Retrosigmoid Craniectomy for Clip Ligation of Flow-Diverted Posterior Inferior Cerebellar Artery Aneurysm.

Flow diversion (FD) has revolutionized the treatment of cerebral aneurysms. Since the introduction of the Pipeline Embolization Device, there has been a significant shift in the management of cerebral aneurysms, with increasing emphasis being placed on use of endoluminal reconstruction as a means of long-term, durable treatment of aneurysms. Increasingly, FD stents are being used as primary treatment for aneurysms, including those that present with subarachnoid hemorrhage.1 Improper use of FD stents, however, may create havoc, as access to the aneurysm sac is blocked with the placement of these devices. Aneurysms that are incompletely treated with FD may continue to grow and rupture. The inability to use coils or endosaccular devices for treatment of these aneurysms means the only options for treatment are placement of additional FD devices, deconstructive strategies with or without bypass, or microsurgical clipping,2 thereby making an aneurysm that may have been straightforward to treat with another strategy a complex lesion to treat with the presence of the FD stent. Although deconstructive techniques can be used for treatment of failed aneurysm occlusion with flow diversion, where possible, surgical clipping can result in the simplest, most durable solution. Herein we present (Video 1) a case of a patient with a posterior inferior cerebellar artery aneurysm treated previously with FD using a single pipeline embolization device without aneurysm occlusion over 1 year of follow-up who was treated with retrosigmoid craniotomy and clipping of aneurysm. Nuances of the approach selection, clipping of the aneurysm, and preservation of the stent are discussed.

Single-Molecule FRET-Based Multiplexed Detection.

Single-molecule multiplexed detection is a high-promise toolkit for the expanding field of biosensing and molecular diagnostics. Among many single-molecule techniques available today for biomarker sensing including fluorescence, force, electrochemical, spectroscopic, barcoding, and other techniques, fluorescence-based approaches are arguably the most widely used methods due to their high sensitivity, selectivity, and readily available fluorophore-labeling schemes for a wide variety of biomolecules. However, multiplexed imaging using fluorescence techniques has proven to be challenging due to the sophisticated labeling schemes often requiring multiple FRET (fluorescence resonance energy transfer) pairs and/or excitation sources, which lead to overlapping signals and complicate data analysis. Here, we describe a single-molecule FRET method that enables multiplexed analysis while still using only one FRET pair, and thus the described approach is a significant step forward from conventional FRET methods.

Exploring the dynamics of adult Axin2 cell lineage integration into dentate gyrus granule neurons.

The Wnt pathway plays critical roles in neurogenesis. The expression of Axin2 is induced by Wnt/ß-catenin signaling, making this gene a reliable indicator of canonical Wnt activity. We employed pulse-chase genetic lineage tracing with the Axin2-CreERT2 allele to follow the fate of Axin2+ lineage in the adult hippocampal formation. We found Axin2 expressed in astrocytes, neurons and endothelial cells, as well as in the choroid plexus epithelia. Simultaneously with the induction of Axin2 fate mapping by tamoxifen, we marked the dividing cells with 5-ethynyl-2'-deoxyuridine (EdU). Tamoxifen induction led to a significant increase in labeled dentate gyrus granule cells three months later. However, none of these neurons showed any EdU signal. Conversely, six months after the pulse-chase labeling with tamoxifen/EdU, we identified granule neurons that were positive for both EdU and tdTomato lineage tracer in each animal. Our data indicates that Axin2 is expressed at multiple stages of adult granule neuron differentiation. Furthermore, these findings suggest that the integration process of adult-born neurons from specific cell lineages may require more time than previously thought.

Retrosigmoid Transmiddle Cerebellar Peduncle Approach for Resection of a Giant Pontine Cavernous Malformation: 2-Dimensional Operative Video.

