Immunotherapy of Metastatic Mouse Breast Cancer by Adherent Splenocytes Pulsed With Extracts of Heated Tumor Cells and Lactobacillus Casei

Introduction: Flask-adherent Splenocytes (SACs) fulfill antigen-presenting cell requirementsof acquired immune responses. This study was done to evaluate the efficacy of newimmunotherapy against breast cancer made by SACs pulsed with the extract of heated 4T1cells and Lactobacillus casei, as a probiotic.Materials and Methods: Mammary carcinoma was induced by injection of 4T1 cell line inthe flank of female Balb/c mice. The first SACs therapy was started on day 11 after tumorinduction when all animals had developed a palpable tumor. SACs therapy was done twice ata 10-day interval.Results: Mice with mammary tumors received SACs pulsed with combined heated 4T1 cellsand L. casei determined a more desirable survival curve and a slower rate of tumor developmentcompared to the other groups. At least 20% of the group receiving combined immunotherapywere alive by day 58. Those mice receiving SACs pulsed with the Lysate of heated tumorcells died by day 45.The maximum survival of other mice was up to 38 days after tumorinduction. Moreover, SAC pulsed with combined agents significantly amplified the secretionof Interferon-γ (IFN-γ), and conversely reduced the secretion of Transforming growth factor-β(TGF-β) and Interleukin 4 (IL-4) in the splenocyte population compared to splenocytes fromother groups. Combined immunotherapy increased the expression of p53 and caspase 3 genesand reduced the exertion of BCL2 more than other immunotherapy protocols.Conclusion: Immunotherapy with SACs pulsed with heated 4T1 cells and L. casei promotesbeneficial outcomes in the mouse model of breast cancer

Učinak različitih biljnih ekstrakata na aktivnost hidrolaze žučnih soli sojeva bakterije Lactobacillus izolirane iz probavnog sustava peradi

The bile salt hydrolysis (BSH) enzyme weakens fat metabolism through bile salt deconjugation and reduces poultry performance, in order to cope with the antibacterial properties of the bile. Therefore, reducing the activity of this enzyme through the use of feed additives is probably a promising alternative to antibiotics for improving poultry performance. Plant extracts have long been used as feed additives for promoting poultry growth. In the current experiment, five Lactobacillus strains including Lactobacillus animalis, Lactobacillus acidophillus, Lactobacillus gallinarum, Lactobacillus lactis, and Lactobacillus returi were obtained from the poultry hindgut and were used as the probiotic application. A plate test and two-step enzymatic reaction method were used for deconjugation activity determination of the Lactobacillus strains. Further, four plant extracts (i.e., the aerial parts of Rosemary (Rosmarinus officinalis), Roselle calyx (Hibiscus sabdariffa), Berberis vulgaris root, and Green tea) were examined in terms of BSH enzyme inhibitors using the cell-free extracts as the potential antibiotic alternative. Furthermore, the gallbladders of the broilers were freshly collected from the poultry slaughterhouses, and their contents were extracted. The results showed that all Lactobacillus strains could hydrolyze the taurocholate acid (TCA) and chicken bile salt mixture (CBSM) to unconjugated bile acid. Moreover, ethanolic extracts of B. vulgaris root and Green tea relatively reduced the activity of the BSH enzyme that could potentially be investigated as an appropriate alternative in poultry feed in vivo. In conclusion, all five Lactobacillus strains were resistant to bile salts (i.e. TCA and CBSM) by BSH activity, and the addition of Green tea and B. vulgaris root extracts to the bacterial medium demonstrated inhibitory effects against the BSH enzyme.Enzimi hidrolaze žučnih soli (BSH) oslabljuju metabolizam masti dekonjugacijom žučnih soli što dovodi do smanjenja proizvodnosti u peradi. Smanjenje aktivnosti ovog enzima, upotrebom dodataka prehrani, mogla bi biti obećavajuća alternativa za primjenu određenih antibiotika u peradarstvu. Biljni ekstrakti dugo se upotrebljavaju kao dodaci prehrani za poticanje rasta. U ovom je istraživanju pet sojeva bakterije Lactobacillus, uključujući Lactobacillus animalis, Lactobacillus acidophillus, Lactobacillus gallinarum, Lactobacillus lactis i Lactobacillus returi, dobiveno iz stražnjeg dijela crijeva peradi te upotrijebljeno kao probiotik. Test na ploči i enzimska reakcija u dva koraka primijenjene su za utvrđivanje aktivnosti dekonjugacije u sojeva Lactobacillus. Nadalje, četiri biljna ekstrakta - nadzemni dijelovi ružmarina (Rosmarinus officinalis), hibiskusa (Hibiscus sabdariffa), korijen obične žutike (Berberis vulgaris) i zeleni čaj - istraživana su s obzirom na inhibitore enzima BSH upotrebom izvanstaničnih ekstrakata kao moguća zamjena antibiotiku. Osim toga, nakon usmrćivanja, prikupljeni su svježi žučni mjehuri brojlera te je izvađen njihov sadržaj. Rezultati su pokazali da svi sojevi bakterije Lactobacillus mogu hidrolizirali tauroholatnu kiselinu i žučne soli pilića (CBSM) u nekonjugiranu žučnu kiselinu. Štoviše, ekstrakti etanola korijena B. vulgaris i zelenog čaja relativno su smanjili aktivnost BSH enzima što bi se moglo istražiti u hranidbi peradi in vivo. Zaključno, svih pet sojeva bakterije Lactobacillus bilo je otporno na žučne soli npr. tauroholičnu kiselinu (TCA) i (CBSM) putem BSH aktivnosti, a dodatak zelenog čaja i ekstrakta korijena B. vulgaris mediju s bakterijama pokazali su inhibitorne učinke protiv BSH enzima

