Healthy Human Individuals Research Articles

Plasmonic nano-aperture label-free imaging of single small extracellular vesicles for cancer detection

BackgroundSmall extracellular vesicle (sEV) analysis can potentially improve cancer detection and diagnostics. However, this potential has been constrained by insufficient sensitivity, dynamic range, and the need for complex labeling.MethodsIn this study, we demonstrate the combination of PANORAMA and fluorescence imaging for single sEV analysis. The co-acquisition of PANORAMA and fluorescence images enables label-free visualization, enumeration, size determination, and enables detection of cargo microRNAs (miRs).ResultsAn increased sEV count is observed in human plasma samples from patients with cancer, regardless of cancer type. The cargo miR-21 provides molecular specificity within the same sEV population at the single unit level, which pinpoints the sEVs subset of cancer origin. Using cancer cells-implanted animals, cancer-specific sEVs from 20 µl of plasma can be detected before tumors were palpable. The level plateaus between 5–15 absolute sEV count (ASC) per µl with tumors ≥8 mm3. In healthy human individuals (N = 106), the levels are on average 1.5 ASC/µl (+/− 0.95) without miR-21 expression. However, for stage I–III cancer patients (N = 205), nearly all (204 out of 205) have levels exceeding 3.5 ASC/µl with an average of 12.2 ASC/µl (±9.6), and a variable proportion of miR-21 labeling among different tumor types with 100% cancer specificity. Using a threshold of 3.5 ASC/µl to test a separate sample set in a blinded fashion yields accurate classification of healthy individuals from cancer patients.ConclusionsOur techniques and findings can impact the understanding of cancer biology and the development of new cancer detection and diagnostic technologies.

Gut microbial metabolite targets HDAC3-FOXK1-interferon axis in fibroblast-like synoviocytes to ameliorate rheumatoid arthritis

Rheumatoid arthritis (RA) is an autoimmune disease. Early studies hold an opinion that gut microbiota is environmentally acquired and associated with RA susceptibility. However, accumulating evidence demonstrates that genetics also shape the gut microbiota. It is known that some strains of inbred laboratory mice are highly susceptible to collagen-induced arthritis (CIA), while the others are resistant to CIA. Here, we show that transplantation of fecal microbiota of CIA-resistant C57BL/6J mice to CIA-susceptible DBA/1J mice confer CIA resistance in DBA/1J mice. C57BL/6J mice and healthy human individuals have enriched B. fragilis than DBA/1J mice and RA patients. Transplantation of B. fragilis prevents CIA in DBA/1J mice. We identify that B. fragilis mainly produces propionate and C57BL/6J mice and healthy human individuals have higher level of propionate. Fibroblast-like synoviocytes (FLSs) in RA are activated to undergo tumor-like transformation. Propionate disrupts HDAC3-FOXK1 interaction to increase acetylation of FOXK1, resulting in reduced FOXK1 stability, blocked interferon signaling and deactivation of RA-FLSs. We treat CIA mice with propionate and show that propionate attenuates CIA. Moreover, a combination of propionate with anti-TNF etanercept synergistically relieves CIA. These results suggest that B. fragilis or propionate could be an alternative or complementary approach to the current therapies.

Probiotic functional gene explorations in the genome of Limosilactobacillus fermentum GD5MG

Limosilactobacillus fermentum is an isolate obtained from oral gingival samples of healthy human individuals. The whole genome of Lb. fermentum GD5MG is composed of a circular DNA molecule containing 1,834,134 bp and exhibits a GC content of 52.80 %. The sequencing effort produced 38.6 million reads, each 150 bp in length, resulting in a sequencing depth of 2912.48x. Our examination unveiled a total of 1961 protein-coding genes, 27 rRNA genes, 24 tRNA genes, 3 non-coding RNA genes, and 63 pseudogenes with the use of gene annotations in NCBI Prokaryotic Genome Annotation tool. RAST revealed 1863 coding genes distributed across 209 subsystems, with a predominant involvement in amino acid, carbohydrate, and protein metabolism. Phylogenetic analysis infers that the Lb. fermentum GD5MG shares 281 gene clusters. Furthermore, the genome features showed a single CRISPR locus of 45 bp in length. Three genes associated with adhesion ability (strA, dltD, and dltA) and 26 genes related to acid tolerance, digestive enzyme secretion, and bile salt resistance were identified. Numerous genes associated with oral probiotic properties, comprising adhesion, acid and bile salt tolerance, oxidative stress tolerance, and sugar metabolism, were identified in the genome. Our findings shed light on the genomic characteristics of Lb. fermentum GD5MG, which are probable probiotics with functional benefits in humans.

