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OBJECTIVE: The analytic solution of the overall pressure and oxygen partial pressure in the scaled module of manned space vehicle was deduced, it can be used by ECLSS overall engineering designer to predict the pressure control parameters when space flight or ground test is operating. METHOD: According to different gas supply situations and the role of pressure control zone prescribed by ECLSS, the flight duration was divided into several segments under the role of pressure control zone, and the differential equations were solved. RESULT: The analytic solutions of the total pressure and partial pressure of oxygen were obtained, and the theoretical curve corresponds well to the test value. CONCLUSION: The analytic solutions obtained were valid, and satisfied the requirement of engineering accuracy.
The subject of the present study is the Monte Carlo path-integral evaluation of the moments of spectral functions. Such moments can be computed by formal differentiation of certain estimating functionals that are infinitely differentiable against time whenever the potential function is arbitrarily smooth. Here, I demonstrate that the numerical differentiation of the estimating functionals can be more successfully implemented by means of pseudospectral methods (e.g., exact differentiation of a Chebyshev polynomial interpolant), which utilize information from the entire interval . The algorithmic detail that leads to robust numerical approximations is the fact that the path-integral action and not the actual estimating functional are interpolated. Although the resulting approximation to the estimating functional is nonlinear, the derivatives can be computed from it in a fast and stable way by contour integration in the complex plane, with the help of the Cauchy integral formula (e.g., by Lyness' method). An interesting aspect of the present development is that Hamburger's conditions for a finite sequence of numbers to be a moment sequence provide the necessary and sufficient criteria for the computed data to be compatible with the existence of an inversion algorithm. Finally, the issue of appearance of the sign problem in the computation of moments, albeit in a milder form than for other quantities, is addressed.
Cartilaginous fish, primarily sharks, rays and skates (elasmobranchs), appeared 450 million years ago. They are the most primitive vertebrates, exhibiting jaws and teeth, adaptive immunity, a pressurized circulatory system, thymus, spleen, and a liver comparable to that of humans. The most used elasmobranch in biomedical research is the spiny dogfish shark, Squalus acanthias. Comparative genomic analysis of the dogfish shark, the little skate (Leucoraja erincea), and other elasmobranchs have yielded insights into conserved functional domains of genes associated with human liver function, multidrug resistance, cystic fibrosis, and other biomedically relevant processes. While genomic information from these animals is informative in an evolutionary framework, experimental verification of functions of genomic sequences depends heavily on cell culture approaches. We have derived the first multipassage, continuously proliferating cell line of a cartilaginous fish. The line was initiated from embryos of the spiny dogfish shark. The cells were maintained in a medium modified for fish species and supplemented with cell type-specific hormones, other proteins and sera, and plated on a collagen substrate. SAE cells have been cultured continuously for three years. These cells can be transfected by plasmids and have been cryopreserved. Expressed Sequence Tags generated from a normalized SAE cDNA library included a number of markers for cartilage and muscle, as well as proteins influencing tissue differentiation and development, suggesting that SAE cells may be of mesenchymal stem cell origin. Examination of SAE EST sequences also revealed a cartilaginous fish-specific repetitive sequence that may be evidence of an ancient mobile genetic element that most likely was introduced into the cartilaginous fish lineage after divergence from the lineage leading to teleosts.
OBJECTIVE: The seat-to-head transfer function of the human body reflects the biodynamic response. Based on measured data, biodynamic models have been proposed to reflect this response. They must satisfy usually the international published mean values of the seat-to-head transfer function. The question arises to what extent mean values reflect individual pattern of biodynamics. METHODS: An experimental study was performed with 39 male subjects sitting on a hard seat without back rest and with supported feet. They were exposed to random whole-body vibration at three intensities with a relaxed and an erect posture. The accelerations in the z-direction were measured at the seat and head. The seat-to-head transfer functions with the associated coherence functions were calculated. RESULTS: The biodynamic response characterised by the maximum of the seat-to-head transmissibility and the frequency of its occurrence is influenced by the posture of the subjects in a dominant way and shows an individual variability of considerable extent. The mean responses suggest a missing effect of vibration intensity, but individually different effects of the intensity were found. Repeated measurements confirmed this result. CONCLUSIONS: The application of a model validated by the comparison with mean values of the transmissibility could cause misleading conclusions, if it is used for the prediction of individual spinal loads. Models prepared for the calculation of individual loads should be validated by a mean individual transmissibility derived from repeated measurements. RELEVANCE: The results illustrate the loss of information by averaging individual transfer functions and the consequence of a limited validity and applicability in occupational health, ergonomics, and design.