Anti-inflammatory and antioxidant potential of Artemisia dracunculus L. aqueousextract against acetic acid induced ulcerative colitis in male Wistar rats

Ulcerative colitis (UC) is one of the two types of inflammatory bowel disease (IBD) which is increasing worldover due to modern life style. Patients with UC are prone to develop colorectal cancer. While the disease severity decides the treatment option, researchers look towards herbal medicines with anti-inflammatory properties for minimal or nil side effects. Artemisia dracunculus L., commonly called Tarragon, is a medicinal herb used in traditional Asian medicine mainly in Iran, India, Pakistan and Azerbaijan due to its special compounds. In this study, we tried to elucidate the effects of aqueous extract of tarragon on acetic acid induced ulcerative colitis (UC) in rats. Male Wistar rats were grouped into four groups of eight each viz., control; experimental control (UC was induced via luminal instillation of 4% acetic acid); and UC induced + aqueous tarragon extract (100 mg/kg) or prednisolone (2 mg/kg) orally for ten consecutive days. Tissue specimens were collected after the experimental period for evaluation of caspase-3 and cyclooxygenase-2 (COX-2) expression by immunohistochemistry. Real-time PCR was used to monitor the levels of IL-1, IL-6 and TNF-α in colonic homogenates. Moreover, the levels of myeloperoxidase, nitric oxide and total antioxidant capacity were measured in colonic homogenates. The results showed that both treatment regimens could similarly reduce the severity of disease symptoms. Treatment with aqueous extract of tarragon caused a better improvement (P <0.05) in the levels of myeloperoxidase enzyme, and total antioxidant capacity of colonic homogenates compared to prednisolone. Nevertheless, the levels of the expression of caspase-3, and COX-2 and TNF-α were reduced in UC rats received prednisolone more than UC rats received aqueous extract of tarragon. The was no statistical difference in the levels of nitric oxide, IL-1 and IL-6 between UC rats received tarragon extract or prednisolone. Overall, these findings suggest that the aqueous extract of tarragon is a promising strategy to control ulcerative colitis. Aqueous extract can also be used as an anti-inflammatory and immune system stimulant in conditions where the immune system is damaged

Altered Immune Responses in Mice After Receiving Nicotine-pulsed Mesenchymal Stem Cell-conditioned Medium

