bluuebunny/crossref_metadata_2025 huggingface dataset:
Vaccine-Induced Protection against Orthopoxvirus Infection Is Mediated through the Combined Functions of CD4 T Cell-Dependent Antibody and CD8 T Cell Responses
Authors: Chaudhri Geeta , Tahiliani Vikas , Eldi Preethi , Karupiah Gunasegaran
Published: November 2014

Antibody production by B cells in the absence of CD4 T cell help has been shown to be necessary and sufficient for protection against secondary orthopoxvirus (OPV) infections. This conclusion is based on short-term depletion of leukocyte subsets in vaccinated animals, in addition to passive transfer of immune serum to naive hosts that are subsequently protected from lethal orthopoxvirus infection. Here, we show that CD4 T cell help is necessary for neutralizing antibody production and virus control during a secondary ectromelia virus (ECTV) infection. A crucial role for CD4 T cells was revealed when depletion of this subset was extended beyond the acute phase of infection. Sustained depletion of CD4 T cells over several weeks in vaccinated animals during a secondary infection resulted in gradual diminution of B cell responses, including neutralizing antibody, contemporaneous with a corresponding increase in the viral load. Long-term elimination of CD8 T cells alone delayed virus clearance, but prolonged depletion of both CD4 and CD8 T cells resulted in death associated with uncontrolled virus replication. In the absence of CD4 T cells, perforin- and granzyme A- and B-dependent effector functions of CD8 T cells became critical. Our data therefore show that both CD4 T cell help for antibody production and CD8 T cell effector function are critical for protection against secondary OPV infection. These results are consistent with the notion that the effectiveness of the smallpox vaccine is related to its capacity to induce both B and T cell memory.

IMPORTANCESmallpox eradication through vaccination is one of the most successful public health endeavors of modern medicine. The use of various orthopoxvirus (OPV) models to elucidate correlates of vaccine-induced protective immunity showed that antibody is critical for protection against secondary infection, whereas the role of T cells is unclear. Short-term leukocyte subset depletion in vaccinated animals or transfer of immune serum to naive, immunocompetent hosts indicates that antibody alone is necessary and sufficient for protection. We show here that long-term depletion of CD4 T cells over several weeks in vaccinated animals during secondary OPV challenge reveals an important role for CD4 T cell-dependent antibody responses in effective virus control. Prolonged elimination of CD8 T cells alone delayed virus clearance, but depletion of both T cell subsets resulted in death associated with uncontrolled virus replication. Thus, vaccinated individuals who subsequently acquire T cell deficiencies may not be protected against secondary OPV infection.
Receptor Recognition Mechanisms of Coronaviruses: a Decade of Structural Studies
Authors: Li Fang
Published: November 2014

Receptor recognition by viruses is the first and essential step of viral infections of host cells. It is an important determinant of viral host range and cross-species infection and a primary target for antiviral intervention. Coronaviruses recognize a variety of host receptors, infect many hosts, and are health threats to humans and animals. The receptor-binding S1 subunit of coronavirus spike proteins contains two distinctive domains, the N-terminal domain (S1-NTD) and the C-terminal domain (S1-CTD), both of which can function as receptor-binding domains (RBDs). S1-NTDs and S1-CTDs from three major coronavirus genera recognize at least four protein receptors and three sugar receptors and demonstrate a complex receptor recognition pattern. For example, highly similar coronavirus S1-CTDs within the same genus can recognize different receptors, whereas very different coronavirus S1-CTDs from different genera can recognize the same receptor. Moreover, coronavirus S1-NTDs can recognize either protein or sugar receptors. Structural studies in the past decade have elucidated many of the puzzles associated with coronavirus-receptor interactions. This article reviews the latest knowledge on the receptor recognition mechanisms of coronaviruses and discusses how coronaviruses have evolved their complex receptor recognition pattern. It also summarizes important principles that govern receptor recognition by viruses in general.
Nonmuscle Myosin Heavy Chain IIB Mediates Herpes Simplex Virus 1 Entry
Authors: Arii Jun , Hirohata Yoshitaka , Kato Akihisa , Kawaguchi Yasushi
Published: November 2014

Nonmuscle myosin heavy chain IIA (NMHC-IIA) has been reported to function as a herpes simplex virus 1 (HSV-1) entry coreceptor by interacting with viral envelope glycoprotein B (gB). Vertebrates have three genetically distinct isoforms of the NMHC-II, designated NMHC-IIA, NMHC-IIB, and NMHC-IIC. COS cells, which are readily infected by HSV-1, do not express NMHC-IIA but do express NMHC-IIB. This observation prompted us to investigate whether NMHC-IIB might associate with HSV-1 gB and be involved in an HSV-1 entry like NMHC-IIA. In these studies, we show that (i) NMHC-IIB coprecipitated with gB in COS-1 cells upon HSV-1 entry; (ii) a specific inhibitor of myosin light chain kinase inhibited cell surface expression of NMHC-IIB in COS-1 cells upon HSV-1 entry as well as HSV-1 infection, as reported with NMHC-IIA; (iii) overexpression of mouse NMHC-IIB in IC21 cells significantly increased their susceptibility to HSV-1 infection; and (iv) knockdown of NMHC-IIB in COS-1 cells inhibited HSV-1 infection as well as cell-cell fusion mediated by HSV-1 envelope glycoproteins. These results supported the hypothesis that, like NMHC-IIA, NMHC-IIB associated with HSV-1 gB and mediated HSV-1 entry.

