Anatomical Proximity Between Sciatic Nerve and Ischial Spine and its Relationship to the Development of Deep Gluteal Pain Syndrome.

OBJECTIVE: Deep gluteal syndrome (DGS) is a medical diagnosis in which the pathoanatomy of the subgluteal space contributes to pain. The growing recognition that gluteal neuropathies can be associated with the presence of a bone-neural conflict with irritation or compression may allow us to shed some light on this pathology. This study aims to determine whether the location of the sciatic nerve in relation to the ischial spine contributes to the development of DGS. METHODS: The sciatic nerve - ischial spine relationship was analyzed based on magnetic resonance imaging (MRI) in 15 surgical patients who underwent piriformis release, and in 30 control patients who underwent MRI of the pelvis for reasons unrelated to sciatica. The sciatic nerve exit from the greater sciatic foramen was classified as either zone A (medial to the ischial spine); zone B (on the ischial spine); or zone C (lateral to the ischial spine). RESULTS: The sciatic nerve was significantly closer to the ischial spine in surgical patients than in MRI controls (P=0.014). When analyzing patients of similar age, sciatic nerves in surgical patients were significantly closer (P=0.0061) to the ischial spine, and located in zone B significantly more (P=0.0216) as compared to MRI controls. Patients who underwent surgery for piriformis release showed a significant decrease in pain postoperatively (P<0.0001). CONCLUSIONS: The results from this study suggest that the relationship between the ischial spine and sciatic nerve may play a role in the development of DGS. This may also help establish which patients would benefit more from surgical intervention.

Applications of navigation in full-endoscopic spine surgery.

PURPOSE: Advancement in all surgery continues to progress towards more minimally invasive surgical (MIS) approaches. One of the platform technologies which has helped drive this trend within spine surgery is the development of endoscopy; however, the limited anatomic view experienced when performing endoscopic spine surgery requires a significant learning curve. The use of intraoperative navigation has been adapted for endoscopic spine surgery, as this provides computer-reconstructed visual data presented in three dimensions, which can increase feasibility of this technique to more surgeons. METHODS: This paper will describe the principles, technical considerations, and applications of stereotactic navigation-guided endoscopic spine surgery. RESULTS: Full-endoscopic spine surgery has advanced in recent years such that it can be utilized in both decompressive and fusion surgeries. One of the major pitfalls to any minimally invasive surgery (including endoscopic) is that the limited surgical view can often complicate the surgery or confuse the surgeon, leading to longer operative times, higher risks, among others. This is the real utility to using navigation in conjunction with the endoscope-when registered correctly and utilized appropriately, navigated endoscopic spine surgery can take some of the guesswork out of the minimally invasive approach. CONCLUSIONS: Using navigation with endoscopy in spine surgery can potentially expand this technique to surgeons who have yet to master endoscopy as the assistance provided by the navigation can alleviate some of the complexities with anatomic understanding and surgical planning.

The Re-Evaluation of Frailty in Predicting Complications after Long-Segment Spinal Fusion for Adult Spinal Deformity.

OBJECTIVE: To evaluate on a national scale how frailty status (stratified using the 5-item Modified Frailty Index (mFI-5)) affects the operative characteristics of and complications after long-segment spinal fusion (LSF) for adult spinal deformity (ASD). METHODS: Adults undergoing LSF of ≥3 vertebrae in the National Surgical Quality Improvement Program database years 2015-2020 were split into 2 cohorts: nonfrail with mFI = 0 or 1; frail with mFI ≥2. Demographics, operative characteristics, and 30-day complications were contrasted between the cohorts using the Student's t-test, the Fisher's exact test, or a multivariate analysis when appropriate. RESULTS: In the 340 LSF cases collected, 268 fell into the nonfrail cohort and 72 into the frail cohort. The frail cohort constituted a high rate of geriatric age (65.3% vs. 38.1%; P < 0.001), higher body mass index (32.9 ± 0.86 vs. 30.2 ± 0.39; P = 0.005), and more comorbidities in 9 of 14 measures. After surgery, the frail cohort experienced more urinary tract infections (odds ratio [OR], 3.33; confidence interval [CI], 1.01-10.94; P = 0.04). However, the frail cohort shared similarities with the nonfrail cohort in terms of length of stay (5.11 ± 0.51 vs. 6.01 ± 1.62 days; P = 0.60), home discharge (OR, 0.76; CI, 0.42-1.39; P = 0.38), readmission (OR, 2.45; CI, 0.87-6.89; P = 0.09), and overall rate of complications (OR, 0.89; CI, 0.50-1.59; P = 0.70). CONCLUSIONS: Despite trends found in past studies of ASD, this analysis showed that the frailty status of mFI ≥2 is a poor predictor of surgical and hospitalization course and overall complications in LSF when examined up to 30 days postoperatively.