BACKGROUND: Preservation of parasympathetic and sympathetic nerves is required to avoid urogenital function disturbances after total mesorectal excision (TME) for rectal carcinoma. This study sought to determine whether intraoperative stimulation of parasympathetic nerves with monitoring of bladder contraction is useful in meeting this demand. STUDY DESIGN: In a prospective pilot study, 17 patients, 11 men and 6 women, underwent TME with pelvic autonomic nerve preservation performed by an experienced surgeon. The parasympathetic nerves were stimulated by an electrostimulation device (Screener 3625, Medronic), and the resulting bladder contraction was measured manometrically in all patients. Variations in pulse rate and voltage were measured to determine optimal stimulation parameters. A standardized questionnaire was used to record urogenital function disturbances. Residual urine volume was measured by ultrasound pre- and postoperatively. Shortterm outcomes data were evaluated to establish a possible association between intraoperative test results and postoperative bladder function. RESULTS: In 15 of 17 patients undergoing TME with pelvic autonomic nerve preservation for rectal carcinoma, the parasympathetic nerves were identified based on nerve stimulation-induced bladder contraction. Two patients with negative results on intraoperative nerve stimulation had persisting bladder dysfunction requiring an indwelling catheter after discharge from hospital. In spite of a short median followup of 2 months (range 1 to 4 months), in 7 of 10 men with intact erectile function prior to surgery, postoperative erectile dysfunction could be excluded. The study showed a pulse rate of 35 Hz and an electric potential of 12 V to be optimal stimulation parameters, associated with a mean intravesical pressure rise of 12.7 cm H(2)O (range 2.8 to 18.0 cm H(2)O). CONCLUSIONS: Intraoperative nerve stimulation with monitoring of intravesical pressure represents a technically simple procedure for the identification and verification of function of pelvic parasympathetic nerves during TME for rectal carcinoma.
Fusarium oxysporum is a significant threat to agriculture and One Health, requiring advanced molecular tools for functional genomic analyses and biological control agent development. Existing gene-editing methods are hampered by costly protoplast preparation protocols and by CRISPR-Cas9 limitations, such as restricted protospacer adjacent motif (PAM) sequences and complex guide RNA requirements. We engineered an efficient CRISPR/Cpf1 system that overcomes these issues through three main innovations: small-scale protoplast generation using filter column-based methods that greatly reduce enzyme consumption while simplifying workflows, a CRISPR/Cpf1 system with shorter guide RNA design and staggered DNA cleavage to promote homologous recombination, and minimal homology arm strategies that significantly decrease cloning complexity. Extensive validation confirms successful gene targeting with molecular verification and functional analysis via standardized pathogenicity assays. This integrated platform offers affordable, accessible tools for systematic F. oxysporum research, enhancing fundamental understanding of plant-pathogen interactions and supporting high-throughput screening vital for agricultural biotechnology and biological agent development.
Prospective nucleic acid tests were carried out for human immunodeficiency virus (HIV) and hepatitis C virus (HCV) using the COBAS Amplicor HIV-1 and HCV tests (Roche Diagnostics, Meylan, France) on potential organ (n=113) and cornea (n=368) donors in France to evaluate their performance and suitability for use as a complement to routine serological tests. Blood samples were collected from organ donors with preserved cardiac function after verification of cerebral death. Blood samples were collected from cornea donors post-mortem within 48 hr after death. An internal control was added to the samples before extraction to monitor each individual polymerase chain reaction (PCR). The nucleic acid tests were always interpretable in organ donors and negative in all except in 2 anti-HCV positive patients. One had an indeterminate HIV p24 antigen but was negative for HIV RNA. HIV and HCV RNA were not found in cornea donors with a negative serology but indeterminate molecular results were frequent in this group (17.6%). Cornea donors also gave significantly more (14.4%) indeterminate serological results than organ donors (1.8%) (P<0.001). This was due to the poor quality of the blood samples collected post-mortem. However, there was no correlation between indeterminate results of serological and molecular tests. There were 16/19 (84%) indeterminate serological results for HIV and 4/4 (100%) for HCV that were negative by PCR. Thus, nucleic acid tests could be useful for qualifying a donor whose serological results are indeterminate. The extraction procedures on post-mortem specimens and/or blood collection must be changed to improve the performance of nucleic acid tests.
We have previously shown that endonucleases present in a protein complex, which has specificity for cyclobutane pyrimidine dimers, locate sites of damage in DNA by a processive mechanism of action in normal human lymphoblastoid cells. In contrast, the endonucleases present in this complex from xeroderma pigmentosum complementation group A (XPA) cells locate damage sites by a distributive or significantly less processive mechanism. Since the XPA protein has been shown to be responsible for the DNA repair defect in XPA cells, this protein was examined for involvement in the mechanism of target site location of these endonucleases. A recombinant XPA protein, produced by expression of the normal XPA cDNA in E. coli, was isolated and purified. The results show that the recombinant XPA protein was able to correct the defect in ability of the XPA endonucleases to act by a processive mechanism of action on UVC irradiated DNA. These studies indicate that the XPA protein, in addition to a role in damage recognition or damage verification, may function as a processivity factor.
