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The molecular signaling cascades that regulate angiogenesis and microvascular remodeling are fundamental to normal development, healthy physiology, and pathologies such as inflammation and cancer. Yet quantifying such complex, fractally branching vascular patterns remains difficult. We review application of NASA’s globally available, freely downloadable VESsel GENeration (VESGEN) Analysis software to numerous examples of 2D vascular trees, networks, and tree-network composites. Upon input of a binary vascular image, automated output includes informative vascular maps and quantification of parameters such as tortuosity, fractal dimension, vessel diameter, area, length, number, and branch point. Previous research has demonstrated that cytokines and therapeutics such as vascular endothelial growth factor, basic fibroblast growth factor (fibroblast growth factor-2), transforming growth factor-beta-1, and steroid triamcinolone acetonide specify unique “fingerprint” or “biomarker” vascular patterns that integrate dominant signaling with physiological response. In vivo experimental examples described here include vascular response to keratinocyte growth factor, a novel vessel tortuosity factor; angiogenic inhibition in humanized tumor xenografts by the anti-angiogenesis drug leronlimab; intestinal vascular inflammation with probiotic protection by Saccharomyces boulardii, and a workflow programming of vascular architecture for 3D bioprinting of regenerative tissues from 2D images. Microvascular remodeling in the human retina is described for astronaut risks in microgravity, vessel tortuosity in diabetic retinopathy, and venous occlusive disease.

The Spaceflight Associated Neuro-ocular Syndrome (SANS), associated with the headward fluid shifts incurred in microgravity during long-duration missions, remains a high-priority health and performance risk for human space exploration. To help characterize the pathophysiology of SANS, NASA’s VESsel GENeration Analysis (VESGEN) software was used to map and quantify vascular adaptations in the retina before and after 70 days of bed rest at 6-degree Head-Down Tilt (HDT), a well-studied microgravity analog. Results were compared to the retinal vascular response of astronauts following 6-month missions to the International Space Station (ISS). By mixed effects modeling, the trends of vascular response were opposite. Vascular density decreased significantly in the 16 retinas of eight astronauts and in contrast, increased slightly in the ten retinas of five subjects after HDT (although with limited significance). The one astronaut retina diagnosed with SANS displayed the greatest vascular loss. Results suggest that microgravity is a major variable in the retinal mediation of fluid shifts that is not reproduced in this HDT bed rest model.

OBJECTIVE To determine whether self-monitoring of blood glucose (SMBG) is associated with lower HbA1c in youth with type 2 diabetes taking oral medications only or after starting insulin for persistently elevated HbA1c. RESEARCH DESIGN AND METHODS Treatment Options for Type 2 Diabetes in Adolescents and Youth (TODAY) study participants (n = 699) taking oral medications were asked to perform SMBG twice daily. After reaching primary outcome (PO) (HbA1c ≥8% [64 mmol/mol]) over 6 months or an inability to wean from temporary insulin because of metabolic decompensation), insulin glargine was started. HbA1c and percent of SMBG (SMBG%) (percent days when the meter was used one or more times) before and after PO were analyzed. RESULTS SMBG declined over time and was inversely related to HbA1c (P < 0.0001). Of 298 youth who reached PO and started insulin, 282 had SMBG data. At PO, mean ± SD age was 15.8 ± 2.3 years, BMI 35.5 ± 7.9 kg/m2, and HbA1c 9.6 ± 2.0% (81 ± 21.9 mmol/mol); 65.3% were female. Median SMBG% was 40% at PO, which increased to 49% after 6 months and fell to 41% after 1 year on insulin. At PO, 22% of youth checked ≥80% of days, which increased to 25% and fell to 19% after 6 and 12 months using insulin, respectively. At PO, compared with those who checked CONCLUSIONS Low SMBG adherence was common and associated with higher HbA1c. Optimal SMBG frequency in youth using or not using insulin, and whether less frequent SMBG is a marker for overall worse self-care, require further study.

Purpose Ocular structural and functional changes, collectively termed spaceflight-associated neuro-ocular syndrome (SANS), have been described in astronauts undergoing long-duration missions in the microgravity environment of the International Space Station. We tested the hypothesis that retinal vascular remodeling, particularly by smaller vessels, mediates the chronic headward fluid shifts associated with SANS. Methods As a retrospective study, arterial and venous patterns extracted from 30° infrared Heidelberg Spectralis retinal images of eight crew members acquired before and after six-month missions were analyzed with NASA's recently released VESsel GENeration Analysis (VESGEN) software. Output parameters included the fractal dimension and overall vessel length density that was further classified into large and small vascular branching generations. Vascular results were compared with SANS-associated clinical ocular measures. Results Significant postflight decreases in Df, Lv, and in smaller but not larger vessels were quantified in 11 of 16 retinas for arteries and veins (P value for Df, Lv, and smaller vessels in all 16 retinas were ≤0.033). The greatest vascular decreases occurred in the only retina displaying clinical evidence of SANS by choroidal folds and optic disc edema. In the remaining 15 retinas, decreases in vascular density from Df and Lv ranged from minimal to high by a custom Subclinical Vascular Pathology Index. Conclusions Together with VESGEN, the Subclinical Vascular Pathology Index may represent a new, useful SANS biomarker for advancing the understanding of SANS etiology and developing successful countermeasures for long duration space exploration in microgravity, although further research is required to better characterize retinal microvascular adaptations.

