115 Invivo confocal laser endomicroscopy visualisation of fresh and frozen bull spermatozoa in the genital tract of dairy heifers

2020 ◽  
Vol 32 (2) ◽  
pp. 184
Author(s):  
C. Richard ◽  
X. Druart ◽  
T. Saint-Beuve ◽  
V. Gelin ◽  
L. Laffont ◽  
...  
Keyword(s):  
Genital Tract ◽  
Visual Analysis ◽  
Semen Quality ◽  
Video Quality ◽  
Small Ruminants ◽  
Electronic System ◽  
Confocal Laser Endomicroscopy ◽  
Confocal Laser ◽  
Control Group ◽  
Uterine Body

Fertility in cattle includes the ability of the uterus to provide the appropriate environment for pregnancy success, including the transport of spermatozoa before fertilization. Confocal laser endomicroscopy technology (Cellvizio) has been previously used in small ruminants to visualise labelled spermatozoa invivo in the uterine horns of ewes (Druart et al. 2009 Reprod. 138, 15-53). Nevertheless, to the best of our knowledge, no invivo study has reported a live visual analysis of the behaviour of spermatozoa in the bovine uterus upon AI. The aim of this study was to develop an experimental procedure to label bull spermatozoa and to visualise their progression in the uterine horns of dairy cows, using Cellvizio and uterine cervix catheterization. Fresh ejaculated bull spermatozoa were double-labelled with octadecyl rhodamine B chloride and MitoTracker Green FM dyes before dilution and freezing processing. At each step of the process, semen quality was evaluated with a semen quality analyzer (SQA-V b, Medical Electronic System) and compared to nonlabelled bull reference semen. The specificity of the labelling was validated invitro. We also developed a specific device to insert the Cellvizio fibre probe into the genital tract, in order to image vagina, cervix, and uterine body, as well as the proximal, middle, and distal parts of the uterine horn, including the utero-tubal junction (UTJ), before and after AI with fresh and frozen labelled spermatozoa. Then, 10 heifers were oestrus synchronized. Video sequences were recorded at oestrus (n=5), just before AI, at the time of AI (7-12h after the onset of oestrus), 30 and 120min after AI, and 14 days after oestrus (n=5; luteal phase), to investigate the effect of the steroid-primed uterine environment on the distribution and progression of spermatozoa in the genital tract. Using our approach, labelled spermatozoa were detected (1) at the day of oestrus and 14 days after oestrus, (2) in the uterine body at the time of AI, (3) in various parts of the genital tract 30min after AI, and (4) in the UTJ, the cervix, and the vagina 120min after AI. In addition, labelling the spermatozoa did not alter the fertilizing capacity: 4 of the 5 oestrus females were pregnant at Day 35. Additionally, an IVF test showed a blastocyst rate of 45.8% for labelled semen vs. 39.4% for the control group (Carvalho et al. 2017 Reprod. 154, 695-710). In conclusion, our study provides a method to phenotype in situ the transport of spermatozoa in the genital tract of heifers. Further investigations are in progress to optimize video quality, to develop an algorithm for an automatic analysis (spermatozoa count, speed, and path) of the recorded sequences.

scholarly journals Immunosuppressant Cyclosporin A-Based Regulation of Lipopolysaccharide-Triggered Pro- and Anti-Inflammatory Cytokines in the Genital Tract of Female Rabbits

2018 ◽  
Author(s):  
Li-Qin Yang ◽  
Qiu-Ying Wu ◽  
Xuan-Yu Chen ◽  
Chun Wang ◽  
Zhang Yan ◽  
...  
Keyword(s):  
Mrna Expression ◽  
Inflammatory Cytokines ◽  
Genital Tract ◽  
Protein Production ◽  
Control Group ◽  
Protein Levels ◽  
Lps Challenge ◽  
Anti Inflammatory ◽  
Uterine Body ◽  
Pro Inflammatory Cytokines