Oral bacterial insights from a comparative study between healthy and comorbid diseased human individuals

The human oral cavity is colonized by a diverse microbial community, which includes both native and transient colonizers. The microbial composition is crucial for maintaining oral homeostasis, but due to overgrowth or imbalances of these microbial communities, dysbiosis can occur. There is a lack of understanding of the research of native and transient colonizers in the oral cavity of the Indian subpopulation Therefore, in our present study, we explored the role and prevalence of transient and native colonizers between healthy and comorbid oral diseased human individuals. Culture-dependent techniques and culture-independent 16S r DNA metagenomic analyses were employed to isolate and study the interactions of native and transient colonizers from human oral samples. Among the 66 human individuals of both healthy and comorbid individuals, the most abundant isolate was found to be Bacillus amyloliquefaciens MCC 4424. In addition, the more prevalent culturable isolate from the healthy samples was Streptococcus salivarius MTCC 13009, whereas in comorbid samples Staphylococcus pasteuri MTCC 13076, Rothia dentocariosa MTCC 13010 and Pseudomonas aeruginosa MTCC 13077 were prevalent to a greater extent. 16S rDNA metagenomic analyses revealed the prevalence and abundance of genera such as Bacteroidetes and Proteobacteria in healthy individuals; consequently, Fusobacteria and Firmicutes were observed mostly in comorbid individuals. The significant differences in bacterial population density were observed in terms of the Shannon index (p = 0.5145) and Simpson index (p = 0.9061) between the healthy and comorbid groups. B. amyloliquefaciens MCC 4424 exhibits antagonistic behavior when grown as a dual-species with native and transient colonizers. This result is very consistent with the findings of antibiofilm studies using confocal laser scanning microscopy, which revealed a significant reduction in biofilm biovolume (73 %) and maximum thickness (80 %) and an increase in the rough coefficient of biofilms (30 %). Our data suggested that B. amyloliquefaciens MCC 4424 can be a native colonizer of Indian sub-populations. It may act as a novel candidate for oral healthcare applications and greatly aids in the regulation of transient species in the oral cavity.

A conserved antigen induces respiratory Th17-mediated broad serotype protection against pneumococcal superinfection

Several vaccines targeting bacterial pathogens show reduced efficacy upon concurrent viral infection, indicating that a new vaccinology approach is required. To identify antigens for the human pathogen Streptococcus pneumoniae that are effective following influenza infection, we performed CRISPRi-seq in a murine model of superinfection and identified the conserved lafB gene as crucial for virulence. We show that LafB is a membrane-associated, intracellular protein that catalyzes the formation of galactosyl-glucosyl-diacylglycerol, a glycolipid important for cell wall homeostasis. Respiratory vaccination with recombinant LafB, in contrast to subcutaneous vaccination, was highly protective against S.pneumoniae serotypes 2, 15A, and 24F in a murine model. In contrast to standard capsule-based vaccines, protection did not require LafB-specific antibodies but was dependent on airway CD4+ T helper 17 cells. Healthy human individuals can elicit LafB-specific immune responses, indicating LafB antigenicity in humans. Collectively, these findings present a universal pneumococcal vaccine antigen that remains effective following influenza infection.

