A tool for mapping microglial morphology, morphOMICs, reveals brain-region and sex-dependent phenotypes.

Environmental cues influence the highly dynamic morphology of microglia. Strategies to characterize these changes usually involve user-selected morphometric features, which preclude the identification of a spectrum of context-dependent morphological phenotypes. Here we develop MorphOMICs, a topological data analysis approach, which enables semiautomatic mapping of microglial morphology into an atlas of cue-dependent phenotypes and overcomes feature-selection biases and biological variability. We extract spatially heterogeneous and sexually dimorphic morphological phenotypes for seven adult mouse brain regions. This sex-specific phenotype declines with maturation but increases over the disease trajectories in two neurodegeneration mouse models, with females showing a faster morphological shift in affected brain regions. Remarkably, microglia morphologies reflect an adaptation upon repeated exposure to ketamine anesthesia and do not recover to control morphologies. Finally, we demonstrate that both long primary processes and short terminal processes provide distinct insights to morphological phenotypes. MorphOMICs opens a new perspective to characterize microglial morphology.

A systematic characterization of microglia-like cell occurrence during retinal organoid differentiation.

Cerebral organoids differentiated from human-induced pluripotent stem cells (hiPSC) provide a unique opportunity to investigate brain development. However, organoids usually lack microglia, brain-resident immune cells, which are present in the early embryonic brain and participate in neuronal circuit development. Here, we find IBA1+ microglia-like cells alongside retinal cups between week 3 and 4 in 2.5D culture with an unguided retinal organoid differentiation protocol. Microglia do not infiltrate the neuroectoderm and instead enrich within non-pigmented, 3D-cystic compartments that develop in parallel to the 3D-retinal organoids. When we guide the retinal organoid differentiation with low-dosed BMP4, we prevent cup development and enhance microglia and 3D-cysts formation. Mass spectrometry identifies these 3D-cysts to express mesenchymal and epithelial markers. We confirmed this microglia-preferred environment also within the unguided protocol, providing insight into microglial behavior and migration and offer a model to study how they enter and distribute within the human brain.

Microglia enable mature perineuronal nets disassembly upon anesthetic ketamine exposure or 60-Hz light entrainment in the healthy brain.

Perineuronal nets (PNNs), components of the extracellular matrix, preferentially coat parvalbumin-positive interneurons and constrain critical-period plasticity in the adult cerebral cortex. Current strategies to remove PNN are long-lasting, invasive, and trigger neuropsychiatric symptoms. Here, we apply repeated anesthetic ketamine as a method with minimal behavioral effect. We find that this paradigm strongly reduces PNN coating in the healthy adult brain and promotes juvenile-like plasticity. Microglia are critically involved in PNN loss because they engage with parvalbumin-positive neurons in their defined cortical layer. We identify external 60-Hz light-flickering entrainment to recapitulate microglia-mediated PNN removal. Importantly, 40-Hz frequency, which is known to remove amyloid plaques, does not induce PNN loss, suggesting microglia might functionally tune to distinct brain frequencies. Thus, our 60-Hz light-entrainment strategy provides an alternative form of PNN intervention in the healthy adult brain.

Two-phase (low-dose) computed tomography is as effective as 4D-CT for identifying enlarged parathyroid glands.

