scholarly journals Effects of Rifaximin on Luminal and Wall-Adhered Gut Commensal Microbiota in Mice

2021 ◽  
Vol 22 (2) ◽  
pp. 500
Author(s):  
Marina Ferrer ◽  
Mònica Aguilera ◽  
Vicente Martinez
Keyword(s):  
Fluorescent In Situ Hybridization ◽  
Fragment Length Polymorphism ◽  
Functional Gastrointestinal Disorders ◽  
Antibacterial Properties ◽  
Broad Spectrum Antibiotic ◽  
Commensal Microbiota ◽  
Bacterial Biodiversity ◽  
Rule Out

Rifaximin is a broad-spectrum antibiotic that ameliorates symptomatology in inflammatory/functional gastrointestinal disorders. We assessed changes in gut commensal microbiota (GCM) and Toll-like receptors (TLRs) associated to rifaximin treatment in mice. Adult C57BL/6NCrl mice were treated (7/14 days) with rifaximin (50/150 mg/mouse/day, PO). Luminal and wall-adhered ceco-colonic GCM were characterized by fluorescent in situ hybridization (FISH) and microbial profiles determined by terminal restriction fragment length polymorphism (T-RFLP). Colonic expression of TLR2/3/4/5/7 and immune-related markers was assessed (RT-qPCR). Regardless the period of treatment or the dose, rifaximin did not alter total bacterial counts or bacterial biodiversity. Only a modest increase in Bacteroides spp. (150 mg/1-week treatment) was detected. In control conditions, only Clostridium spp. and Bifidobacterium spp. were found attached to the colonic epithelium. Rifaximin showed a tendency to favour their adherence after a 1-week, but not 2-week, treatment period. Minor up-regulation in TLRs expression was observed. Only the 50 mg dose for 1-week led to a significant increase (by 3-fold) in TLR-4 expression. No changes in the expression of immune-related markers were observed. Rifaximin, although its antibacterial properties, induces minor changes in luminal and wall-adhered GCM in healthy mice. Moreover, no modulation of TLRs or local immune systems was observed. These findings, in normal conditions, do not rule out a modulatory role of rifaximin in inflammatory and or dysbiotic states of the gut.

scholarly journals Biofilms from a Brazilian water distribution system include filamentous fungi

2013 ◽  
Vol 59 (3) ◽  
pp. 183-188 ◽  
Author(s):  
V.M. Siqueira ◽  
H.M.B. Oliveira ◽  
C. Santos ◽  
R.R.M. Paterson ◽  
N.B. Gusmão ◽  
...  
Keyword(s):  
Filamentous Fungi ◽  
Distribution System ◽  
Fluorescent In Situ Hybridization ◽  
Distribution Systems ◽  
Water Distribution ◽  
Water Distribution System ◽  
Sampling Device ◽  
Calcofluor White

Filamentous fungi in drinking water can block water pipes, can cause organoleptic biodeterioration, and are a source of pathogens. There are increasing reports of the involvement of the organisms in biofilms. This present study describes a sampling device that can be inserted directly into pipes within water distribution systems, allowing biofilm formation in situ. Calcofluor White M2R staining and fluorescent in situ hybridization with morphological analyses using epifluorescent microscopy were used to analyse biofilms for filamentous fungi, permitting direct observation of the fungi. DAPI (4′,6-diamidino-2-phenylindole) was applied to detect bacteria. Filamentous fungi were detected in biofilms after 6 months on coupons exposed to raw water, decanted water and at the entrance of the water distribution system. Algae, yeast, and bacteria were also observed. The role of filamentous fungi requires further investigations.

Megafaunal Butchering at Lubbock Lake, Texas: A Taphonomic Reanalysis

1988 ◽  
Vol 30 (2) ◽  
pp. 221-231 ◽  
Author(s):  
Lee Ann Kreutzer
Keyword(s):  
Human Behavior ◽  
Late Pleistocene ◽  
Meat Processing ◽  
Faunal Remains ◽  
Point Bar ◽  
Feature Structure ◽  
Bone Accumulation ◽  
Rule Out

Five seasons of excavation in Feature Area 2-1 of the Lubbock Lake Landmark, Texas, exposed a megafaunal bone accumulation in sands and gravels deposited by a late Pleistocene meandering stream. Many bone specimens exhibit evidence of alteration, supporting interpretation of the feature as an in situ, secondary meat-processing area; the gravels are interpreted as the point bar of a meandering stream. Faunal remains lying stratigraphically above the point bar have been considered to form a separate, noncultural feature produced by stream flooding. However, rose diagrams and analysis of adjusted residuals demonstrate that a statistically significant amount of bone in each feature is aligned along axes of preferred orientation. Further, the orientation patterns and statistical analyses of both features exhibit the same trends, suggesting that the same processes affected both. Although the evidence does not rule out a role of human behavior, it does demonstrate that stream currents significantly influenced feature structure.