IMPORTANCE
Herpes simplex virus 1 (HSV-1) was reported to utilize nonmuscle myosin heavy chain IIA (NMHC-IIA) as an entry coreceptor associating with gB. Vertebrates have three genetically distinct isoforms of NMHC-II. In these isoforms, NMHC-IIB is of special interest since it highly expresses in neuronal tissue, one of the most important cellular targets of HSV-1
in vivo
. In this study, we demonstrated that the ability to mediate HSV-1 entry appeared to be conserved in NMHC-II isoforms. These results may provide an insight into the mechanism by which HSV-1 infects a wide variety of cell types
in vivo
.
Management of traumatic wound dehiscence of a functional graft 34 years after penetrating keratoplasty
Authors: Gunasekaran Srilathaa , Sharma Namrata , Titiyal Jeewan S
Published: December 2014

A functional corneal graft 34 years after penetrating keratoplasty (PKP) is rare and even rarer is its survival after wound dehiscence. We present a case of a 69-year-old man who had presented to the emergency services in January 2012 with corneal wound dehiscence OS extending to 8 o’clock following blunt trauma. His vision was perception of light. The best corrected visual acuity (BCVA) of the patient prior to graft dehiscence was 20/30. The patient had undergone optical penetrating keratoplasty for healed keratitis in OS in 1978 followed by phacoemulsification with intraocular lens implantation in 2009. Suturing of the graft was performed under topical anaesthesia. At 18 months follow-up, the BCVA was 20/60 with clear graft centrally and specular count was 865 cells/mm2. This case highlights that early and appropriate management of corneal graft dehiscence can achieve good functional outcome even 34 years following PKP.
Factors associated with the age of the onset of diabetes in women aged 50 years or more: a population-based study
Authors: Valadares Ana L R , Machado Vanessa S S , Costa-Paiva Lúcia S , de Sousa Maria H , Pinto-Neto Aarão M
Published: November 2014

Objective

Investigate factors associated with the onset of diabetes in women aged more than 49 years.

Design and methods

Cross-sectional, population-based study using self-reports with 622 women. The dependent variable was the age of occurrence of diabetes using the life table method. Cox multiple regression models were adjusted to analyse the onset of diabetes according to predictor variables. Sociodemographic, clinical and behavioural factors were evaluated.

Results

Of the 622 women interviewed, 22.7% had diabetes. The mean age at onset was 56 years. The factors associated with the age of occurrence of diabetes were self-rated health (very good, good) (coefficient=−0.792; SE of the coefficient=0.215; p=0.0001), more than two individuals living in the household (coefficient=0.656, SE of the coefficient=0.223; p=0.003), and body mass index (BMI) (kg/m2) at 20–30 years of age (coefficient= 0.056, SE of the coefficient=0.023; p=0.014).

Conclusions

Self-rated health considered good or very good was associated with a higher rate of survival without diabetes. Sharing a home with two or more other people and a weight increase at 20–30 years of age was associated with the onset of type 2 diabetes.
A pilot study of a digital drainage system in pneumothorax
Authors: Tunnicliffe Georgia , Draper Adrian
Published: December 2014

Over recent years there has been increasing usage of digital systems within cardiothoracic surgery to quantify air leaks and aid in clinical decision-making regarding the removal of chest drains postoperatively. The literature suggests improved agreement on timing of removal of chest drains and a reduced length of stay of patients. It could be that such devices could be useful tools for the clinician managing cases of pneumothorax.

Methods

This pilot study recruited adults admitted under the medical team with a pneumothorax requiring a chest drain. Participants had the underwater seal device changed for a digital device (Thopaz) which allowed continuous monitoring of the air leak. Drains were removed when either there was no ongoing air leak and the lung had expanded, or surgery was deemed necessary.

Results

Thirteen patients with pneumothorax (four primary, nine secondary) used the device during their admission including one patient treated in the community (the device has internal suction). Data were used to aid the clinician in management of the pneumothorax including the timing of surgery/ removal of drain and commencement of suction.

Discussion

Digital devices appear to be safe and effective and may prove to be a useful tool in the management of pneumothorax.
A case-crossover study of transient risk factors influence on occupational injuries: a study protocol based on a review of previous studies
Authors: Oesterlund Anna H , Lander Flemming , Lauritsen Jens
Published: November 2014

Background

The occupational injury incident rate remains relatively high in the European Union. The case-crossover study gives a unique opportunity to study transient risk factors that normally would be very difficult to approach. Studies like this have been carried out in both America and Asia, but so far no relevant research has been conducted in Europe.