Update on Spinal Fusion.

Spinal fusion is frequently performed for a variety of indications. It is performed to treat instability due to trauma, infection, or neoplasm. It may be used to treat regional or global spinal deformity. There are even occasions when it is appropriate as a treatment of low back pain without overt instability or deformity. One common indication for fusion is as an adjunct to decompression for patients with neurogenic claudication or radiculopathy caused by stenosis associated with spondylolisthesis. There have been a number of high-quality publications in high-quality journals that have reported conflicting results regarding the utility of fusion in this patient population. The existence of conflicting data from seemingly similarly designed trials has resulted in some confusion as to when a fusion should be used. This chapter will describe the controversy, discuss the likely basis for the disparate results reported in the literature, and recommend a reasonable treatment strategy. Going forward, the SLIP II study is an ongoing randomized, controlled trial designed to help clarify the situation. Preliminary findings drawn from this study will be discussed.

Hydrocephalus Following Giant Transosseous Vertex Meningioma Resection.

Introduction Meningiomas are among the most common primary intracranial tumors. While well-described, there is limited information on the outcomes and consequences following treatment of giant-sized vertex-based meningiomas. These meningiomas have specific risks and potential complications due to their size, location, and involvement with extracalvarial soft tissue and dural sinuses. Herein, we present four giant-sized vertex transosseous meningioma cases with involvement and occlusion of the sagittal sinus, that postoperatively developed external hydrocephalus and ultimately required shunting. Methods A retrospective chart review identified patients with large vertex meningiomas that were: (1) large (>6 cm) with hemispheric (no skull base) location, (2) involvement of the superior sagittal sinus resulting in complete sinus occlusion, (3) involvement of dura resulting in a large duraplasty area, (4) transosseous involvement requiring a 5 cm or larger craniectomy for resection of invaded calvarial bone. Results Tumors were resected in all four cases, with all patients subsequently developing external hydrocephalus which required shunting within 2 weeks to 6 months postsurgery. Conclusion We believe this may be the first report of the development of hydrocephalus following surgical resection of these large lesions. Based on our observations, we propose that a combination of superior sagittal sinus occlusion and changes in brain elasticity and compliance affect the brain's CSF absorptive capacity, which ultimately lead to hydrocephalus development. We suggest that neurosurgeons be aware that postoperative hydrocephalus can quickly develop following treatment of giant-sized vertex-based meningiomas, and that correction of hydrocephalus with shunting can readily be achieved.

Upper Thoracic Spine Synovial Cyst Resulting in Paraplegia Following Transient Hypotension.

Development of synovial cysts in the rigid thoracic spine is rare. Additionally, synovial cysts with compression of nerve roots typically cause subacute or chronic radiculopathy. We present a patient who had a new diagnosis of upper thoracic (T1-2) synovial cyst that caused acute paraplegia while hospitalized for therapies and surgical planning. The patient is a 56-year-old male with a history of congestive heart failure secondary to alcoholic cardiomyopathy. He presented with a progressive bilateral lower extremity discoordination, urinary incontinence, and altered perineal sensation. His examination revealed intact strength to bedside assessment, intact rectal tone, but upgoing toes on Babinski testing. Given concern for myelopathy, MRI thoracic spine was obtained and demonstrated large T1-2 synovial cyst causing severe compression with associated T2 signal change within the spinal cord. He underwent expedited cardiac optimization that included resumption of outpatient antihypertensive medications and the addition of a single dose of intravenous diuretic. The patient had subsequent transient hypotension following significant diuresis and developed acute paraplegia in his bilateral lower extremities. Fluids and vasopressors were initiated, and he underwent emergent surgery for decompression and synovial cyst resection. The patient did very well and had normalization of his neurological exam within 24 hours. We present a case of acute paraplegia secondary to hypotension and spinal cord hypoperfusion in a patient with upper thoracic synovial cyst. This is rare pathology with an even more unique presentation. The authors recommend careful perioperative hemodynamic monitoring to help avoid acute worsening in this patient population.

