Full-length 16S rRNA gene amplicon analysis of human gut microbiota using MinION™ nanopore sequencing confers species-level resolution.

BACKGROUND: Species-level genetic characterization of complex bacterial communities has important clinical applications in both diagnosis and treatment. Amplicon sequencing of the 16S ribosomal RNA (rRNA) gene has proven to be a powerful strategy for the taxonomic classification of bacteria. This study aims to improve the method for full-length 16S rRNA gene analysis using the nanopore long-read sequencer MinION™. We compared it to the conventional short-read sequencing method in both a mock bacterial community and human fecal samples. RESULTS: We modified our existing protocol for full-length 16S rRNA gene amplicon sequencing by MinION™. A new strategy for library construction with an optimized primer set overcame PCR-associated bias and enabled taxonomic classification across a broad range of bacterial species. We compared the performance of full-length and short-read 16S rRNA gene amplicon sequencing for the characterization of human gut microbiota with a complex bacterial composition. The relative abundance of dominant bacterial genera was highly similar between full-length and short-read sequencing. At the species level, MinION™ long-read sequencing had better resolution for discriminating between members of particular taxa such as Bifidobacterium, allowing an accurate representation of the sample bacterial composition. CONCLUSIONS: Our present microbiome study, comparing the discriminatory power of full-length and short-read sequencing, clearly illustrated the analytical advantage of sequencing the full-length 16S rRNA gene.

Lower Cervical Dural Arteriovenous Fistula with a "Skip Lesion" in the Brainstem: A Case Report.

Spinal dural arteriovenous fistulas (SDAVFs) are rare vascular malformations that can occur anywhere in the spine. Most SDAVFs lead to slow aggressive myelopathy due to venous congestion at a level adjacent to the shunt point. However, rare cases of localized brainstem edema without spinal cord lesions have been reported. In this study, we present a case of a lower cervical SDAVF that showed localized congestive edema of the medulla in the absence of an edematous change in the cervical spinal cord. The patient was a 57-year-old woman who experienced vertigo and vomiting without myelopathy that did not improve with conservative treatment. Magnetic resonance imaging (MRI) revealed high signal intensity in the left medulla on T2-weighted imaging (T2WI), while angiography revealed an SDAVF at the right C8 segmental level supplied by the right thyrocervical trunk. She underwent surgical interruption of the draining vein, which led to a rapid improvement in her symptoms. A subsequent follow-up MRI confirmed resolution of both the medullary edema and the dilated draining vein. SDAVFs may cause vertigo and vomiting, which are brainstem symptoms. Early diagnosis and surgical intervention are crucial for successful treatment outcomes.

Preventing Post-incisional Dural Shrink in Craniotomy: Introducing the "Roll-up Technique".

Dural dryness makes suturing difficult during dural closure after craniotomy. In this case, dural plasty is often performed using a membrane taken from the surrounding tissue (e.g., fascia or periosteum) or an artificial replacement membrane. Herein, we introduce our novel "roll-up technique" to reduce the utilization of substitute membranes and explore its effectiveness in dural closure. We retrospectively examined the medical records of 50 patients who underwent craniotomy for the first time for supratentorial intracranial lesions between 2015 and 2022. Furthermore, we divided them into two groups: (1) the conventional technique group, which consisted of patients in whom the dura mater was flipped after incision and protected with a moistened gauze (n = 23), and (2) the roll-up technique group, which consisted of patients in whom the dura mater was incised in a U shape, rolled up, and protected with a moist gauze (n = 27). After surgery, we compared the success rates of primary closure, operating time, craniotomy area, and percentage of complications (e.g., cerebrospinal fluid [CSF] leakage or infection) between the groups. Dural closure without dural substitutes using the roll-up technique had a higher success rate than that using the conventional technique (26/27 [96.3%] cases vs. 14/23 [60.9%] cases; P = 0.003). Postoperative CSF leakage or infection did not occur, and no statistically significant difference was observed in the operating time between the groups (P = 0.247). The roll-up technique for dural closure may effectively prevent post-incisional dural shrink after craniotomy.