Introduction: Previous investigations have documented that nicotine-pulsed mesenchymalstem cells (MSCs) can induce an anti-inflammatory phenotype in some immune cells in vitro.This study aimed to assess the effects of nicotine-pulsed MSCS in the function of immunecells, macrophages, and lymphocytes of mice receiving these cels.Materials and Methods: Bone marrow-derived MSCs (1.5×106) were seeded in a T75flaskand incubated with 0, .1, .5, or 1 μM nicotine until the cells reached 90% confluency.Afterwards, immunophenotyping change, vitality, concentration of TGF-β, IL-10, and IDOlevels of the MSC-conditioned medium were examined. Correspondent to in vitro results, theC57BL/6 mice intravenously received 400 μL of the conditioned medium of MSCs (CM),conditioned medium of nicotine (.5 μM)-pulsed MSCs (CMN), or medium. After 12 h, thelymphocytes, neutrophils, and peritoneal macrophages of the mice were isolated and theirfunction was evaluated ex vivo.Results: The least effective dose concentration of nicotine that led to an anti-inflammatoryenvironment by the MSC-conditioned medium was 0.5 μM. Nicotine at this concentrationprompted a higher level of TGF-β, IDO concentration in the conditioned medium. However,this concentration did not affect the MScs’ markers expressions or MScs’ vitality. T lymphocytesisolated from the mice receiving CMN showed a significant decrease in proliferation rate. Theratio of the IFN-γ gene expression to IL-4 gene expression in splenocytes was significantlyreduced in the mice receiving CMN compared to the mice receiving CM. The neutral reduptake, respiratory burst, and nitric oxide production of the peritoneal macrophage onlydecreased in the mice treated with CMN. These factors also decreased in neutrophils isolatedfrom mice receiving CM or CMN. However, these decreases were more prominent in the micetreated with CMN.Conclusion: Treatment of MSCs by nicotine may be a promising strategy to enhance theimmunomodulatory properties of these cells

Effect of Nicotine on Immune System Function

As a parasympathetic alkaloid and the main substance in cigarette smoke, nicotine modulates the immune system, inhibits innate and acquired immunity and is used in treating many autoimmune diseases. It often stimulates the α7 receptor and causes an anti-inflammatory state in the body. This study is designed to evaluate the role of nicotine treatment on immune system. The results showed that nicotine affects many cells in immune system, alters the downstream intracellular mechanisms and changes lymphocytes polarization. This substance alters TLRs and STATs gene expression and thus changes in the innate immune system. All these events inhibit the secretion of pro-inflammatory cytokines and chemokines which increase angiogenesis and metastasis and exacerbates tumors due to increasing survival and cell growth. Nicotine can aggravate tumors in cancer patients, with many positive effects observed in the treating autoimmune disease, Nicotine treatment function in different conditions depends on factors such as concentration, how it is employed, treatment duration and other conditions such as body conditions affecting the immune system, hence, further studies and review of all conditions are required

Effects of two plant extracts and native Lactobacillus culture on immune response, lymphoid organs and antioxidant properties of broiler chickens

Probiotics and phytogenics have been evaluated as potential alternatives to antibiotic growth promoters (AGP) in poultry feeds in terms of their ability to improve growth performance in commercial poultry production through improving growth performance, feed conversion ratio and immune response efficiency. This study investigated the benefits of Lactobacillus culture (LC), green tea extracts (GTE) and Berberis vulgaris extracts (BVRE) have been investigated on the immune response, lymphoid organs, and antioxidant properties of broiler chickens. A total of 320 one-day-old Ross 308 chicks were randomly allotted to 8 treatment groups, each including 4 replicates of 10 chicks. A 2×2×2 factorial arrangement of 8 dietary treatments was used to appraise the effects of the mixture of five LC (none vs. 1-5 × 108 cfu/g), GTE (none vs. 2500 ppm) and BVRE (none vs. 2500 ppm). The relative weight of lymphoid organs (spleen, thymus and bursa of Fabricius), antioxidant parameters of malondialdehyde (MDA) content and total antioxidant capacity (T-AOC) and immune response indices (white blood cells, antibody response to sheep red blood cell, respiratory burst and splenocytes proliferation) were assessed. According to the results of the current experiment, the relative weights of the spleen and bursa were significantly higher than the control group in broilers fed the LC diet (P ˂ 0.01). The combination of LC and GTE significantly decreased MDA as compared to broilers fed the control diet (P ˂ 0.05). Moreover, the GTE diet markedly increased the T-AOC compared to the control (P ˂ 0.01). The LC and plant extract treatments significantly improved the humoral and cellular immune systems (P ˂ 0.01). Based on obtained results, plant extracts in combination with Lactobacillus strains can improve the immune responses of broiler chickens

Global burden of 288 causes of death and life expectancy decomposition in 204 countries and territories and 811 subnational locations, 1990–2021: a systematic analysis for the Global Burden of Disease Study 2021