Method

Case-crossover studies of occupational injuries were collected from PubMed and Embase and read through. Previous experiences concerning method, exposure and outcome, time-related measurements and construction of the questionnaire were taken into account in the preparation of a pilot study. Consequently, experiences from the pilot study were used to design the study protocol. Approximately 2000 patients with an occupational injury will be recruited from the emergency departments in Herning and Odense, Denmark. A standardised questionnaire will be used to collect basic demographic data and information on eight transient risk factors. Based on previous studies and knowledge on occupational injuries the transient risk factors we chose to examine were: time pressure, performing a task with a different method/using unaccustomed technique, change in working surroundings, using a phone, disagreement, feeling ill, being distracted and using malfunctioning machinery/tools or work material. Exposure time ‘just before the injury’ will be compared with two control periods, ‘previous day at the same time of the injury’ (pair match) and the previous work week (usual frequency).

Discussion

This study protocol describes a unique opportunity to calculate the effect of transient risk factors on occupational injuries in a European setting.
Acquired immunodeficiency similar to Gulf War illness in a dead former serviceman
Authors: Roncati Luca , Gatti A M , Pusiol T , Barbolini G , Maiorana A
Published: November 2014

A 38-year-old non-commissioned officer was certified unfit for military duty several months before his death. The forensic autopsy revealed a severe bone marrow aplasia and a pulmonary angioinvasive aspergillosis. Moreover, the presence of inorganic foreign particles in the pulmonary macrophages and intestinal endothelia was observed. The microanalysis implemented on these last selected specimens revealed the presence of silica particles microimpregnated by lanthanides and steel. The patient's acquired immunodeficiency appears comparable with that of Iraqi civilians suffering from Gulf War illness. This is the first report in the literature of the presence of intestinal endothelia engulfed by foreign war particulates; the silica particles may have entered the intestinal endothelia via the blood stream or by ingestion of impregnated fruit and vegetable foodstuffs. This finding provides new perspectives in the assessment of war-associated diseases and includes electron probe microanalysis among the new techniques of military and forensic medicine.
Patient-reported outcome measures for patients with cerebral aneurysms acquired via social media: data from a large nationwide sample
Authors: Chen Michael , Mangubat Erwin , Ouyang Bichun
Published: December 2014

Background

With greater survival rates, patient-reported outcome measures (PROMs) among survivors of ruptured cerebral aneurysm should be an increasing concern among neurointerventionalists. Prior studies were limited in scale and generalizability. Our study aims were to (1) evaluate the validity of cerebral aneurysm PROMs obtained from social media; (2) determine the persistence of PROMs over time; and (3) determine what PROMs still exist in those with no physical impairments.

Methods

By engaging national brain aneurysm support groups and using an online questionnaire modeled after the generic EQ-5D instrument, we asked respondents to classify their health in five dimensions including mobility, self-care, usual activities, pain/discomfort, and anxiety/depression using a 3-point Likert scale.

Results

In 2 months we received 604 responses from 46 states in the USA. Our cohort of ruptured aneurysm respondents reported PROMs similar to previously published series. Over time, headache and anxiety improved while depression, level of exercise, and return to work remained unchanged. We found that memory worsened after 2 years. Among those without any physical impairment, rates of 20.6%, 14.9%, 12.6%, and 23% were seen for significant headaches, significant memory loss, significant depression, and sense of life being negatively affected, respectively.

Conclusions

Despite this novel study design, we obtained results comparable to prior studies. These results suggest that many patients with ruptured cerebral aneurysms, regardless of whether they are >2 years after the event and/or free of physical impairment, struggle with a poor quality of life. The latency, scale, and low cost of this study design may accelerate future cerebral aneurysm PROM research.
Focus on: Hendra virus in Australia
Authors: Hughes Kristopher
Published: November 2014

Cases of Hendra virus infection in horses in Australia have been seen regularly since the virus was first isolated in 1994. Kristopher Hughes, associate professor of equine medicine at Charles Sturt University in Australia, gives an overview of how knowledge of the virus has developed in the past 20 years
Ten‐minute chat
Authors: Hallas David
Published: December 2014

David Hallas qualified from Liverpool vet school in 1990 and is general manager at Intervet/Schering‐Plough Animal Health.
Developing an interest in wildlife
Authors: Mullineaux Liz
Published: December 2014

Liz Mullineaux developed an interest in British wildlife through working with local rehabilitators in Somerset. In 2004, she travelled to New Zealand on a Frank Beattie Travel Scholarship to investigate wildlife rehabilitation projects there. On her return, she developed her interest further by registering for a part‐time doctorate at the University of Edinburgh, which she completed earlier this year
Bringing history alive
Authors: Scott M C
Published: December 2014

M. C. Scott was senior clinical anaesthetist at Cambridge vet school before turning to writing. Of the 10 novels she has published, the first four were contemporary crime thrillers. She now writes historical novels and has set up the Historical Writers' Association
A career in conservation
Authors: Leader‐Williams Nigel
Published: December 2014

Nigel Leader‐Williams is Director of Conservation Leadership in the Department of Geography at the University of Cambridge, where he works to build capacity in conservation through interdisciplinary research and teaching, with a focus on large mammals that conflict with human interests
Working on the wild side
Authors: Sainsbury Tony
Published: December 2014

The Zoological Society of London and the Royal Veterinary College jointly run Masters courses on wild animals – one in wild animal health and another in wild animal biology. Senior lecturer Tony Sainsbury explains
Developing an interest in behaviour
Authors: Corridan Claire
Published: December 2014

Claire Corridan studied veterinary medicine because she was interested in animal welfare. She has since built her career on behavioural medicine
Enjoying the company of healthy animals
Authors: Carr John
Published: December 2014

