Transcriptome analysis of blastoderms exposed to prolonged egg storage and short periods of incubation during egg storage.

BACKGROUND: Cool temperature egg storage prior to incubation is a common practice in the broiler industry; however, prolonged egg storage causes increased embryonic mortality and decreased hatchability and growth in surviving chicks. Exposing eggs to short periods of incubation during egg storage (SPIDES) reduces the adverse consequences of prolonged storage. SPIDES increases blastodermal cell viability by reducing apoptosis, though the counteracting mechanisms are unclear. To define the impact of prolonged storage and SPIDES, transcriptome analysis compared gene expression from blastoderms isolated from eggs exposed to the following treatments: control (CR, stored at 17 °C for 4 days), prolonged storage (NSR, stored at 17 °C for 21 days), SPIDES (SR, stored at 17 °C for 21 days with SPIDES), and incubated control (C2, stored at 17 °C for 4 days followed by incubation to HH (Hamburger-Hamilton) stage 2, used as the ideal standard development) (n = 3/group). Data analysis was performed using the CLC Genomics Workbench platform. Functional annotation was performed using DAVID and QIAGEN Ingenuity Pathway Analysis. RESULTS: In total, 4726 DEGs (differentially expressed genes) were identified across all experimental group comparisons (q < 0.05, FPKM> 20, |fold change| > 1.5). DEGs common across experimental comparisons were involved in cellular homeostasis and cytoskeletal protein binding. The NSR group exhibited activation of ubiquitination, apoptotic, and cell senescence processes. The SR group showed activation of cell viability, division, and metabolic processes. Through comparison analysis, cellular respiration, tRNA charging, cell cycle control, and HMBG1 signaling pathways were significantly impacted by treatment and potential regulatory roles for ribosomal protein L23a (RPL23A) and MYC proto-oncogene, BHLH transcription factor (MYC) were identified. CONCLUSIONS: Prolonged egg storage (NSR) resulted in enriched cell stress and death pathways; while SPIDES (SR) resulted in enriched basic cell and anti-apoptotic pathways. New insights into DNA repair mechanisms, RNA processing, shifts in metabolism, and chromatin dynamics in relation to egg storage treatment were obtained through this study. Although egg storage protocols have been examined through targeted gene expression approaches, this study provided a global view of the extensive molecular networks affected by prolonged storage and SPIDES and helped to identify potential upstream regulators for future experiments to optimize egg storage parameters.

Incidence of brain metastasis from esophageal cancer.

We investigated whether the incidence of brain metastasis (BM) from primary esophageal and esophagogastric cancer is increasing. A single-institution retrospective review identified 583 patients treated from January 1997 to January 2016 for stages I through IV cancer of the esophagus and esophagogastric junction (follow-up, ≥3 months). Collected data included demographic information, date and staging at primary diagnosis, histologic subtype, treatment regimen for primary lesion, date of BM diagnosis, presence or absence of central nervous system symptoms, presence or absence of extracranial disease, treatment regimen for intracranial lesions, and date of death. The overall cohort included 495 patients (85%) with adenocarcinoma and 82 (14%) with squamous cell carcinoma (492 [84%] were male; median age at diagnosis, 68 years [range: 26-90 years]). BM was identified in 22 patients (3.8%) (median latency after primary diagnosis, 11 months). Among patients with BM, the primary histology was adenocarcinoma in 21 and squamous cell carcinoma in 1 (P = 0.30). BM developed in 12 who were initially treated for locally advanced disease and in 10 stage IV patients who presented with distant metastases. Overall survival (OS) after BM diagnosis was 18% at 1 year (median, 4 months). No difference in OS after BM diagnosis was observed in patients initially treated for localized disease compared to patients who presented with stage IV disease; however, OS was superior for patients who initially had surgical resection compared to patients treated with whole brain radiotherapy or stereotactic radiosurgery alone (1-year OS, 67% vs. 0%; median OS, 13.5 vs. 3 months; P = 0.003). The incidence of BM is low in patients with esophageal cancer. Outcomes were poor overall for patients with BM, but patients who underwent neurosurgical resection had improved survival.

Self-Referenced Coherent Diffraction X-Ray Movie of Ångstrom- and Femtosecond-Scale Atomic Motion.

