Experimental comparison of trauma in lateral (+Gy), rearward facing (+Gx), and forward facing (-Gx) body orientations when restrained by lap belt only.
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The processing of pseudorabies virus glycoprotein gX in infected cells and in an uninfected cell line.
Pseudorabies virus (PRV) produces a glycoprotein, gX, that accumulates in the medium of infected cells. The gX gene was expressed in Chinese hamster ovary cells (CHOgX cells) using the cytomegalovirus Towne major immediate early promoter. Like PRV-infected cells, CHOgX cells produced gX and exported it into the medium. Tunicamycin reduced the molecular weight of the gX in the medium to 89 kDa, compared with 99 kDa for gX made in the absence of drug. In the presence of tunicamycin gX produced by both PRV-infected cells and CHOgX cells was still glycosylated, as indicated by incorporation of [14C]glucosamine. The most likely form of this glycosylation is O-linked. In a pulse-chase experiment, gX first appeared in a 90-kDa form, then a 115-kDa form. This 115-kDa form is probably cleaved to give the 99-kDa form of gX that is released into the medium. The 115-kDa form was much more persistent in the PRV-infected Vero cells than in the CHOgX cells. In both cell types, gX was labeled by [35S]sulfate in the presence and absence of tunicamycin.
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Characterization of the human gene for Gx alpha, a pertussis toxin-insensitive regulatory GTP-binding protein.
We have cloned the human chromosomal gene coding for the alpha subunit of Gx (Gx alpha), a heterotrimeric signal-transducing GTP-binding protein (G protein) that is insensitive to pertussis toxin. Gx alpha cDNA has been cloned both from rat brain (Matsuoka, M., Itoh, H., Kozasa, T., and Kaziro, Y. (1988) Proc. Natl. Acad. Sci. U.S.A. 85, 5384-5388) and from human retina (referred to as Gz alpha, Fong, H.K.W., Yoshimoto, K.K., Eversole-Cire, P., and Simon, M.I. (1988) Proc. Natl. Acad. Sci. U.S.A. 85, 3066-3070). In this paper, we have analyzed the structure of human gene for Gx alpha, which spans more than 60 kilobases. Comparison of the nucleotide sequence of the human chromosomal gene with that of the human retinal Gx alpha cDNA revealed that the gene is composed of three exons and two introns. The first exon contains about 170 base pairs of the 5'-noncoding sequence. The second exon contains the 5'-noncoding and the N-terminal coding sequences, and the third exon contains the C-terminal and the 3'-noncoding sequences. S1 mapping and primer extension analysis have identified the presence of multiple transcription initiation sites, upstream of which were found 12 SP1 binding sites and one CAAT sequence but no TATA sequence. Southern blot analysis indicated that a single copy of the Gx alpha gene is present per haploid human genome. RNA blot hybridization analysis revealed that Gx alpha mRNA is expressed mainly in brain.
Base Sequence↗
A glycoprotein gX-beta-galactosidase fusion gene as insertional marker for rapid identification of pseudorabies virus mutants.
We describe the isolation and characterization of infectious pseudorabies virus (PrV) mutants expressing functional beta-galactosidase. To obtain high level expression of the enzyme, sequences of the bacterial beta-galactosidase gene starting with codon 8 were inserted in frame behind the promoter and the first seven codons of the nonessential PrV glycoprotein gX-gene. Cotransfection of this construct with viral DNA yielded PrV mutants that could be easily identified after plaque staining with a chromogenic substrate. These mutants carry the gX-beta galactosidase fusion gene inserted into the authentic gX-gene leading to loss of gX-expression. The gX-beta galactosidase fusion gene could be excised as an expression cassette and placed into other non-essential PrV genomic regions, such as the thymidine kinase gene and the glycoprotein gI-gene, resulting in inactivation of the target genes. The fusion gene remains stably integrated in the viral genome at all three locations tested. It therefore appears ideal as an insertional and easily identifiable marker and greatly facilitates isolation and purification of PrV mutants.
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Replication and virulence of pseudorabies virus mutants lacking glycoprotein gX.