Inspired by the television series ‘Daktari’ in the 1960s, John Carr decided he wanted to be a vet and, having worked on a large farm as a boy, he found he particularly liked pigs. He is now an international consultant in pig medicine
Embarking on a surgical internship
Authors: Egan Padraig
Published: December 2014

Internships are popular with newly graduated vets as a stepping stone to specialisation, so to enrol as an intern after four years in mixed practice was a decision that Padraig Egan did not take lightly
New graduate's diary
Authors: Hall Jen
Published: December 2014

Jennifer Hall is three months into her first job and is quickly gaining more knowledge and experience.
Nursing exotic animals and wildlife
Authors: Whitlock Emma
Published: December 2014

Emma Whitlock knew she wanted to be a veterinary nurse after seeing the postoperative care of patients during work experience at her local practice. She is now senior avian and exotic veterinary nurse at Great Western Exotics. Here, she describes the route her career has taken
Following the food production pathway
Authors: Nute Jill
Published: December 2014

Few veterinary students entertain the idea that their career might involve public health. Jill Nute has devised a week‐long course that aims to provide interested students with a unique opportunity to get to grips with an underrated area of veterinary science
Ten‐minute chat
Authors: O'Flynn Ann
Published: December 2014

In her final year at vet school, Ann O'Flynn realised she wanted something a bit different from the normal vet jobs. She was already playing rugby for England and needed an employer who would understand that sporting excellence was just as important to her as her professional career. She found that the Army offered just what she wanted.
Nonlinear refractive index change and optical rectification in a GaN-based step quantum wells with strong built-in electric field
Authors: Zhang Li
Published: November 2014

Based on the compact density matrix approach, the linear and nonlinear refractive index change (RIC) and optical rectification (OR) coefficients in a GaN -based step QW with strong built-in electric field (BEF) have been theoretically deduced and investigated in detail. The analytical electronic state is derived by the two airy functions. And the band nonparabolicity is taken into account by using an energy dependence effective mass (EDEM) method. Numerical calculations on a four-layer AlN / GaN / Al x Ga 1-x N / AlN step QW are performed, and the curves for the geometric factors, the linear, the nonlinear, the total RICs and the OR coefficients as functions of the structural parameters of the step QW are discussed. The features for these curves were specified and reasons for the features were explained reasonably. It is found that the decreasing of well width Lw, and step barrier width Lb and the doped concentration x in step barrier will result in the significant enhancement of the RICs. With the decrease of Lw, Lb and x, the resonant photon energies of RIC and OR coefficients have obvious blue-shift. Moreover, the RIC and OR coefficients behave different dependence on the structural parameters of the GaN -based step QWs. The profound physical reasons are also analyzed.
Energy-Efficient and Area-Efficient QC-LDPC with RS Decoders Using 2M-LMSA
Authors: Yao Chang-Kun , Tang Yun-Ching , Lin Hongchin
Published: November 2014

This study proposes an energy-efficient and area-efficient dual-path low-density parity-check (LDPC) with Reed–Solomon (RS) decoder for communication systems. Hardware complexity is reduced by applying a dual-path 2-bit modified layered min-sum algorithm (2M-LMSA) to a (2550, 2040) quasi-cyclic LDPC (QC-LDPC) code with the column and row weights of 3 and 15, respectively. The simplified check node units (CNUs) reduce memory and routing complexity as well as the energy needed to decode each bit. A throughput of 11 Gb/s is achieved by using 90-nm CMOS technology at a clock frequency of 208 MHz at 0.9 V with average power of 244 mW on a chip area of 3.05 mm2. Decoding performance is further improved by appending the (255, 239) RS decoder after the LDPC decoder. The LDPC plus RS decoder consumes the power of 434 mW on the area of 3.45 mm2.
Free Vibration of Two-Directional Functionally Graded Annular Plates Using Chebyshev Collocation Technique and Differential Quadrature Method
Authors: Kumar Yajuvindra
Published: November 2014

This article deals with the free axisymmetric vibration of two-directional functionally graded annular plates. Ceramic and metal are considered two constituents of the functionally graded material (FGM), which are graded through thickness and radial directions of the plate. The Chebyshev collocation technique and differential quadrature method are employed to derive the frequency equations for an annular plate with both edges clamped and another one with both edges simply supported. The results for nonhomogeneous isotropic annular plates are also presented. The accuracy and efficiency of the present approach are confirmed through comparison of the frequencies obtained for homogeneous isotropic annular plates. Identical results are obtained for the two methods used. The effects of volume fraction index, coefficient of radial variation, exponent of power law, inner to outer radii ratio, and boundary conditions are discussed on the first three natural frequencies. It is found that the frequency of a functionally graded annular plate is greater than that of a homogeneous annular plate.
Asymmetric Solutions of SDOF System with Wire Rope Vibration Isolator Subjected to Harmonic Excitation
Authors: Zou Keguan , Nagarajaiah Satish , Dick Andrew
Published: November 2014