Meaningful assessment and comparisons on the injury potentials of various types of ammunition and credible forensic reconstruction require an agreement on standard tissue simulant. Gelatine blocks are relatively inexpensive and easy to produce and they meet most of the criteria for a good terminal ballistic simulant. This research looks into the variables of preparing ballistic gelatine and their effects on penetration resistance. It was found out that consistent quality gelatine blocks are easy to make and that the variables like water temperature have a far smaller effect than previously thought. Also water acidity variances allowed by European Council directive on drinking water do not have any measurable effect. A proposed standard method for gelatine preparation is presented together with penetration function for verification of gelatine quality.
The effect of endothelium removal on pial artery constriction in response to norepinephrine (NE) was studied in vitro using a perfused vessel setup in which pressure increases indicate vasoconstriction. In deenodothelialized rabbit arteries, the reaction to extraluminal NE was found to be characterized by a much higher Emax (2.0 times) and a slight (but significant) leftward shift of the concentration-response curve (lower EC50) compared with control vessels. In cat arteries subjected to either extra- or intraluminal NE, the Emax was also substantially higher in deendothelialized preparations (4.4 and 5.1 times, respectively), but there was no significant difference in the EC50 values. Anatomical verification and functional tests (acetylcholine-induced dilatation) confirmed the presence and the absence of the endothelium in control and lesioned arteries, respectively. This modulatory influence of the endothelium may be of importance in cerebrovascular pathology.
We have established a cartilaginous fish cell line [Squalus acanthias embryo cell line (SAE)], a mesenchymal stem cell line derived from the embryo of an elasmobranch, the spiny dogfish shark S. acanthias. Elasmobranchs (sharks and rays) first appeared >400 million years ago, and existing species provide useful models for comparative vertebrate cell biology, physiology, and genomics. Comparative vertebrate genomics among evolutionarily distant organisms can provide sequence conservation information that facilitates identification of critical coding and noncoding regions. Although these genomic analyses are informative, experimental verification of functions of genomic sequences depends heavily on cell culture approaches. Using ESTs defining mRNAs derived from the SAE cell line, we identified lengthy and highly conserved gene-specific nucleotide sequences in the noncoding 3' UTRs of eight genes involved in the regulation of cell growth and proliferation. Conserved noncoding 3' mRNA regions detected by using the shark nucleotide sequences as a starting point were found in a range of other vertebrate orders, including bony fish, birds, amphibians, and mammals. Nucleotide identity of shark and human in these regions was remarkably well conserved. Our results indicate that highly conserved gene sequences dating from the appearance of jawed vertebrates and representing potential cis-regulatory elements can be identified through the use of cartilaginous fish as a baseline. Because the expression of genes in the SAE cell line was prerequisite for their identification, this cartilaginous fish culture system also provides a physiologically valid tool to test functional hypotheses on the role of these ancient conserved sequences in comparative cell biology.
Atherosclerosis is an inflammatory disease, occurring preferentially in branched or curved arterial regions exposed to disturbed flow conditions including oscillatory shear stress (OS). In contrast, straight portions exposed to undisturbed laminar shear stress (LS) are relatively lesion free. The opposite effects of atheroprotective LS and proatherogenic OS are likely to be determined by differential expression of genes and proteins, including redox regulating factors. OS induces inflammation via mechanisms involving increased reactive oxygen species (ROS) production from the NADPH oxidases. Through a transcript profiling study and subsequent verification and functional studies, the authors discovered that OS induces inflammation by producing bone morphogenic protein 4 (BMP4) in endothelial cells. BMP4 stimulates expression and activity of NADPH oxidase requiring p47phox and Nox-1 in an autocrine-like manner. The NADPH oxidase activation by BMP4 then leads to ROS production, NF-kappaB activation, intercellular adhesion molecule 1 (ICAM-1) expression, and subsequent increased monocyte adhesivity of endothelial cells. It is proposed that endothelial NADPH oxidases play a critical role in disturbed flow- and BMP4-dependent inflammation, which is the critical early atherogenic response occurring in atheroprone areas. This emerging field of shear stress, BMP4, NADPH oxidases, inflammation, and atherosclerosis is reviewed.
A crucial aim upon the completion of the human genome is the verification and functional annotation of all predicted genes and their protein products. Here we describe the mapping of peptides derived from accurate interpretations of protein tandem mass spectrometry (MS) data to eukaryotic genomes and the generation of an expandable resource for integration of data from many diverse proteomics experiments. Furthermore, we demonstrate that peptide identifications obtained from high-throughput proteomics can be integrated on a large scale with the human genome. This resource could serve as an expandable repository for MS-derived proteome information.