Pioneering work in defining brainstem safe entry zones and development of surgical techniques has made microsurgical removal of brainstem cavernous malformation possible.1,2 The middle cerebellar peduncle (MCP) is a robust route for access to deep-seated lesions within the pons. The peduncle can be accessed by splitting the petrosal fissure of the cerebellum to obtain a more direct view to the MCP through a retrosigmoid approach. The author presents a case of a female with a history of 3 bleeds attributable to a giant pontine cavernous malformation. The patient consented to the procedure. Given the aggressive natural history of deep cavernous malformations, especially in the year following a bleed, surgical removal was recommended. Step-by-step description of access to the MCP through splitting the petrosal fissure, and key steps for removal of a pontine cavernous malformation are described.

Transcervical Vertebral Artery Origin Endarterectomy for Vertebrobasilar Insufficiency.

Vertebral artery (VA) stenosis is a cause of vertebrobasilar insufficiency (VBI) and disabling posterior circulation stroke,1 accounting for up to 30% of all strokes.2 Although the natural history of VBI is not as well delineated as that of carotid stenosis, strokes in the basilar circulation can be more disabling than their anterior circulation counterparts. Stenosis exceeding 30% at the origin of the vertebral artery is associated with increased risk of stroke.3 The authors present a case of a female patient with significant peripheral vascular disease who presented with concerns for VBI. The patient was on antiplatelet and anticoagulative medications and a statin at the time of her presentation. Angiography demonstrated bilateral vertebral artery origin stenosis. The left VA was diminutive and arose directly from the arch (Video 1). The right VA demonstrated critical stenosis at its origin. Attempts at endovascular access of the right VA for placement of a balloon-mounted stent were unsuccessful. The patient underwent a transcervical approach for endarterectomy of the VA origin. The VA can be readily accessed using a small supraclavicular incision to isolate the V1 segment of the vessel. The procedure was performed with the patient heparinized and on antiplatelet medications. Alternatives to this strategy include patch grafting in addition to the endarterectomy or use of a short vein graft to bypass the stenosis of the VA beyond the stenotic segment.

Keyhole Retrosigmoid Craniotomy for Lateral Pontine Cavernous Malformation.

With improvements in anesthesia, monitoring, and peroperative care, the surgical removal of intrinsic brainstem pathology has become a possibility.1 Although surgical removal of deep-seated lesions continues to have significant morbidity, at least temporarily, associated with it, removal of exophytic lesions can be accomplished with little disability for the patient. The key to a good outcome, when removing cerebral cavernous malformation, is preservation of adjacent neurovascular bundles, use of sharp dissection over blunt pulling, judicious use of cautery in and around the brainstem, and preservation of the developmental venous anomaly, when present. The authors present a case of a lateral pontine cerebral cavernous malformation that was exophytic at the lateral and peritrigeminal safe entry zones.2 Neuromonitoring was used an adjunct to ensure safety of the procedure. The lesion is accessed using a keyhole retrosigmoid craniotomy (Video 1). We do not routinely use lumbar drains for these procedures as careful arachnoid dissection can result in adequate cerebrospinal fluid release. The window of access to this area is between CN 5 and the CN 7/8 complex. The arachnoid over the nerves is preserved, but the layer between the nerves is exposed to gain access to the lateral pons. The lesion is sharply dissected from the lateral pons, taking care to save the developmental venous anomaly, from which this lesion arises.

Contralateral Interhemispheric Transfalcine Approach to the Basal Ganglia.

The contralateral interhemispheric approach provides a robust path into the mesial frontal lobe and basal ganglia structures.1 The use of gravity to retract the dominant frontal lobe allows the surgeon to avoid injury caused by exposure of the dominant hemisphere. The transfalcine corridor, however, is long and often not well illuminated, necessitating the use of lighted instruments. Within the path of approach lie the anterior cerebral arteries, which must be carefully dissected and preserved. Upon opening the falx, the entire mesial frontal lobe and deep basal ganglia structures can be readily accessed. Herein, we present a patient with familial cerebral cavernous malformation-1 syndrome who presented after an acute hemorrhage from a deep basal ganglia cerebral cavernous malformation (Video 1). The patient consented to the procedure. The patient was hemiparetic and aphasic, likely secondary to mass effect from the bleed. The lesion was approached from a contralateral interhemispheric approach and removed completely. The patient's examination improved with removal of the mass lesion. This case demonstrates the utility of this approach for accessing deep corridors within the cerebral cortices.

