Human Natural Killer Cells Cryopreserved without DMSO Sustain Robust Effector Responses.

Natural killer (NK) cell-based immunotherapy has benefitted from the multiple strengths that NK cells offer in adoptive transfer settings, not the least of which is their safety and potential for allogeneic use. Such use, however, necessitates the cryopreservation of NK cell-based therapy products to support logistical efforts in deploying these cells in different locations, decentralized from the point of collection or manufacturing. DMSO, the most commonly used cryoprotective agent (CPA), has been effective in protecting immune cells during freezing and thawing, but its ability to induce molecular and genetic changes to immune cells as well as its toxicity has stimulated interest in alternative CPAs. However, replacing DMSO's ability to act intracellularly has been difficult, and the sensitivity of human peripheral blood-derived NK cells to freezing and thawing-induced damage has meant that investigations into the potential of replacing DMSO are lacking. As a first step toward establishing the feasibility of cryopreserving human NK cells with CPAs' alternative to DMSO, we investigate the potential of using noncell-penetrating and cell-penetrating CPAs to recover NK cells post-thaw without DMSO. Here, we find that cryoprotection using cell-penetrating CPAs can retain the viability of human peripheral blood-derived NK cells to a comparable degree to DMSO. In addition, non-DMSO-cryopreserved human NK cells were as cytotoxic as those cryopreserved with DMSO and displayed a comparable level of surface markers of activation. In summary, we present the first example of the potential of developing non-DMSO CPA formulations that could be deployed in future cell therapy regimens.

Randomized Controlled Trial to Study the Acute Effects of Strength Exercise on Insulin Sensitivity in Obese Adults.

An acute session of strength exercise (SE) ameliorates insulin sensitivity (IS) for several hours; however, the effects of SE volume (i.e., number of sets) have not been studied thoroughly. Although it is intuitive that some SE is better than none, and more is better than some for the improvement of IS, high-volume sessions might be challenging for diseased populations to complete, especially obese adults, for whom even a brisk walk can be challenging. This protocol details a randomized clinical trial to assess the acute effects of SE on IS in obese adults. The inclusion criteria are body mass index >30 kg/m2, central obesity (waist circumference >88 cm and >102 cm for women and men, respectively), and age >40 years. Participants will be familiarized with the SE (7 exercises targeting major muscle groups) and then will perform three sessions in a randomized order: session 1 - high-volume session (3 sets/exercise); session 2 - low-volume session (1 set/exercise); session 3 - control session (no exercise). Diet will be controlled the day before and on the day of the sessions. Sessions will be completed at night, and an oral glucose tolerance test will be performed the next morning, from which several indexes of IS will be derived, such as the area under the curve (AUC) of glucose and insulin, the Matsuda index, the Cederholm index, the muscle IS index, and the Gutt index. Based on pilot studies, we expect ~15% improvement in IS (insulin AUC, and Matsuda and Cederholm indexes) after the high-volume session, and ~8% improvement after the low-volume session compared to the control session. This study will benefit individuals who find high-volume SE sessions challenging but still aim to improve their IS by investing 1/3 of their time and effort.

Three-Dimensional-Printed Titanium Versus Polyetheretherketone Cages for Lumbar Interbody Fusion: A Systematic Review of Comparative In Vitro, Animal, and Human Studies.

Interbody fusion is a workhorse technique in lumbar spine surgery that facilities indirect decompression, sagittal plane realignment, and successful bony fusion. The 2 most commonly employed cage materials are titanium (Ti) alloy and polyetheretherketone (PEEK). While Ti alloy implants have superior osteoinductive properties they more poorly match the biomechanical properties of cancellous bones. Newly developed 3-dimensional (3D)-printed porous titanium (3D-pTi) address this disadvantage and are proposed as a new standard for lumbar interbody fusion (LIF) devices. In the present study, the literature directly comparing 3D-pTi and PEEK interbody devices is systematically reviewed with a focus on fusion outcomes and subsidence rates reported in the in vitro, animal, and human literature. A systematic review directly comparing outcomes of PEEK and 3D-pTi interbody spinal cages was performed. PubMed, Embase, and Cochrane Library databases were searched according to PRISMA (Preferred Reporting Items for Systematic Reviews and Meta-Analysis) guidelines. Mean Newcastle-Ottawa Scale score for cohort studies was 6.4. A total of 7 eligible studies were included, comprising a combination of clinical series, ovine animal data, and in vitro biomechanical studies. There was a total population of 299 human and 59 ovine subjects, with 134 human (44.8%) and 38 (64.4%) ovine models implanted with 3D-pTi cages. Of the 7 studies, 6 reported overall outcomes in favor of 3D-pTi compared to PEEK, including subsidence and osseointegration, while 1 study reported neutral outcomes for device related revision and reoperation rate. Though limited data are available, the current literature supports 3D-pTi interbodies as offering superior fusion outcomes relative to PEEK interbodies for LIF without increasing subsidence or reoperation risk. Histologic evidence suggests 3D-Ti to have superior osteoinductive properties that may underlie these superior outcomes, but additional clinical investigation is merited.