Pseudorabies virus (PRV) glycoprotein gX accumulates in the medium of infected cells. In an attempt to study the function of gX, two viruses were constructed that lacked a functional gX gene. One virus, PRV delta GX1, was derived by insertion of the herpes simplex virus thymidine kinase gene into the gX-coding region. The other virus, PRV delta GXTK-, was derived by subsequent deletion of the inserted herpes simplex virus thymidine kinase gene. Both viruses replicated in cell cultures but produced no gX. Furthermore, PRV delta GX1 was capable of killing mice with a 50% lethal dose of less than 100 PFU.
Animals↗
Immune response in pigs to Aujeszky's disease viruses defective in glycoprotein g1 or gX.
Two Aujeszky's disease virus glycoprotein genes, gX and g1, have been used to produce deletion mutants which have then been developed into vaccines. These deletions then allow differentiation between pigs infected with wild type virus and those given the vaccine. It is not clear whether the glycoproteins encoded for by these genes are needed to induce a full protective immune response, in which case deletion mutants would suffer from lack of potency. To test this, commercially available Aujeszky's virus vaccines which lacked either gX or g1 were compared and isogenic constructs were made which differed only in the absence or presence of gX and, or, g1. These constructs and vaccines were used to vaccinate the natural host of Aujeszky's disease, the pig, and potency was measured using challenge with wild type virus. In all cases vaccines which lacked g1 performed significantly less well than those in which g1 was present, whereas deletions of gX had no significant effect on vaccine performance.
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[Resistance to +Gz and +Gx loads of subjects in older age groups, healthy and with initial signs of atherosclerosis].
Tolerance to +Gz and +Gx acceleration of healthy and atherosclerotic subjects, aged 40-49, was investigated during 256 centrifugation tests in Bulgaria and the USSR. As compared to the healthy people, the atherosclerotic subjects showed a lower tolerance: when exposed to 7 +Gz the tolerance threshold decreased from 5.93 +/- 0.57 g to 5.7 +/- 0.44 g (i. e. by 0.23 g). Visual disorders in the healthy and atherosclerotic subjects were recorded in 29 and 50%, respectively, and loss of consciousness in 9.56 and 15.15%, respectively (Bulgarian data). When exposed to 5 +Gz and 6 +Gx no subjects displayed visual disorders or loss of consciousness (Soviet data). However, lower tolerance of 5 +Gz due to cardiac arrhythmias was observed in 4.8% atherosclerotic patients and in none healthy subjects. Lower tolerance to 6 +Gx associated with cardiac arrhythmias was recorder in 23.8% atherosclerotic versus 11.8% healthy subjects (i. e. 12% more). Nevertheless, in most subjects, both healthy and atherosclerotic, acceleration tolerance was good. These findings indicate that individual assessment of acceleration tolerance is of great importance for people older than 40 years with health abnormalities.
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Studies of the orientation of the mitochondrial redox carriers. III. Orientation of the gx and gy axes of the hemes of cytochrome oxidase with respect to the plane of the membrane in oriented membrane multilayers.
The EPR absorption properties of the hemes of cytochrome oxidase and their liganded derivatives were examined in oriented multilayers from isolated oxidase, mitochondrial membranes and membrane fragments of a bacterium, Paracoccus denitrificans. The hemes of the oxidase in all the systems investigated were oriented normal to the plane of the multilayers. The directions of the g signals corresponding to the gx and gy axes of the g tensor were found to be different in low-spin ferric heme in fully oxidized oxidase and in half-reduced liganded oxidase. It is suggested that this different orientation of gx and gy in fully oxidized oxidase and half-reduced liganded oxidase arises because the respective EPR signals belong to two different hemes, those of cytochrome a and a3.
Animals↗
[Effect of gravitational loads (+Gx) of the training schedule on the structure of the main arteries of dogs].
In dogs subjected to gravitational overloadings (+Gx) according to a special training schedule the wall structure of the main arteries (carotid, humeral, femoral and aorta) was studied by histological methods. The amount of catecholamines (adrenaline and noradrenaline) in the venous blood plasma and in the adrenal tissue was the index of the sympathoadrenal system condition. It was demonstrated that in trained animals the tolerance threshold to continuously increasing overloadings (+Gx) rose. It was accompanied by an elevated amount of catecholamines in blood and the adrenals In the walls of the main arteries studied a moderate hyperelastosis and a slight collagenization of the adventitia was noted. All parts of the microcirculatory bed were moderately dilated, which favoured a better blood outflow. The data obtained demonstrate adaptational changes, which can be interpreted as indices of training and reliability of the organism.