Dynamics of Adult Axin2 Cell Lineage Integration in Granule Neurons of the Dentate Gyrus.

The Wnt pathway plays critical roles in neurogenesis. The expression of Axin2 is induced by Wnt/ß-catenin signaling, making this gene a sensitive indicator of canonical Wnt activity. We employed pulse-chase genetic lineage tracing with the Axin2-CreERT2 allele to follow the fate of Axin2 -positive cells in the adult hippocampal formation. We found Axin2 expressed in astrocytes, neurons and endothelial cells, as well as in the choroid plexus epithelia. Simultaneously with tamoxifen induction of Axin2 fate mapping, the dividing cells were marked with 5-ethynyl-2'-deoxyuridine (EdU). Tamoxifen induction resulted in significant increase of dentate gyrus granule cells three months later; however, none of these neurons contained EdU signal. Conversely, six months after the tamoxifen/EdU pulse-chase labeling, EdU-positive granule neurons were identified in each animal. Our data imply that Axin2 is expressed at several different stages of adult granule neuron differentiation and suggest that the process of integration of the adult-born neurons from certain cell lineages may take longer than previously thought.

Recurrence of a large intracranial fusiform aneurysm treated with overlapping Pipeline embolization devices: illustrative case.

BACKGROUND: Flow diversion, specifically with the Pipeline embolization device (PED), represents a paradigm shift in the treatment of intracranial aneurysms. Several studies have demonstrated its efficacy and at times superiority to conventional treatment modalities for aneurysms with a fusiform morphology, giant size, or wide neck. However, there may be a nonsignificant risk of recurrence after flow diversion of these historically difficult-to-treat aneurysms, relative to aneurysms with a more favorable morphology and size (i.e., saccular, narrow necked). To date, only three papers in the literature have demonstrated the recurrence of a completely occluded aneurysm on follow-up. OBSERVATIONS: The authors describe a patient with a giant middle cerebral artery fusiform aneurysm treated with multiple telescoping PEDs. On the 3-month follow-up angiogram, there was complete occlusion of the aneurysm. The patient was lost to follow-up and presented 4 years later with a recurrence of the aneurysm between PED segments, requiring retreatment. The patient represented 3 years posttreatment with the need for repeat treatment of the fusiform aneurysm due to separation of the existing PEDs along with stent reconstruction. At the 20-month follow-up after the third treatment, the initial aneurysm target was found to be occluded. LESSONS: This case illustrates the need for long-term follow-up, specifically for patients with giant wide-necked or fusiform aneurysms treated with overlapping PEDs.

Intraarterial Transplantation of Mitochondria After Ischemic Stroke Reduces Cerebral Infarction.

Background-: Transplantation of autologous mitochondria into ischemic tissue may mitigate injury caused by ischemia and reperfusion. Methods-: Using murine stroke models of middle cerebral artery occlusion, we sought to evaluate feasibility of delivery of viable mitochondria to ischemic brain parenchyma. We evaluated the effects of concurrent focused ultrasound activation of microbubbles, which serves to open the blood-brain barrier, on efficacy of delivery of mitochondria. Results-: Following intra-arterial delivery, mitochondria distribute through the stroked hemisphere and integrate into neural and glial cells in the brain parenchyma. Consistent with functional integration in the ischemic tissue, the transplanted mitochondria elevate concentration of adenosine triphosphate in the stroked hemisphere, reduce infarct volume and increase cell viability. Additional of focused ultrasound leads to improved blood brain barrier opening without hemorrhagic complications. Conclusions-: Our results have implications for the development of interventional strategies after ischemic stroke and suggest a novel potential modality of therapy after mechanical thrombectomy.

Computational generation and characterization of IsdA-binding aptamers with single-molecule FRET analysis.