Impacts of Austerity on Urogenital Infection Rates in Active Duty Servicewomen.

INTRODUCTION: Female warfighters are at increased risk of urogenital infections (UGIs) when serving in austere environments with varying levels of access to water, sanitation, and hygiene resources. Urogenital infections among servicewomen were among the top five reported medical encounters and one of the top seven reasons for medical evacuation from deployed locations between 2008 and 2013. In this study, we examine the use of water and sanitation resources across three environments (home duty station, field training, and deployment) and analyze associations between water, sanitation, and hygiene resource access and UGIs during training or deployment. The analyses are based on answers from 751 diverse active duty servicewomen (ADSW) at a large military installation in the southeastern USA. MATERIALS AND METHODS: Data for this population-based, prospective, cross-sectional survey design were collected during the administration of the Military Women's Readiness Urogenital Health Questionnaire. This research study was guided by two specific research questions derived from self-reported data in sections 2 and 3 of the questionnaire. RESULTS: The sample consisted of an extremely diverse pool of U.S. Army ADSW attached to highly operational units with robust field training and deployment tempos. Over one-half of the participants reported being diagnosed with one or more UGIs before military service. Of that participant group, more than 76.0% experienced one or more infections since joining the military. We found that the majority of UGIs occurred while U.S. Army ADSW were at their home duty stations rather than during field training or deployment. Of the three types of UGIs, bacterial vaginosis is more often associated with water and sanitation constraints. None of the water factors were significantly associated with UGIs during deployment. Intentional dehydration was not associated with UGIs in either field training or deployment, but intentionally delaying urination was associated with a significant increase in the odds of developing urinary tract infection during deployment. We identified trends in hygiene practices that may put ADSW at a higher risk for UGIs. CONCLUSIONS: Austere conditions, which may exist in any service environment, pose risks to the urogenital health and wellness of a female warfighter. Access to clean water and sanitation resources in military settings is essential for optimal health and operational readiness. It is necessary to identify and investigate critical research and policy gaps in need of investment and support for successful, evidence-based integration of female warfighters into military combat roles and to optimize their performance.

Patterns in Urogenital Health in Active Duty Servicewomen: A Prospective Cross-Sectional Survey Evaluating Impacts of Water, Sanitation, and Hygiene Resources Across Three Military Environments.

INTRODUCTION: Female warfighters are at risk for sex-specific and frequently unrecognized urogenital health challenges that may impede their ability to serve. The constraints on water, sanitation, and hygiene (WASH) resources by austere environments, whether at home or abroad, require women to practice unhealthy hygiene behaviors that jeopardize their urogenital health. In this manuscript, we examine the use of WASH resources by U.S. Army active duty servicewomen (ADSW) across three settings-home duty, field training, and deployment-to determine how the changing availability of WASH resources alters hygiene and sanitation practices in austere environments. Additionally, we report findings from a previously developed theoretical framework for determining the impacts of austere environments on ADSW's hygiene knowledge, behaviors, and outcomes. MATERIALS AND METHODS: This study used a prospective, cross-sectional survey design with population sampling and generalized ordered logit regression models. We recruited ADSW assigned to a large military installation in the southeastern United States after conducting a structured field hygiene training for female readiness. RESULTS: We surveyed a highly diverse sample of 751 ADSW and found that the use of WASH resources significantly differs between field training and deployment. Clean running water significantly differed among all settings, with nearly 70% reporting clean running water to be rarely or never available during field training. Bathing facility types significantly differed in each setting, with Cadillac bathrooms more often used during deployment than during field training and wipe baths more often used during field training than during deployment. We found that women were significantly less likely to wash their private areas more than 2 days a week during field training, compared to deployment. Women reported changing their underwear ≤2 times per week in field training environments as compared to during deployment. Soap and water were reported as the most common ways of washing in both home duty stations and during deployment, followed by wipes at home duty stations and in deployed settings. Participants reported wipes as their primary bathing method during field training, followed by soap and water. Participants used intentional dehydration or delayed urination more frequently in field training than during deployment. Women reported significant differences in holding their urine between field training and deployment, with principal reasons reported as "dirty facilities" and "distance to the nearest restroom." Significantly, they reported safety concerns when bathing, using the bathroom, or toileting facilities across the three settings. CONCLUSIONS: This is the first study to characterize sex-specific challenges by a large sample of operational ADSW and to explore the use of WASH resources in home duty, field training, and deployment settings. The results show that field training is more austere than deployed settings, indicating that austerity, not deployment, increases the urogenital infection risk for ADSW. With women more integrated into the military strategy than at any time in U.S. history, military leaders can use our results to develop interventions that ameliorate the unique challenges that influence the military readiness and overall health of female warfighters.

