Morphology of blood microbiota in healthy individuals assessed by light and electron microscopy

IntroductionThe blood microbiome is still an enigma. The existence of blood microbiota in clinically healthy individuals was proven during the last 50 years. Indirect evidence from radiometric analysis suggested the existence of living microbial forms in erythrocytes. Recently targeted nucleic acid sequencing demonstrated rich microbial biodiversity in the blood of clinically healthy individuals. The morphology and proliferation cycle of blood microbiota in peripheral blood mononuclear cells (PBMC) isolated from freshly drawn and cultured whole blood are obscure.MethodsTo study the life cycle of blood microbiota we focused on light, and electron microscopy analysis. Peripheral blood mononuclear cells isolated from freshly drawn blood and stress-cultured lysed whole blood at 43°C in presence of vitamin K from healthy individuals were studied.ResultsHere, we demonstrated that free circulating microbiota in the PMBC fraction possess a well-defined cell wall and proliferate by budding or through a mechanism similar to the extrusion of progeny bodies. By contrast, stress-cultured lysed whole blood microbiota proliferated as cell-wall deficient microbiota by forming electron-dense or electron-transparent bodies. The electron-dense bodies proliferated by fission or produce in chains Gram-negatively stained progeny cells or enlarged and burst to release progeny cells of 180 – 200 nm size. On the other hand, electron-transparent bodies enlarged and emitted progeny cells through the membrane. A novel proliferation mechanism of blood microbiota called by us “a cell within a cell” was observed. It combines proliferation of progeny cells within a progeny cell which is growing within the “mother” cell.DiscussionThe rich biodiversity of eukaryotic and prokaryotic microbiota identified in blood by next-generation sequencing technologies and our microscopy results suggest different proliferation mechanisms in whole and cultured blood. Our documented evidence and conclusions provide a more comprehensive view of the existence of normal blood microbiota in healthy individuals

ZN(II)/AU(I) AND ZN(II)/AG(I) COMPLEXES WITH SALEN SCHIFF BASE EXPRESS PROMISING CYTOTOXIC ACTIVITY IN HUMAN CANCER CELLS

Objective: The aim of our study was to evaluate the influence of two complexes of Zn(II)/Au(I) and Zn(II)/Ag(I) with Schiff base ligand (H2Salen) obtained from the condensation reaction between salicylaldehyde and ethylenediamine (abbreviated ZnSalenAu, ZnSalenAg) on viability and proliferation of cultured human cancer cells.Methods: The following cell lines were used as model systems: Human cervical carcinoma (cervical carcinoma), A549 (non-small cell lung cancer [NSCLC]), glioblastoma multiforme (8MGBA), and A431 (squamous cell carcinoma) and its multidrug-resistant (MDR) clones A431-MDR, A431-MRP, and A431-ABCG2 that express mdr1, mrp1, or abcg2 gene, respectively. The investigations were performed by thiazolyl blue tetrazolium bromide test, neutral red uptake cytotoxicity assay, crystal violet staining, hematoxylin and eosin staining, double staining with acridine orange, and propidium iodide in short-term experiments (12–72 h, with monolayer cell cultures) as well as colony-forming method in long-term experiments (25 days, with three dimensional cancer cell colonies).Results: The results obtained revealed that ZnSalenAu and ZnSalenAg decreased significantly viability and proliferation of the treated cells in a time- and concentration-dependent manner being more active as compared to the free ligand H2Salen.Conclusion: The present study demonstrates for the 1st time the ability of two heterometallic complexes ZnSalenAu and ZnSalenAg to decrease significantly viability and proliferation of cultured cell lines established from some of the most common and aggressive human cancers (NSCLC, carcinoma of uterine cancer, 8MGBA, and squamous cell carcinoma) as well as MDR cancer cells

Lycopene: total-scale literature landscape analysis of a valuable nutraceutical with numerous potential applications in the promotion of human and animal health

