BackgroundFourier Transform Infrared Imaging (FTIRI) is used to investigate the amide anisotropies at different surfaces of a three-dimensional cartilage or tendon block. With the...Full Text Available
We recently found that injection of 2 mCi of yttrium 90 (90Y; approximately 23,000 rads) into normal canine knees stimulated glycosaminoglycan (GAG) synthesis by femoral condylar cartilage. The present investigation was conducted to determine whether radiation affects cartilage metabolism directly. Rates of GAG synthesis and degradation in normal canine articularcartilage were studied following irradiation. Cultured synovium from the same knees was treated similarly, to determine the effects of irradiation on hyaluronic acid synthesis. Twenty-four hours after exposure to 1,000 rads, 10,000 rads, or 50,000 rads, 35S-GAG synthesis by the cartilage was 93%, 69%, and 37%, respectively, of that in control, nonirradiated cartilage. The effect was not rapidly reversible: 120 hours after exposure to 50,000 rads, GAG synthesis remained at only 28% of the control level. ...
Hyaluronic acid (HA), a principal matrix molecule in many tissues, is present in high amounts in articularcartilage. HA contributes in unique ways to the physical behavior of the tissue, and has been shown to have beneficial effects on chondrocyte activity. The goal of this study was to incorporate graduated amounts of HA into type I collagen scaffolds for the control of chondrocyte-mediated contraction and chondrogenesis in vitro. The results demonstrated that the amount of contraction of HA/collagen scaffolds by adult canine articular chondrocytes increased with the HA content of the scaffolds. The greatest amount of chondrogenesis after two weeks was found in the scaffolds which had undergone the most contraction. HA can play a useful role in adjusting the mechanical behavior of tissue engineering scaffolds and chondrogenesis in chondrocyte-seeded scaffolds.
In the case of injuries of the ankle joint, an imaging technique is expected to reveal the state of the bones, ligaments, and articularcartilage. Irrespective of novel imaging techniques, a plain radiography will be made in order to obtain a first picture of the injury. Examples are given, showing that further orientation along the Lauge-Hansen classification scheme of this type of injury helps to completely ascertain the damage to the Articulatio talocruralis. Further examinations applying specific methods such as arthrography will detect possible, isolated syndesmosis ruptures; lesions of the ligaments can also be directly detected by CT or MRT. The articularcartilage and its lesions can be visualized by a combination of CT and arthrography, but are better shown by the completely non-invasive MRT. This latter method also gives insight into pathological processes with the bone. It allows to detect ...
The three main objectives of this study were: (1) to determine if depletion of proteoglycans from the cartilage matrix that occurs during osteoarthritis causes a measurable increase of cartilage proteoglycan components in the synovial fluid and sera, (2) to observe what effect intracellular cAMP has on the expression of matrix components by chondrocytes, and (3) to determine if freshly isolated chondrocytes contain detectable levels of mRNA for fibronectin. Canine serum keratan sulfate and hyaluronate were measured to determine if there was an elevation of these serum glycosaminoglycans in a canine model of osteoarthritis. A single intra-articular injection of chymopapain into a shoulder joint increased serum keratan sulfate 10 fold and hyaluronate less than 2 fold in 24 hours. Keratan sulfate concentrations in synovial fluids of dogs about one year old were unrelated to the presence of spontaneous ...
Objective: To determine the reproductibility of three-dimensional volume and thickness measurements of the knee joint cartilage with MRI in volunteers. Methods: The knees of 7 healthy individuals (ages 23 to 58 yrs.) were sagitally imaged with a resolution of 2x0.31x0.31 mm{sup 3}, using a fat-suppressed FLASH-3 D sequence. The knee was repositioned in between replicate acquisitions, 6 data sets being obtained in each case. After semiautomatic segmentation and three-dimensional reconstruction of the cartilage, the thickness was determined independent of the original section orientation. The coefficient of variation for repeated volume measurements and the deviations of the maximal cartilage thickness values were calculated subsequently. Results: The mean variation of the cartilage volumes of the replicate measurements was 1.4% ({+-}0.8%) in the patella, 1.7% ({+-}1.5%) in the femur, 3.0% ({+-}1.2%) in ...