Technical Nuances and Outcomes of Telescoping Pipeline Flow Diverters: A Multicenter Study.

BACKGROUND: "Telescoping" multiple overlapping Pipeline Embolization Devices (PEDs; Medtronic) has increased their utility by allowing for more impermeable coverage and providing the ability to off-set landing zone sites and extend treatment constructs. OBJECTIVE: To consider the technical nuances and challenges of telescoping PEDs for the treatment of intracranial aneurysms. METHODS: Databases from 3 U.S. academic neurovascular centers were retrospectively queried to identify patients with intracranial aneurysms treated with multiple PED constructs. Data on patient and aneurysm characteristics, as well as outcomes including Raymond-Roy occlusion classification, modified Rankin Scale score, and complications, were gathered. RESULTS: Forty-six patients had 48 intracranial aneurysms treated, including 16 (33%) in whom placement of telescoping PEDs was planned. Fourteen (30%) patients presented with a ruptured aneurysm. Twenty-one aneurysms (44%) were treated with proximal extension, 13 (27%) with distal extension, and 14 (29%) with PED placement inside one another. Thirty (70%) patients had complete aneurysm occlusion at follow-up. Two (4%) patients had to be retreated. Three patients with unruptured and 1 with ruptured aneurysm had a permanent intraprocedural complication. We present descriptive cases illustrating PEDs that were placed inside one another, proximally, distally, and to improve wall apposition because of vessel tortuosity. CONCLUSION: Our data indicate a higher than expected complication rate that is likely because of the technical complexity of these cases. The case illustrations presented demonstrate the indications and challenging aspects of telescoping PEDs.

Tendon Healing in a Mouse Model of Loeys-Dietz Syndrome: Controlled Study Using a Patellar Tendon Transection Model.

BACKGROUND: Loeys-Dietz syndrome (LDS) is a rare autosomal-dominant connective tissue disorder caused by genetic mutations in the transforming growth factor-ß (TGFß) signaling pathway. In addition to vascular malformations, patients with LDS commonly present with bone and tendon abnormalities, including joint laxity. While TGFß signaling dysregulation has been implicated in many of these clinical manifestations, the degree to which it influences the tendinopathy and tendon healing issues in LDS has not been determined. METHODS: Wound healing after patellar tendon transection was compared between wild-type (WT) and Tgfbr2-mutant (LDS) mice (7 mice per group). In all mice, the right patellar tendon was transected at midsubstance, while the left was untouched to serve as a control. Mice were euthanized 6 weeks after surgery. Tendon specimens were harvested for histopathologic grading according to a previously validated scoring metric, and gene expression levels of Mmp2, Tgfb2, and other TGFß-signaling genes were assayed. Between-group comparisons were made using 1-way analysis of variance with post hoc Tukey honestly significant difference testing. RESULTS: Expression levels of assayed genes were similar between LDS and WT tendons at baseline; however, at 6 weeks after patellar tendon transection, LDS tendons showed sustained elevations in Mmp2 and Tgfb2 compared with baseline values; these elevations were not seen in normal tendons undergoing the same treatments. Histologically, untreated LDS tendons had significantly greater cellularity and cell rounding compared with untreated WT tendons, and both WT and LDS tendons had significantly worse histologic scores after surgery. CONCLUSION: We present the first mechanistic insight into the effect of LDS on tendons and tendon healing. The morphologic differences between LDS and WT tendons at baseline may help explain the increased risk of tendon/ligament dysfunction in patients with LDS, and the differential healing response to injury in LDS may account for the delayed healing and weaker repair tissue. LEVEL OF EVIDENCE: Level V.