A new modification of homotopy analysis method (HAM) is proposed for capturing asymmetric solutions of wire rope isolation systems. Analytical expressions of asymmetric solutions to wire rope isolation systems are obtained. A dynamic system with quadratic polynomial restoring force is investigated specifically. Then the analytical results are applied to a single-degree-of-freedom (SDOF) system with wire rope vibration isolator to investigate the response curve and other dynamic characteristics. The analytical approximations match satisfactorily with the numerical results. The presented analytical approximation is a useful method to derive the response curves and examine limit cycles without resorting to numerical simulations.
Worker/wrapper/makes it/faster
Authors: Hackett Jennifer , Hutton Graham
Published: December 2014

Much research in program optimization has focused on formal approaches to correctness: proving that the meaning of programs is preserved by the optimisation. Paradoxically, there has been comparatively little work on formal approaches to efficiency: proving that the performance of optimized programs is actually improved. This paper addresses this problem for a general-purpose optimization technique, the worker/wrapper transformation. In particular, we use the call-by-need variant of improvement theory to establish conditions under which the worker/wrapper transformation is formally guaranteed to preserve or improve the time performance of programs in lazy languages such as Haskell.
There is no fork
Authors: Marlow Simon , Brandy Louis , Coens Jonathan , Purdy Jon
Published: December 2014

We describe a new programming idiom for concurrency, based on Applicative Functors, where concurrency is implicit in the Applicative <*> operator. The result is that concurrent programs can be written in a natural applicative style, and they retain a high degree of clarity and modularity while executing with maximal concurrency. This idiom is particularly useful for programming against external data sources, where the application code is written without the use of explicit concurrency constructs, while the implementation is able to batch together multiple requests for data from the same source, and fetch data from multiple sources concurrently. Our abstraction uses a cache to ensure that multiple requests for the same data return the same result, which frees the programmer from having to arrange to fetch data only once, which in turn leads to greater modularity.

While it is generally applicable, our technique was designed with a particular application in mind: an internal service at Facebook that identifies particular types of content and takes actions based on it. Our application has a large body of business logic that fetches data from several different external sources. The framework described in this paper enables the business logic to execute efficiently by automatically fetching data concurrently; we present some preliminary results.
How to keep your neighbours in order
Authors: McBride Conor Thomas
Published: December 2014

I present a datatype-generic treatment of recursive container types whose elements are guaranteed to be stored in increasing order, with the ordering invariant rolled out systematically. Intervals, lists and binary search trees are instances of the generic treatment. On the journey to this treatment, I report a variety of failed experiments and the transferable learning experiences they triggered. I demonstrate that a
total
element ordering is enough to deliver insertion and flattening algorithms, and show that (with care about the formulation of the types) the implementations remain as usual. Agda's
instance arguments
and
pattern synonyms
maximize the proof search done by the typechecker and minimize the appearance of proofs in program text, often eradicating them entirely. Generalizing to indexed recursive container types, invariants such as
size
and
balance
can be expressed in addition to
ordering
. By way of example, I implement insertion and deletion for 2-3 trees, ensuring both order and balance by the discipline of type checking.
Using formal methods to enable more secure vehicles
Authors: Fisher Kathleen
Published: December 2014

Networked embedded systems are ubiquitous in modern society. Examples include SCADA systems that manage physical infrastructure, medical devices such as pacemakers and insulin pumps, and vehicles such as airplanes and automobiles. Such devices are connected to networks for a variety of compelling reasons, including the ability to access diagnostic information conveniently, perform software updates, provide innovative features, and lower costs. Researchers and hackers have shown that these kinds of networked embedded systems are vulnerable to remote attacks and that such attacks can cause physical damage and can be hidden from monitors [1, 4].

DARPA launched the HACMS program to create technology to make such systems dramatically harder to attack successfully. Specifically, HACMS is pursuing a clean-slate, formal methods-based approach to the creation of high-assurance vehicles, where high assurance is defined to mean functionally correct and satisfying appropriate safety and security properties. Specific technologies include program synthesis, domain-specific languages, and theorem provers used as program development environments. Targeted software includes operating system components such as hypervisors, microkernels, file systems, and device drivers as well as control systems such as autopilots and adaptive cruise controls. Program researchers are leveraging existing high-assurance software including NICTA's seL4 microkernel and INRIA's CompCert compiler.

Although the HACMS project is less than halfway done, the program has already achieved some remarkable success. At program kick-off, a Red Team easily hijacked the baseline open-source quadcopter that HACMS researchers are using as a research platform. At the end of eighteen months, the Red Team was not able to hijack the newly-minted "SMACCMCopter" running high-assurance HACMS code, despite being given six weeks and full access to the source code of the copter. An expert in penetration testing called the SMACCMCopter "the most secure UAV on the planet."

