neutrophil count at various chronic endometritis severity and its dependency on the functional and metabolic activity in neutrophilic leucocytes in peripheral blood.Methods. This prospective non-randomised controlled trial estimated the CD45+ leucocyte and activated CD16b+ neutrophil counts in inflammation infiltrate using immunohistochemistry protocols. Cell counts per section 1 mm2 were measured with computer morphometry. The content of and NADPH oxidase activity in activated neutrophilic leucocytes in venous blood were estimated with a nitroblue tetrazolium reduction test.Results. The study included 40 women with a history of chronic endometritis (CE) divided in two cohorts by endometrial biopsy data, with inactive (n = 25) and active CE (n = 15). A control cohort comprised 20 women with no signs of CE. The inactive CE cohort had higher counts of CD45+ leucocytes and activated CD16b+ neutrophils in infiltrate compared to control. Higher content of activated neutrophilic leucocytes with higher NADPH oxidase activity were found in peripheral blood. Morphological exacerbation markers of EC were associated with sharper peaks of CD45+ and CD16b+ cell counts in infiltrate and an elevated functional metabolic activity in circulating neutrophilic leucocytes. A strong direct correlation was revealed between blood activated neutrophil and endometrial CD16b+ neutrophil counts, as well as NADPH oxidase activity in blood neutrophils and infiltrate CD16b+ cell counts.Conclusion. Even minor morphological markers of exacerbated endometrial inflammation are accompanied by the elevated infiltrate counts of both total CD45+ leucocytes and activated CD16b+ neutrophils. The functional metabolic activity of peripheral blood neutrophilic leucocytes is interlinked with the inflammatory infiltrate cell composition and reflects severity of chronic endometrial inflammation.

The screen herbal immunopotentiator and research on its effect on the innate immune system and disease resistance of Nile tilapia (Oreochromis niloticus) against Streptococcus agalactiae

To screen potential herbal immunopotentiators, the effects of 20 herbal aqueous extracts (HAEs) on the oxygen respiratory burst activity (ORBA) and phagocytic activity of tilapia blood leucocytes were determined using nitroblue tetrazolium (NBT) reduction and phagocytizing latex beads methods, respectively. The screened-out herbal aqueous extract (HAE) supplemented diet (10 g/kg, dry herb/dry feed) was consecutively fed to healthy tilapia for in vivo evaluating its effects on the ORBA and phagocytic activity of head-kidney leucocytes, the acid phosphatase (ACP), lysozyme (LZM), catalase (CAT) and superoxide dismutase (SOD) activities of serum, and the mRNA expression levels of IL-β, IL-8, Hsp70, and TNF-α in the spleen. For evaluating the efficacy of the screened-out HAE on preventing streptococcosis, the tilapia were respectively and consecutively fed the control diet and the screened-out HAE supplemented diet for seven and fourteen days and subsequently infected with Streptococcus agalactiae. Their survival cures were compared using the log-rank test. Additionally, the components in the screened-out HAE were identified using ultra-performance liquid chromatography-Q exactive orbitrap-mass spectrometry (UPLC-QE-MS). Results showed that, among the candidate HAEs, Scutellaria baicalensis HAE showed more ability in terms of simultaneously enhancing the ORBA and phagocytic activity of tilapia blood leucocytes as compared to the other candidate HAEs. Oral administration of S. baicalensis HAE significantly elevated the ORBA and phagocytic activity of head-kidney leucocytes, the ACP, LZM, and SOD activities of sera, the mRNA expression levels of IL-8 and Hsp 70, and the cumulative survival rate of tilapia with S. agalactiae infections. The negative and positive models of UPLC-QE-MS identified 256 and 596 components of S. baicalensis HAE, respectively. Taken together, the results of this study indicate that S. baicalensis HAE could prove useful in the prevention of tilapia streptococcosis and suggest that it could be further developed as immunostimulants.