While the bee simile in book 1 of Paradise Lost has garnered much critical attention, the significant image of the hive in Milton’s corpus remains largely overlooked. Deriving from an ancient literary tradition, the bee metaphor was enlisted in seventeenth-century England as a divine symbol of monarchical and ecclesiastical power structures. From his polemic to his epic, Milton, by contrast, consistently uses apian imagery to register harsh critiques of earthly monarchy, feminine influence, and Catholic superstition. Beginning with an unexamined bee passage in Eikonoklastes , this article traces Milton’s revisionist reading of the apiological kingdom, which culminates in his representation of insect and related serpentine imagery in Paradise Lost . I argue that the Miltonic hive serves as a symbol of a natural corruption of the divine hierarchy, an inevitable failure to replicate on earth or in hell the benevolent design of heaven.

The significance of William Shakespeare’s use of the bee kingdom in his drama and poetry lies in the way that he simultaneously observes and challenges entrenched conventions in the literary tradition. Shakespeare’s apiological references—bees, drones, honey, the hive—reveal a nuanced understanding of how hierarchy governs and impedes human interactions and how the division of labor, both human and nonhuman, so often begets violence and betrayal. Bees pollinate farmers’ and gardeners’ crops, and the honey and wax the hive yields are viewed as valuable products for human’s consumption and commodities for their financial gain. For female characters in Shakespeare, the honey trope consistently indicates heroines’ observance of ideals of feminine virtue, particularly chastity and innocence. By the time Shakespeare is writing, virgin bees are reframed in England as chaste and wholesome. In his tragedies, histories, and poetry, Shakespeare promotes a dichotomy of virtue where honeyed descriptions generally indicate masculine treachery and feminine purity.

We developed laboratory exercises using zebrafish (Danio rerio) and nematodes (Caenorhabditis elegans) for a sophomore-level Integrative Biology Laboratory course. Students examined live wildtype zebrafish at different stages of development and noted shifts occurring in response to fgf8a deficiency. Students were introduced to development in other fish species to demonstrate how variation in developmental systems affects phenotype. Finally, students cultured glp-1(bn18ts) C. elegans mutants under different conditions to illustrate how the environment and genetics act concurrently to modulate development. Undergraduate students responded positively to both the fish and C. elegans laboratory modules. These novel laboratory exercises are intended to promote an integrative view of biology and to help prepare undergraduate students for independent research with faculty.

The contentious inclusion of an imprisoned princess's speech from Sir Philip Sidney's The Countess of Pembroke's Arcadia (1593) in the Eifon Basilifye (1649) has long overshadowed the most significant contribution of a woman's voice to the "King's book." In three poignant passages of the TLikpn Basilifye, the purported book of Charles I's final prayers and meditations before his execution, Princess Elizabeth, Charles I's daughter, authorizes the account of her martyred father's final day. The critical neglect of Elizabeth's role in memorializing the executed king might at first seem surprising given that during Charles's life the young princess was beloved and respected by Royalists and Parliamentarians alike. After the regicide, however, Elizabeth's life and death as a Stuart princess registered varied reactions that fell along party lines. For Royalists, she became an emblem of the suffering Stuart monarchy; accordingly, they emphasized her misfortune in being held captive in a cold, remote castle by tyrants and refused permission to be reunited with her sister. They mourned Elizabeth's death of illness at Carisbrooke Castle on 8 September 1650 as a renewal of their grief for Charles. For Parliamentarians, by contrast, respecting her status as the sovereign's child would appear inconsistent with their decision to abolish the English monarchy on 7 February 1649. ' This political motivation also provides insight into John Milton's significant but previously overlooked suppression of the Lady Elizabeth passages of the Eifypn in his own commissioned response in Eifypnofyastes (1649).2 In the 165Os, in other words, Elizabeth's writing had a polarizing effect: Royalists used it as propaganda, whereas Milton and the Parliamentarians sought to silence her authorial voice to combat this Royalist strategy.In the EikonBasilifye, a supplementary section to the text called "A Perfect Copy of Prayers Used by His Majestic in the Time of His Sufferings Delivered to Dr. Juxon Bishop of London" contains the three Lady Elizabeth passages and a paraphrase of Pamela's prayer, "A Prayer in Time of Captivitie," from Sidney's Arcadia. The first of several popular editions containing the supplementary section was printed by William Dugard on 15 March 1649.3 The first passage concerning Elizabeth, "A True Relation of the King[']s Speech to the Lady Elisabeth, and the Duke of Glo[u]cester, the Day before his Death," contains an observation of the royal family recorded by an anonymous observer.4 However, the next two passages, "Another Relation from the Lady Elisabeth[']s Own Hand" and "Another Relation from the Lady Elisabeth," are written from the princess's first-person perspective of "the last time I had the happiness to see [Charles I]" (281). In these three excerpts, the anonymous observer and Elizabeth herself firmly establish the young girl's authority as a witness to her father's grace under persecution.Although it may seem counterintuitive to view the words of a child as privileged speech, Elizabeth's age and social status actually made her uniquely equipped to move an authence sympathetic to the plight of the Stuarts and of the Episcopacy that they supported. As Michael Mascuch notes in his study of the young, seventeenth-century prophet Sarah Wight, "[T]he 'child' symbolizes not so much a body's physical immaturity as an intellectual disposition toward religious knowledge and authority."5 In the case of Elizabeth, her defense of Charles's martyrdom would bear a sincerity that accounts by Doctor Juxon or Charles's attendants might not; her words are not those of a politically motivated supporter or, from a Parliamentarian perspective, a sycophant, but rather those of a daughter determined to serve the memory of her father.What is at stake in the polemic between Eikon Basilifye and Eikonoklastes is the collective memory and popular perception of Charles I and the English monarchy. Is the Eikon, as his supporters would contend, a monument to Charles and a record of his martyrdom? …