Staphylococcus aureus is a major foodborne bacterial pathogen. Early detection of S. aureus is crucial to prevent infections and ensure food quality. The iron-regulated surface determinant protein A (IsdA) of S. aureus is a unique surface protein necessary for sourcing vital iron from host cells for the survival and colonization of the bacteria. The function, structure, and location of the IsdA protein make it an important protein for biosensing applications relating to the pathogen. Here, we report an in-silico approach to develop and validate high-affinity binding aptamers for the IsdA protein detection using custom-designed in-silico tools and single-molecule Fluorescence Resonance Energy Transfer (smFRET) measurements. We utilized in-silico oligonucleotide screening methods and metadynamics-based methods to generate 10 aptamer candidates and characterized them based on the Dissociation Free Energy (DFE) of the IsdA-aptamer complexes. Three of the aptamer candidates were shortlisted for smFRET experimental analysis of binding properties. Limits of detection in the low picomolar range were observed for the aptamers, and the results correlated well with the DFE calculations, indicating the potential of the in-silico approach to support aptamer discovery. This study showcases a computational SELEX method in combination with single-molecule binding studies deciphering effective aptamers against S. aureus IsdA, protein. The established approach demonstrates the ability to expedite aptamer discovery that has the potential to cut costs and predict binding efficacy. The application can be extended to designing aptamers for various protein targets, enhancing molecular recognition, and facilitating the development of high-affinity aptamers for multiple uses.

Preclinical models of middle cerebral artery occlusion: new imaging approaches to a classic technique.

Stroke remains a major burden on patients, families, and healthcare professionals, despite major advances in prevention, acute treatment, and rehabilitation. Preclinical basic research can help to better define mechanisms contributing to stroke pathology, and identify therapeutic interventions that can decrease ischemic injury and improve outcomes. Animal models play an essential role in this process, and mouse models are particularly well-suited due to their genetic accessibility and relatively low cost. Here, we review the focal cerebral ischemia models with an emphasis on the middle cerebral artery occlusion technique, a "gold standard" in surgical ischemic stroke models. Also, we highlight several histologic, genetic, and in vivo imaging approaches, including mouse stroke MRI techniques, that have the potential to enhance the rigor of preclinical stroke evaluation. Together, these efforts will pave the way for clinical interventions that can mitigate the negative impact of this devastating disease.

FRET-Based Single-Molecule Detection of Pathogen Protein IsdA Using Computationally Selected Aptamers.

Iron-regulated surface determinant protein A (IsdA) is a key surface protein found in the foodborne bacteria─Staphylococcus aureus (S. aureus)─which is known to be critical for bacterial survival and colonization. S. aureus is pathogenic and has been linked to foodborne diseases; thus, early detection is critical to prevent diseases caused by this bacterium. Despite IsdA being a specific marker for S. aureus and several detection methods have been developed for sensitive detection of this bacteria such as cell culture, nucleic acids amplification, and other colorimetric and electrochemical methods, the detection of S. aureus through IsdA is underdeveloped. Here, by combining computational generation of target-guided aptamers and fluorescence resonance energy transfer (FRET)-based single-molecule analysis, we presented a widely applicable and robust detection method for IsdA. Three different RNA aptamers specific to the IsdA protein were identified and their ability to switch a FRET construct to a high-FRET state in the presence of protein was verified. The presented approach demonstrated the detection of IsdA down to picomolar levels (×10-12 M, equivalent to ∼1.1 femtomoles IsdA) with a dynamic range extending to ∼40 nM. The FRET-based single-molecule technique that we reported here is capable of detecting the foodborne pathogen protein IsdA with high sensitivity and specificity and has a broader application in the food industry and aptamer-based sensing field by enabling quantitative detection of a wide range of pathogen proteins.

Magnetic Resonance Imaging of Mouse Cerebral Cavernomas Reveal Differential Lesion Progression and Variable Permeability to Gadolinium.