Effective removal of Cd2+, Zn2+ by immobilizing the non-absorbent active catalyst by packed bed column reactor for industrial wastewater treatment

Cadmium and zinc are leading heavy metal pollutants causing serious health problems when discharged into the aquatic environments. The present investigation focused on the bioaccumulation of Cd2+ and Zn2+depending on the sorption process by Bacillus amyloliquefaciens HM28. The selected bacterium was multi-metal (Zn2+, Pb2+, Cd2+, Cu2+ and Li+) and antibiotic (cefotaxime, ampicilin, nalidixic acid, ceftazidime, penicillin and kanamycin) resistance was resolved. The identified strain showed maximum resistance onCd2+ (2575 ppm) and Zn2+ (1300 ppm). The sorption of Cd2+ and Zn2+ by a dried bacterium was investigated. Biosorption of Cd2+ was maximum (98.4 ± 5.2%) at 100 mg/L concentration and maximum Zn2+ (98.3 ± 1.5%) was detected in the medium containing 150 mg/L metal ion. Bioremoval was maximum after 30 min contact time with dried biomass and the absorption rate improved. The optimum Cd2+ and Zn2+ bioremoval yield of 93 ± 4.4% and 89.8 ± 4.3% were observed, at pH 7.0 and 7.5, respectively. Despite the significant reduction in growth rate, heavy metals increased nitro-blue tetrazolium reduction from 11 ± 1.3 to 67 ± 3.3%. Dehydrogenase activity elevated due to heavy metal stress. Bacterial biomass was immobilized in a glass column (20 cm × 2 cm). Biosorption of Cd2+ and Zn2+ ions were performed in a packed bed column. The breakthrough time of Cd2+ was 210 min at 1 mL/min flow rate and it decreased 94 min at 5 mL/min flow rate, whereas 240 min at 1 mL/min, and 90 min at 5 mL/min, respectively. The absorption capacity was 4.87 ± 0.8 to 5.43 ± 0.5 mg/g for Cd2+ and 3.85 ± 0.3 to 4.53 ± 0.4 mg/g for Zn2+. The present findings revealed the potential of B. amyloliquefaciens HM28 biomass in Cd2+ and Zn2+ biosorption, with feasibility in the bioremediation of Cd2+ and Zn2+ contaminated water.

English

BACKGROUND Oxidative stress plays a role in the pathogenesis of type 2 diabetes either by affecting insulin secretion or increasing insulin resistance. Oxidative stress is dealt by the body with the help of several antioxidant systems. The antioxidant levels in disorders causing oxidative stress such as diabetes mellitus are found to be low. Total antioxidant capacity (TAC), superoxide dismutase (SOD) and glutathione peroxide (GPx) levels are found to be depleted in diabetic groups when compared to control group comprising of healthy individuals. The activities of SOD and GPx are significantly low in diabetic patients. This study was conducted to determine the total antioxidant capacity and levels of antioxidants such as SOD and GPx in patients with type 2 diabetes and association between antioxidant levels and type 2 diabetics as well as type 2 diabetes with its associated complications. METHODS The study population comprised of 150 individuals, 50 subjects belonging to each subgroup of type 2 diabetics, and its associated complications and healthy subjects. The study was conducted in a tertiary care centre in Mangalore, Karnataka from September 2016 to December 2016. The antioxidant capacity of RBC, fasting blood levels of SOD and GPx were estimated by phospho molybdenum method. Nitroblue tetrazolium (NBT) reduction method and Rotruck method were also used. RESULTS The total antioxidant capacity, SOD and GPx were lower among diabetics and diabetic patients with complications as compared to normal nondiabetic subjects. CONCLUSIONS There is no significant difference in oxidative stress between patients with and without chronic complications of diabetes. KEY WORDS Antioxidant Levels, Glutathione Peroxidase, Oxidative Stress, Superoxide Dismutase, Nitroblue tetrazolium Reduction

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.

Immunomodulatory effect of d-carvone in swiss albino mice with benzo(a)pyrene-induced lung cancer