The cholesterol-depleting drug methyl-b-cyclodextrin (Me-bCD) was tested for its effects on amphibian oocyte maturation, cholesterol depletion, and low-density membrane recovery. Progesterone-induced oocyte maturation was accelerated by pretreatment of cells with 5‐50 mM Me-b-CD in a dose-dependent manner. Treatment of oocytes with 50 mM Me-b-CD alone was sufficient to induce germinal vesicle breakdown, stimulate formation of meiotic spindles, and stimulate phosphorylation of mitogen-activated protein kinase over time courses longer than those observed after progesterone treatment. After short-term (30 min) labeling of oocytes with [ 3 H]cholesterol, 30‐90 min of treatment with 5‐50 mM Me-b-CD removed 50%‐70% of cellassociated label, and cholesterol depletion was not observed with a-cyclodextrin. After long-term (20‐23 h) labeling of oocytes with [ 3 H]cholesterol, Me-b-CD treatment resulted in dosedependent cholesterol depletion in the 5‐50 mM range, and 50 mM Me-b-CD removed approximately 50% of cell-associated label after 9 h. Treatment of oocytes with 5‐50 mM Me-b-CD also decreased recovery of low-density membrane by detergentfree sucrose gradient centrifugation. These results implicate cholesterol and low-density membrane domains in the signaling mechanisms leading to germinal vesicle breakdown in amphibian oocytes. gametogenesis, meiosis, progesterone, signal transduction, steroid hormones

Somitogenesis is a complex process during early vertebrate development involving interactions between many factors to form a bilateral somite series. A role for chromatin remodelers in somitogenesis has not yet been demonstrated. Here, we investigate the function of chromodomain helicase DNA binding protein 7 (chd7) during zebrafish somitogenesis. We show that Chd7 deficiency leads to asymmetric segmentation of the presomitic mesoderm (PSM), as revealed by expression of the somitogenesis genes, cdx1a, dlc, her7, mespa, and ripply1. Moreover, we show that abrogation of Chd7 results in the loss of asymmetric expression of spaw in the lateral plate mesoderm, which is consistent with more general laterality defects. Based on the observation that insufficient Chd7 leads to left–right asymmetry defects during PSM segmentation, and because CHD7 has been linked to human spinal deformities, we suggest that zebrafish chd7 morphants may be a good in vivo model to examine the pathophysiology of these diseases. Developmental Dynamics 240:2272–2280, 2011. © 2011 Wiley-Liss, Inc.

