A Novel Approach Towards Less Invasive Multi ‘Omics Gut Analyses: A Pilot Study

Newer ‘omics approaches such as metatranscriptomics and metabolomics allow functional assessments of the interaction(s) between the gut microbiome and the human host. In order to generate meaningful data with these approaches, though, the method of sample collection is critical. Prior studies have relied upon expensive and invasive means towards sample acquisition such as intestinal biopsy, while other studies have relied upon easier methods of collection such as fecal samples that do not necessarily represent those microbes in contact with the host. In this pilot study, we attempt to characterize a novel, minimally invasive method towards sampling the human microbiome using mucosal cytology brush sampling compared to intestinal gut biopsy on 5 healthy participants undergoing routine screening colonoscopy. We compared metatranscriptomic analyses between the two collection methods, identifying increased taxonomic evenness and beta diversity in the cytology brush samples, and similar community transcriptional profiles between the two methods. Metabolomics assessment demonstrated striking differences between the two methods, implying a difference in bacterial-derived versus human absorbed metabolites. Put together, this study supports the use of a less invasive method of microbiome sampling with cytology brushes, but caution must be exercised when performing metabolomics assessment as this represents differential metabolite production but not absorption by the host.

PSV-4 Peri-estrus ovarian, uterine, and hormonal variables influence the uterine luminal fluid metabolome in beef heifers

In cattle, uterine luminal fluid (ULF) is the main source of molecules that support embryo development and survival during the peri-implantation period. Overarching hypothesis was that peri-estrus changes in ULF volume through accumulation and resorption mechanisms influence ULF composition during the estrous cycle and early pregnancy. Objectives were (1) to characterize individual temporal and spatial changes in ULF volume, endometrial and luteal vascularity, endometrial and luteal size, and progesterone (P4) concentrations during the peri-estrus period in beef heifers and, (2) associate such changes with the metabolite composition in the ULF, four days after estrus. Fourteen Bos indicus heifers that presented a PGF2α responsive CL received 500 µg PGF2α analog i.m. and were examined daily by rectal B-mode and pulse-wave color-Doppler ultrasonography until the fifth day after estrus (estrus = d 0). Plasma P4 was measured daily. On d 4, the uterine body was sampled using a cytology brush for targeted metabolomic analysis by mass spectrometry. Multivariate analyses clustered heifers according to ovarian, uterine, and hormonal variables in clusters A (n = 5) and B (n = 8 heifers). Individual metabolite concentrations were compared between clusters A and B by univariate analysis using t-test after FDR correction. Concentrations of Pro, Ala, Leu, Gly, Val, Lys, Ile, Phe, Asp, Orn, Tyr, Arg, Trp, Suc, Cit, ADMA, the sum of essential Amino Acids (AA), sum of non-essential AA, sum of aromatic AA, and total AA were greater in cluster A (FDR ≤ 0.05). ULF volume dynamics and associated uterine, ovarian, and hormonal variables during the peri-estrus period presented a concerted variation among heifers, which was associated with the ULF composition four days after estrus. Potential implications for embryo receptivity and reproductive outcomes are the focus of the current investigation.

Endoscopic and fluoroscopic-guided closure of the eustachian tube using a biliary cytology brush and liquid embolic agent for a persistent CSF leak after schwannoma resection

Vestibular schwannoma is a known cause of progressive sensorineural hearing loss. Treatment options include observation, radiation therapy and surgical resection. Cerebrospinal fluid (CSF) fistula is a known postsurgical complication that can lead to CSF otorrhoea, rhinorrhoea or CSF leakage from the surgical wound. We present a case report of a patient who underwent vestibular schwannoma resection and postoperatively developed CSF rhinorrhoea, which was refractory to multiple attempts at surgical repair. This was successfully treated under endoscopic and fluoroscopic guidance using a biliary cytology brush to disrupt the surface of the eustachian tube followed by injection of n-Butyl cyanoacrylate.

Comparison of the Cytology Brush and Low Volume Lavage Techniques to Evaluate Endometrial Cytology for Diagnosing Endometritis in Chronically Infertile Mares

Endometritis is the most common cause of subfertility and infertility in mares that leads to great economical loss. Endometrial cytology (EC) is a reliable diagnostic tool, widely used to assess the health of the reproductive tract in both humans and domestic mammals. The aim of this study was to compare two techniques for the collection of EC samples in chronically infertile mares. Ten mares aged 4 to 20 were used. In each mare, samples for EC were collected first by cytology brush (CB) and then by low volume lavage (LVL) and stained using the Diff Quick stain. Each sample was assessed for the following parameters: background of the slides, quality and quantity of the cells obtained, number of inflammatory cells. Also, we assessed the number of operators, time and costs needed for each technique. More debris was noticed in the LVL samples, as well as a higher percentage of distorted cells. However, inflammatory cells were detected in all positive samples. Sampling by LVL required two to three operators and recovery of the lavage fluid was sometimes difficult. In conclusion, both techniques are accurate for the detection of uterine inflammation.