Interobserver variance and patient heterogeneity influencing the treatment of grade I spondylolisthesis.

BACKGROUND CONTEXT: Despite well done randomized clinical trials, the role of fusion as an adjunct to decompression for the treatment of patients with degenerative spondylolisthesis remains controversial. There is substantial variation in the use of fusion as well as the techniques used for fusion for a population of patients all described by a single ICD10 code. PURPOSE: We sought to investigate the source of the variation in the perceived role of fusion by looking at surgeon as well as patient-specific factors. STUDY DESIGN: Prospective cohort study examining the variability of recommendations from an expert panel of surgeons-based imaging and clinical vignettes. PATIENT SAMPLE: Patients with degenerative spondylolisthesis and stenosis. OUTCOME MEASURES: A six-category treatment schema based on level of invasiveness of proposed surgeries with one through three representing nonfusion strategies and categories four through six representing fusion strategies. METHODS: The authors are conducting the ongoing spinal laminectomy vs instrumented pedicle screw II study in which patients with grade one degenerative spondylolisthesis and stenosis are randomized to two groups: a review group in which patients are treated as per recommendations of an expert panel and a nonreview group in which patients are treated as per the referring surgeon's preference. In the former (review group), clinical vignettes and radiographic studies were evaluated by an expert panel of spine surgeons. The panel then provided these recommendations to the referring surgeon. We investigated the underlying variability by looking both at the number of similar or different recommendations received by an individual patient (surgeon-related variability) as well as the number of similar or different recommendations offered by individual surgeons across the population of patients (patient heterogeneity). Agreement between surgeons for fusion vs nonfusion (Categories 1-3 vs 4-6) was calculated using a Kappa value from a mixed effects logistic regression model. We looked at Kappa for agreement and weighted Kappa for association of ratings on the ordinal 1 to 6 scale with a mixed effects linear regression model. Additionally, we analyzed the summary of data between patients after averaging the rater scores within patients. Similarly, we summarized the data between surgeons after averaging their scores over the patients that each surgeon reviewed. RESULTS: One hundred and fourteen patients received 1,463 treatment recommendations. On average, fusion was recommended 58.5% of the time. Overall agreement was low, and perfect agreement on the need for fusion was seen in only 24 (21.1%) of patients. Kappa statistic for agreement on fusion was 0.378 (95% confidence interval 0.324-0.432). The average score across surgeons was 4.2 (0.6) with a range of 3 to 5.3. The most common single recommendation was for fusion with interbody fusion (40.8%) and the lowest was for decompression with noninstrumented fusion (0.5%). CONCLUSIONS: We demonstrated variability in surgical approach when individual patients were evaluated by a panel of surgeons indicating that even "expert" surgeons disagree with each other regarding the need for fusion in individual patients. We were also able to demonstrate that individual patients received consistent recommendations that were very different from those received by other individuals evaluated by the same surgeons. This indicates that there is patient-related heterogeneity driving variability independent of surgeon factors.

Computed Tomography Guidance for Percutaneous Glycerol Rhizotomy for Trigeminal Neuralgia.