Whole mount in situ hybridization (WISH) is a common technique in molecular biology laboratories used to study gene expression through the localization of specific mRNA transcripts within whole mount specimen. This technique (adapted from Albertson and Yelick, 2005) was used in an upper level undergraduate Comparative Vertebrate Biology laboratory classroom at Syracuse University. The first two thirds of the Comparative Vertebrate Biology lab course gave students the opportunity to study the embryology and gross anatomy of several organisms representing various chordate taxa primarily via traditional dissections and the use of models. The final portion of the course involved an innovative approach to teaching anatomy through observation of vertebrate development employing molecular techniques in which WISH was performed on zebrafish embryos. A heterozygous fibroblast growth factor 8 a (fgf8a) mutant line, ace, was used. Due to Mendelian inheritance, ace intercrosses produced wild type, heterozygous, and homozygous ace/fgf8a mutants in a 1:2:1 ratio. RNA probes with known expression patterns in the midline and in developing anatomical structures such as the heart, somites, tailbud, myotome, and brain were used. WISH was performed using zebrafish at the 13 somite and prim-6 stages, with students performing the staining reaction in class. The study of zebrafish embryos at different stages of development gave students the ability to observe how these anatomical structures changed over ontogeny. In addition, some ace/fgf8a mutants displayed improper heart looping, and defects in somite and brain development. The students in this lab observed the normal development of various organ systems using both external anatomy as well as gene expression patterns. They also identified and described embryos displaying improper anatomical development and gene expression (i.e., putative mutants). For instructors at institutions that do not already own the necessary equipment or where funds for lab and curricular innovation are limited, the financial cost of the reagents and apparatus may be a factor to consider, as will the time and effort required on the part of the instructor regardless of the setting. Nevertheless, we contend that the use of WISH in this type of classroom laboratory setting can provide an important link between developmental genetics and anatomy. As technology advances and the ability to study organismal development at the molecular level becomes easier, cheaper, and increasingly popular, many evolutionary biologists, ecologists, and physiologists are turning to research strategies in the field of molecular biology. Using WISH in a Comparative Vertebrate Biology laboratory classroom is one example of how molecules and anatomy can converge within a single course. This gives upper level college students the opportunity to practice modern biological research techniques, leading to a more diversified education and the promotion of future interdisciplinary scientific research.

Congenital lymphangiectasia is a rare disease of the lymphatic system often causing respiratory symptoms from chylous pleural effusions. Treatment options for these effusions include drainage by multiple thoracenteses or chest thoracostomy tube placement. Previous studies have reported the efficacy of surgical intervention with adjunctive nutrition therapy using total parenteral nutrition (TPN) in decreasing chylous effusions. Few reports, however, describe the use of very low-fat diets with medium chain triglycerides (MCT) as a therapeutic option. Objective: In order to help manage recurrent pleural effusions in a 28-year-old woman with a history of congenital lymphangiectasia, an oral diet restricted in fat using MCT was administered. Methods: The patient was treated with a very low-fat diet, with fat consumption approximately 5% of total caloric intake using food sources and MCT oil. Her diet was supplemented with a multivitamin and linoleic essential fatty acid. Results: After implementation of this d...

The detection of the prodynorphin gene in anuran amphibians and lungfishes may indicate that this gene arose as a result of the duplication of the proenkephalin gene early during the divergence of the Sarcopterygii, or that this gene may predate the divergence of the ray-finned fish and the lobe-finned fish. The cloning of prodynorphin-related genes from the pufferfish and zebrafish supports the latter hypothesis. This study analyzes trends in the radiation of the prodynorphin gene in teleosts. Prodynorphin cDNAs were cloned from the brain of the eel Anguilla rostrata and the Nile tilapia, Oreochromis niloticus. These teleost prodynorphin sequences have distinct alpha-neoendorphin, dynorphin A, and dynorphin B sequences, and a novel opioid sequence, YGGFI. The relationship of these teleost prodynorphin sequences to other actinopterygian and sarcopterygian prodynorphin sequences will be discussed.

The cholesterol-depleting drug methyl-beta-cyclodextrin (Me-beta-CD) was tested for its effects on amphibian oocyte maturation, cholesterol depletion, and low-density membrane recovery. Progesterone-induced oocyte maturation was accelerated by pretreatment of cells with 5-50 mM Me-beta-CD in a dose-dependent manner. Treatment of oocytes with 50 mM Me-beta-CD alone was sufficient to induce germinal vesicle breakdown, stimulate formation of meiotic spindles, and stimulate phosphorylation of mitogen-activated protein kinase over time courses longer than those observed after progesterone treatment. After short-term (30 min) labeling of oocytes with [(3)H]cholesterol, 30-90 min of treatment with 5-50 mM Me-beta-CD removed 50%-70% of cell- associated label, and cholesterol depletion was not observed with alpha-cyclodextrin. After long-term (20-23 h) labeling of oocytes with [(3)H]cholesterol, Me-beta-CD treatment resulted in dose- dependent cholesterol depletion in the 5-50 mM range, and 50 mM Me-beta-CD removed approximately 50% of cell-associated label after 9 h. Treatment of oocytes with 5-50 mM Me-beta-CD also decreased recovery of low-density membrane by detergent-free sucrose gradient centrifugation. These results implicate cholesterol and low-density membrane domains in the signaling mechanisms leading to germinal vesicle breakdown in amphibian oocytes.