Application and biomolecular monitoring of psychrophilic anaerobic digestion

2006 ◽  
Vol 54 (2) ◽  
pp. 41-47 ◽  
Author(s):  
G. Collins ◽  
S. Connaughton ◽  
A.-M. Enright ◽  
C. Scully ◽  
T. Mahony ◽  
...  
Keyword(s):  
Anaerobic Digestion ◽  
Fluorescent In Situ Hybridization ◽  
Fragment Length Polymorphism ◽  
Physiological Activity ◽  
Treatment Plant ◽  
Granular Sludge ◽  
Operating Conditions ◽  
Methanogenic Activity ◽  
Plant Monitoring

Thirteen anaerobic hybrid expanded granular sludge bed-anaerobic filter bioreactors were used for psychrophilic (15–18 °C) anaerobic digestion of a variety of synthetic and non-synthetic wastewaters, including: food-processing, dairy, aromatic- and aliphatic-containing and brewery discharges. Specific methanogenic activity assays were employed to assess temporal physiological activity dynamics. Terminal restriction fragment length polymorphism genetic fingerprinting and fluorescent in situ hybridization were used to monitor shifts in the structure of the microbial communities in the bioreactors in response to operating conditions. Treatment efficiencies obtained were comparable to previous mesophilic (37 °C) trials. Methanogenic activity developed under psychrophilic conditions and putative psychrophilic populations were detected within otherwise psychrotolerant mesophilic communities. Shifts in the population structure of archaeal (methanogenic) communities were more indicative of process disruption than bacterial communities. Biomolecular techniques were demonstrated as valuable tools for anaerobic wastewater treatment plant monitoring.

scholarly journals A role of territory formation in disruption of heterologous pairings in Drosophila male meiosis

2020 ◽  
Author(s):  
Christopher A. Hylton ◽  
John E. Tomkiel Dean
Keyword(s):  
In Situ Hybridization ◽  
Fluorescent In Situ Hybridization ◽  
Fruit Fly ◽  
Male Meiosis ◽  
Chromosome 3 ◽  
Prophase I ◽  
Territory Formation ◽  
Nuclear Domains

Pairings between heterologous chromosomes in meiosis can lead to nondisjunction and the production of aneuploid gametes. To minimize these aberrant outcomes, organisms have evolved mechanisms to disrupt such improper pairings prior to orientation and segregation. In the male fruit fly, Drosophila melanogaster, bivalents segregate to distinct nuclear domains in prophase I, and it has been proposed that the formation of these distinct territories may play a role in disrupting interactions between limited homologies on heterologous chromosomes. To test this, we used fluorescent in situ hybridization to examine pairing between the X chromosome and Dp(1;3) chromosomes in which a segment of the X had been transposed to chromosome 3. We found that 120kb of homology was sufficient to insure nearly complete pairing but was not sufficient to direct merotelic segregation of the paired elements, suggesting that such pairings were being disrupted. We compared the perdurance of X / Dp(1;3) pairings to that of X / Dp(1;Y) pairings (in which homologs are paired),and found that heterologous pairings were disrupted at a higher frequency at the S2b stage of prophase I, the stage at which territory formation is initiated. Our results support the model that movement of bivalents into distinct domains in prophase I provides a mechanism to disrupt pairings between limited regions of homology, and thus may be one means of preventing improper segregation of heterologs in this organism.

scholarly journals Role of Fluorescent In Situ Hybridization, Cholangioscopic Biopsies, and EUS-FNA in the Evaluation of Biliary Strictures

2018 ◽  
Vol 63 (3) ◽  
pp. 636-644 ◽  
Author(s):  
Christian Brooks ◽  
Valerie Gausman ◽  
Chanthel Kokoy-Mondragon ◽  
Khushboo Munot ◽  
Sunil P. Amin ◽  
...  
Keyword(s):  
In Situ Hybridization ◽  
Fluorescent In Situ Hybridization ◽  
Biliary Strictures

scholarly journals Antenatal Microbial Colonization of Mammalian Gut

2018 ◽  
Vol 26 (8) ◽  
pp. 1045-1053 ◽  
Author(s):  
Elisa Borghi ◽  
Valentina Massa ◽  
Marco Severgnini ◽  
Grazia Fazio ◽  
Laura Avagliano ◽  
...  
Keyword(s):  
In Situ Hybridization ◽  
Amniotic Fluid ◽  
Fluorescent In Situ Hybridization ◽  
Microbial Colonization ◽  
Bacterial Dna ◽  
Commensal Microbiota ◽  
High Interindividual Variability ◽  
Next Generation Sequencing Ngs ◽  
Generation Sequencing