BACKGROUND: Percutaneous glycerol rhizotomy (PGR) is a well-described treatment for trigeminal neuralgia; however, the technique in using surface landmarks and fluoroscopy has not drastically changed since being first introduced. In this paper, we describe a protocol for PGR using computed tomography (CT) guidance based on an experience of over 7 yr and 200 patients. OBJECTIVE: To introduce an approach for PGR using CT guidance and, in doing so, demonstrate possible benefits over the traditional fluoroscopic technique. METHODS: Using a standard CT scanner, patients are placed supine with head in extension. Barium paste and a CT scout image are used to identify and plan a trajectory to the foramen ovale. A laser localization system built into the CT scanner helps to guide placement of the spinal needle into the foramen ovale. The needle position in the foramen is confirmed with a short-sequence CT scan. RESULTS: CT-guided PGR provides multiple benefits over standard fluoroscopy, including improved visualization of the skull base and significant reduction in radiation exposure to the surgeon and staff. Side benefits include improved procedure efficiency, definitive imaging evidence of correct needle placement, and potentially increased patient safety. We have had no significant complications in over 200 patients. CONCLUSION: CT-guided PGR is a useful technique for treating trigeminal neuralgia based on better imaging of the skull base, better efficiency of the procedure, and elimination of radiation exposure for the surgeon and staff compared to traditional fluoroscopic based techniques.

Deadly Proliferation and Transformation of Pilocytic Astrocytoma in Pregnancy.

BACKGROUND: Intracranial tumor growth associated with pregnancy is not an uncommon phenomenon. Pilocytic astrocytoma is typically considered to be an indolent tumor with little to no risk of progression to higher-grade lesion. We present a rare case of cerebellar pilocytic astrocytoma transformation to hemorrhagic high-grade glioma during pregnancy. CASE DESCRIPTION: Patient EK was a 32-year-old female with neurofibromatosis type 1 and known cerebellar pilocytic astrocytoma. For nearly a decade before her pregnancy, her cerebellar tumor was stable on imaging. Routine magnetic resonance imaging (MRI) of the head obtained at 20 weeks' gestation continued to demonstrate tumor stability. At 24 weeks' gestation, the patient had sudden, severe headaches. MRI of the head showed evidence of significant tumor expansion. The following day, the patient was found unresponsive. Computed tomography of the head demonstrated hemorrhage within the tumor and tonsillar herniation. Her neurologic examination revealed no brainstem reflexes; however, given her age and pregnancy, she underwent emergent decompression and tumor debulking. Unfortunately, she never improved neurologically. Final pathology identified the lesion as high-grade glioma with anaplastic changes and hemorrhagic conversion. CONCLUSIONS: This is a unique case of indolent cerebellar pilocytic astrocytoma that transformed to high-grade glioma during pregnancy, proven by tumor growth on MRI and anaplasia on pathology. We hypothesize that increased levels of pregnancy hormones (progesterone, vascular endothelial growth factor, placental growth factor, among others) likely contributed to tumor growth. We recommend that all glial tumors be monitored extremely closely throughout pregnancy, and perhaps one should consider surgical treatment (if possible) before patients become pregnant.

Use of a Laryngeal Mask Airway Decreases Radiation Exposure During Computed Tomography-Guided Percutaneous Glycerol Rhizotomy for Trigeminal Neuralgia.

BACKGROUND: We have been using computed tomography (CT) guidance for percutaneous glycerol rhizotomy (PGR) for the last 7 years. As a quality improvement exercise, we recently began using general anesthesia (GA) with the use of a laryngeal mask airway (LMA) because of our perception that the procedure went faster and that there was less radiation exposure because of less patient movement. We aim to compare PGR radiation exposure and procedural time between patients receiving local anesthetic with sedation and those receiving GA/LMA. METHODS: A single-center historical cohort study was performed using patients treated with PGR between 2017 and 2019. Ninety-two surgeries were conducted during the study period: 64 surgeries had local anesthetic with intravenous sedation, and 28 surgeries had deeper anesthetic with LMA. Data analyzed included the number of CT sequences obtained, needle placement time, and total radiation dose. RESULTS: Use of GA/LMA resulted in a 23% decrease in mean radiation dose (565.5 vs. 436.1 µGy × cm, P = 0.014), number of CT sequences required (7.4 vs. 5.7, P = 0.003), and needle placement time (12.8 vs. 9.8 minutes, P = 0.006). Additionally, 10 patients underwent multiple glycerol rhizotomies during the collection period with both anesthetic types being used at least once. Seven of 10 patients (70.0%) had a reduction in total radiation dose, number of CT sequences obtained, and needle placement time when GA/LMA was used. There were no procedure- or anesthetic-related complications in this patient cohort. CONCLUSIONS: The use of GA/LMA during PGR is associated with decreased radiation exposure without increased anesthetic complications.