BACKGROUND: Cerebral cavernous malformations, also known as cavernous angiomas, are blood vessel abnormalities comprised of clusters of grossly enlarged and hemorrhage-prone capillaries. The prevalence in the general population, including asymptomatic cases, is estimated to be 0.5%. Some patients develop severe symptoms, including seizures and focal neurological deficits, whereas others remain asymptomatic. The causes of this remarkable presentation heterogeneity within a primarily monogenic disease remain poorly understood. METHODS: We established a chronic mouse model of cerebral cavernous malformations, induced by postnatal ablation of Krit1 with Pdgfb-CreERT2, and examined lesion progression in these mice with T2-weighted 7T magnetic resonance imaging (MRI). We also established a modified protocol for dynamic contrast-enhanced MRI and produced quantitative maps of gadolinium tracer gadobenate dimeglumine. After terminal imaging, brain slices were stained with antibodies against microglia, astrocytes, and endothelial cells. RESULTS: These mice develop cerebral cavernous malformations lesions gradually over 4 to 5 months of age throughout the brain. Precise volumetric analysis of individual lesions revealed nonmonotonous behavior, with some lesions temporarily growing smaller. However, the cumulative lesional volume invariably increased over time and after about 2 months followed a power trend. Using dynamic contrast-enhanced MRI, we produced quantitative maps of gadolinium in the lesions, indicating a high degree of heterogeneity in lesional permeability. MRI properties of the lesions were correlated with cellular markers for endothelial cells, astrocytes, and microglia. Multivariate comparisons of MRI properties of the lesions with cellular markers for endothelial and glial cells revealed that increased cell density surrounding lesions correlates with stability, whereas denser vasculature within and surrounding the lesions may correlate with high permeability. CONCLUSIONS: Our results lay a foundation for better understanding individual lesion properties and provide a comprehensive preclinical platform for testing new drug and gene therapies for controlling cerebral cavernous malformations.

Reperfusion injury in acute ischemic stroke: Tackling the irony of revascularization.

Reperfusion injury is an unfortunate consequence of restoring blood flow to tissue after a period of ischemia. This phenomenon can occur in any organ, although it has been best studied in cardiac cells. Based on cardiovascular studies, neuroprotective strategies have been developed. The molecular biology of reperfusion injury remains to be fully elucidated involving several mechanisms, however these mechanisms all converge on a similar final common pathway: blood brain barrier disruption. This results in an inflammatory cascade that ultimately leads to a loss of cerebral autoregulation and clinical worsening. In this article, the authors present an overview of these mechanisms and the current strategies being employed to minimize injury after restoration of blood flow to compromised cerebral territories.

Comparison the effects of probiotic and prebiotic as antibiotic alternatives on Salmonella colonization, performance, and egg quality in laying hens challenged with Salmonella enterica serotype Enteritidis.

Background: Consumption of contaminated eggs with Salmonella enterica serotype Enteritidis (SE) cause gastroenteritis in human. Aims: The present study examined the effect of probiotic and prebiotic compared to antibiotic on the colonization of SE in the ceca, and the quantity and quality of produced eggs in the laying hens challenged with SE. Methods: One hundred Hy-Line W-36 laying hens with 44-week-olds were studied for 13 weeks in a randomized complete block design containing five treatments and four replicates with five birds in each replicate. Treatments included: negative control, positive control, and antibiotic: diets containing antibiotic (Oxytetracycline 0.15 g/kg diet), probiotic (Bactocell® 0.1 g/kg diet), and prebiotic (Diamond V Original XPCTM 1.25 g/kg diet). All experimental groups except negative control were challenged with 1 ml of suspension solution containing 1 × 107 CFU/ml SE by oral gavage at the start of the ninth week of the experiment. Laying performance traits and cecal bacterial population were measured at the end of each week. Results: Probiotic and prebiotic showed a greater effect in the reduction of yolk cholesterol and blood cholesterol level before and after challenge with SE, respectively (P<0.05). In pre-challenge period, treatments had no effect on the cecal bacterial population; but after the challenge, three dietary supplements decreased the colonization of SE in the ceca of laying hens, and prebiotic showed more preventive effect (P<0.05). Conclusion: The result of this study showed that the prebiotic can be effective in reducing and preventing SE colonization in laying hens and act as an alternative to antibiotics.