BACKGROUND: Protoplasts have emerged as a powerful model system in plant functional genomics, offering significant utility in functional gene analysis, protein interaction studies, and transient expression platforms for gene editing. Despite their versatility, inherent limitations restrict their broader application, highlighting the need for systematic investigations into their responses to abiotic stressors, such as temperature fluctuations and saline-alkali conditions (200 mM saline mixture: 170mM NaCl and 30mM Na2CO3, pH = 9.1). RESULTS: In this study, we comprehensively examined the effects of varying temperatures and saline-alkali stress on the integrity, viability, and reactive oxygen species (ROS) metabolism of maize protoplasts. Key markers of oxidative stress-including ROS accumulation, lipid peroxidation (measured as malondialdehyde, MDA), antioxidant enzyme activity (superoxide dismutase, SOD), and hydrogen peroxide (H2O2) levels-were quantified to assess the oxidative stress response. Protoplasts maintained at 4 °C demonstrated enhanced stability and antioxidant capacity, preserving cell viability and endogenous protein integrity for up to 16 h. Conversely, exposure to 37 °C significantly compromised protoplast viability, while incubation at 28 °C exerted minimal effects within 16 h. CONCLUSIONS: Our study investigated the effects of various temperature stresses and salt-alkali stress on maize protoplasts. The results demonstrated that both temperature and salt-alkali stress significantly impacted protoplast production, viability, and the expression of endogenous proteins. These findings not only characterize the redox response of maize protoplasts, but also provide guidance for protoplast isolation and other procedures: 4 °C is suitable for short-term maintenance, 25-28 °C for routine functional assays, and 37 °C should be avoided. These findings provide valuable insights into the stress responses of protoplasts and establish a foundation for future research aimed at improving plant stress tolerance through protoplast-based techniques.
A crucial component of lithographic modeling is the resist. Resists typically used at extreme-ultraviolet (EUV) wavelengths are derivatives of deep-ultraviolet chemically amplified resists. Models that describe these resists are often very complicated and are dependent on a large number of free parameters. Point-spread-function-based resist modeling serves as a simple alternative. I show this type of modeling to be a viable technique at EUV wavelengths by directly comparing modeling results with a variety of printing metrics, including process windows and isodense bias.
Ten cases of meningioma of the pineal region were histologically verified at a single institution during an 8-year period. These represented 0.3% of 3061 intracranial meningiomas, as well as 6.2% of 164 pineal tumors. Patients were predominantly women. Symptoms and signs of increased intracranial pressure were most common, whereas Parinaud's syndrome was observed in only one case. Computerized tomography (CT) was the main diagnostic test used in the present series. It usually revealed a hyperdense mass with high-intensity contrast enhancement and a calcified mass eccentrically located at the periphery, which returned to its central location postsurgically and was likely to represent a calcified pineal gland. Magnetic resonance imaging was performed in the more recent cases and confirmed the suspicion raised by CT of a meningioma with a calcified mass at its periphery. Surgery was performed using an occipital transtentorial microsurgical approach with the patient placed in the semisitting position. Gross-total tumor removal was achieved in all cases, although this required sacrifice of the vein of Galen in six patients. Venous occlusion was performed only after adequate intraoperative verification of functional occlusion and blood flow diversion from the galenic system and had no consequences in any case but one. This latter patient had an infiltrative meningioma that was removed at the expense of intraoperative sacrifice of the two internal cerebral veins, as well as unavoidable trauma to the region of the quadrigeminal plate. All the remaining patients improved postsurgery, and only one had a permanent visual field deficit as a result of the surgical approach. Pineal meningiomas represent a rare subgroup of pineal tumors that can be treated surgically with reasonably good results. Sacrifice of major basal veins may be required to achieve radical tumor removal and can be tolerated by the patient provided that functional occlusion of the galenic system, as indicated by preoperative angiography, is verified during surgery.
The results of training of chronic obstructive bronchitis (COB) patients according to the educational program "COB-school" are presented. The program was developed basing on the results of examination of 132 COB patients (80 males and 52 females) which included assessment of clinical picture, external respiration function, histological verification, standard multifactorial personality investigation, Luscher's test, specially designed questionnaire to diagnose social status. The findings demonstrate that COB patients respond to their disease with inadequate emotions, their psychological reactions are destructive, they suffer from social maladaptation. It is also shown that COB belong to diseases with abnormal pulmonary ventilation which is stressogenic and, consequently, treatment discipline of such patients is low. Methodology of the educational program rests on psychological laws of coping with stressogenic situation. The informative blocks of the program were constructed basing on these laws while the analysis of personality disorders provided grounds for optimal information presentation. The program covered 64 female and 50 male learners with COB. The effect of the program was evaluated by several clinical indices and reached 66.8 and 66.0% in females and males, respectively and a total of 66.4%. Thus, educational program with psychological and social correction noticeably improves discipline of COB patients' treatment.