In this talk, I will describe the HACMS program: its motivation, the underlying technologies, current results, and future directions.
From Ada 9x to spaceport America
Authors: Anderson Christine
Published: December 2014

Ada 95, aka Ada9X at the time because we didn't know when we would be done, was a labor of love for most of us. A spectacular team was assembled from all over the world. I had the distinct pleasure and honor of being the Department of Defense Ada 9X Project Manager. The lessons I learned and the experience I gained allowed me to do many other things after Ada 95 was completed. I ran an Air Force space technology laboratory. I was responsible for building military satellites and launching them from the Cape. Currently, I am responsible for developing and operating the first purpose-built commercial spaceport. Looking back, the common thread through all of these endeavors is innovation, dedication, strong team work, and a passion for the job at hand. That
can do
spirit and boundless energy is a must for success and I have been fortunate to work on unprecedented projects with colleagues who possessed these qualities. Anecdotes from the Ada9X Project to today's emerging commercial space industry will be provided from my experiences and observations.
TRA2β controls Mypt1 exon 24 splicing in the developmental maturation of mouse mesenteric artery smooth muscle
Authors: Zheng Xiaoxu , Reho John J. , Wirth Brunhilde , Fisher Steven A.
Published: November 2014

Diversity of smooth muscle within the vascular system is generated by alternative splicing of exons, yet there is limited understanding of its timing or control mechanisms. We examined splicing of myosin phosphatase regulatory subunit (Mypt1) exon 24 (E24) in relation to smooth muscle myosin heavy chain (Smmhc) and smoothelin (Smtn) alternative exons (Smmhc E6 and Smtn E20) during maturation of mouse mesenteric artery (MA) smooth muscle. The role of transformer 2β (Tra2β), a master regulator of splicing in flies, in maturation of arterial smooth muscle was tested through gene inactivation. Splicing of alternative exons in bladder smooth muscle was examined for comparative purposes. MA smooth muscle maturation began after postnatal week 2 and was complete at maturity, as indicated by switching to Mypt1 E24+ and Smtn E20− splice variants and 11-fold induction of Smmhc. Similar changes in bladder were complete by postnatal day 3. Splicing of Smmhc E6 was temporally dissociated from Mypt1 E24 and Smtn E20 and discordant between arteries and bladder. Tamoxifen-induced smooth muscle-specific inactivation of Tra2β within the first week of life but not in maturity reduced splicing of Mypt1 E24 in MAs. Inactivation of Tra2β causing a switch to the isoform of MYPT1 containing the COOH-terminal leucine zipper motif (E24−) increased arterial sensitivity to cGMP-mediated relaxation. In conclusion, maturation of mouse MA smooth muscle begins postnatally and continues until sexual maturity. TRA2β is required for specification during this period of maturation, and its inactivation alters the contractile properties of mature arterial smooth muscle.
Influence of ZIP14 (slc39A14) on intestinal zinc processing and barrier function
Authors: Guthrie Gregory J. , Aydemir Tolunay B. , Troche Catalina , Martin Alyssa B. , Chang Shou-Mei , Cousins Robert J.
Published: November 2014

ZIP14 is a zinc transport protein with high expression in the small intestine and liver. Zip14 is upregulated during endotoxemia and leads to increased liver zinc content and transient hypozinemia. Since body zinc status and inflammation are associated with changes in intestinal permeability, we hypothesized that ZIP14 may influence intestinal permeability. Wild-type (WT) and Zip14 knockout (KO) mice were used to determine ZIP14-associated intestinal zinc metabolism and effects on permeability. Fractionation of plasma membranes revealed that ZIP14 is localized to the basolateral membrane of enterocytes. Studies utilizing 65Zn administered by subcutaneous injection revealed greater zinc accumulation in the SI of Zip14 KO mice compared with WT mice. Isolation of endosomes confirmed the presence of ZIP14. Quantification of endosomal zinc concentration by FluoZin-3AM fluorescence demonstrated that zinc is trapped in endosomes of Zip14 KO mice. Intestinal permeability assessed both by plasma FITC-dextran following gavage and by serum endotoxin content was greater in Zip14 KO mice. Threonine phosphorylation of the tight junction protein occludin, which is necessary for tight junction assembly, was reduced in KO mice. Claudin 1 and 2, known to have an inverse relationship in regards to tight junction integrity, reflected impaired barrier function in KO jejunum. These data suggest involvement of ZIP14 in providing zinc for a regulatory role needed for maintenance of the intestinal barrier. In conclusion, ZIP14 is a basolaterally localized protein in enterocytes and is involved in endosomal trafficking of zinc and is necessary for proper maintenance of intestinal tight junctions.
Role of protein phosphorylation in excitation-contraction coupling in taurine deficient hearts
Authors: Ramila K. C. , Jong Chian Ju , Pastukh Viktor , Ito Takashi , Azuma Junichi , Schaffer Stephen W.
Published: November 2014

Taurine is a beta-amino acid found in very high concentration in the heart. Depletion of these intracellular stores results in the development of cardiomyopathy, thought to be mediated by abnormal sarcoplasmic reticular (SR) Ca2+transport. There is also evidence that taurine directly alters the Ca2+sensitivity of myofibrillar proteins. Major regulators of SR Ca2+ATPase (SERCA2a) are the phosphorylation status of a regulatory protein, phospholamban, and SERCA2a expression, which are diminished in the failing heart. The failing heart also exhibits reductions in myofibrillar Ca2+sensitivity, a property regulated by the phosphorylation of the muscle protein, troponin I. Therefore, we tested the hypothesis that taurine deficiency leads to alterations in SR Ca2+ATPase activity related to reduced phospholamban phosphorylation and expression of SERCA2a. We found that a sequence of events, which included elevated protein phosphatase 1 activity, reduced autophosphorylation of CaMKII, and reduced phospholamban phosphorylation, supports the reduction in SR Ca2+ATPase activity. However, the reduction in SR Ca2+ATPase activity was not caused by reduced SERCA2a expression. Taurine transporter knockout (TauTKO) hearts also exhibited a rightward shift in the Ca2+dependence of the myofibrillar Ca2+ATPase, a property that is associated with an elevation in phosphorylated troponin I. The findings support the observation that taurine deficient hearts develop systolic and diastolic defects related to reduced SR Ca2+ATPase activity, a change mediated in part by reduced phospholamban phosphorylation.
Enhancing hepatic mitochondrial fatty acid oxidation stimulates eating in food-deprived mice
Authors: Mansouri Abdelhak , Pacheco-López Gustavo , Ramachandran Deepti , Arnold Myrtha , Leitner Claudia , Prip-Buus Carina , Langhans Wolfgang , Morral Núria
Published: November 2014