BACKGROUND: Four-dimensional computed tomography (4D-CT) is often used for patients with primary hyperparathyroidism and non-definitive localization after Sestamibi scan (MIBI) and ultrasound (US), but may expose patients to unnecessary radiation, typically between 10 and 26 millisieverts (mSv). We hypothesize that a simpler two-phase CT protocol would have a similar sensitivity, specificity and accuracy to those published for 4D-CT, while exposing the patient to less radiation. METHODS: We reviewed 54 patients with primary hyperparathyroidism and non-definitive localization studies who had a two-phase CT between 2009 and 2012 at our tertiary referral center. RESULTS: The mean radiation dose of two-phase CT over the course of the study was 5.2 mSv (range 3.5 mSv-9.1 mSv). Two-phase CT had a 77% (CI = 65%-86%) sensitivity and an 87% (CI = 73%-95%) specificity to lateralize enlarged parathyroid glands to the correct side of the neck and a 58% (CI = 45%-68%) sensitivity and 91% (CI = 83%-94%) specificity to localize parathyroid tumors to the correct quadrant of the neck. The overall accuracy of two-phase CT to lateralize enlarged parathyroids was 81% (CI = 73%, 88%) and the accuracy to localize enlarged parathyroids was 79% (CI = 73%, 84%). DISCUSSION: As a second line investigation two-phase CT has a similar sensitivity, specificity and accuracy to those published for 4D-CT in patients with non-localized, enlarged parathyroids with less radiation exposure. Two-phase CT can help localize enlarged parathyroid glands not definitively identified using MIBI and US. CONCLUSION: Two-phase CT allows clinicians to accurately identify enlarged parathyroid glands while exposing the patient to less radiation than 4D-CT.

Extended neoadjuvant chemotherapy for borderline resectable pancreatic cancer demonstrates promising postoperative outcomes and survival.

BACKGROUND: The optimum approach to neoadjuvant therapy for patients with borderline resectable pancreatic cancer is undefined. Herein we report the outcomes of an extended neoadjuvant chemotherapy regimen in patients presenting with borderline resectable adenocarcinoma of the pancreatic head. METHODS: Patients identified as having borderline resectable pancreatic head cancer by American Hepato-Pancreato-Biliary Association/Society of Surgical Oncology consensus criteria from 2008 to 2012 were tracked in a prospectively maintained registry. Included patients were initiated on a 24-week course of neoadjuvant chemotherapy. Medically fit patients who completed neoadjuvant treatment without radiographic progression were offered resection with curative intent. Clinicopathologic variables and surgical outcomes were collected retrospectively and analyzed. RESULTS: Sixty-four patients with borderline resectable pancreatic cancer started neoadjuvant therapy. Thirty-nine (61 %) met resection criteria and underwent operative exploration with curative intent, and 31 (48 %) were resected. Of the resected patients, 18 (58 %) had positive lymph nodes, 15 (48 %) required en-bloc venous resection, 27 (87 %) had a R0 resection, and 3 (10 %) had a complete pathologic response. There were no postoperative deaths at 90 days, 16 % of patients had a severe complication, and the 30-day readmission rate was 10 %. The median overall survival of all 64 patients was 23.6 months, whereas that of unresectable patients was 15.4 months. Twenty-five of the resected patients (81 %) are still alive at a median follow-up of 21.6 months. CONCLUSIONS: Extended neoadjuvant chemotherapy is well tolerated by patients with borderline resectable pancreatic head adenocarcinoma, selects a subset of patients for curative surgery with low perioperative morbidity, and is associated with favorable survival.

Use the duodenum, it's right there: a retrospective cohort study comparing biliary reconstruction using either the jejunum or the duodenum.