Minimally Invasive, Supine, Lateral Supracerebellar Infratentorial Approach for Resection of a Pineal Cyst.

Most pineal cysts are not surgical lesions and should be conservatively managed. Select lesions, notably those that present with hydrocephalus and Parinaud syndrome and lesions presenting with symptoms consistent with intermittent blockage of cerebrospinal fluid pathways and/or neurovascular compression, however, can be considered for surgical intervention. Two workhorse surgical approaches to the pineal region include the occipital interhemispheric transtentorial and supracerebellar infratentorial approaches. Each approach provides unique benefits and drawbacks and is associated with morbidities. In this patient, we demonstrate the use of a minimally invasive, supine, lateral supracerebellar infratentorial approach to the pineal region (Video 1). The approach makes use of early access to the foramen magnum to release cerebrospinal fluid for cerebellar relaxation, followed by navigation-guided, minimal dissection of the supracerebellar potential space to arrive at the pineal cyst. Opening of the arachnoid membranes ventrolateral to the confluence of the deep cerebral veins allows for direct access to the cyst with minimal manipulation of the venous complex and risk for injury. The cyst is then fenestrated, and the cyst capsule is removed. It should be noted that surgical fenestration does not guarantee relief of headache symptoms, and patients should be consoled about this preoperatively.

Modified Orbitozygomatic Craniotomy for Clipping of a Ruptured Thrombotic A1-A2 Aneurysm.

Ruptured thrombotic aneurysms pose a dual challenge of subarachnoid hemorrhage and local mass effect on neurovascular structures causing cranial nerve palsies or other neurologic symptoms. Although many thrombotic aneurysms can be treated with endovascular techniques, the benefit of surgical treatment of these aneurysms is the fact that clipping can be followed by removal of the clot and decompression of the contents of the aneurysm sac, thereby relieving local mass effect. In Video 1 we present the case of a young man with a ruptured thrombotic anterior cerebral artery (first segment of anterior cerebral artery-second segment of anterior cerebral artery) aneurysm who presented with bilateral vision loss. The aneurysm was clipped via an orbitozygomatic approach, although an interhemispheric approach could have been an alternative, and the contents of the aneurysm sac, which were compressing the optic apparatus, were removed. The patient remained blind in the right eye after the operation, likely due to the initial subarachnoid hemorrhage being directed into the optic nerve. He did, however, have improvement of vision in his left eye. Microsurgical clipping of thrombotic aneurysms allows for exclusion of the aneurysm from the circulation, thereby protecting the patient from repeated hemorrhage, and simultaneous decompression of the local mass effect caused by the rapid increase in the size of the aneurysm due to the clot burden.

A Novel Tissue Atlas and Online Tool for the Interrogation of Small RNA Expression in Human Tissues and Biofluids.

One promising goal for utilizing the molecular information circulating in biofluids is the discovery of clinically useful biomarkers. Extracellular RNAs (exRNAs) are one of the most diverse classes of molecular cargo, easily assayed by sequencing and with expressions that rapidly change in response to subject status. Despite diverse exRNA cargo, most evaluations from biofluids have focused on small RNA sequencing and analysis, specifically on microRNAs (miRNAs). Another goal of characterizing circulating molecular information, is to correlate expression to injuries associated with specific tissues of origin. Biomarker candidates are often described as being specific, enriched in a particular tissue or associated with a disease process. Likewise, miRNA data is often reported to be specific, enriched for a tissue, without rigorous testing to support the claim. Here we provide a tissue atlas of small RNAs from 30 different tissues and three different blood cell types. We analyzed the tissues for enrichment of small RNA sequences and assessed their expression in biofluids: plasma, cerebrospinal fluid, urine, and saliva. We employed published data sets representing physiological (resting vs. acute exercise) and pathologic states (early- vs. late-stage liver fibrosis, and differential subtypes of stroke) to determine differential tissue-enriched small RNAs. We also developed an online tool that provides information about exRNA sequences found in different biofluids and tissues. The data can be used to better understand the various types of small RNA sequences in different tissues as well as their potential release into biofluids, which should help in the validation or design of biomarker studies.