Hepatic fatty acid oxidation (FAO) has long been implicated in the control of eating. Nevertheless, direct evidence for a causal relationship between changes in hepatic FAO and changes in food intake is still missing. Here we tested whether increasing hepatic FAO via adenovirus-mediated expression of a mutated form of the key regulatory enzyme of mitochondrial FAO carnitine palmitoyltransferase 1A (CPT1mt), which is active but insensitive to inhibition by malonyl-CoA, affects eating and metabolism in mice. CPT1mt expression increased hepatocellular CPT1 protein levels. This resulted in an increase in circulating ketone body levels in fasted CPT1mt-expressing mice, suggesting an increase in hepatic FAO. These mice did not show any significant changes in cumulative food intake, energy expenditure, or respiratory quotient after 4-h food deprivation. After 24-h food deprivation, however, the CPT1mt-expressing mice displayed increased food intake. Thus expression of CPT1mt in the liver increases hepatic FAO capacity, but does not inhibit eating. Rather, it may even stimulate eating after prolonged food deprivation. These data do not support the hypothesis that an increase in hepatic FAO decreases food intake.
Reflex neuromodulation of bladder function elicited by posterior tibial nerve stimulation in anesthetized rats
Authors: Kovacevic Mario , Yoo Paul B.
Published: November 2014

Although posterior tibial nerve stimulation (PTNS) has been shown in both clinical and animal studies to elicit bladder-inhibitory reflexes, our understanding of the role of posterior tibial nerve (PTN) afferents that elicit these responses is significantly limited. To this end, we investigated the effects of frequency-dependant PTNS in urethane-anesthetized rats undergoing repeated urodynamic fills. Nerve stimulation trials (10 min) resulted in statistically significant inhibition of the urinary bladder, both during and after nerve stimulation ( P < 0.05). PTNS applied at 5 Hz resulted in both acute and prolonged changes that corresponded to 38.0% and 34.1% reductions in the bladder contraction frequency, respectively. In contrast, PTNS applied at 10 Hz could only elicit an acute decrease (22.9%) in bladder activity. Subsequent electrical activation of individual PTN branches (lateral or medial plantar nerves) confirmed that these bladder reflexes are mediated by specific subsets of the PTN trunk. Both acute and prolonged inhibition of the bladder were achieved by electrical stimulation of the lateral plantar (10 and 20 Hz) and medial plantar (5 and 10 Hz) nerves. Finally, we report a bladder-excitatory reflex that is elicited by electrical activation of either the PTN trunk or lateral plantar nerve at 50 Hz. This study shows that multiple bladder reflexes are tuned to specific subsets of nerve afferents and stimulation frequencies, each of which provide novel insights into the physiological effects of PTNS.
Activation of AMP-activated protein kinase inhibits ER stress and renal fibrosis
Authors: Kim Hyosang , Moon Soo Young , Kim Joon-Seok , Baek Chung Hee , Kim Miyeon , Min Ji Yeon , Lee Sang Koo
Published: November 2014

It has been suggested that endoplasmic reticulum (ER) stress facilitates fibrotic remodeling. Therefore, modulation of ER stress may serve as one of the possible therapeutic approaches to renal fibrosis. We examined whether and how activation of AMP-activated protein kinase (AMPK) suppressed ER stress induced by chemical ER stress inducers [tunicamycin (TM) and thapsigargin (TG)] and also nonchemical inducers in tubular HK-2 cells. We further investigated the in vivo effects of AMPK on ER stress and renal fibrosis. Western blot analysis, immunofluorescence, small interfering (si)RNA experiments, and immunohistochemical staining were performed. Metformin (the best known clinical activator of AMPK) suppressed TM- or TG-induced ER stress, as shown by the inhibition of TM- or TG-induced upregulation of glucose-related protein (GRP)78 and phosphorylated eukaryotic initiation factor-2α through induction of heme oxygenase-1. Metformin inhibited TM- or TG-induced epithelial-mesenchymal transitions as well. Compound C (AMPK inhibitor) blocked the effect of metformin, and 5-aminoimidazole-4-carboxamide-1β riboside (another AMPK activator) exerted the same effects as metformin. Transfection with siRNA targeting AMPK blocked the effect of metformin. Consistent with the results of cell culture experiments, metformin reduced renal cortical GRP78 expression and increased heme oxygenase-1 expression in a mouse model of ER stress-induced acute kidney injury by TM. Activation of AMPK also suppressed ER stress by transforming growth factor-β, ANG II, aldosterone, and high glucose. Furthermore, metformin reduced GRP78 expression and renal fibrosis in a mouse model of unilateral ureteral obstruction. In conclusion, AMPK may serve as a promising therapeutic target through reducing ER stress and renal fibrosis.
Oxidative stress contributes to orthopedic trauma-induced acute kidney injury in obese rats
Authors: Mittwede Peter N. , Xiang Lusha , Lu Silu , Clemmer John S. , Hester Robert L.
Published: November 2014