BACKGROUND Regular, detailed reporting on population health by underlying cause of death is fundamental for public health decision making. Cause-specific estimates of mortality and the subsequent effects on life expectancy worldwide are valuable metrics to gauge progress in reducing mortality rates. These estimates are particularly important following large-scale mortality spikes, such as the COVID-19 pandemic. When systematically analysed, mortality rates and life expectancy allow comparisons of the consequences of causes of death globally and over time, providing a nuanced understanding of the effect of these causes on global populations. METHODS The Global Burden of Diseases, Injuries, and Risk Factors Study (GBD) 2021 cause-of-death analysis estimated mortality and years of life lost (YLLs) from 288 causes of death by age-sex-location-year in 204 countries and territories and 811 subnational locations for each year from 1990 until 2021. The analysis used 56 604 data sources, including data from vital registration and verbal autopsy as well as surveys, censuses, surveillance systems, and cancer registries, among others. As with previous GBD rounds, cause-specific death rates for most causes were estimated using the Cause of Death Ensemble model-a modelling tool developed for GBD to assess the out-of-sample predictive validity of different statistical models and covariate permutations and combine those results to produce cause-specific mortality estimates-with alternative strategies adapted to model causes with insufficient data, substantial changes in reporting over the study period, or unusual epidemiology. YLLs were computed as the product of the number of deaths for each cause-age-sex-location-year and the standard life expectancy at each age. As part of the modelling process, uncertainty intervals (UIs) were generated using the 2·5th and 97·5th percentiles from a 1000-draw distribution for each metric. We decomposed life expectancy by cause of death, location, and year to show cause-specific effects on life expectancy from 1990 to 2021. We also used the coefficient of variation and the fraction of population affected by 90% of deaths to highlight concentrations of mortality. Findings are reported in counts and age-standardised rates. Methodological improvements for cause-of-death estimates in GBD 2021 include the expansion of under-5-years age group to include four new age groups, enhanced methods to account for stochastic variation of sparse data, and the inclusion of COVID-19 and other pandemic-related mortality-which includes excess mortality associated with the pandemic, excluding COVID-19, lower respiratory infections, measles, malaria, and pertussis. For this analysis, 199 new country-years of vital registration cause-of-death data, 5 country-years of surveillance data, 21 country-years of verbal autopsy data, and 94 country-years of other data types were added to those used in previous GBD rounds. FINDINGS The leading causes of age-standardised deaths globally were the same in 2019 as they were in 1990; in descending order, these were, ischaemic heart disease, stroke, chronic obstructive pulmonary disease, and lower respiratory infections. In 2021, however, COVID-19 replaced stroke as the second-leading age-standardised cause of death, with 94·0 deaths (95% UI 89·2-100·0) per 100 000 population. The COVID-19 pandemic shifted the rankings of the leading five causes, lowering stroke to the third-leading and chronic obstructive pulmonary disease to the fourth-leading position. In 2021, the highest age-standardised death rates from COVID-19 occurred in sub-Saharan Africa (271·0 deaths [250·1-290·7] per 100 000 population) and Latin America and the Caribbean (195·4 deaths [182·1-211·4] per 100 000 population). The lowest age-standardised death rates from COVID-19 were in the high-income super-region (48·1 deaths [47·4-48·8] per 100 000 population) and southeast Asia, east Asia, and Oceania (23·2 deaths [16·3-37·2] per 100 000 population). Globally, life expectancy steadily improved between 1990 and 2019 for 18 of the 22 investigated causes. Decomposition of global and regional life expectancy showed the positive effect that reductions in deaths from enteric infections, lower respiratory infections, stroke, and neonatal deaths, among others have contributed to improved survival over the study period. However, a net reduction of 1·6 years occurred in global life expectancy between 2019 and 2021, primarily due to increased death rates from COVID-19 and other pandemic-related mortality. Life expectancy was highly variable between super-regions over the study period, with southeast Asia, east Asia, and Oceania gaining 8·3 years (6·7-9·9) overall, while having the smallest reduction in life expectancy due to COVID-19 (0·4 years). The largest reduction in life expectancy due to COVID-19 occurred in Latin America and the Caribbean (3·6 years). Additionally, 53 of the 288 causes of death were highly concentrated in locations with less than 50% of the global population as of 2021, and these causes of death became progressively more concentrated since 1990, when only 44 causes showed this pattern. The concentration phenomenon is discussed heuristically with respect to enteric and lower respiratory infections, malaria, HIV/AIDS, neonatal disorders, tuberculosis, and measles. INTERPRETATION Long-standing gains in life expectancy and reductions in many of the leading causes of death have been disrupted by the COVID-19 pandemic, the adverse effects of which were spread unevenly among populations. Despite the pandemic, there has been continued progress in combatting several notable causes of death, leading to improved global life expectancy over the study period. Each of the seven GBD super-regions showed an overall improvement from 1990 and 2021, obscuring the negative effect in the years of the pandemic. Additionally, our findings regarding regional variation in causes of death driving increases in life expectancy hold clear policy utility. Analyses of shifting mortality trends reveal that several causes, once widespread globally, are now increasingly concentrated geographically. These changes in mortality concentration, alongside further investigation of changing risks, interventions, and relevant policy, present an important opportunity to deepen our understanding of mortality-reduction strategies. Examining patterns in mortality concentration might reveal areas where successful public health interventions have been implemented. Translating these successes to locations where certain causes of death remain entrenched can inform policies that work to improve life expectancy for people everywhere. FUNDING Bill & Melinda Gates Foundation