Lycopene intake from tomatoes and other food sources has multiple potential health benefits. This report aimed to evaluate the current research literature on lycopene concerning human and animal health. The electronic Web of Science Core Collection database was searched with (lycopene*) AND (health* OR illness* OR disease* OR medic* OR pharma* OR drug* OR therap*). The resulted 3972 papers were analyzed with the aid of bibliometric software. Besides the United States, the lycopene papers received global contributions, particularly from China, Italy, India, and Spain. Examples of frequently mentioned chemicals/chemical classes were carotenoid, beta carotene, alpha carotene, beta cryptoxanthin, and alpha tocopherol. Examples of frequently mentioned medical conditions were prostate cancer, cardiovascular disease, and obesity. Published scientific articles reveal the diverse potential of lycopene in prompting human and animal health, and the knowledge on the bioactivities of this phytoche(undefined)info:eu-repo/semantics/publishedVersio

Zeolite and Neurodegenerative Diseases

Neurodegenerative diseases (NDs), characterized by progressive degeneration and death of neurons, are strongly related to aging, and the number of people with NDs will continue to rise. Alzheimer’s disease (AD) and Parkinson’s disease (PD) are the most common NDs, and the current treatments offer no cure. A growing body of research shows that AD and especially PD are intricately related to intestinal health and the gut microbiome and that both diseases can spread retrogradely from the gut to the brain. Zeolites are a large family of minerals built by [SiO4]4− and [AlO4]5− tetrahedrons joined by shared oxygen atoms and forming a three-dimensional microporous structure holding water molecules and ions. The most widespread and used zeolite is clinoptilolite, and additionally, mechanically activated clinoptilolites offer further improved beneficial effects. The current review describes and discusses the numerous positive effects of clinoptilolite and its forms on gut health and the gut microbiome, as well as their detoxifying, antioxidative, immunostimulatory, and anti-inflammatory effects, relevant to the treatment of NDs and especially AD and PD. The direct effects of clinoptilolite and its activated forms on AD pathology in vitro and in vivo are also reviewed, as well as the use of zeolites as biosensors and delivery systems related to PD

Culturable and Non-Culturable Blood Microbiota of Healthy Individuals

Next-generation sequencing (NGS) and metagenomics revolutionized our capacity for analysis and identification of the microbial communities in complex samples. The existence of a blood microbiome in healthy individuals has been confirmed by sequencing, but some researchers suspect that this is a cell-free circulating DNA in blood, while others have had isolated a limited number of bacterial and fungal species by culture. It is not clear what part of the blood microbiota could be resuscitated and cultured. Here, we quantitatively measured the culturable part of blood microbiota of healthy individuals by testing a medium supplemented with a high concentration of vitamin K (1 mg/mL) and culturing at 43 °C for 24 h. We applied targeted sequencing of 16S rDNA and internal transcribed spacer (ITS) markers on cultured and non-cultured blood samples from 28 healthy individuals. Dominant bacterial phyla among non-cultured samples were Proteobacteria 92.97%, Firmicutes 2.18%, Actinobacteria 1.74% and Planctomycetes 1.55%, while among cultured samples Proteobacteria were 47.83%, Firmicutes 25.85%, Actinobacteria 16.42%, Bacteroidetes 3.48%, Cyanobacteria 2.74%, and Fusobacteria 1.53%. Fungi phyla Basidiomycota, Ascomycota, and unidentified fungi were 65.08%, 17.72%, and 17.2% respectively among non-cultured samples, while among cultured samples they were 58.08%, 21.72%, and 20.2% respectively. In cultured and non-cultured samples we identified 241 OTUs belonging to 40 bacterial orders comprising 66 families and 105 genera. Fungal biodiversity accounted for 272 OTUs distributed in 61 orders, 105 families, and 133 genera. Bacterial orders that remained non-cultured, compared to blood microbiota isolated from fresh blood collection, were Sphingomonadales, Rhizobiales, and Rhodospirillales. Species of orders Bacillales, Lactobacillales, and Corynebacteriales showed the best cultivability. Fungi orders Tremellales, Polyporales, and Filobasidiales were mostly unculturable. Species of fungi orders Pleosporales, Saccharomycetales, and Helotiales were among the culturable ones. In this study, we quantified the capacity of a specific medium applied for culturing of blood microbiota in healthy individuals. Other culturing conditions and media should be tested for optimization and better characterization of blood microbiota in healthy and diseased individuals