Time-resolved femtosecond x-ray diffraction patterns from laser-excited molecular iodine are used to create a movie of intramolecular motion with a temporal and spatial resolution of 30 fs and 0.3 Å. This high fidelity is due to interference between the nonstationary excitation and the stationary initial charge distribution. The initial state is used as the local oscillator for heterodyne amplification of the excited charge distribution to retrieve real-space movies of atomic motion on ångstrom and femtosecond scales. This x-ray interference has not been employed to image internal motion in molecules before. Coherent vibrational motion and dispersion, dissociation, and rotational dephasing are all clearly visible in the data, thereby demonstrating the stunning sensitivity of heterodyne methods.

Observations of turkey eggs stored up to 27 days and incubated for 8 days: embryo developmental stage and weight differences and the differentiation of fertilized from unfertilized germinal discs.

For logistical reasons, egg storage prior to incubation is a growing practice in the commercial turkey industry. Yet the consequence of increasing egg storage over 7 d is a progressive increase in embryo mortality. The objective of this study was to provide the information necessary to differentiate an early dead embryo from an unfertilized egg after 8 days of incubation (DOI). Five groups of eggs each from inseminated and virgin hens were stored for progressively increasing periods of time (5-d or less, 6 to 10 d, 11 to 15 d, 16 to 20 d, and 21 to 27 d) and incubated. At 8 DOI, eggs were examined and the stage of development (Hamburger and Hamilton, 1951) and embryo weights in normally developed eggs were determined. There was a significant negative correlation between the stage of development and embryo weight with increasing storage periods. All remaining eggs from the inseminated and virgin hens were broken-out and the appearance of the yolk and the fertilized and unfertilized germinal discs examined. The yolks of both hen groups with unfertilized ova maintained a homogeneous uniform yellow-orange color. In contrast, yolks of ova that had been fertilized, with or without early-dead embryos, and yolks from virgin hens that showed evidence of parthenogenetic development (3%) had a heterogeneous appearance. Using fluorescence microscopy, the heterogeneous appearance was due to sheets of aberrant cells and less frequently dispersed cells and folds of the perivitelline layer. It was concluded that clear egg breakouts need to be performed to more accurately assess the impact of egg storage on embryonic mortality. Furthermore, such breakouts should be performed with a high intensity light directed across the surface of the germinal disc to clearly differentiate the subtle differences between an early-dead embryo and an unfertilized germinal disc.

Understanding administrative abdominal aortic aneurysm mortality data.

OBJECTIVE: Administrative data in the form of Hospital Episode Statistics (HES) and the Scottish Morbidity Record (SMR) have been used to describe surgical activity. These data have also been used to compare outcomes from different hospitals and regions, and to corroborate data submitted to national audits and registries. The aim of this observational study was to examine the completeness and accuracy of administrative data relating to abdominal aortic aneurysm (AAA) repair. METHODS: Administrative data (SMR-01 returns) from a single health board relating to AAA repair were requested (September 2007 to August 2012). A complete list of validated procedures; termed the reference data set was compiled from all available sources (clinical and administrative). For each patient episode electronic health records were scrutinised to confirm urgency of admission, diagnosis, and operative repair. The 30-day mortality was recorded. The reference data set was used to systematically validate the SMR-01 returns. RESULTS: The reference data set contained 608 verified procedures. SMR-01 returns identified 2433 episodes of care (1724 patients) in which a discharge diagnosis included AAA. This included 574 operative repairs. There were 34 missing cases (5.6%) from SMR-01 returns; nine of these patients died within 30 days of the index procedure. Omission of these cases made a statistically significant improvement to perceived 30-day mortality (p < .05, chi-square test). If inconsistent SMR-01 data (in terms of ICD-10 and OPCS-4 codes) were excluded only 81.9% of operative repairs were correctly identified and only 30.9% of deaths were captured. DISCUSSION: The SMR-01 returns contain multiple errors. There also appears to be a systematic bias that reduces apparent 30-day mortality. Using these data alone to describe or compare activity or outcomes must be done with caution.

Cell versus protoplasm: revisionist history.

Recent investigations give reason to question anew the historical status of the 'cell theory' as the ultimate driving force in the development of our understanding of life's processes at the most fundamental level. A revisitation of critical research papers and commentaries from the 19th Century shows that the disregarded (and historically maligned) 'protoplasmic theory of life' played a more deterministic role in the early advancement of knowledge on cell structure and function.

"Fuzziness" in the celular interactome: a historical perspective.