The widely accepted dogma of intrauterine sterility and initial colonization of the newborn during birth has been blurred by recent observations of microbial presence in meconium, placenta, and amniotic fluid. Given the importance of a maternal-derived in utero infant seeding, it is crucial to exclude potential environmental or procedural contaminations and to assess fetal colonization before parturition. To this end, we analyzed sterilely collected intestinal tissues, placenta, and amniotic fluid from rodent fetuses and tissues from autoptic human fetuses. Total bacterial DNA was extracted from collected samples and analyzed by Next Generation Sequencing (NGS) techniques using hypervariable 16S ribosomal RNA (rRNA) regions (V3-V4). Colonizing microbes were visualized in situ, using labeled probes targeting 16S ribosomal DNA by fluorescent in situ hybridization. The NGS analysis showed the presence of pioneer microbes in both rat and human intestines as well as in rodent placentas and amniotic fluids. Microbial communities showed fetus- and dam-dependent clustering, confirming the high interindividual variability of commensal microbiota even in the antenatal period. Fluorescent in situ hybridization analysis confirmed the microbes’ presence in the lumen of the developing gut. These findings suggest a possible antenatal colonization of the developing mammalian gut.

scholarly journals Genetic and Cytogenetic Mapping of DMI1, DMI2, and DMI3 Genes of Medicago truncatula Involved in Nod Factor Transduction, Nodulation, and Mycorrhization

2002 ◽  
Vol 15 (11) ◽  
pp. 1108-1118 ◽  
Author(s):  
Jean-Michel Ané ◽  
Julien Lévy ◽  
Philippe Thoquet ◽  
Olga Kulikova ◽  
Françoise de Billy ◽  
...  
Keyword(s):  
Medicago Truncatula ◽  
Fluorescent In Situ Hybridization ◽  
Fragment Length Polymorphism ◽  
Bulked Segregant Analysis ◽  
Linkage Groups ◽  
Cytogenetic Mapping ◽  
Nod Factor ◽  
Amplification Fragment Length Polymorphism ◽  
Cytogenetic Studies

The DMI1, DMI2, and DMI3 genes of Medicago truncatula, which are required for both nodulation and mycorrhization, control early steps of Nod factor signal transduction. Here, we have used diverse approaches to pave the way for the map-based cloning of these genes. Molecular amplification fragment length polymorphism markers linked to the three genes were identified by bulked segregant analysis. Integration of these markers into the general genetic map of M. truncatula revealed that DMI1, DMI2, and DMI3 are located on linkage groups 2, 5, and 8, respectively. Cytogenetic studies using fluorescent in situ hybridization (FISH) on mitotic and pachytene chromosomes confirmed the location of DMI1, DMI2, and DMI3 on chromosomes 2, 5, and 8. FISH-pachytene studies revealed that the three genes are in euchromatic regions of the genome, with a ratio of genetic to cytogenetic distances between 0.8 and 1.6 cM per μm in the DMI1, DMI2, and DMI3 regions. Through grafting experiments, we showed that the genetic control of the dmi1, dmi2, and dmi3 nodulation phenotypes is determined at the root level. This means that mutants can be transformed by Agrobacterium rhizogenes to accelerate the complementation step of map-based cloning projects for DMI1, DMI2, and DMI3.

HPV nonrelated endocervical adenocarcinoma in hereditary cancer syndromes

2020 ◽  
Vol 106 (6) ◽  
pp. NP67-NP72
Author(s):  
Ileana Carnevali ◽  
Eleonora Di Lauro ◽  
Valeria Pensotti ◽  
Nora Sahnane ◽  
Eleonora Leoni ◽  
...  
Keyword(s):  
Fluorescent In Situ Hybridization ◽  
Cancer Susceptibility ◽  
Cancer Genetic Counseling ◽  
Correct Identification ◽  
Inherited Cancer ◽  
Lynch Syndrome Family ◽  
Cancer Syndromes ◽  
The Relationship

Introduction: The relationship between endocervical cancer and cancer susceptibility syndromes is not yet fully understood. We present 2 cases of endocervical cancer: 1 arising in a patient carrier with a pathogenic BRCA1 variant and the second detected in a Lynch syndrome family carrying the MSH2 germline pathogenic variant. Case description: Somatic analyses including loss of heterozygosity and fluorescent in situ hybridization demonstrated that the second hit in patient 1 is BRCA1-related. Mismatch repair somatic analyses in the second family demonstrated that the endocervical cancers of patient 2 and of her sister are MSH2-related. These data confirm the relationship between the pathogenesis of endocervical cancer and the presence of germline BRCA1 and MSH2 mutations. Conclusions: Our study confirms that gynecologic cancers including rare entities such as non–human papillomavirus–related endocervical cancer (NHPVA) are sentinels for inherited cancer syndromes. Endocervical cancer NHPVAs might be considered for cancer genetic counseling in order to improve cancer prevention. For this reason, the role of pathologists is particularly important for the correct identification of the cervical tumor site.

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