Adaptation, Physiological↗
Marine Vibrio Biocatalysts as Unique Green Transformation (GX) Tools at the Time to Sustainable Development Goals (SDGs).
Vibrios have sustained various types of ocean ecosystems, being key players in marine mineral cycles and essential partners in specific groups of marine life. Observed genome plasticity and metabolic versatility are some of the unique biological features of vibrios, and these traits could contribute in expanding their ecological niche in marine environments. Vibrios are now recognized as ecophysiologically essential microbial species for our planet. At the time to "Sustainable Development Goals" (SDGs), their genome plasticity and metabolic versatility have also been studied with the aim of solving global issues such as energy production and plastic pollution by creating new microbial biocatalysts. Here, we introduce recent progress on the application of vibrios aiming towards green transformation (GX).
Vibrio↗
Open reading frames encoding a protein kinase, homolog of glycoprotein gX of pseudorabies virus, and a novel glycoprotein map within the unique short segment of equine herpesvirus type 1.
DNA sequence analysis of the unique short (Us) segment of the genome of equine herpesvirus type 1 Kentucky A strain (EHV-1) by our laboratory and strains Kentucky D and AB1 by other workers identifies a total of nine open reading frames (ORF). In this report, we present the DNA sequence of three of these newly identified ORFs, designated EUS 2, EUS 3, and EUS 4. The EUS 2 ORF is 1146 nucleotides (nt) in length and encodes a potential protein of 382 amino acids. Cis-regulatory sequences upstream of the putative ATG start codon include a G/C box 112 nt upstream and two potential TATA-like elements located between 15 and 90 nt before the ATG. The EUS 2 translation product exhibits significant homology to Ser/Thr protein kinases encoded within the Us segments of other herpesviruses, such as herpes simplex virus (26% homology) and pseudorabies virus (PRV), (45% homology), and possesses sequence domains conserved in protein kinases of cellular and viral origin. The EUS 3 ORF begins 127 nt downstream from the EUS 2 stop codon and ends at a stop codon 1119 nt further downstream. A single TATA-like element maps 61 nt upstream of the ORF. This ORF encodes a potential protein of 373 amino acids and is a homolog of glycoprotein gX of PRV, as judged by overall homology of amino acid residues, cysteine displacement, and presence of potential glycosylation sites and signal sequence. Interestingly, the EUS 4 ORF encodes a potential membrane glycoprotein that does not exhibit homology to any reported protein sequence. The EUS 4 ORF encodes a 383 amino acid polypeptide with a sequence indicative of a signal sequence at its amino terminal end, glycosylation sites for N-linked oligosaccharides, and a transmembrane domain near its carboxyl terminus. Several cis-acting regulatory sequences lie upstream of this ORF. These findings support the observation that the short region of alphaherpesviruses show considerable variation in their genetic content and gene organization.
Amino Acid Sequence↗
[Functional state of the human hearing analyzer exposed to +Gx acceleration].
The functional state of the acoustic analyzer of man was investigated during an exposure to +Gx acceleration of 4-14 g applied at an angle of 78 degrees to the long axis of the body. During an exposure to 8-10 g the hearing state began to deteriorate. This included an increase in the tonic thresholds of hearing sensitivity with respect to the aerial and bone conduction, and an increase in the differential thresholds of hearing with respect to the strength and pitch. With an increase of the acceleration value these changes grew, reaching maximum at 14 g. It is suggested that possible mechanisms of changes in the hearing sensitivity are associated with disorders in the systems of sound conveyance and perception.
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Crystallization and preliminary x-ray diffraction studies of subtilisin GX from Bacillus sp. GX6644.
Subtilisin GX, a serine protease from Bacillus species GX6644, has been crystallized by the vapor diffusion method using ammonium sulfate as the precipitant. The space group is P212121 with a = 38.4 A, b = 70.3 A, c = 73.5 A, and one molecule in the asymmetric unit. The crystals diffract to beyond 2.0-A resolution and are suitable for a high resolution three-dimensional structure determination. All x-ray data used in the preliminary crystallographic study were collected with an electronic area detector.