Some historical background is given for appreciating the impact of the empirical construct known as the cellular protein-protein interactome, which is a seemingly de novo entity that has arisen of late within the context of postgenomic systems biology. The approach here builds on a generalized principle of "fuzziness" in protein behavior, proposed by Tompa and Fuxreiter.(1) Recent controversies in the analysis and interpretation of the interactome studies are rationalized historically under the auspices of this concept. There is an extensive literature on protein-protein interactions, dating to the mid-1900s, which may help clarify the "fuzziness" in the interactome picture and, also, provide a basis for understanding the physiological importance of protein-protein interactions in vivo.

Early carotid endarterectomy following thrombolysis in the hyperacute treatment of stroke.

BACKGROUND: Thrombolysis with intravenous recombinant tissue plasminogen activator improves the probability of complete neurological recovery if given promptly following the onset of acute ischaemic stroke. Carotid endarterectomy (CEA) can reduce the risk of further embolic stroke in selected patients and is most effective within 14 days of the incident event. The safety of surgery so soon after thrombolysis is unknown. The aim of this study was to report the immediate outcomes of this management strategy early in the unit experience and to encourage pooling of data, recognizing that this will be an uncommon procedure even in busy stoke units with an active lysis programme. METHODS: Data were extracted from two prospectively collected databases, and included patient demographics, type of stroke, type and timing of surgical procedure, and immediate outcome. On presentation with a stroke, all patients underwent urgent computed tomography (CT) of the brain. Those eligible received thrombolysis according to the unit protocol. They underwent CT angiography 24 h after thrombolysis and patients with a severe carotid stenosis had surgery. RESULTS: Ten of a cohort of 450 patients who had received lysis underwent CEA. Seven of these were women and eight of the procedures were carried out under local anaesthetic. Surgery was performed a median of 8 (range 2-23) days after the index event; there were no major complications. CONCLUSION: Few patients with acute stroke are eligible, but CEA performed soon after thrombolytic therapy for stroke appears to be safe.

From protoplasmic theory to cellular systems biology: a 150-year reflection.

Present-day cellular systems biology is producing data on an unprecedented scale. This field has generated a renewed interest in the holistic, "system" character of cell structure-and-function. Underlying the data deluge, however, there is a clear and present need for a historical foundation. The origin of the "system" view of the cell dates to the birth of the protoplasm concept. The 150-year history of the role of "protoplasm" in cell biology is traced. It is found that the "protoplasmic theory," not the "cell theory," was the key 19th-century construct that drove the study of the structure-and-function of living cells and set the course for the development of modern cell biology. The evolution of the "protoplasm" picture into the 20th century is examined by looking at controversial issues along the way and culminating in the current views on the role of cytological organization in cellular activities. The relevance of the "protoplasmic theory" to 21st-century cellular systems biology is considered.

Radiological physics characteristics of the extracted heavy ion beams of the bevatron.

Studies of the depth-ionization properties and the biological effects of heavy ion beams produced at the bevatron have extended work previously done with less energetic beams from other sources. Results indicate that heavy ion beams are suitable for tumor therapy, studies relating to space biology, and fundamental radiobiology.

An overview of the surgical aspects of lower limb venous disease.

Lower limb venous disease encompasses a wide spectrum of pathology, the importance of which relates to high prevalence rather than mortality. The complications of chronic venous insufficiency (CVI), namely lipodermatosclerosis and chronic venous ulceration, represent a major burden to healthcare providers and a high degree of personal morbidity for patients. Management is based upon accurate clinical diagnosis supported by non-invasive imaging. Open surgical and minimally invasive techniques are used to treat varicose veins. Chronic skin complications of CVI require a multidisciplinary approach.

Metabolic channeling versus free diffusion: transition-time analysis.

Metabolic channeling is the term used to describe the restricted flow of substrates and products in multienzyme systems. It has been argued for some time that free diffusion is sufficiently rapid to obviate the need for channeling and, furthermore, that it is also fast enough to prevent competing side reactions from interfering with the metabolic flow. In this article we argue that a thorough consideration of the temporal behavior of metabolite pools suggests that channeling is important in many cases.

Missed opportunities for the detection of abdominal aortic aneurysms.