Human scent as a first-line defense against disease

Individuals may have a different body odor, when they are sick compared to healthy. In the non-human animal literature, olfactory cues have been shown to predict avoidance of sick individuals. We tested whether the mere experimental activation of the innate immune system in healthy human individuals can make an individuals’ body odor be perceived as more aversive (intense, unpleasant, and disgusting). Following an endotoxin injection (lipopolysaccharide; 0.6 ng/kg) that creates a transient systemic inflammation, individuals smelled more unpleasant compared to a placebo group (saline injection). Behavioral and chemical analyses of the body odor samples suggest that the volatile components of samples from “sick” individuals changed qualitatively rather than quantitatively. Our findings support the hypothesis that odor cues of inflammation in axillary sweat are detectable just a few hours after experimental activation of the innate immune system. As such, they may trigger behavioral avoidance, hence constituting a first line of defense against pathogens of infected conspecifics.

Boosting serotonin increases information gathering by reducing subjective cognitive costs.

Serotonin is implicated in the valuation of aversive costs, such as delay or physical effort. However, its role in governing sensitivity to cognitive effort, for example deliberation costs during information gathering, is unclear. We show that treatment with a serotonergic antidepressant in healthy human individuals of either sex enhances a willingness to gather information when trying to maximize reward. Using computational modelling, we show this arises from a diminished sensitivity to subjective deliberation costs during the sampling process. This result is consistent with the notion that serotonin alleviates sensitivity to aversive costs in a domain-general fashion, with implications for its potential contribution to a positive impact on motivational deficits in psychiatric disorders.SIGNIFICANCE STATEMENT:Gathering information about the world is essential for successfully navigating it. However, sampling information is costly, and we need to balance between gathering too little and too much information. The neurocomputational mechanisms underlying this arbitration between a putative gain, such as reward, and the associated costs, such as allocation of cognitive resources, remain unclear. In this study, we show that week-long daily treatment with a serotonergic antidepressant enhances a willingness to gather information when trying to maximize reward. Computational modelling indicates this arises from a reduced perception of aversive costs, rendering information gathering less cognitively effortful. This finding points to a candidate mechanism by which serotonergic treatment might help alleviate motivational deficits in a range of mental illnesses.

The human preBCR uses CDR-H3 amino acid composition to control early B cell selection and prevent oncogenesis

Abstract Human B cell repertoire selection depends on the composition of the complementary determining heavy chain (CDR-H3), which lies at the center of the antigen binding site. During B cell development, preB cells express a preB cell receptor (preBCR), which is composed of μH chain bound to surrogate light chain (SLC). In mouse, the VpreB R 101D 57R 51motif of the SLC selects against μH chains with hydrophobic CDR-H3s. Abnormal selection at the preBCR stage predisposes to autoreactive B cell survival and leukemia. We sought to study preBCR selection in healthy human individuals versus acute lymphoblastic leukemia (ALL) patients to dissect the mechanism of formation of the human protective CDR-H3 repertoire. We hypothesized that expression of a preBCR with an abnormal amino acid content prevents differentiation of the ALL blasts into a late preB and then a mature B cell, promoting oncogenic survival. We sorted early B cells from bone marrow swabs of healthy controls and ALL patients. We extracted RNA, deep sequenced preB cells and leukemic blasts and then analyzed their CDR-H3 based on amino acid content using bioinformatics and structure modeling. Our results suggest that in healthy individuals, similar to mouse, the human preBCR selects for CDR-H3 based on amino acid content with a preference for tyrosine (Y) and negative selection of hydrophobic antigen binding sites. The CDR-H3s of ALL leukemic blasts have amino acid characteristics similar to the negatively selected antigen binding sites of healthy individuals. In conclusion, this type of analysis might be used to monitor optimal immunoglobulin repertoire diversification. R21AI163555

Elevation of truncated (48 kDa) form of unconventional myosin 1C in blood serum correlates with severe Covid-19