After trauma, obese patients have an increased risk of developing acute kidney injury (AKI). We have demonstrated that obese Zucker (OZ) rats, but not lean Zucker (LZ) rats, develop AKI 24 h after orthopedic trauma. ROS have been implicated in the pathophysiology of AKI in models of critical illness. However, the contribution of ROS to trauma-induced AKI in the setting of obesity has not been determined. We hypothesized that AKI in OZ rats after trauma is mediated by increased oxidative stress. Male LZ and OZ rats were divided into control and trauma groups, with a subset receiving treatment after trauma with the antioxidant apocynin (50 mg/kg ip, 2 mM in drinking water). The day after trauma, glomerular filtration rate, plasma creatinine, urine kidney injury molecule-1, and albumin excretion as well as renal oxidant and antioxidant activity were measured. After trauma, compared with LZ rats, OZ rats exhibited a significant decrease in glomerular filtration rate along with significant increases in plasma creatinine and urine kidney injury molecule-1 and albumin excretion. Additionally, oxidative stress was significantly increased in OZ rats, as evidenced by increased renal NADPH oxidase activity and urine lipid peroxidation products (thiobarbituric acid-reactive substances), and OZ rats also had suppressed renal superoxide dismutase activity. Apocynin treatment significantly decreased oxidative stress and AKI in OZ rats but had minimal effects in LZ rats. These results suggest that ROS play an important role in AKI in OZ rats after traumatic injury and that ROS may be a potential future therapeutic target in the obese after trauma.
mTOR contributes to ER stress and associated apoptosis in renal tubular cells
Authors: Dong Guie , Liu Yu , Zhang Lei , Huang Shuang , Ding Han-Fei , Dong Zheng
Published: November 2014

ER stress has been implicated in the pathogenesis of both acute and chronic kidney diseases. However, the molecular regulation of ER stress in kidney cells and tissues remains poorly understood. In this study, we examined tunicamycin-induced ER stress in renal proximal tubular cells (RPTC). Tunicamycin induced the phosphorylation and activation of PERK and eIF2α within 2 h in RPTC, which was followed by the induction of GRP78 and CHOP. Consistently, tunicamycin also induced apoptosis in RPTC. Interestingly, mTOR was activated rapidly during tunicamycin treatment, as indicated by phosphorylation of both mTOR and p70S6K. Inhibition of mTOR with rapamycin partially suppressed the phosphorylation of PERK and eIF2a and the induction of CHOP and GRP78 induction during tunicamycin treatment. Rapamycin also inhibited apoptosis during tunicamycin treatment and increased cell survival. Collectively, the results suggest that mTOR plays a regulatory role in ER stress, and inhibition of mTOR may have potential therapeutic effects in ER stress-related renal diseases.
Inhaled nitric oxide alters the distribution of blood flow in the healthy human lung, suggesting active hypoxic pulmonary vasoconstriction in normoxia
Authors: Asadi Amran K. , Sá Rui Carlos , Kim Nick H. , Theilmann Rebecca J. , Hopkins Susan R. , Buxton Richard B. , Prisk G. Kim
Published: November 2014

Hypoxic pulmonary vasoconstriction (HPV) is thought to actively regulate ventilation-perfusion (V̇a/Q̇) matching, reducing perfusion in regions of alveolar hypoxia. We assessed the extent of HPV in the healthy human lung using inhaled nitric oxide (iNO) under inspired oxygen fractions (FiO2) of 0.125, 0.21, and 0.30 (a hyperoxic stimulus designed to abolish HPV without the development of atelectasis). Dynamic measures of blood flow were made in a single sagittal slice of the right lung of five healthy male subjects using an arterial spin labeling (ASL) MRI sequence, following a block stimulus pattern (3 × 60 breaths) with 40 ppm iNO administered in the central block. The overall spatial heterogeneity, spatiotemporal variability, and regional pattern of pulmonary blood flow was quantified as a function of condition (FiO2× iNO state). While spatial heterogeneity did not change significantly with iNO administration or FiO2, there were statistically significant increases in Global Fluctuation Dispersion,(a marker of spatiotemporal flow variability) when iNO was administered during hypoxia (5.4 percentage point increase, P = 0.003). iNO had an effect on regional blood flow that was FiO2dependent ( P = 0.02), with regional changes in the pattern of blood flow occurring in hypoxia ( P = 0.007) and normoxia ( P = 0.008) tending to increase flow to dependent lung at the expense of nondependent lung. These findings indicate that inhaled nitric oxide significantly alters the distribution of blood flow in both hypoxic and normoxic healthy subjects, and suggests that some baseline HPV may indeed be present in the normoxic lung.