[Hemitrapeziectomy of the carpometacarpal joint of the thumb]. / Hemiresektionsarthroplastik am Daumensattelgelenk.

BACKGROUND: Hemiresection arthoplasty or partial trapeziectomy of the carpometacarpal joint of the thumb is a surgical option in stages 1-3 according to Eaton-Littler classification. Preservation of the intact scaphotrapeziotrapezoidal (STT) joint and maintenance of the length of the thumb is the advantage of this technique. TECHNIQUE: Technically, partial trapeziectomy is only slightly more complex compared to trapeziectomy with or without suspension. An interposition of autologous or allogeneic material is recommended; suspension arthroplasty is also possible in almost all common variants. Despite these theoretical advantages, this technique has not proven advantageous in practice, neither in terms of function nor in terms of time, compared to other techniques. The great advantage of partial trapeziectomy, however, is the wide range of options for revision surgery in the case of persistent complaints. RESULTS: The risk of proximalization of the thumb due to an inadequate interposition is generally minor and is usually caused by implant sintering. This can easily be compensated for by implanting a larger graft. Persistent instabilities are the most common cause of persistence of symptoms and can usually be remedied by autologous or allogeneic suspension arthroplasty. In addition, the entire range of endoprosthetics is also possible with a previous spare resection of the trapezium and the possibility of a complete trapeziectomy remains in any case. The obvious theoretical advantages of arthroscopic hemitrapeziectomy need to be confirmed in practice in future.

Effect of HIV/HAART and Other Clinical Variables on the Oral Mycobiome Using Multivariate Analyses.

The oral microbiome is considered an important factor in health and disease. We recently reported significant effects of HIV and several other clinical variables on the oral bacterial communities in a large cohort of HIV-positive and -negative individuals. The purpose of the present study was to similarly analyze the oral mycobiome in the same cohort. To identify fungi, the internal transcribed spacer 2 (ITS2) of the fungal rRNA genes was sequenced using oral rinse samples from 149 HIV-positive and 88 HIV-negative subjects that had previously undergone bacterial amplicon sequencing. Quantitative PCR was performed for total fungal content and total bacterial content. Interestingly, samples often showed predominance of a single fungal species with four major clusters predominated by Candida albicans, Candida dubliniensis, Malassezia restricta, or Saccharomyces cerevisiae Quantitative PCR analysis showed the Candida-dominated sample clusters had significantly higher total fungal abundance than the Malassezia or Saccharomyces species. Of the 25 clinical variables evaluated for potential influences on the oral mycobiome, significant effects were associated with caries status, geographical site of sampling, sex, HIV under highly active antiretroviral therapy (HAART), and missing teeth, in rank order of statistical significance. Investigating specific interactions between fungi and bacteria in the samples often showed Candida species positively correlated with Firmicutes or Actinobacteria and negatively correlated with Fusobacteria, Proteobacteria, and Bacteroidetes Our data suggest that the oral mycobiome, while diverse, is often dominated by a limited number of species per individual; is affected by several clinical variables, including HIV positivity and HAART; and shows genera-specific associations with bacterial groups.IMPORTANCE The oral microbiome is likely a key element of homeostasis in the oral cavity. With >600 bacterial species and >160 fungal species comprising the oral microbiome, influences on its composition can have an impact on both local and systemic health. We recently reported significant effects of HIV and several other clinical variables on the oral bacterial community in a large cohort of HIV-positive and -negative subjects. We describe here a comprehensive analysis of the oral mycobiome in the same cohort. Similar to the bacterial community, HIV under highly active antiretroviral therapy (HAART) had a significant impact on the mycobiome composition, but with less impact compared to other clinical variables. Additionally, unlike the oral bacterial microbiome, the oral mycobiome is often dominated by a single species with 4 major clusters of fungal communities. Together, these results suggest the oral mycobiome has distinct properties compared with the oral bacterial community, although both are equally impacted by HIV.

Modern dolomite formation caused by seasonal cycling of oxygenic phototrophs and anoxygenic phototrophs in a hypersaline sabkha.