Antioxidant mechanism in the preventive effect of myrtenal on Alzheimer`s disease progression on experimental mouse model

Introduction: Alzheimer’s disease (AD) is the most common form of dementia causing problems with mem-ory, thinking and behavior. So far there is no unified theory for AD pathogenesis and effective treatment. Scientific reports indicate many natural substances possessing neuroprotective properties. New studies demonstrated that natural monoterpen myrtenal combines antioxidant and anti-acetylcholinesterase activ-ity. Our unpublished data reveal significant improving effect of myrtenal on cognitive function of rodents. Aim: Goal of this study is to examine the effect of myrtenal on AD progression using animal model. Materials and Methods: Experimental model of dementia from AD type was produced on male Albino mice via scopolamine treatment (1 mg/kg i.p., 11 days) and was verified with cognitive test (Step through) and biochemical markers: lipid peroxidation and glutathione content in brain. Dement animals were treat-ed simultaneously with myrtenal (20 mg/kg i.p., 11 days). Its preventive effect was evaluated when compared with the effect of lipoic acid (30mg/kg i.p., 11 days) and galantamine (1 mg/kg i.p., 11 days) as referents. Data were analyzed using t-test of Student-Fisher.Results: Myrtenal produced a significant restoration of cognitive function (with 33%) in dement mice in comparison to scopolamine controls. In healthy rodents, myrtenal had antioxidant activity and decreased significantly brain lipid peroxidation, but in dement animals showed pro-oxidant activity. Administered to-gether myrtenal and lipoic acid demonstrated even better prevention on memory and also decreased estab-lished pro-oxidant activity of myrtenal in dement mice. Conclusion: Analyzed changed parameters (cognitive and biochemical) suggest antioxidant mechanism in myrtenal preventive effect on AD progression

Newly synthesized neuropeptides with central nervous activity in mice

Aim: Object of present study are two newly synthesized neuropeptides with short chains: analogues of Tyr-MIF – 1 with code P1 and of Nociceptine with code P2. Materials and Methods: On male albino mice we studied the changes in the cognitive functions of animals after 3, 7 and 14-days pretreatment with both compounds (5 mg/kg intraperitoneally- i.p.) via: step through test (for learning and memory), Rot-a-rod test (for muscular coordination) and Hole board test (for exploratory activity). Their potential analgesic effect was evaluated by Acetic acid test and their activity on the cen-tral nervous system (CNS) was evaluated via interaction with hexobarbital (HB- 100 mg/kg i.p). Statistics were performed with Student – Fisher test.Results: On the 3rd day after treatment daily both compounds had no effect on cognitive functions of animals, but on the 7th day the analogue of Tyr- MIF –1 (peptide P1) significantly improved the memory (by 60%) and decreased also the exploratory activity of treated animals. The analogue of Nociceptine-P2 demonstrated significant dose-dependent analgesic effect. On the 14th day both compounds improved neuro-muscular coordination of animals. In single doses two compounds shorten significantly duration of hexobarbital narcosis (Р1 by 40% and Р2 by 50%) via unknown mechanism, probably related to functional antagonism between the neuropeptides and hexobarbital on CNS level. Conclusion: Newly synthesized neuropeptides are promising biological active substances with effect on CNS. The analogue of Tyr-MIF–1 improves cognitive function of animals and the analogue of Nociceptine has significant dose-dependent analgesic effect

Gene Co-Expression Network Modular Analysis Reveals Altered Immune Mechanisms in HIV-HAND

Although the introduction of HAART has completely changed the natural course of HIV infection, the number of chronic forms of HIV-associated neurocognitive disorder (HAND) has risen. It is estimated that up to half of subjects undergoing HAART therapy exhibit mild cognitive impairments. In the current study, we apply the gene co-expression network modular analysis, a well-established system biology approach, to the gene expression profiles of cases from the National NeuroAIDS Tissue Consortium (NNTC). We observed a negative enrichment for genes associated with the control of immune responses and putatively regulated by the transcription factors IRF8 and SPI1 and by both type I and II interferons. Our study provides evidence of altered immune responses, which are likely associated with the occurrence of HAND in the absence of HIV encephalitis (HIVE)