In Covid-19 and autoimmune patients, there are several similarities revealed in the immune responses (Liu et al., 2021; Woodruff et al., 2020). Earlier, we firstly detected a truncated (48 kDa) form of the unconventional Myosin 1C (48/Myo1C) in a fraction of proteins soluble in 10% 2,2,2-trichloroacetic acid (TCA). These proteins were obtained from blood serum of patients with autoimmune diseases, such as multiple sclerosis, systemic lupus erythematosus, and rheumatoid arthritis (Kit et al., 2018). Here, we demonstrated that content of 48/Myo1C was also elevated in blood serum of the severe Covid-19 patients. Whereas in blood of 28 clinically healthy human individuals regularly tested for Covid-19 infection, the amount of this protein was undetectable or very low, in blood of 16 of 28 patients hospitalized with severe course of this disease, its amount was significantly increased. Dexamethasone, steroid hormone which is widely used for treatment of severe Covid-19 patients, induced time-dependent elevation of the 48/Myo1C in blood of such patients. The 48/Myo1C dose-dependently suppressed the viability of anti-CD3-activated lymphocytes of human peripheral blood. Recently, we used affinity chromatography on the magnetic poly(glycidyl-methacrylate) (mag-PGMA–NH2) microparticles functionalized with Myo1C and MALDI–TOF mass spectrometry with molecular modeling in silico in order to identify potential molecular partners of the 48/Myo1C. It was found that 48/Myo1C might bind to component 3 of the complement system and the anti-thrombin-III (Starykovych et al., 2021). Thus, the mechanisms of the pathogenic action of truncated form of Myo1C in severe COVID-19 patients may involve a suppression of the immune cells, as well as modulation of complement and coagulation cascades.

Increased cerebral vascular resistance underlies preserved cerebral blood flow in response to orthostasis in humans on a high-salt diet.

Cerebral blood flow autoregulation protects brain tissue from blood pressure variations and maintains cerebral perfusion pressure by changes in vascular resistance. High salt (HS) diet impairs endothelium-dependent vasodilation in many vascular beds, including cerebral microcirculation, and may affect vascular resistance. The aim of present study was to determine if 7-day HS diet affected the reactivity of middle cerebral artery (MCA) to orthostatic challenge in healthy human individuals, and if autoregulatory mechanisms and sympathetic neural regulation were involved in this phenomenon.Twenty-seven persons participated in study (F:21, M:6, age range 19-24). Participants consumed 7-day low-salt (LS) diet (< 2.3g kitchen salt/day) and afterwards 7-day HS diet (> 11.2g kitchen salt/day). Blood and urine analysis and anthropometric measurements were performed after each diet. Arterial blood pressure, heart rate and heart rate variability, and cerebral and systemic hemodynamic parameters were recorded simultaneously with transcranial Doppler ultrasound and The Task Force®Monitor in response to orthostatic test.Participants remained normotensive during HS diet. Following both, the LS and HS dietary protocols, mean cerebral blood flow (CBF), as well as the velocity time integral and diastolic blood pressure decreased, and cerebral pulsatility index increased after rising up. Importantly, cerebrovascular resistance significantly increased in response to orthostasis only after HS diet. Urine concentration of noradrenaline and vanillylmandelic acid, baroreflex sensitivity (BRS), and sympathetic neural control was significantly decreased in HS diet.Results suggest that CBF in response to orthostatic test was preserved in HS condition due to altered vascular reactivity of MCA, with increased cerebrovascular resistance and blunted BRS and sympathetic activity.

Post-COVID-19 impacts on male sexual and reproductive functions

Background: Assessment of the male sexual and reproductive health status with and without severe acute respiratory syndrome coronavirus 2infected COVID19 disease could create awareness about the lateral impact of those patients recovered from COVID19 infection. Aim and Objectives: Therefore, the main focus of the current study was to determine the postCOVID19 infection on male sexual and reproductive functions. Materials and Methods: A crosssectional study by involving two groups of the study individuals namely: Group 1: age and sexmatched normal healthy control human individuals. Group 2: postCOVIDtreated patients, at least 912 months after treatment success. The International Index of Erectile Function (IIEF), a questionnaire developed by Rosen et al. to question on sexual functions has been used in the present study. Results: Based on analysis of the 15point International Index of Erectile Function analysis, it was found that overall there was a statistically significant difference in (P &lt; 0.05) in sexual behaviors in both the groups. It was also found that marital status, place of living, and vaccination status are statistically significant associated with sexual behaviors, whereas occupation, religion, and education are statistically insignificant. Conclusion: The current study showed significant differences after the COVID19 infection and treatment in male fertility from a single tertiary care hospitalbased study in Western Uttar Pradesh. However, involving a large population from different parts of the country could pave the way to show what needs to be done by the scientific and clinical community to tackle our limited understanding of the disease. Keywords: Erectile dysfunction, International Index of Erectile Function, male sexual and reproductive functions, postCOVID19.