Background: Globally, lung cancer is the second most cause of deaths, which accounts for approximately 29% of the cancer cases worldwide. d-Carvone is considered a vital constituent of many essential oils with immense pharmacological benefits. Objective: In this study, we examined the immunomodulatory effect of d-carvone in the Swiss albino mice model against benzo(a)pyrene (BaP)-induced lung carcinoma. Materials and Methods: BaP was orally administered to the mice (50 mg/kg body weight [bw] for 4 weeks [twice a week]). Post-BaP administration, d-carvone (20 mg/kg bw) was orally administered to the mice (20 mg/kg bw). We calculated the tumor incidence and performed the following measurements: lung and bw, hematological parameters, immune complexes (phagocytic and avidity indexes, nitroblue tetrazolium reduction, soluble immune complex levels, immunocompetent cells (leukocytes, lymphocytes, and neutrophils), immunoglobin (Ig) levels (IgG, IgA, and IgM), level of xenobiotics and enzymes that point toward hepatic dysfunction, carcinoembryonic antigen (CEA), and proinflammatory cytokines (PICs) in experimental and normal mice. The level of oxidative stress in the experimental animals was investigated. The lung tissues of investigational animals were examined for the histopathological changes. Results: According to the results, there was an increased level of lipid peroxidation and decreased level of antioxidant activity in the lung tumor samples. The counts of lymphocytes, polymorphonuclear cells, and macrophages were notably decreased and increased, respectively, by the d-carvone post-treatment. Moreover, BaP-induced inflammation that is indicated by the increased in the level of PICs and CEA proteins in lung samples. d-Carvone attenuated the levels of PICs and CEA. Conclusion: The results of this study reveal that d-carvone prevented the cells against BaP-induced inflammation in lung cancer without causing adverse effects.

Thioguanine restoration through type I photosensitization-superoxide oxidation-glutathione reduction cycles.

UVA-induced deleterious effect of thiopurine prodrugs including azathioprine, 6-mercaptopurine and 6-thioguanine (6-TG) increases the risk of cancer development due to the incorporation of 6-TG in patients' DNA. The catalytic mechanism by which thiobases act as a sustained oxidant producer has yet to be explored, especially through the Type I electron transfer pathway that produces superoxide radicals (O2˙-). Under Fenton-like conditions O2˙- radicals convert to extremely reactive hydroxyl radicals (˙OH), thus carrying even higher risk of biological damage than that induced by the well-studied type II reaction. By monitoring 6-TG/UVA-induced photochemistry in mass spectra and superoxide radicals (O2˙-) via nitro blue tetrazolium (NBT) reduction, this work provides two new findings: (1) in the presence of reduced glutathione (GSH), the production of O2˙-via the type I reaction is enhanced 10-fold. 6-TG thiyl radicals are identified as the primary intermediate formed in the reaction of 6-TG with O2˙-. The restoration of 6-TG and concurrent generation of O2˙- occur via a 3-step-cycle: 6-TG type I photosensitization, O2˙- oxidation and GSH reduction. (2) In the absence of GSH, 6-TG thiyl radicals undergo oxygen addition and sulfur dioxide removal to form carbon radicals (C6) which further convert to thioether by reacting with 6-TG molecules. These findings help explain not only thiol-regulation in a biological system but chemoprevention of cancer.

Saikosaponin a contributed to CCIN treatment by promoting neutrophil bactericidal activity via activation CBL-dependent ERK pathway

BackgroundDespite granulocyte colony-stimulating factor (GCSF) is widely used in clinical, cancer chemotherapy induced neutropenia (CCIN) infection and infection-related mortality is high for lack of functionally mature neutrophils. Generating functional neutrophils is new therapeutic approaches to reduce CCIN-associated infection and mortality. Saikosaponin a (SSA) is one of the major bioactive components of Radix Bupleuri (RB) and exerts immunoregulatory effects. PurposeThe present study aims to investigate the efficacy and mechanism of SSA in CCIN therapy. MethodsSSA was applied both in vitro and in vivo to assess the efficacy of CCIN therapy. The differentiation of neutrophils was measured by Nitroblue tetrazolium (NBT) reduction assay and Giemsa staining assay. The neutrophil differentiation related real-time transcription factors were detected by quantitative PCR (RT-qPCR) and Western Blot. Bacteria killing assay was used to assess the ability of fighting infection. Network pharmacology was employed to explore the mechanism network, and the predicted pathways were validated by Western Blot. ResultsWe found that SSA contributed to generate functional mature neutrophils which capable of fighting infection both in vitro and in vivo. Network pharmacology prediction showed 55 pathways were predicted involved in SSA against CCIN. Further validation showed that CBL-ERK1/2 pathway was activated by SSA, which could upregulate PU.1 and CEBPβ expression leading to neutrophil differentiation. ConclusionsOur findings suggest a natural regimen SSA regenerates microbicidal neutrophils to effectively reduce CCIN-associated infection via activating CBL-ERK1/2 pathway, providing a rationale for future therapeutic approaches.