Clostridioides difficile uses amino acids associated with gut microbial dysbiosis in a subset of patients with diarrhea.

The gut microbiota plays a critical role in pathogen defense. Studies using antibiotic-treated mice reveal mechanisms that increase susceptibility to Clostridioides difficile infection (CDI), but risk factors associated with CDI in humans extend beyond antibiotic use. Here, we studied the dysbiotic gut microbiota of a subset of patients with diarrhea and modeled the gut microbiota of these patients by fecal transplantation into germ-free mice. When challenged with C. difficile, the germ-free mice transplanted with fecal samples from patients with dysbiotic microbial communities showed increased gut amino acid concentrations and greater susceptibility to CDI. A C. difficile mutant that was unable to use proline as an energy source was unable to robustly infect germ-free mice transplanted with a dysbiotic or healthy human gut microbiota. Prophylactic dietary intervention using a low-proline or low-protein diet in germ-free mice colonized by a dysbiotic human gut microbiota resulted in decreased expansion of wild-type C. difficile after challenge, suggesting that amino acid availability might be important for CDI. Furthermore, a prophylactic fecal microbiota transplant in mice with dysbiosis reduced proline availability and protected the mice from CDI. Last, we identified clinical risk factors that could potentially predict gut microbial dysbiosis and thus greater susceptibility to CDI in a retrospective cohort of patients with diarrhea. Identifying at-risk individuals and reducing their susceptibility to CDI through gut microbiota-targeted therapies could be a new approach to preventing C. difficile infection in susceptible patients.

Hypertrophic Olivary Degeneration: Neurosurgical Perspective and Literature Review.

BACKGROUND: Hypertrophic olivary degeneration (HOD) occurs because of posterior fossa or brainstem lesions that disrupt the dentato-rubro-olivary tract, well known as the Guillain-Mollaret triangle. Clinical and radiologic hallmarks of this condition are palatal myoclonus and T2 hyperintensity of the inferior olivary complex on magnetic resonance imaging (MRI), respectively. Because symptomatic HOD can complicate the recovery of patients with posterior fossa or brainstem lesions, the purpose of this study is to evaluate clinical and imaging findings of patients with HOD. METHODS: Sixteen patients (8 female and 8 male) with a mean age of 40.7 years, (range, 5-83 years) years were included in this study based on clinical symptoms and MRI findings. RESULTS: We reviewed the clinical and imaging findings in 16 cases of HOD at our institution. Seven patients (43.7%) had posterior fossa tumors, 6 patients (37.5%) had cavernoma, 2 patients (12.5%) sustained traumatic brain injury, and only 1 patient (6.2%) had cerebellar infarction. Posterior fossa surgery was performed in 13 (81.2%) of these patients. HOD was detected a mean of 7.2 months (range, 0.5-18 months) after surgery or primary neurologic insult. Unilateral HOD was observed in 10 patients (62.5%), while bilateral HOD was observed in only 6 patients (37.5%). Seven patients (43.7%) were asymptomatic for HOD, whereas 5 patients (31.2%) had symptoms attributable to HOD. Two patients died because of primary tumors, although mean follow-up after detection of HOD on MRI was 52.2 months (range, 1-120 months) in the remaining 14 patients. In these cases, no change in clinical symptoms or imaging findings was detected during follow-up. CONCLUSIONS: In this series, posterior fossa tumors and cavernomas were the most common causes of HOD. Although most of the patients with HOD remained asymptomatic, HOD complicated the course of recovery in almost one quarter of the patients included in this study. Neurosurgeons should be aware of HOD, which has characteristic clinical and imaging findings. In addition, HOD can complicate the recovery of patients with disruption to the dentato-rubro-olivary tract.

Homoleptic Trivalent Tris(alkyl) Rare Earth Compounds.