The "Dolomite Problem" has been a controversy for over a century, owing to massive assemblages of low-temperature dolomite in ancient rocks with little dolomite forming today despite favorable geochemical conditions. Experiments show that microbes and their exopolymeric substances (EPS) nucleate dolomite. However, factors controlling ancient abundances of dolomite can still not be explained. To decode the enigma of ancient dolomite, we examined a modern dolomite forming environment, and found that a cyclic shift in microbial community between cyanobacteria and anoxygenic phototrophs creates EPS suited to dolomite precipitation. Specifically, EPS show an increased concentration of carboxylic functional groups as microbial composition cycles from cyanobacterial to anoxygenic phototroph driven communities at low-and high- salinity, respectively. Comparing these results to other low-T forming environments suggests that large turnover of organic material under anoxic conditions is an important driver of the process. Consequently, the shift in atmospheric oxygen throughout Earth's history may explain important aspects of "The Dolomite Problem". Our results provide new context for the interpretation of dolomite throughout Earth's history.

A spectrum of (Dis)Belief: Coronavirus frames in a rural midwestern town in the United States.

Community responses to the SARS-CoV-2, or "coronavirus" outbreaks of 2020 reveal a great deal about society. In the absence of government mandates, debates over issues such as mask mandates and social distancing activated conflicting moral beliefs, dividing communities. Policy scholars argue that such controversies represent fundamental frame conflicts, which arise from incommensurable worldviews, such as contested notions of "liberty" versus "equity". This article investigates frames people constructed to make sense of coronavirus and how this affected social behavior in 2020. We conducted an interpretive framing analysis using ethnographic data from a predominately white, conservative, and rural midwestern tourist town in the United States from June to August 2020. We collected semi-structured interviews with 87 community members, observed meetings, events, and daily life. We identified four frames that individuals constructed to make sense of coronavirus: Concern, Crisis, Constraint, and Conspiracy. Concern frames illustrated how some individuals are uniquely affected and thus protect themselves. Crisis frames recognized coronavirus as a pervasive and profound threat requiring unprecedented action. Constraint frames emphasized the coronavirus response as a threat to financial stability and personal growth that should be resisted. Conspiracy frames denied its biological basis and did not compel action. These four conflicting frames demonstrate how social fragmentation, based on conflicting values, led to an incomplete pandemic response in the absence of government mandates at the national, state, and local levels in rural America. These findings provide a social rationale for public health mandates, such as masking, school/business closures, and social distancing, when contested beliefs impede collective action.

Inter-Rater Reliability of Web-Based Calibrated Peer Review within a Pharmacy Curriculum.

Objective. To assess the inter-rater reliability of student evaluations submitted through a web-based calibrated peer review system versus faculty evaluations in a Doctor of Pharmacy (PharmD) curriculum. Methods. Two calibrated peer review assignments were integrated into two sequential pharmacotherapy-based courses in the first professional year of the PharmD curriculum. To assess the inter-rater reliability of student peer and self-evaluations compared to faculty evaluations, 50 student submissions from each assignment were randomly selected for faculty evaluation, using the same evaluation criteria used by students. Statistical analysis was performed to assess the difference between the scores given by students versus faculty members and the correlation between the combined and individual assignment scores given by students versus faculty members. Results. Mean faculty scores did not significantly differ from student peer or self-evaluation scores. When the assignments were combined, there was a significant fair correlation between scores assigned by peers and those assigned by faculty members, and between scores assigned by students (self-evaluation) and faculty members. For individual assignments, one showed significant positive correlations for both peer and self-assessment compared to faculty, while the other assignment did not. Conclusion. Incorporating longitudinal calibrated peer review activities into the PharmD curriculum demonstrated fair inter-rater reliability between scores assigned by pharmacy student and faculty members. Further research on calibrated peer review is needed in pharmacy education to evaluate its reliability and utility. However, the current study suggests that the use of a web-based calibrated peer review system within a pharmacy curriculum could provide additional opportunities for critical evaluation and student self-assessment, as well as encourage more open-ended communication activities without increasing the grading burden on faculty members.

Intraosseous Cannulation of a Calvarial Diploic Vein for Embolization of a Symptomatic Dural Arteriovenous Fistula: A Technical Case Report.