NADPH diaphorase detects S-nitrosylated proteins in aldehyde-treated biological tissues

NADPH diaphorase is used as a histochemical marker of nitric oxide synthase (NOS) in aldehyde-treated tissues. It is thought that the catalytic activity of NOS promotes NADPH-dependent reduction of nitro-blue tetrazolium (NBT) to diformazan. However, it has been argued that a proteinaceous factor other than NOS is responsible for producing diformazan in aldehyde-treated tissues. We propose this is a NO-containing factor such as an S-nitrosothiol and/or a dinitrosyl-iron (II) cysteine complex or nitrosated proteins including NOS. We now report that (1) S-nitrosothiols covalently modify both NBT and TNBT, but only change the reduction potential of NBT after modification, (2) addition of S-nitrosothiols or β- or α-NADPH to solutions of NBT did not elicit diformazan, (3) addition of S-nitrosothiols to solutions of NBT plus β- or α-NADPH elicited rapid formation of diformazan in the absence or presence of paraformaldehyde, (4) addition of S-nitrosothiols to solutions of NBT plus β- or α-NADP did not produce diformazan, (5) S-nitrosothiols did not promote NADPH-dependent reduction of tetra-nitro-blue tetrazolium (TNBT) in which all four phenolic rings are nitrated, (6) cytoplasmic vesicles in vascular endothelial cells known to stain for NADPH diaphorase were rich in S-nitrosothiols, and (7) procedures that accelerate decomposition of S-nitrosothiols, markedly reduced NADPH diaphorase staining in tissue sections subsequently subjected to paraformaldehyde fixation. Our results suggest that NADPH diaphorase in aldehyde-fixed tissues is not enzymatic but is due to the presence of NO-containing factors (free SNOs or nitrosated proteins such as NOS), which promote NADPH-dependent reduction of NBT to diformazan.

Antimicrobial Activity of Vanadium and Germanium Nanoaquacitrates in vitro and their Physiological Effect on Wheat Plants in vivo