Homoleptic tris(alkyl) rare earth complexes Ln{C(SiHMe2)3}3 (Ln = La, 1a; Ce, 1b; Pr, 1c; Nd, 1d) are synthesized in high yield from LnI3THFn and 3 equiv of KC(SiHMe2)3. X-ray diffraction studies reveal 1a-d are isostructural, pseudo-C3-symmetric molecules that contain two secondary Ln↼HSi interactions per alkyl ligand (six total). Spectroscopic assignments are supported by comparison with Ln{C(SiDMe2)3}3 and DFT calculations. The Ln↼HSi and terminal SiH exchange rapidly on the NMR time scale at room temperature, but the two motifs are resolved at low temperature. Variable-temperature NMR studies provide activation parameters for the exchange process in 1a (ΔH⧧ = 8.2(4) kcal·mol-1; ΔS⧧ = -1(2) cal·mol-1K-1) and 1a-d9 (ΔH⧧ = 7.7(3) kcal·mol-1; ΔS⧧ = -4(2) cal·mol-1K-1). Comparisons of lineshapes, rate constants (kH/kD), and slopes of ln(k/T) vs 1/T plots for 1a and 1a-d9 reveal that an inverse isotope effect dominates at low temperature. DFT calculations identify four low-energy intermediates containing five ß-Si-H⇀Ln and one γ-C-H⇀Ln. The calculations also suggest the pathway for Ln↼HSi/SiH exchange involves rotation of a single C(SiHMe2)3 ligand that is coordinated to the Ln center through the Ln-C bond and one secondary interaction. These robust organometallic compounds persist in solution and in the solid state up to 80 °C, providing potential for their use in a range of synthetic applications. For example, reactions of Ln{C(SiHMe2)3}3 and ancillary proligands, such as bis-1,1-(4,4-dimethyl-2-oxazolinyl)ethane (HMeC(OxMe2)2) give {MeC(OxMe2)2}Ln{C(SiHMe2)3}2, and reactions with disilazanes provide solvent-free lanthanoid tris(disilazides).

Changes in microbial ecology after fecal microbiota transplantation for recurrent C. difficile infection affected by underlying inflammatory bowel disease.

BACKGROUND: Gut microbiota play a key role in maintaining homeostasis in the human gut. Alterations in the gut microbial ecosystem predispose to Clostridium difficile infection (CDI) and gut inflammatory disorders such as inflammatory bowel disease (IBD). Fecal microbiota transplantation (FMT) from a healthy donor can restore gut microbial diversity and pathogen colonization resistance; consequently, it is now being investigated for its ability to improve inflammatory gut conditions such as IBD. In this study, we investigated changes in gut microbiota following FMT in 38 patients with CDI with or without underlying IBD. RESULTS: There was a significant change in gut microbial composition towards the donor microbiota and an overall increase in microbial diversity consistent with previous studies after FMT. FMT was successful in treating CDI using a diverse set of donors, and varying degrees of donor stool engraftment suggesting that donor type and degree of engraftment are not drivers of a successful FMT treatment of CDI. However, patients with underlying IBD experienced an increased number of CDI relapses (during a 24-month follow-up) and a decreased growth of new taxa, as compared to the subjects without IBD. Moreover, the need for IBD therapy did not change following FMT. These results underscore the importance of the existing gut microbial landscape as a decisive factor to successfully treat CDI and potentially for improvement of the underlying pathophysiology in IBD. CONCLUSIONS: FMT leads to a significant change in microbial diversity in patients with recurrent CDI and complete resolution of symptoms. Stool donor type (related or unrelated) and degree of engraftment are not the key for successful treatment of CDI by FMT. However, CDI patients with IBD have higher proportion of the original community after FMT and lack of improvement of their IBD symptoms and increased episodes of CDI on long-term follow-up.

Human-derived gut microbiota modulates colonic secretion in mice by regulating 5-HT3 receptor expression via acetate production.