Integrated View of Baseline Protein Expression in Human Tissues.

The availability of proteomics datasets in the public domain, and in the PRIDE database, in particular, has increased dramatically in recent years. This unprecedented large-scale availability of data provides an opportunity for combined analyses of datasets to get organism-wide protein abundance data in a consistent manner. We have reanalyzed 24 public proteomics datasets from healthy human individuals to assess baseline protein abundance in 31 organs. We defined tissue as a distinct functional or structural region within an organ. Overall, the aggregated dataset contains 67 healthy tissues, corresponding to 3,119 mass spectrometry runs covering 498 samples from 489 individuals. We compared protein abundances between different organs and studied the distribution of proteins across these organs. We also compared the results with data generated in analogous studies. Additionally, we performed gene ontology and pathway-enrichment analyses to identify organ-specific enriched biological processes and pathways. As a key point, we have integrated the protein abundance results into the resource Expression Atlas, where they can be accessed and visualized either individually or together with gene expression data coming from transcriptomics datasets. We believe this is a good mechanism to make proteomics data more accessible for life scientists.

Antibody Levels Poorly Reflect on the Frequency of Memory B Cells Generated following SARS-CoV-2, Seasonal Influenza, or EBV Infection.

The scope of immune monitoring is to define the existence, magnitude, and quality of immune mechanisms operational in a host. In clinical trials and praxis, the assessment of humoral immunity is commonly confined to measurements of serum antibody reactivity without accounting for the memory B cell potential. Relying on fundamentally different mechanisms, however, passive immunity conveyed by pre-existing antibodies needs to be distinguished from active B cell memory. Here, we tested whether, in healthy human individuals, the antibody titers to SARS-CoV-2, seasonal influenza, or Epstein-Barr virus antigens correlated with the frequency of recirculating memory B cells reactive with the respective antigens. Weak correlations were found. The data suggest that the assessment of humoral immunity by measurement of antibody levels does not reflect on memory B cell frequencies and thus an individual's potential to engage in an anamnestic antibody response against the same or an antigenically related virus. Direct monitoring of the antigen-reactive memory B cell compartment is both required and feasible towards that goal.

A library of induced pluripotent stem cells from clinically well-characterized, diverse healthy human individuals.

SummaryA library of well-characterized human induced pluripotent stem cell (hiPSC) lines from clinically healthy human subjects could serve as a useful resource of normal controls for in vitro human development, disease modeling, genotype-phenotype association studies, and drug response evaluation. We report generation and extensive characterization of a gender-balanced, racially/ethnically diverse library of hiPSC lines from 40 clinically healthy human individuals who range in age from 22 to 61 years. The hiPSCs match the karyotype and short tandem repeat identities of their parental fibroblasts, and have a transcription profile characteristic of pluripotent stem cells. We provide whole-genome sequencing data for one hiPSC clone from each individual, genomic ancestry determination, and analysis of mendelian disease genes and risks. We document similar transcriptomic profiles, single-cell RNA-sequencing-derived cell clusters, and physiology of cardiomyocytes differentiated from multiple independent hiPSC lines. This extensive characterization makes this hiPSC library a valuable resource for many studies on human biology.