In 10 cases of femoral neck fracture treated with prosthetic replacement, the preoperative sup(99m)Tc-MDP scintigraphs were investigated and then compared with each histological findings of the extracted femoral head. The appearances of the scintigraphs were classified into following 3 patterns; increased activity pattern of sup(99m)Tc-MDP uptake in the overall femoral head (type I) in 4 cases, deficient activity pattern at the lateral side correspond to weight-bearing area (type II) in 2 cases and considerably deficient activity pattern in the whole femoral head (type III) in remaining 4 cases. Histological examination of the extracted femoral heads revealed the narrow part of ischemic necrosis in type I. However type II showed extensive necrosis in the portion of deficient activity by scintigraphs and in type III, the femoral heads except articularcartilage were entirely necrotic. This classification of sup(99m)Tc-MDP scintigraphy is ...
In IO cases of femoral neck fracture treated with prosthetic replacement, the preopera tive sup(99m)Tc-MDP scintigraphs were investigated and then compared with each histological findings of the extracted femoral head. The appearances of the scintigraphs were classified into following 3 patterns; increased activity pattern of sup(99m)Tc-MDP uptake in the overall femoral head (type I) in 4 cases, deficient activity pattern at the lateral side correspond to weight-bearing area (type II) in 2 cases and considerably deficient activity pattern in the whole femoral head (type III) in remaining 4 cases. Histological examination of the extracted femoral heads revealed the narrow part of ischemic necrosis in type I. However type II showed extensive necrosis in the portion of deficient activity by scintigraphs and in type III, the femoral heads except articularcartilage were entirely necrotic. This classification of sup(99m)Tc-MDP scintigraphy is ...
Objective. The aim of this study was to assess the capability of high-resolution images obtained with a commercially available pelvic phased-array surface coil to demonstrate normal hip anatomy. Design. We retrospectively analyzed the oblique coronal magnetic resonance (MR) images of hips of 36 consecutive patients acquired on a 1.5-T clinical imager using a pelvic phased-array coil as a receiver, a 16-20 cm field of view, and 5 mm slice thickness. Patients. Thirty-six patients were studied, age 15-81 years. There were 20 males and 16 females. Results and conclusions. The articularcartilage, cortex, superior labrum, and iliofemoral ligament were well visualized on proton density weighted fat saturation (PDF) images. The femoral and obturator vessels, obturator nerve, and various muscles were easily seen on T1-weighted images. High-resolution imaging of the hip is achievable in a reasonable amount of time using newer phased-array surface coils ...
Magnetic resonance (MR) imaging and histopathology were used to diagnose an articular process synovial myxoma in a dog. On MR images, the tumor was characterized by distortion of the left L1-L2 articular process, widening of the articular process joint, and the presence of a mass contiguous with the synovium of the articular process that displaced the spinal cord. The tumor was T2-hyperintense, T1-hypointense, relative to muscle, and had mild contrast enhancement. The MR features of articular process synovial myxoma may be distinct from other diseases of the articular process joint.
The large aggregating proteoglycan aggrecan is a major structural component of the extracellular matrix of articularcartilage. Recent cDNA cloning of the human aggrecan gene (AGC1) reveals a core protein of at least 2316 amino acids characterized by several distinct structural domains. Two globular domains, termed G1 and G2, are present at the amino terminus of the molecule and a third, termed G3, is present at the carboxy terminus. The G1 domain is homologous in structure to the cartilage link protein and accounts for the aggregating potential of aggrecan through its ability to interact with hyaluronic acid. The aggrecan gene is known to consist of 15 exons, with each exon encoding a distinct functional region of the mature protein. However, while the link protein gene is known to reside on chromosome 5 in the human, the location of the aggrecan gene is currently undetermined in any species. The probe (pAGG2) for the ...
Background We developed a tissue-engineered biphasic cartilage bone substitute construct which has been shown to integrate with host cartilage and differs from autologous osteochondral transfer in which integration with host cartilage does not occur. Questions/purposes (1) Develop a reproducible in vitro model to study the mechanisms regulating tissue-engineered cartilage integration with host cartilage, (2) compare the integrative properties of tissue-engineered cartilage with autologous cartilage and (3) determine if chondrocytes from the in-vitro formed cartilage migrate across the integration site. Methods A biphasic construct was placed into host bovine osteochondral explant and cultured for up to 8 weeks (n?=?6 at each time point). Autologous osteochondral implants served as controls...
Splenectomy or pretreatment of adult hamsters with cyclophosphamide (Cytoxan) increased the lethality of the TC-83 vaccine strain of Venezuelan encephalitis virus (VEE), inoculated subcutaneously, from 12% for normal hamsters to 75 and 76% respectively. A...
Effect of single and chronic irradiation on factors of hamsters natural immunity such as complement, lysozyme and antibodies in norm and in the cestode invasion has been investigated. Significant phase changes in the complement activity level, lysozyme content and specific antibodies tite have been shown to occur in hamster blood under the influence of ionizing radiation. Cestode invasion in irradiated host organism increases the negative effect of ionizing radiation on the hamster immyne system.