In this study, we evaluated the effects of Cyclosporine A (CsA) on Lipopolysaccharide (LPS)-induced cytokine production in the genital tract of female rabbits. Twelve sexually mature and healthy female rabbits were randomly divided into four groups (n = 3 each). The rabbits in the LPS group were given an intrauterine infusion of Escherichia coli LPS (4 mg/kg body weight (BW)). Rabbits in the CsA group were given CsA (20 mg/kg BW). Rabbits in the LPS + CsA group were given LPS (4 mg/kg BW) and CsA (20 mg/kg BW). The control group received only LPS and CsA carrier. The gene expression and protein levels of pro- and anti-inflammatory cytokines were observed using qRT-PCR and immuno-histochemical (IHC) assay, respectively. Our study showed that IL-1β, IL-6, IL-8, TNF-α, IFN-γ, IL-4, IL-10, IL-13, and TGF-β were expressed in female genital organs. The LPS challenge increased the mRNA expression of IL-6 and TNF-α in the uterine body and IL-1β in the uterotubal junction compared to the control group. CsA increased the basal mRNA expression of anti-inflammatory cytokines (i.e., IL-4 in the uterine body, uterotubal junction, and oviductal ampulla; IL-10 in the cervix, oviductal isthmus, and ampulla; and TGF-β in the uterotubal junction and oviductal ampulla) and pro-inflammatory cytokines (i.e., IL-6 and IL-8 in the cervix; IL-1β in the oviductal isthmus; TNF-α in the oviductal ampulla; and IFN-γ in the uterine body compared to the control group). In addition, CsA inhibited the mRNA expression of pro-inflammatory cytokines, such as IL-6 in the uterine body, uterotubal junction, and oviductal isthmus; TNF-α in the uterine body; and IFN-γ in the uterotubal junction and oviductal isthmus induced by the LPS challenge. The IHC assay showed the LPS-induced increase in protein production of IL-6 in the uterine body and oviductal isthmus. CsA increased the protein production of IL-10 in the cervix, uterine body, oviductal ampulla, and isthmus. Moreover, CsA decreased the protein production of IL-6 in the uterine body and oviductal isthmus induced by LPS.

scholarly journals Confocal laser endomicroscopy detects colonic inflammation in patients with irritable bowel syndrome: a prospective study

2020 ◽  
Vol 08 (04) ◽  
pp. E550-E557 ◽  
Author(s):  
Carlos Robles-Medranda ◽  
Roberto Oleas ◽  
Manuel Valero ◽  
Miguel Puga-Tejada ◽  
Miguel Soria-Alcívar ◽  
...  
Keyword(s):  
Irritable Bowel Syndrome ◽  
Histological Evaluation ◽  
Confocal Laser Endomicroscopy ◽  
Confocal Laser ◽  
Control Group ◽  
Tertiary Institution ◽  
Predictive Values ◽  
Colorectal Mucosa ◽  
Irritable Bowel ◽  
Inflammatory Changes

Abstract Background and aims Irritable bowel syndrome (IBS) is considered to be a functional disease, but recent data indicate measurable organic alterations. We aimed to determine the presence of colorectal mucosa microinflammation in vivo via probe-confocal laser endomicroscopy (pCLE) and histological evaluation in IBS patients. Methods This was a prospective, controlled, nonrandomized single-blind diagnostic trial performed in a tertiary institution. pCLE images and targeted biopsy of each colon segment obtained during colonoscopies of IBS patients and controls were analyzed for inflammatory changes. Biopsies were classified using the Geboes scale, and the odds ratio and overall diagnostic accuracy were calculated. Results During the 15-month study period, 37 patients were allocated to each group. The mean age was 53.1 ± 14.3 years; 64.9 % were female. Signs of colonic mucosa inflammation were evident on 65.8 % of pCLE images from IBS patients compared to 23.4 % of images from controls (OR 6.28; 4.14–9.52; P < 0.001). In total, 20/37 patients had microinflammation via pCLE in ≥ 3 colon segments in the IBS group, compared to 1/37 in the control group. A Geboes score > 0 was attributed to 60.8 % of biopsies from patients in the IBS group compared to 27.5 % of biopsies from the control group. The sensitivity, specificity, positive and negative predictive values, observed and interrater agreement of pCLE-detected inflammatory changes in IBS using histology as gold standard were 76 %, 91 %, 76 %, 91 %, 86.5 %, and 66.8 %, respectively. Conclusions Patients with IBS have a six-fold higher prevalence of colorectal mucosa microinflammation than healthy controls. pCLE might be a reliable method to detect colorectal mucosa microinflammation in IBS patients.