IMPORTANCE: This is the largest series to date comparing end-to-side biliary reconstruction for all indications performed using either the duodenum or jejunum and with at least 2-year follow-up. OBJECTIVE: To demonstrate that duodenal anastomoses for biliary reconstruction are at least as safe and effective as Roux-en-Y jejunal anastomoses, with the benefits of operative simplicity and ease of postoperative endoscopic evaluation. DESIGN, SETTING, AND PARTICIPANTS: Retrospective record review with telephone survey of patients undergoing nonpalliative biliary reconstruction in the hepatopancreatobiliary surgery division of a high-volume tertiary care facility. INTERVENTIONS: Biliary reconstruction via either end-to-side Roux-en-Y jejunal anastomosis or direct duodenal anastomosis. MAIN OUTCOMES AND MEASURES: The primary end points were anastomosis-related complications (leak, cholangitis, bile gastritis, or stricture), and the secondary end points were overall complications, endoscopic or radiologic interventions, readmissions, and death. RESULTS: Ninety-six nonpalliative biliary reconstructions were performed between February 1, 2000, and November 23, 2011 for bile duct injury, cholangiocarcinoma, choledochal cysts, or benign strictures; the procedures included 59 duodenal reconstructions and 37 Roux-en-Y jejunal reconstructions. The groups were similar with regard to demographics, operative indications, postoperative length of stay, and mortality rates. However, anastomosis-related complications (leaks, cholangitis, or strictures) were fewer in the duodenal than the jejunal cohort (7 patients [12%] vs 13 [35%]; P = .009). Of patients with stricture, 5 of 9 in the jejunal cohort required percutaneous transhepatic access for management compared with only 1 of 2 in the duodenal cohort. CONCLUSIONS AND RELEVANCE: Duodenal anastomosis is a safe, simple, and often preferable method for biliary reconstruction. This anastomosis can successfully be performed to all levels of the biliary tree with low rates of leak, stricture, cholangitis, and bile gastritis. When anastomotic complications do occur, there is less need for transhepatic intervention because of easier endoscopic access.

One-gland exploration for mediastinal parathyroid adenomas: cervical and thoracoscopic approaches.

BACKGROUND: In patients with sporadic primary hyperparathyroidism, preoperative localization studies may discover a solitary mediastinal parathyroid adenoma. In this circumstance a 1-gland mediastinal exploration, either cervical or thoracoscopic, may be curative. METHODS: In an 18-month period, 5 of 120 consecutive patients underwent an initial 1-gland mediastinal exploration for a solitary mediastinal parathyroid adenoma and 2 patients had a 1-gland mediastinal exploration for persistent hyperparathyroidism. Clinical presentation, imaging studies, surgical techniques, and outcomes were reviewed. RESULTS: Sestamibi scans showed a mediastinal parathyroid adenoma in all 7 patients. Computed tomography provided anatomic localization of middle mediastinal parathyroid adenomas. A cervical approach was used in 4 patients who had a superior mediastinal parathyroid adenoma. Thoracoscopic excision was performed in 3 patients with a middle mediastinal parathyroid adenoma. No complications occurred. Calcium and parathyroid hormone levels normalized in all patients. CONCLUSIONS: Sporadic primary hyperparathyroidism caused by a solitary mediastinal parathyroid adenoma can be treated successfully with 1-gland mediastinal exploration either by a cervical or a thoracoscopic approach as indicated by localization imaging.

Maximizing outcomes while minimizing exploration in hyperparathyroidism using localization tests.

HYPOTHESIS: Preoperative localization (ultrasonography and scintigraphy) can be used to limit operative exploration in primary hyperparathyroidism while providing a high rate of success. DESIGN: Prospective cohort analysis of 3 types of exploration (1-gland, unilateral, or 4-gland), as directed by localization. RESULTS: In 185 consecutive patients who underwent operations, the final diagnoses were solitary adenoma in 87% and multigland disease in 13%. Ultrasonography (75%) and scintigraphy (83%) demonstrated an enlarged parathyroid gland and, together with operative findings, resulted in 61 1-gland, 63 unilateral, and 61 4-gland explorations, with an initial success rate of 96% and an ultimate success rate of 99%. Limiting exploration resulted in a significant decrease in operative time and hospitalization. CONCLUSION: Localization can limit exploration with success.

Outcomes after resection of cholangiocellular carcinoma.