Ammonium Sulfate↗
Mathematical modeling of the head and neck response to -Gx impact accelration (minimum articulation requirements).
Data on the dynamic response of the living human head and neck to -Gx impact acceleration was analyzed. The Calspan 3-D Computer Simulator of a Motor Vehicle Crash Victim was used to provide estimates of the head and neck response to be expected for the very specific deceleration profiles simulated. Two segments connected via a pivot were used to represent the head-neck system. The monitored T1 deceleration profile was used to drive this system and the simulation head-neck response was checked for accuracy in replicating motional characteristics and trends in the response mechanism. Two head pivot locations were considered. The first was the occipital condylar point and the second was a theorized hingepoint in the head which minimized the distance from this point to the T1 anatomical coordinate system over the range of body orientations observed in the photographic data. For the two geometrical representations, a successive approximation technique was employed to analyze the resulting data. This approach initially removed all constraints to head and neck motion, and the resulting simulation results were compared to the human data. Restrictions to head and neck motion were then successively added until an adequate replication of the human data was obtained. This approach made it possible to correlate specific events, such as loading of the head-to-neck dorsiflexion limiting angle, to head deceleration profile characteristics. Results were compared to calculations obtained for eight additional subjects and proved to be in good agreement.
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Prediction of head/neck dynamic response of selected military subjects to -Gx acceleration.
Eighteen young male subjects with NAMRL sled test experience to 15 G in --Gx acceleration were measured for physical characteristics of the head and neck and general body anthropometry. Measurements taken include head/neck range of motion, neck muscle stretch reflex time, neck muscle isometric strength capabilities, and seated and standard anthropometry. Data from these tests were tabulated and five subjects whose physical characteristics were most similar were selected for use in simulations. Experimental data from NAMRL sled tests were obtained for the five subjects in 6- and 15-G test runs. Measurements data from the five subjects were used to establish a data set for the MVMA-2D Crash Victim Simulator and acceleration profiles for 6- and 15-G sled runs were used as input to the model. Simulation results for head angular acceleration, head angular velocity, head angular position, head resultant acceleration, and T1 resultant acceleration were compared with the averaged experimental curves for the five subjects. In general, excellent agreement between simulation and experimental results was obtained although some consistent differences were noted. Effects of varying levels of muscle activation were investigated. Variations in muscle tension level were found to have significant effects on simulation results at both 6 and 15, G, especially on head angular position. The effects were noticeably greater at lower acceleration's, however. The model was also used to investigate some of the biomechanical mechanisms behind observed response characteristics.
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Predictive model of dynamic response of the human head/neck system to -Gx impact acceleration.
This paper describes the mathematical framework, underlying an empirical model, that predicts human head response using only the motion present at vertebra T1. Based on this framework, a model for --Gx impact acceleration was developed from data obtained on six volunteer subjects participating in the NAMRL impact acceleration experiments. Model performance was evaluated by comparing the errors in the predicted head responses with the normal variations observed between the responses of different subjects under identical impact accelerations. Independent sets of data were used for building and testing the model. The results of the evaluation indicate that the model will be useful in subsequent studies of human response to impact acceleration.
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Mechanism of head and neck response to -Gx impact acceleration: a math modeling approach.
Mathematical modeling has attained wider acceptance in recent years. In particular, the use of computer programs to simulate the dynamic response of a human in a crash situation has become an attractive alternative to full-scale experimental testing. This paper analyzes data on the dynamic response of the living human head and neck to -Gx impact acceleration, where the motion of the subject's head and neck in the midsagittal plane was monitored with inertial instrumentation and high-speed photography for confirmation. The Calspan "3D Computer Simulator of Motor Vehicle Crash Victims" was used to predict expected responses for the deceleration pulses employed. These estimates were compared to the fully instrumented human test runs. The standard 15-segment and 14-joint representation of the occupant was modified to include two sternoclavicular joints, increasing the articulation in the upper torso. Analysis of the data indicated that muscular activity in the head and neck seemed to be evident and does influence motion of the head, even at relatively high (10-G peak, 530 G/s onset) acceleration levels. Simulation of muscular contraction, using a spring-damper arrangement, improved the results significantly. Additionally, possible limitations to head-to-neck motion, such as ligament restrictions, were also modeled.
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The Gx system in relation to C-type viruses and heredity.
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Animals↗