BACKGROUND AND IMPORTANCE: Dural arteriovenous fistulas (DAVFs) may present unique challenges for treatment depending on the anatomy and pattern of venous drainage. If endovascular techniques are to be employed, the DAVF must be amenable to transvenous or transarterial therapy. When access of peripheral vasculature does not provide a straightforward path, less conventional options may be available. This case highlights a novel, technically simple, and effective approach for the treatment of a subset of DAVFs, with venous drainage through calvarial diploic veins, that would make endovascular treatment otherwise challenging or impossible. CLINICAL PRESENTATION: We present a case of a 66-yr-old female patient who was diagnosed with a symptomatic DAVF located along the sphenoid ridge with a large intraosseous channel containing the draining vein of the fistula. This lesion was successfully treated with transcranial endovascular embolization via direct intraosseous cannulation of the calvarial diploic vein. This novel approach obviated the need for a full-thickness craniotomy, afforded only minimal bone loss, and preserved the integrity of the dura. A 3-mo follow-up angiogram confirmed complete cure of the DAVF with no residual arteriovenous shunt. At 20 mo postembolization, the patient was symptom free, with no reported neurologic deficits. Complete diagnostic work-up, treatment planning in a multidisciplinary environment, and a novel approach for endovascular embolization utilizing a hybrid operating suite played key roles in the successful implementation of this technique. CONCLUSION: This is the first report of direct intraosseous cannulation of a calvarial diploic vein for successful transcranial endovascular embolization of a symptomatic DAVF.

All-fiber widely tunable ultrafast laser source for multimodal imaging in nonlinear microscopy.

In this Letter, we present an all-fiber, easy-to-use, wavelength-tunable, ultrafast laser, based on soliton self-frequency shifting in an Er-doped polarization-maintaining very large mode area fiber. The system is tunable over 370 nm, starting at 1620 nm, with an average power of up to 1.5 W that emits 120 fs short laser pulses directly out of the fusion-spliced fiber without using bulky pulse compression optics. The output is subsequently frequency doubled to a wavelength range covering 810 nm up to almost 1000 nm with more than 500 mW average power and 120 fs pulse width. Having a polarization extinction ratio better than 40 dB and a M2 of 1.1, the fully computer-controlled laser system presents a robust and compact laser source. These parameters make the laser perfectly suited for multimodal imaging in nonlinear microscopy.

Proximal interphalangeal joint arthrodesis using a compression wire: A comparative biomechanical study.

Arthrodesis of the proximal interphalangeal joint is a proven technique for treating of a range of pathological conditions, including osteoarthritis. There are multiple surgical procedures. A biomechanical study was conducted to compare the stability of a compression wire to intraosseous wiring for the arthrodesis. Seventeen formalin-fixed human fingers were randomly assigned into two groups and the bone mineral density was determined. Arthrodesis in 20° flexion was performed using an oblique compression wire (n=8) or intraosseous wiring (n=9). The stability of the arthrodesis was tested by applying a tensile bending force until failure. The mean force needed to fail the compression wire arthrodesis and intraosseous wire arthrodesis was not significantly different (76.2N, SD 31N and 63.0N, SD 28N). There was no correlation between bone density and force to failure. The compression wire was within the approximate range achieved by intraosseous wiring in withstanding substantial force before failure. From a biomechanical point of view, a compression wire is feasible for PIP arthrodesis.

Microbial community composition and dolomite formation in the hypersaline microbial mats of the Khor Al-Adaid sabkhas, Qatar.

The Khor Al-Adaid sabkha in Qatar is among the rare extreme environments on Earth where it is possible to study the formation of dolomite-a carbonate mineral whose origin remains unclear and has been hypothetically linked to microbial activity. By combining geochemical measurements with microbiological analysis, we have investigated the microbial mats colonizing the intertidal areas of sabhka. The main aim of this study was to identify communities and conditions that are favorable for dolomite formation. We inspected and sampled two locations. The first site was colonized by microbial mats that graded vertically from photo-oxic to anoxic conditions and were dominated by cyanobacteria. The second site, with higher salinity, had mats with an uppermost photo-oxic layer dominated by filamentous anoxygenic photosynthetic bacteria (FAPB), which potentially act as a protective layer against salinity for cyanobacterial species within the deeper layers. Porewater in the uppermost layers of the both investigated microbial mats was supersaturated with respect to dolomite. Corresponding to the variation of the microbial community's vertical structure, a difference in crystallinity and morphology of dolomitic phases was observed: dumbbell-shaped proto-dolomite in the mats dominated by cyanobacteria and rhombohedral ordered-dolomite in the mat dominated by FAPB.

Expanding the endoscopic transpterygoid corridor to the petroclival region: anatomical study and volumetric comparative analysis.