Probe-based confocal laser endomicroscopy (pCLE): a preclinical investigation of the male genital tract

2015 ◽  
Vol 31 (1) ◽  
pp. 57-65
Author(s):  
Matthias Trottmann ◽  
Ronald Sroka ◽  
Herbert Stepp ◽  
Bernhard Liedl ◽  
Armin J. Becker ◽  
...  
Keyword(s):  
Genital Tract ◽  
Confocal Laser Endomicroscopy ◽  
Confocal Laser ◽  
Male Genital Tract ◽  
Preclinical Investigation ◽  
Male Genital

Effects of Fluid Therapy on Mesenteric Microcirculation Using New Probe-Based Confocal Laser Endomicroscopy (Cellvizio®) in a Porcine Model of Endotoxic Shock

2021 ◽  
pp. 1-11
Author(s):  
Charlotte Daniere ◽  
Guillaume Louart ◽  
Benjamin Louart ◽  
Marylène Bacle ◽  
Florian Bazalgette ◽  
...  
Keyword(s):  
Fluid Resuscitation ◽  
Fluid Therapy ◽  
Sham Group ◽  
Porcine Model ◽  
Endotoxic Shock ◽  
Confocal Laser Endomicroscopy ◽  
Confocal Laser ◽  
Control Group ◽  
Significant Difference ◽  
Mesenteric Microcirculation

<b><i>Background:</i></b> Microcirculatory alterations have been observed at the early phase of sepsis, although macrocirculation seems preserved. The aim of this study was to analyze the effect of crystalloid fluid therapy on mesenteric microcirculation, assessed by using the confocal laser endomicroscope Cellvizio®, in an endotoxic porcine model. <b><i>Methods:</i></b> It is a prospective endotoxic shock (lipopolysaccharide infusion) experimental trial. Piglets were divided into 3 groups: 6 in the sham group (no LPS injection, no fluid), 9 in the control group (LPS infusion, no fluid), and 6 in the crystalloids group (LPS infusion and fluid resuscitation with crystalloids). Fluid resuscitation consisted in a fluid bolus of 20 mL/kg 0.9% saline over 30 min followed by a 10 mL/kg/h fluid rate over 4 h. Mesenteric microcirculation was assessed using a confocal laser endomicroscope (Cellvizio®). Blood flow within capillaries was visually assessed according to the point of care microcirculation (POEM) score. <b><i>Results:</i></b> At baseline, the 3 groups were similar regarding hemodynamic, biological, and microcirculatory parameters. At T360, the POEM score significantly decreased in the control and crystalloids groups, whereas it remained unchanged in the sham group (respectively, 1.62 ± 1.06, 1.2 ± 0.45, and 5.0 ± 0, <i>p</i> = 0.011). There was no significant difference in cardiac output at T360 between the sham and crystalloids groups (3.1 ± 0.8 vs. 2.3 ± 0.6, <i>p</i> = 0.132) or between the control and crystalloids groups (2.0 ± 0.6 vs. 2.3 ± 0.6, <i>p</i> = 0.90). <b><i>Conclusion:</i></b> There was no significant improvement of microcirculatory alterations after crystalloids resuscitation despite improvement in macrocirculatory parameters in early experimental sepsis.

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