Assessment of subtle changes in the primary macromolecular components of cartilage, proteoglycan (PG) and collagen, is critical for the diagnosis of early stages of osteoarthritis (OA), but...Full Text Available
In laboratory studies, hamsters (Mesocricetus spp.) exhibit intense male-male aggression, thus making them an excellent model system for studies of the functional and mechanistic...Full Text Available
In Syrian hamsters (Mesocricetus auratus), the expression of reproductive behavior requires the perception of social odors. The behavioral response to these odors is mediated...Full Text Available
The study was conducted to determine potential target organ toxicity and its reversibility in hamsters following twice daily oral doses of NSC-D629243 for 28 consecutive days. Male and female hamsters were administered NSC-D629243 dissolved in sesame oil ...
To determine the accuracy of the intra-articular location of hyaluronic acid injection using a blind approach and to establish the usefulness of fluoroscopy-guided intra-articular injection. A fluoroscopy unit was used for 368 intra-articular injections of hyaluronic acid to 93 knees in 65 patients. Initially, blind needle positioning was conducted on the fluoroscopy table. The failure rate of the blind approach among the 368 injections was evaluated, and a relationship between the Kellgren-Lawrence grade (K-L grade) and the incidence of repeated failures using the blind approach was determined for injections to 52 knees in 37 patients who received a complete cycle of injections (five consecutive injections with a one-week interval between injections). Using a blind approach, 298 of 368 trials (81.2%) resulted in a needle tip being placed in an intra-articular location, while 70 of 368 trials resulted ...
ObjectiveTo achieve osteochondral regeneration utilizing transplantation of cartilage-lineage cells and adequate scaffolds, it is essential to characterize the behavior of transplanted...Full Text Available
The structure of newly synthesized proteoglycans from explant cultures of cartilage from joints subjected to transection of the anterior cruciate ligament (osteoarthritic) and from normal (non- or sham-operated) joints was examined. The structure of the products of proteoglycan turnover was also examined using explants of normal and osteoarthritic cartilage maintained in culture for a 48 h chase period. The findings were as follows: Newly synthesized (/sup 35/S)-proteoglycans extracted from cartilage explants from osteoarthritic joints whether examined 3 weeks, 3 months, or 6 months after surgery were larger than those from corresponding normal cartilage. This can be explained by the synthesis in osteoarthritic cartilage of abnormally long chondroitin sulfate chains on newly synthesised proteoglycans. The extracts also contained a newly formed small proteoglycan species that was ...
Rheumatoid arthritis (RA) is a debilitating autoimmune disease of global prevalence. The disease is characterized by synovial inflammation leading to cartilage and bone damage. Most of the conventional...Full Text Available
Matrilin-1 is expressed predominantly in cartilage and co-localizes with matrilin-3 with which it can form hetero-oligomers. We recently described novel structural and functional features of the matrilin-3...Full Text Available
In many rodent species, such as Syrian hamsters, reproductive behavior requires neural integration of chemosensory information and steroid hormone cues. The medial amygdala processes both of...Full Text Available
Hamsters breathed, nose-only, for 13 weeks, 5 days/week, 6 hr/day, either man-made vitreous fiber (MMVF)10a, MMVF33, or long amosite asbestos at approximately 300 World Health Organization (WHO) fibers/cc...Full Text Available
Mesenchymal stem cells (MSCs) are multipotent cells that can differentiate into chondrocytes, osteoblasts, myocytes, adipocytes, and a variety of other cell types. Several studies have been directed toward using MSCs from patients with osteoarthritis (OA) for cartilage repair, not only because these are the ones that will require a source of autologous stem cells if biological repair of cartilage lesions is to be a therapeutic option, but also to further an understanding of stem cell differentiation. Previous studies have shown that a major drawback of current cartilage and intervertebral disc tissue repair is that human MSCs from OA patients express type X collagen (COL X). COL X, a marker of late-stage chondrocyte hypertrophy, is implicated in endochondral ossification. However, those st...
To address the functions of FGFR2 and FGFR3 signaling during mandibular skeletogenesis, we over-expressed in the developing chick mandible, replication-competent retroviruses carrying truncated FGFR2c or FGFR3c that function as dominant negative receptors (RCAS-dnFGFR2 and RCAS-dnFGFR3). Injection of RCAS-dnFGFR3 between HH15 and 20 led to reduced proliferation, increased apoptosis, and decreased differentiation of chondroblasts in Meckel's cartilage. These changes resulted in the formation of a hypoplastic mandibular process and truncated Meckel's cartilage. This treatment also affected the proliferation and survival of osteoprogenitor cells in osteogenic condensations, leading to the absence of five mandibular bones on the injected side. Injection of RCAS-dnFGFR2 between HH15 and 20 or R...