OBJECTIVE The endoscopic endonasal transmaxillary transpterygoid (TMTP) approach has been the gateway for lateral skull base exposure. Removal of the cartilaginous eustachian tube (ET) and lateral mobilization of the internal carotid artery (ICA) are technically demanding adjunctive steps that are used to access the petroclival region. The gained expansion of the deep working corridor provided by these maneuvers has yet to be quantified. METHODS The TMTP approach with cartilaginous ET removal and ICA mobilization was performed in 5 adult cadaveric heads (10 sides). Accessible portions of the petrous apex were drilled during the following 3 stages: 1) before ET removal, 2) after ET removal but before ICA mobilization, and 3) after ET removal and ICA repositioning. Resection volumes were calculated using 3D reconstructions generated from thin-slice CT scans obtained before and after each step of the dissection. RESULTS The average petrous temporal bone resection volumes at each stage were 0.21 cm3, 0.71 cm3, and 1.32 cm3 (p < 0.05, paired t-test). Without ET removal, inferior and superior access to the petrous apex was limited. Furthermore, without ICA mobilization, drilling was confined to the inferior two-thirds of the petrous apex. After mobilization, the resection was extended superiorly through the upper extent of the petrous apex. CONCLUSIONS The transpterygoid corridor to the petroclival region is maximally expanded by the resection of the cartilaginous ET and mobilization of the paraclival ICA. These added maneuvers expanded the deep window almost 6 times and provided more lateral access to the petroclival region with a maximum volume of 1.5 cm3. This may result in the ability to resect small-to-moderate sized intradural petroclival lesions up to that volume. Larger lesions may better be approached through an open transcranial approach.

CREBBP knockdown enhances RAS/RAF/MEK/ERK signaling in Ras pathway mutated acute lymphoblastic leukemia but does not modulate chemotherapeutic response.

Relapsed acute lymphoblastic leukemia is the most common cause of cancer-related mortality in young people and new therapeutic strategies are needed to improve outcome. Recent studies have shown that heterozygous inactivating mutations in the histone acetyl transferase, CREBBP, are particularly frequent in relapsed childhood acute lymphoblastic leukemia and associated with a hyperdiploid karyotype and KRAS mutations. To study the functional impact of CREBBP haploinsufficiency in acute lymphoblastic leukemia, RNA interference was used to knock down expression of CREBBP in acute lymphoblastic leukemia cell lines and various primagraft acute lymphoblastic leukemia cells. We demonstrate that attenuation of CREBBP results in reduced acetylation of histone 3 lysine 18, but has no significant impact on cAMP-dependent target gene expression. Impaired induction of glucocorticoid receptor targets was only seen in 1 of 4 CREBBP knockdown models, and there was no significant difference in glucocorticoid-induced apoptosis, sensitivity to other acute lymphoblastic leukemia chemotherapeutics or histone deacetylase inhibitors. Importantly, we show that CREBBP directly acetylates KRAS and that CREBBP knockdown enhances signaling of the RAS/RAF/MEK/ERK pathway in Ras pathway mutated acute lymphoblastic leukemia cells, which are still sensitive to MEK inhibitors. Thus, CREBBP mutations might assist in enhancing oncogenic RAS signaling in acute lymphoblastic leukemia but do not alter response to MEK inhibitors.

Self-frequency-shifted solitons in a polarization-maintaining, very-large-mode area, Er-doped fiber amplifier.

We demonstrate soliton self-frequency-shifted, femtosecond-pulse amplification in a newly-developed, polarization-maintaining, Er-doped, very-large-mode-area fiber amplifier. The PM-VLMA Er fiber had a core diameter of 50 µm, an effective area of ~1050 µm2, and Er absorption of 50 dB/m. The measured birefringence beat length of the PM-VLMA Er fiber was 14.1 mm. The soliton wavelength could be shifted by more than 90 nm. The soliton generation process resulted in remarkably clean, 86 fs pulses with 21 nJ energy at 1650 nm and 244 kW peak power from an all-fiber, fusion spliced system without bulk-optics for pulse compression. The polarization extinction ratio of the soliton was greater than 40 dB, and the M2 was 1.1. The fully polarization-maintaining fiber laser system provides robust and stable soliton generation. Peak-to-peak variation in the soliton wavelength, measured over the course of an hour was only 0.03% and pulse energy variation was only 0.5%.