Neurofeedback for Attention-Deficit/Hyperactivity Disorder: Evidence Base and Results of a Large RCT

Conflict adaptation refers to the improved conflict control induced after experiencing conflict and is a prominent index of adaptive cognitive control. Reversal of conflict adaptation may be maladaptive and predictive of certain mental disorders. Here, we employed real-time functional near infrared spectroscopy-based neurofeedback training, with the left dorsolateral prefrontal cortex as the target brain area, to investigate whether reversal of conflict adaptation during a word-color Stroop task could be recovered to be normal. Healthy human individuals with reversal pattern of conflict adaptation in the pretest were randomly assigned into the experimental or control groups. Distributed training for 80 min led to greater improvements in the experimental group who received real neurofeedback compared to those in the control group who received sham neurofeedback. These results indicated causal evidence for understanding the generation of conflict adaptation and heighten the prospects of clinical application of neurofeedback training.

An investigation on gastric emptying behavior of apple in the dynamic digestion model ARK® and its validation using MRI of human subjects – A pilot study

The employment of dynamic digestion models for understanding the digestion behavior of foods has become an emerging trend in recent times, as the in vivo techniques used currently pose safety and ethical concerns. On the other hand, it is equally important to validate the mentioned digestion models, so that they can aid in assessing various aspects of digestion. Hence, this study was planned to investigate the gastric emptying behavior of apples in a 3D printed stomach in vitro dynamic digestion model artificial stomach response kit. Further, the obtained results are validated by studying the gastric emptying behavior of apples in healthy and adult human individuals. Gastric emptying trends in both the in vivo and in vitro trials showed close half emptying times, i.e., 51.00 ± 7.54 and 57.84 ± 4.41 min respectively. Also, Tlag, K, β obtained were similar in both the in vivo and in vitro experiments. In addition to that, a strong positive correlation is noticed between the in vivo and in vitro trials, r = 0.99. Upon successful validation with foods having diversified properties, the ARK® model can be complementary for in vivo techniques to understand the gastric emptying behavior and other digestion aspects of foods.

Maxillary Sinus Dimensions of Different Human Age Groups by CT Scan Imaging

Background: The maxillary sinus is an air-filled space, situated in maxillary bones and may be recognized with different shapes and sizes, its walls are obviously thin, and its apex can extend to zygomatic processes of these bones and can occupy most of zygomatic bone. Materials and methods: A prospective study of 330 healthy human individuals who attending to radiology section during the period from December 2018 to October 2019 to do CT scans for maxillary sinuses. Various CT images were taken to calculate the three dimensions of the maxillary sinuses. Results: The mean value for right maxillary human sinus anteroposterior length, width, and height for males were 40.2 ± 4.2mm, 25.2 ± 4.2mm, and 45.0 ± 5.1mm respectively; while for the left was 39.2 ± 3.9mm, 24.5 ± 4.5mm, and 47.2 ± 4.5mm respectively. Whereas, the mean value for right maxillary human sinus length, width, and height in females were 38.7 ± 4.0mm, 24.3 ± 4.0mm, and 42.6 ± 5.0mm respectively; while for the left was 38.3 ± 4.0mm, 23.6 ± 4.2mm, and 44.1 ± 4.7mm respectively. High significant differences (p≤0.05) recorded among age groups for length of both sides of both genders, width in left sinus of both genders, height of right side of both genders, and finally height of left side in females. Conclusion: Maxillary sinus dimensions measured by CT scans revealed a precise recognition of 3D configuration for the sinus that may be useful as a specific identification feature of any human individual.

From reversal to normal: Robust improvement in conflict adaptation through real-time functional near infrared spectroscopy-based neurofeedback training

Conflict adaptation refers to the improved conflict control induced after experiencing conflict and is a prominent index of adaptive cognitive control. Reversal of conflict adaptation may be maladaptive and predictive of certain mental disorders. Here, we employed real-time functional near infrared spectroscopy-based neurofeedback training, with the left dorsolateral prefrontal cortex as the target brain area, to investigate whether reversal of conflict adaptation during a word-color Stroop task could be recovered to be normal. Healthy human individuals with reversal pattern of conflict adaptation in the pretest were randomly assigned into the experimental or control groups. Distributed training for 80 min led to greater improvements in the experimental group who received real neurofeedback compared to those in the control group who received sham neurofeedback. These results indicated causal evidence for understanding the generation of conflict adaptation and heighten the prospects of clinical application of neurofeedback training.