The Effects of Nicotine and Caffeine Treatment on the Vitality and Regenerative Potentials of Mesenchymal Stem Cells

Background & Objective: Previous studies indicated that Mesenchymal stem cells (MSCs) express some of the nicotinic receptor subunits and adenosine receptors. Therefore, MSCs function may be controlled by two very consuming environmental substance like nicotine (nicotinic receptor-agonist) and caffeine (adenosine antagonist). The purpose of the present study is to determine the role of nicotine or caffeine on the some of the function of MSCs. Materials & methods: The mesenchymal stem cells were isolated from the bone marrow of NMRI. The cells were incubated with caffeine (0.1mM) and or with nicotine (0.1µM) for 48 hours. In a set of experiments, the cells were trypsinized and the vitality, the biological activity of cell membranes and the potentials mitochondrial redaction of them were assumed. In another experiments, the cutler plates were scratched with tip of the sampler, and the regeneration velocity of scar was evaluated by an optical camera. Results: Treatment of MSCs with nicotine caused a significant increase in the absolute number of MSCS, concurrent with an increase in mitochondrial function. While caffeine decreased the vitality of cells, it also promoted a significant reduction in mitochondrial activity of remnant live cells. Nicotine induced a significant reduction in membrane activity of cells, while caffeine treatment exhibited a marked raise in neutral red uptake by MSCs compared to neutral red uptake by control cells. Regenerative assay showed that nicotine markedly promoted the regenerative potential of MSCs, while caffeine markedly reversed the regenerative potential of MSCs. Conclusion: It seems that nicotine at used dose promoted the regenerative potential of MSCs, while caffeine had a revers effects

Anti-inflammatory and antioxidant potential of Artemisia dracunculus L. aqueous extract against acetic acid induced ulcerative colitis in male Wistar rats

42-50Ulcerative colitis (UC) is one of the two types of inflammatory bowel disease (IBD) which is increasing worldover due to modern life style. Patients with UC are prone to develop colorectal cancer. While the disease severity decides the treatment option, researchers look towards herbal medicines with anti-inflammatory properties for minimal or nil side effects. Artemisia dracunculus L., commonly called Tarragon, is a medicinal herb used in traditional Asian medicine mainly in Iran, India, Pakistan and Azerbaijan due to its special compounds. In this study, we tried to elucidate the effects of aqueous extract of tarragon on acetic acid induced ulcerative colitis (UC) in rats. Male Wistar rats were grouped into four groups of eight each viz., control; experimental control (UC was induced via luminal instillation of 4% acetic acid); and UC induced + aqueous tarragon extract (100 mg/kg) or prednisolone (2 mg/kg) orally for ten consecutive days. Tissue specimens were collected after the experimental period for evaluation of caspase-3 and cyclooxygenase-2 (COX-2) expression by immunohistochemistry. Real-time PCR was used to monitor the levels of IL-1, IL-6 and TNF-α in colonic homogenates. Moreover, the levels of myeloperoxidase, nitric oxide and total antioxidant capacity were measured in colonic homogenates. The results showed that both treatment regimens could similarly reduce the severity of disease symptoms. Treatment with aqueous extract of tarragon caused a better improvement (P <0.05) in the levels of myeloperoxidase enzyme, and total antioxidant capacity of colonic homogenates compared to prednisolone. Nevertheless, the levels of the expression of caspase-3, and COX-2 and TNF-α were reduced in UC rats received prednisolone more than UC rats received aqueous extract of tarragon. The was no statistical difference in the levels of nitric oxide, IL-1 and IL-6 between UC rats received tarragon extract or prednisolone. Overall, these findings suggest that the aqueous extract of tarragon is a promising strategy to control ulcerative colitis. Aqueous extract can also be used as an anti-inflammatory and immune system stimulant in conditions where the immune system is damaged