Serotonin [5-hydroxytryptamine (5-HT)], an important neurotransmitter and a paracrine messenger in the gastrointestinal tract, regulates intestinal secretion by its action primarily on 5-HT3 and 5-HT4 receptors. Recent studies highlight the role of gut microbiota in 5-HT biosynthesis. In this study, we determine whether human-derived gut microbiota affects host secretory response to 5-HT and 5-HT receptor expression. We used proximal colonic mucosa-submucosa preparation from age-matched Swiss Webster germ-free (GF) and humanized (HM; ex-GF colonized with human gut microbiota) mice. 5-HT evoked a significantly greater increase in short-circuit current (ΔIsc) in GF compared with HM mice. Additionally, 5-HT3 receptor mRNA and protein expression was significantly higher in GF compared with HM mice. Ondansetron, a 5-HT3 receptor antagonist, inhibited 5-HT-evoked ΔIsc in GF mice but not in HM mice. Furthermore, a 5-HT3 receptor-selective agonist, 2-methyl-5-hydroxytryptamine hydrochloride, evoked a significantly higher ΔIsc in GF compared with HM mice. Immunohistochemistry in 5-HT3A-green fluorescent protein mice localized 5-HT3 receptor expression to enterochromaffin cells in addition to nerve fibers. The significant difference in 5-HT-evoked ΔIsc between GF and HM mice persisted in the presence of tetrodotoxin (TTX) but was lost after ondansetron application in the presence of TTX. Application of acetate (10 mM) significantly lowered 5-HT3 receptor mRNA in GF mouse colonoids. We conclude that host secretory response to 5-HT may be modulated by gut microbiota regulation of 5-HT3 receptor expression via acetate production. Epithelial 5-HT3 receptor may function as a mediator of gut microbiota-driven change in intestinal secretion.NEW & NOTEWORTHY We found that gut microbiota alters serotonin (5-HT)-evoked intestinal secretion in a 5-HT3 receptor-dependent mechanism and gut microbiota metabolite acetate alters 5-HT3 receptor expression in colonoids.View this article's corresponding video summary at https://www.youtube.com/watch?v=aOMYJMuLTcw&feature=youtu.be.

Gut microbiome predictors of treatment response and recurrence in primary Clostridium difficile infection.

BACKGROUND: Clostridium difficile infection (CDI) may not respond to initial therapy and frequently recurs, but predictors of response and recurrence are inconsistent. The impact of specific alterations in the gut microbiota determining treatment response and recurrence in patients with CDI is unknown. AIM: To assess microbial signatures as predictors of treatment response and recurrence in CDI. METHODS: Pre-treatment stool samples and clinical metadata including outcomes were collected prospectively from patients with their first CDI episode. Next generation 16s rRNA sequencing using MiSeq Illumina platform was performed and changes in microbial community structure were correlated with CDI outcomes. RESULTS: Eighty-eight patients (median age 52.7 years, 60.2% female) were included. Treatment failure occurred in 12.5% and recurrence after response in 28.5%. Patients who responded to treatment had an increase in Ruminococcaceae, Rikenellaceae, Clostridiaceae, Bacteroides, Faecalibacterium and Rothia compared to nonresponders. A risk-index built from this panel of microbes differentiated responders (mean 0.07 ± 0.24) from nonresponders (0.52 ± 0.42; P = 0.0002). Receiver operating characteristic (ROC) curve demonstrated that risk-index was a strong predictor of treatment response with an area under the curve (AUC) of 0.85. Among clinical parameters tested, only proton pump inhibitor use predicted recurrent CDI (OR 3.75, 95% CI 1.27-11.1, P = 0.01). Patients with recurrent CDI had statistically significant increases in Veillonella, Enterobacteriaceae, Streptococci, Parabacteroides and Lachnospiraceae compared to patients without recurrence and a risk index was able to predict recurrence (AUC = 0.78). CONCLUSION: Gut microbiota signatures predict treatment response and recurrence potentially, allowing identification of patients with Clostridium difficile infection that may benefit from early institution of alternate therapies.

Netrin-1 Peptide Is a Chemorepellent in Tetrahymena thermophila.