BACKGROUND: Cholangiocellular carcinoma (CCC) is a rare primary liver malignancy that arises from intrahepatic bile duct canaliculi and presents as a liver mass. Our purpose is to report operative morbidity and mortality and to determine long-term survival after resection for CCC. METHODS: Retrospective review of 31 consecutive patients who underwent resection during a 20-year period. RESULTS: Thirty-day hospital mortality was 3%, and postoperative morbidity was 38%. Kaplan-Meier 5-year survival was 35%; mean survival was 37 months; absolute 5-year survival was 33%. Mean survival in stages I, II, IIIA, and IIIC were 57, 33, 26, and 14 months, respectively (P = 0.03 comparing I to >I). Recurrence occurred in 18 patients; 89% were in the liver. Carbohydrate antigen 19-9 >100 U/mL was found to be an indicator of poor prognosis (P = 0.009). CONCLUSIONS: Resection for CCC can be performed with acceptable morbidity and mortality rates and results in good survival and cure. Hepatic recurrence is common. Carbohydrate antigen 19-9 may be useful in determining prognosis.

Surgical treatment of hyperparathyroidism improves health-related quality of life.

HYPOTHESIS: The surgical treatment of primary hyperparathyroidism results in an improved health-related quality of life. DESIGN: Prospective cohort analysis of consecutive patients with primary hyperparathyroidism analyzed preoperatively and 1 year postoperatively. SETTING: Academic multispecialty referral clinic. PATIENTS: We prospectively evaluated 74 consecutive patients who underwent parathyroid exploration for primary hyperparathyroidism during a 15-month period. INTERVENTIONS: The Medical Outcomes Study Short-Form Health Survey (SF-36) was administered before consultation with a surgeon. Patients were categorized based on reason for referral as either asymptomatic (group 1; n = 43) or symptomatic (group 2; n = 29). All patients underwent parathyroid exploration and normalization of calcium levels postoperatively. The SF-36 was then re-administered after 1 year. MAIN OUTCOME MEASURES: Statistical analysis of preoperative and postoperative SF-36 scores, and comparisons with national norms. RESULTS: The SF-36 was completed preoperatively and 1 year postoperatively by 72 (97%) of 74 patients. When the results were compared with published national norms, the preoperative population was significantly impaired in 5 of 8 domains, whereas the postoperative one had improved and was nearly indistinguishable from the norm. In 7 of 8 domains, the postoperative scores were significantly improved compared with preoperative scores. Group 1 patients showed significant preoperative impairment in 3 domains and significantly improved in 2, whereas group 2 patients showed significant impairment and improvement in 7 domains. CONCLUSION: The surgical treatment of primary hyperparathyroidism is associated with durable, statistically significant improvements in health-related quality of life.

Long-term biliary function after reconstruction of major bile duct injuries with hepaticoduodenostomy or hepaticojejunostomy.

HYPOTHESIS: Normal biliary function can be achieved after reconstruction for major bile duct injuries using either hepaticoduodenostomy (HD) or Roux-en-Y hepaticojejunostomy (HJ). DESIGN: Retrospective analysis of consecutive patients requiring biliary enteric reconstructions from February 1, 1993, through January 1, 2002, for bile duct injuries. SETTING: Academic multispecialty referral clinic. PATIENTS: Twenty-seven consecutive patients were evaluated who underwent biliary enteric reconstruction for bile duct injury caused during cholecystectomy. Patients were reconstructed either by HD (18 patients) or HJ (9 patients). INTERVENTIONS: Patients' medical records were reviewed and long-term evaluations were obtained via telephone questionnaire by 2 separate observers (R.J.M. and F.T.L.). Biliary function was evaluated in all using symptoms and liver function test results. Cholangiography was obtained, if indicated clinically. These were reviewed for stricture or dilatation. Any biliary interventions were recorded. MAIN OUTCOME MEASURES: Comparison of long-term biliary function after HD vs HJ reconstructions. RESULTS: All patients were contacted after a median postoperative time of 54 months. Excellent or good results were observed for biliary function in 25 (92%) of the 27 patients. These results were obtained regardless of the type of reconstruction-HD (18 patients) or HJ (9 patients). CONCLUSIONS: We found biliary function to be normal at more than 4 years after biliary-enteric reconstruction for bile duct injury. When surgically feasible, we prefer HD to HJ.