Phytopathogenic bacteria and phytoplasmas, in particular Pseudomonas syringae pv. atrofaciens and Acholeplasma laidlawii var. granulum are causative agents of cereal crops diseases. Nanoparticles and biologically active compounds created with the use of nanotechnologies are considered as alternative modern measures to control phytopathogenic microorganisms. Antimicrobial activity is already established for gold, silver, copper, magnesium, iron, zinc, aluminum, cerium, titanium, cadmium, nickel and vanadium. At the same time, antimicrobial activity of germanium and its citrates as well as citrates of others nanoparticles, including vanadium, are studied insufficiently. The physiological effect of vanadium and germanium nanoaquacitrates on wheat plants in vivo, both on state and photochemical activity of the photosynthetic apparatus and activity of superoxide dismutase of plant tissue are poorly known too. Aim. The aim of our work was to study the antimicrobial activity of vanadium and germanium nanoaquacitrates in vitro and their physiological influence on wheat plants in vivo. Methods. A. laidlawii var. granulum 118 (UCM BM-34) was cultured in the liquid nutrient medium CM IMV-72 (pH=7.8) in thermostat at 320 C for 72 hours. P. syringae pv. atrofaciens D13 was cultured on potato agar in thermostat at 26–280°C. For artificial infection a bacterial suspension with a density of 1×109 CFU/ml according to the turbidity standard was prepared. Nanoaquacitrates of V (30 mg/dm3) and Ge (5 mg/dm3) were used in our study. After a short-term (15 minutes) action of the test compounds, morphological features of A. laidlawii var. granulum and P. syringae pv. atrofaciens were studied using a transmission electron microscope (TEM) JEOL JSM 1400 (Zabolotny IMV NASU). The plants of wheat of Pecheryanka variety were grown in field condition on 50 m2 area plot. Pre-sowing treatment of spring wheat seeds before sowing was carried out with solutions of V (30 mg/dm3) and Ge (5 mg/dm3) nanoaquacitrates according to the experimental scheme. The experiment was carried out in three repetitions. In addition, biological preparation (B.P.) Extrakon, which consists from natural consortium of beneficial microorganisms inoculated into a peat-like substrate, was applied into the soil before sowing. Artificial inoculation by phytoplasma A. laidlawii var. granulum 118 was performed using Clement`s method (subepidermal injection into the stem) in the tillering phase of wheat plants growth. Artificial infection of wheat plants with P. syringae pv. atrofaciens D13 was performed by injecting a bacterial suspension into the stem in the booting phase of plants growth. The activity of superoxide dismutase (SOD) was determined by the ability of the enzyme to inhibit the photochemical reduction of nitro blue tetrazolium. Changes in the following fluorescent parameters were analyzed: Fv/Fp and Rfd. For statistical processing of data, calculations of the arithmetic mean and its standard error were performed. Results. It was shown using electron microscopy that contact of Ge and V nanoaquacitrates with P. syringae pv. atrofaciens D13 and A. laidlawii var. granulum 118 resulted in cytomorphological changes of cells, homeostatic disruption and death. It was found in the field studies that SOD activity level increased in the tissues of wheat leaves to different values in plants infected with phytopathogenic microorganisms without any treatment and in cases of pre-sowing treatment with Ge and V nanoaquacitrates and B.P. Extrakon. Field investigations showed that combined use of consortium of soil microorganisms (B.P. Extrakon) and pre-sowing treatment of wheat seeds with V and Ge nanoaquacitrates decreased negative impact of infection caused by A. laidlawii var. granulum 118 and P. syringae pv. atrofaciens D13 on the state and photochemical activity of the photosynthetic apparatus of Triticum aestivum: the level of quantum efficiency of photosystem II (PSII) and the assimilation activity coefficient Rfd increased. This effect of studied biologically active substances is due to both antimicrobial action and activation of the antioxidant defense system of plant cells. Conclusions. It was shown in our investigations that SOD activity increased in leaf tissues after application of biologically active agents in the next order: pre-sowing treatment with Ge nanoaquacitrates &lt; pre-sowing treatment with V nanoaquacitrates &lt; biological product (B.P.) Extrakon + pre-treatment with Ge nanoaquacitrates &lt; B.P. Extrakon + pre-sowing treatment with V nanoaquacitrates. A significant increase of SOD activity of wheat leaves (in the tubing phase) was noticed at both types of infection – caused by phytopathogenic bacteria and phytoplasma, especially for the last one. A significant increase in the surface area of assimilation apparatus of wheat plants (in the tillering phase) in cases of wheat seeds pre-sowing treatment with Ge and V nanoaquacitrates before planting was observed. Pre-sowing treatment of wheat seeds with Ge and V nanoaquacitrates in combination with B.P. Extrakon use on wheat plants infected with A. laidlawii var. granulum and P. syringae pv. atrofaciens improved photosynthesis and status of photosynthetic apparatus of plants.

Assessment of immunohematological, hematological and biochemical responses in cultivable fish Cyprinus carpio exposed to an antibiotic sulfamethoxazole (SMX)

Abstract Sulfamethoxazole (SMX) is a member of the sulfonamides group of antibiotics which is used extensively in aquaculture throughout the world. In this study, common carp (Cyprinus carpio) was used as the bioindicator to assess the toxicity potential of SMX. Effects were based on chronic toxicity of environmentally relevant dosages of 25, 50, 100, and 200 μg/L of SMX for 28 days. Cytotoxicity through hematology and biochemistry showed a dose–response relationship. Numerous variations were recorded in blood profile and biochemical parameters in SMX-exposed groups when compared to control. Hemoglobin, platelet, and erythrocyte levels were significantly decreased. Leukocyte level was significantly increased with values ranging from 131 to 303 (×103/μL). Changes in biochemical indices: glucose, total protein, and triglycerides showed biphasic trend, but alanine transaminase secretion was significantly increased from 25.13 to 204 U/L at higher concentration compared to control, suggesting liver damage. Spectrophotometric nitroblue tetrazolium reduction assay showed that respiratory burst activity increased as a function of SMX dose and exposure time (0.48–1.33 absorbance) ultimately leading to reduction in immunity. The present study highlights that prolonged exposure of SMX affects biochemistry, hematology, and immunohematology of fish and these biomarkers act as an effective tool for environmental risk assessment of drugs in the aquatic environment.