OBJECTIVE: To determine whether patients attending as emergencies with ruptured AAA could have been detected opportunistically prior to rupture. DESIGN: Retrospective analysis. METHODS: The notes of patients attending hospital with ruptured abdominal aortic aneurysms to four City and DGH hospital in the West of Scotland were examined for previous assessments or investigations with the potential to discover an AAA. RESULTS: In this series 77% of patients were not previously known to have and AAA. Of these patients 76% had been reviewed in hospital during the preceding 5 years on a combined total of 355 occasions. 56% of patients had been seen in hospital during the year preceding rupture on a total of 80 occasions, only undergoing 17 abdominal examinations. CONCLUSION: Clinical examination is not frequently considered in routine practice as a screening tool for AAA but patients who subsequently go on to attend as an emergency ruptured AAA are likely to have consulted medical staff during the preceding years. A large number of patients have missed a prior opportunity for detection.

Mitochondrial function and reactive oxygen species action in relation to boar motility.

Flow cytometric assays of viable boar sperm were developed to measure reactive oxygen species (ROS) formation (oxidization of hydroethidine to ethidium), membrane lipid peroxidation (oxidation of lipophilic probe C(11)-BODIPY(581/591)), and mitochondrial inner transmembrane potential (DeltaPsi(m); aggregation of mitochondrial probe JC-1) during hypothermic liquid storage and freeze-thawing of boar semen and to investigate relationships among ROS, motility, DeltaPsi(m), and ATP production. Basal ROS formation and membrane lipid peroxidation were low in viable sperm of both fresh and frozen-thawed semen, affecting < or =4%. Sperm in fresh, liquid-stored and frozen-thawed semen appeared to be equally susceptible to the activity ROS generators xanthine/xanthine oxidase, FeSO(4)/ascorbate, and hydrogen peroxide (H(2)O(2)). Of the ROS generators tested, FeSO(4)/ascorbate was specific for membrane lipid peroxidation, whereas menadione, xanthine/xanthine oxidase, and H(2)O(2) were specific for oxidization of hydroethidine. Menadione (30microM) and H(2)O(2) (300microM) decreased (P<0.05) motility by 90% during 60min of incubation. Menadione decreased (P<0.05) the incidence of sperm with high DeltaPsi(m) by 95% during 60min of the incubation, although ATP content was not decreased (P>0.05) until 120min. In contrast, H(2)O(2) did not affect DeltaPsi(m) or ATP at any time. The formation of ROS was not associated with any change in viability (90%) for either menadione or H(2)O(2) through 120min. Overall, the inhibitory affects of ROS on motility point to a mitochondrial-independent mechanism. The reduction in motility may have been due to an ROS-induced lesion in ATP utilization or in the contractile apparatus of the flagellum.

Effects of osmolality on inner mitochondrial transmembrane potential and ATP content in spermatozoa recovered from the testes of striped bass (Morone saxatilis).

The objective was to determine the effects of osmolality on the energy status of testicular spermatozoa of striped bass incubated in a TRIS free base-NaCl medium (pH 8) adjusted to either 300 (T300) or 600 mOsm/kg (T600) with NaCl. High mitochondrial inner transmembrane potential (DeltaPsim) was assessed (flow cytometry) with the mitochondrial probe 5, 5', 6, 6'-tetrachloro-1, 1', 3, 3'-tetraethylbenzimidazolyl- carbocyanine iodide (JC-1) and ATP was measured with a luciferin-luciferase assay. Spermatozoa maintained on ice were equally viable (>95% for T300 and T600) for up to 80 min, whereas sperm viability in artificial fresh water (FW) at 27 mOsm/kg decreased (P<0.05) to 67% after 5 min, with only 3.5% viability at 25 min. After 20 min of staining, more spermatozoa (P<0.05) maintained a high DeltaPsim in T300 than in T600 (80 and 50%, respectively). Sperm JC-1 aggregate (Jagg) fluorescence intensity was also greater (P<0.05) in T300 than in T600 (10 and 5 channel number). The Jagg fluorescence was a function of oxidative phosphorylation; the percentage of cells containing Jagg fluorescence decreased to 3% in the presence of carbonyl cyanide 3-chlorophenylhydrazone (CCCP), an uncoupler of cell respiration and oxidative phosphorylation. After incubation for 30 min in the absence of CCCP, sperm ATP concentration was greater (P<0.05) in T300 than in T600 (2.0 vs. 0.2 pmol/10(6) cells), but was below detectability in the presence of CCCP in either medium. In conclusion, we developed a unique approach to assess the energetic status of striped bass spermatozoa during storage and after activation, and concluded that the effects of osmolality must be considered in the design of activating and storage extenders to maintain striped bass sperm motility, viability, and fertility in vitro.

Glutathione peroxidase potentiates the inhibition of platelet function by S-nitrosothiols.