Netrin-1 is a highly conserved, pleiotropic signaling molecule that can serve as a neuronal chemorepellent during vertebrate development. In vertebrates, chemorepellent signaling is mediated through the tyrosine kinase, src-1, and the tyrosine phosphatase, shp-2. Tetrahymena thermophila has been used as a model system for chemorepellent signaling because its avoidance response is easily characterized under a light microscope. Our experiments showed that netrin-1 peptide is a chemorepellent in T. thermophila at micromolar concentrations. T. thermophila adapts to netrin-1 over a time course of about 10 minutes. Netrin-adapted cells still avoid GTP, PACAP-38, and nociceptin, suggesting that netrin does not use the same signaling machinery as any of these other repellents. Avoidance of netrin-1 peptide was effectively eliminated by the addition of the tyrosine kinase inhibitor, genistein, to the assay buffer; however, immunostaining using an anti-phosphotyrosine antibody showed similar fluorescence levels in control and netrin-1 exposed cells, suggesting that tyrosine phosphorylation is not required for signaling to occur. In addition, ELISA indicates that a netrin-like peptide is present in both whole cell extract and secreted protein obtained from Tetrahymena thermophila. Further study will be required in order to fully elucidate the signaling mechanism of netrin-1 peptide in this organism.

Error-free transmission of microring-modulated BPSK.

We demonstrate the generation of error-free binary-phase-shift-keyed (BPSK) data at 5 Gb/s using a silicon microring modulator. The microring-modulated BPSK signal is propagated at fiber lengths up to 80 km, maintaining error-free performance, while demonstrating resilience to chromatic dispersion. Bit-error-rate measurements and eye diagrams show near equivalent performance of a microring-based BPSK modulator as compared to commercial LiNbO3 phase modulators.

In vivo imaging of ligand receptor binding with Gaussia luciferase complementation.

Studies of ligand-receptor binding and the development of receptor antagonists would benefit greatly from imaging techniques that translate directly from cell-based assays to living animals. We used Gaussia luciferase protein fragment complementation to quantify the binding of chemokine (C-X-C motif) ligand 12 (CXCL12) to chemokine (C-X-C motif) receptor 4 (CXCR4) and CXCR7. Studies established that small-molecule inhibitors of CXCR4 or CXCR7 specifically blocked CXCL12 binding in cell-based assays and revealed differences in kinetics of inhibiting chemokine binding to each receptor. Bioluminescence imaging showed CXCL12-CXCR7 binding in primary and metastatic tumors in a mouse model of breast cancer. We used this imaging technique to quantify drug-mediated inhibition of CXCL12-CXCR4 binding in living mice. We expect this imaging technology to advance research in areas such as ligand-receptor interactions and the development of new therapeutic agents in cell-based assays and small animals.

Noninvasive imaging reveals inhibition of ovarian cancer by targeting CXCL12-CXCR4.

Patients with metastatic ovarian cancer continue to have a dismal prognosis, emphasizing the need for new strategies to identify and develop new molecular targets for therapy. Chemokine CXCL12 and its receptor CXCR4 are upregulated in metastatic ovarian cancer cells and the intraperitoneal tumor microenvironment. CXCL12-CXCR4 signaling promotes multiple steps in proliferation and dissemination of ovarian cancer cells, suggesting that targeted inhibition of this pathway will limit tumor progression. To investigate CXCL12-CXCR4 signaling in ovarian cancer and establish effects of inhibiting this pathway on tumor progression and survival, we designed a Gaussia luciferase complementation imaging reporter system to detect CXCL12 binding to CXCR4 in ovarian cancer cells. In cell-based assays, we established that the complementation imaging reporter could detect CXCL12 binding to CXCR4 and quantify specific inhibition of ligand-receptor interaction. We monitored CXCL12-CXCR4 binding and inhibition in a mouse xenograft model of metastatic human ovarian cancer by imaging Gaussia luciferase complementation and assessed tumor progression with firefly luciferase. Bioluminescence imaging studies in living mice showed that treatment with AMD3100, a clinically approved inhibitor of CXCL12-CXCR4, blocked ligand-receptor binding and reduced growth of ovarian cancer cells. Treatment with AMD3100 also modestly improved overall survival of mice with metastatic ovarian cancer. The Gaussia luciferase complementation imaging reporter system will facilitate further preclinical development and optimization of CXCL12-CXCR4 targeted compounds for treatment of ovarian cancer. Our research supports clinical translation of existing CXCR4 inhibitors for molecular therapy for ovarian cancer.