Much ado about nothing? Off-target amplification can lead to false-positive bacterial brain microbiome detection in healthy and Parkinson\u2019s disease individuals

BackgroundRecent studies suggested the existence of (poly-)microbial infections in human brains. These have been described either as putative pathogens linked to the neuro-inflammatory changes seen in Parkinson’s disease (PD) and Alzheimer’s disease (AD) or as a “brain microbiome” in the context of healthy patients’ brain samples.MethodsUsing 16S rRNA gene sequencing, we tested the hypothesis that there is a bacterial brain microbiome. We evaluated brain samples from healthy human subjects and individuals suffering from PD (olfactory bulb and pre-frontal cortex), as well as murine brains. In line with state-of-the-art recommendations, we included several negative and positive controls in our analysis and estimated total bacterial biomass by 16S rRNA gene qPCR.ResultsAmplicon sequencing did detect bacterial signals in both human and murine samples, but estimated bacterial biomass was extremely low in all samples. Stringent reanalyses implied bacterial signals being explained by a combination of exogenous DNA contamination (54.8%) and false positive amplification of host DNA (34.2%, off-target amplicons). Several seemingly brain-enriched microbes in our dataset turned out to be false-positive signals upon closer examination.We identified off-target amplification as a major confounding factor in low-bacterial/high-host-DNA scenarios. These amplified human or mouse DNA sequences were clustered and falsely assigned to bacterial taxa in the majority of tested amplicon sequencing pipelines. Off-target amplicons seemed to be related to the tissue’s sterility and could also be found in independent brain 16S rRNA gene sequences.ConclusionsTaxonomic signals obtained from (extremely) low biomass samples by 16S rRNA gene sequencing must be scrutinized closely to exclude the possibility of off-target amplifications, amplicons that can only appear enriched in biological samples, but are sometimes assigned to bacterial taxa. Sequences must be explicitly matched against any possible background genomes present in large quantities (i.e., the host genome). Using close scrutiny in our approach, we find no evidence supporting the hypothetical presence of either a brain microbiome or a bacterial infection in PD brains.4jqTESRfTS1PdZkk_4BnPMVideo abstract

Isolation of Novel Probiotic Lactobacillus and Enterococcus Strains From Human Salivary and Fecal Sources.

Probiotics are non-pathogenic microorganisms that can interact with the gastrointestinal microbiota. They have numerous beneficial health effects that include enhancement of the host immune response, antiallergic, antimicrobial, anti-cancer, and anti-inflammatory properties. Probiotics are capable of restoring the impaired microbiome of a dysbiotic gut. They can be isolated from different environments. However, it is frequently suggested that probiotics for human use should come from human sources. The objective of this study was to isolate and characterize novel probiotic strains from the saliva and feces of healthy human individuals. To meet the criteria for probiotic attributes, the isolates were subjected to numerous standard morphological and biochemical tests. These tests included Gram staining, catalase tests, antibiotic susceptibility testing, hemolytic and antagonistic evaluation, tolerance tests involving temperature, NaCl levels, pH and bile salts, adherence ability assays, and genotypic characterization involving 16S rRNA gene sequencing. From 26 saliva and 11 stool samples, 185 microbial strains were isolated. Based on morphological and biochemical characteristics, 14 potential probiotic candidates were selected and identified genotypically. The new strains belonged to Lactobacillus fermentum, Enterococcus faecium, and Enterococcus hire. The selected strains were non-hemolytic, showed high tolerance to low pH and bile salts, and strong adherence abilities. Furthermore, the strains displayed a wide range of antimicrobial activities, particularly against antibiotic-resistant pathogens such as methicillin resistant Staphylococcus aureus (MRSA). Moreover, five of the selected isolates demonstrated antiproliferative features against human colon cancer cell line (Caco-2). The results of this investigation confirm the diversity of microbial populations in the human gut and saliva, and since these strains are of human origin, they will highly likely display maximal activities in food and drugs set for human use. Hence, the new strains of this study require additional in vivo experiments to assess their health-promoting effects.