GSH peroxidase (Px) catalyzes the reduction of lipid hydroperoxides (LOOH), known metabolic products of platelets and vascular cells. Because interactions between these cells are modulated by nitric oxide (NO) and LOOH inactivate NO, we investigated the effect of GSH-Px on the inhibition of platelet function by the naturally occurring S-nitrosothiol, S-nitroso-glutathione (SNO-Glu). Concentrations of SNO-Glu that alone did not inhibit platelet function (subthreshold inhibitory concentrations) were added to platelet-rich plasma together with GSH-Px (0.2-20 U/ml); this led to a dose-dependent inhibition of platelet aggregation with an IC50 of 0.6 U/ml GSH-Px. In the presence of subthreshold inhibitory concentrations of SNO-Glu, the LOOH, 5-hydroperoxy-6,8,11,14-eicosatetraenoic acid, increased platelet aggregation, an effect reversed by GSH-Px. Glutathione and SNO-Glu were equally effective as cosubstrates for GSH-Px. Incubation of SNO-Glu with GSH-Px for 1 min led to a 48.5% decrease in the concentration of SNO-Glu. Incubation of SNO-Glu with serum albumin led to the formation of S-nitroso-albumin, an effect enhanced by GSH-Px. These observations suggest that GSH-Px has two functions: reduction of LOOH, thereby preventing inactivation of NO, and metabolism of SNO-Glu, thereby liberating NO and/or supporting further transnitrosation reactions.

In vivo transfer of nitric oxide between a plasma protein-bound reservoir and low molecular weight thiols.

Plasma albumin reacts with nitric oxide (NO) to form the bioactive adduct, S-nitroso-albumin (S-NO-albumin). The limited intracellular access of S-NO-albumin suggests the need for a vascular transfer mechanism of NO from a large plasma S-NO-albumin pool to effect biologic function. To study the role of low molecular weight (LMW) thiols in NO transfer in vivo, we administered intravenous S-NO-albumin (1-300 nmol/kg) to rabbits before and after an intravenous infusion of L-cysteine or N-acetyl-L-cysteine. S-NO-albumin produced dose-dependent hypotension that was significantly augmented by prior infusion of either LMW thiol. LMW thiol infusion significantly accelerated the rate of onset and reduced the duration of action of the hypotension induced by S-NO-albumin. The hemodynamic effects of S-NO-albumin after pretreatment with LMW thiols were mimicked by administration of the corresponding LMW S-nitrosothiol. The transfer of NO from albumin to L-cysteine was directly measured in rabbit plasma using a novel technique that couples high performance liquid chromatography to electrochemical detection. These data demonstrate that NO exchange between plasma protein thiol-bound NO and available LMW thiol pools (transnitrosation) occurs in vivo.

Inhibition of neointimal proliferation in rabbits after vascular injury by a single treatment with a protein adduct of nitric oxide.

Endothelium-derived relaxing factor is important for vascular homeostasis and possesses qualities that may modulate vascular injury, including vasodilation, platelet inhibition, and inhibition of smooth muscle proliferation. S-nitrososerum albumin is a naturally occurring adduct of nitric oxide (NO) with a prolonged biologic half-life and is a potent vasodilator and platelet inhibitor. Given the avidity of serum albumin for subendothelial matrix and the antiproliferative effects of NO, we investigated the effects of locally delivered S-nitroso-bovine serum albumin (S-NO-BSA) and a polythiolated form of bovine serum albumin (pS-BSA) modified to carry several S-nitrosothiol groups (pS-NO-BSA) on neointimal responses in an animal model of vascular injury. Locally delivered S-NO-BSA bound preferentially to denuded rabbit femoral vessels producing a 26-fold increase in local concentration compared with uninjured vessels (P = 0.029). pS-NO-BSA significantly reduced the intimal/medial ratio (P = 0.038) and did so in conjunction with elevations in platelet (P < 0.001) and vascular cGMP content (P < or = 0.001). pS-NO-BSA treatment also inhibited platelet deposition (P = 0.031) after denuding injury. Comparison of BSA, S-NO-BSA, pS-NO-BSA, and control revealed a dose-response relationship between the amount of displaceable NO delivered and the extent of inhibition of neointimal proliferation at 2 wk (P < or = 0.001). Local administration of a stable protein S-nitrosothiol inhibits intimal proliferation and platelet deposition after vascular arterial balloon injury. This strategy for the local delivery of a long-lived NO adduct has potential for preventing restenosis after angioplasty.