Abstract Studies of the megamouth shark, one of three planktivorous sharks, can provide information about their evolutionary history. Megamouth shark feeding has never been observed in life animals, but two alternative hypotheses on biomechanics suggest either feeding, i.e., ram feeding or suction feeding. In this study, the second moment of area of the ceratohyal cartilages, which is an indicator of the flexural stiffness of the cartilages, is calculated for 21 species of ram- and suction-feeding sharks using computed tomography. The results indicate that suction-feeding sharks have ceratohyal cartilages with a larger second moment of area than ram-feeding sharks. The result also indicates that the ram-suction index, which is an indicator of relative contribution of ram and suction behavi...
Adolescent exposure to anabolic androgenic steroids (AAS) alters the development and activity of the glutamate neural system in the latero-anterior hypothalamus (LAH) in hamsters (Mesocricetus auratus); that is, an important neural component of the adolescent AAS-induced aggressive response. In this article, we used retrograde tracing to investigate glutamate-specific alterations in the connections between the LAH and several other nuclei implicated in adolescent AAS-induced aggression. Briefly, hamsters were treated with AAS or sesame-oil control during adolescence and then microinjected with retrograde tracer into the medial amygdala (MeA), lateral septum (LS), or bed nucleus of the stria terminalis (BNST). Brains were then processed for vesicular glutamate transporter 2 (VGLUT2) and examined for AAS-induced changes in the number VGLUT2 cells containing retrograde tracer (VGLUT2/tracer) within the LAH. It is interesting to note that while ...
Flow cytometric techniques were used to measure the cytological and biochemical damage to respiratory tract cells in animals exposed to particulates. Hamsters were exposed to raw and spent oil shale particulates and silica by intratracheal instillation. Exfoliated lung cells were obtained by sacrificing the animals and lavaging the respiratory tract posterior to the trachea with saline. Cell samples were fixed in ethanol and stained with mithramycin for fluorescence analysis of DNA content. DNA content distributions from hamsters exposed to spent oil shale and silica particulates showed atypical changes 28 to 35 days later. Cell counts and total numbers of macrophages, leukocytes, and epithelial cells in the lavage fluid also showed marked changes related to time after exposure.
The distribution of /sup 35/S-sulfate and /sup 3/H-glucosamine in the angular region of the hamster was studied by light and electron microscopic autoradiography following intraperitoneal injection of these compounds to hamsters. Exposed silver grains of /sup 35/S-sulfate were concentrated in the trabecular meshwork, sclera, and cornea, and grains of /sup 3/H-glucosamine were localized in the trabecular region. The radioactivity of both isotopes was observed in the Golgi apparatuses of the endothelial cells of the angular aqueous plexus and the trabecular meshwork. The grains were noted over the entire cytoplasm, except for the nucleus, and then were incorporated into the amorphous substance and collagen fibers in the region adjacent to the angular aqueous sinus. These results suggest that endothelial cells in the angular region synthesize and secrete the sulfated glycosaminoglycans and hyaluronic acid.
Simultaneous exposures of rats and hamsters to inhaled radon daughters, with and without uranium ore dust, were performed daily for five months. Pulmonary pathology developing in 6 to 13 mo after cessation of daily exposures included interstitial fibrosis, emphysema, epithelial hyperplasia, squamous metaplasia, and malignant neoplasia. Rats showed a greater variety and more severe response to these uranium mine inhalation exposures than did hamsters. Inhalation of radon daughters with uranium ore dust displayed the site of greatest damage, including squamous carcinoma, from the nasopharynx to the lungs. Sixty percent of the rats exposed to radon daughters with ore dust developed primary pulmonary carcinomas, providing an appropriate short-term experimental animal model for investigation of respiratory tract carcinogenesis in uranium miners.
Allen's Rule documents a century-old biological observation that strong positive correlations exist among latitude, ambient temperature, and limb length in mammals. Although genetic selection for thermoregulatory...Full Text Available
IntroductionPrimary tumors of the breast containing bone and cartilage are extremely rare, and an osteogenic sarcoma arising from a cystosarcoma phyllodes is exceptional.Case...Full Text Available
A high performance liquid chromatography (HPLC) procedure suitable for the simultaneous determination of the molecular size and concentration of macromolecular hyaluronate and proteoglycans in synovial...Full Text Available
Craniofacial injuries require a variety of different cell types to repopulate areas of bone, cartilage, tendon, and fat. Mesenchymal stem cells (MSCs) provide a multipotent cell source for tissue...Full Text Available
Damage to the anterior talofibular ligament (ATFL) and cacaneofibular ligament (CFL) during ankle sprain may be linked to the development of osteoarthritis. Altered tibiotalar kinematics have...Full Text Available
IntroductionAutologous chondrocyte transplantation (ACT) is a routine technique to regenerate focal cartilage lesions. However, patients with osteoarthritis (OA) are lacking an appropriate...Full Text Available
Mice homozygous for the recessive gene achondroplasia (cn) aged 16 and 17 days and some homozygotes aged 22-34 days have disruptions in the growth of the proximal tibial growth plate which are due solely...Full Text Available
The study of blocking factors requires in vitro assay of cell mediated immunity that parallels the in vivo response. By microcytotoxicity testing, progressor and immune peripheral blood lymphocytes...Full Text Available
The in vitro response of melatonin synthesis was assessed by incubation of individual whole pineal glands for 4 hr without or with different concentrations of isoproterenol in the medium. Pineals were taken either at the end of the 14-hr light phase (day)...
The mutagenic and cytotoxic activity of two newly synthesized doxorubicin derivatives and of one daunorubicin derivative were studied in V79 Chinese hamster cells and bacteria (Salmonella typhimurium...Full Text Available
Sexual behavior is a natural reward for many rodent species, and it often includes chemosensory-directed components. Chemosensory stimuli themselves may also be rewarding. Conditioned place...Full Text Available
In recent years it has become evident from epidemiological and experimental data that a large number of environmental factors, including diet, play a role in modifying the incidence of cancer. Cell culture systems in which oncogenic transformation serves as an end point are powerful tools for evaluating these questions. Using such systems it has been shown recently that pyrolysis products from charred surfaces of broiled meat and fish can transform hamster embryo cells in vitro as well as produce tumors in the animal. Our studies in vitro have demonstrated the oncogenic potential of ionizing radiation in both hamster and human cells and have established in hamster cells the dose response relationship at doses ranging from 1 to 600 rad for x-rays and 0.1 to 150 rad for neutrons. The present work was aimed at evaluating whether there exists a cocarcinogenic interaction between a pyrolysis product and x-rays in their ability ...
The cellular receptor for murine coronavirus mouse hepatitis virus (MHV)-A59 is a member of the carcinoembryonic antigen (CEA) family of glycoproteins in the immunoglobulin superfamily. We isolated...Full Text Available
Purpose: The aim of this study was to evaluate the accuracy of MRI in the measurement of cartilage thickness of the ankle joint in comparison to pathologic and histologic specimens. Patients and methods: The ankle joints of four fresh cadaver feet were imaged on a 1.5 T MR-unit in the coronal plane. Standard T{sub 1}-weighted spin echo (SE) and a T{sub 1}-weighted 3D-GE (FLASH-3D) sequence with fat saturation were applied. Following MR imaging, the talus was explanted and cut parallel to the MR images for macroscopic evaluation and histologic specimens were prepared. MRI measurements of the cartilage thickness of the talus were carried out in two ways: Without and with consideration of a transition zone of intermediate signal intensity between hyperintense cartilage and hypointense cortical bone. The data were compared with the anatomic specimens as the gold standard expressing the difference as a percentage value. On ...
Purpose: To evaluate micro computed tomography (micro-CT) for the assessment of osteochondritis dissecans in comparison with histology. Material and Methods: Osteochondritis dissecans lesions of 3 patients were evaluated using micro-CT (0.125 mA, 40 keV, 60 m slice thickness, 60 m isotropic resolution, entire sample) and light microscopy (toluidine blue, 3-5 m slice thickness). The methods were compared regarding preparation time, detectability of tissue types and morphologic features of bone and cartilage. Results: Non-destructive micro-CT imaging of the entire sample was faster than histologic preparation of a single slice for light microscopy. Morphologic features of bone and cartilage could be imaged in a comparable way to histology. It was not possible to image cells or different tissue types of bone and cartilage with micro-CT. Conclusion: Micro-CT is a fast, non-destructive tool that may be a supplement or, if ...
Purpose: To evaluate micro computed tomography (micro-CT) for the assessment of osteochondritis dissecans in comparison with histology. Material and Methods: Osteochondritis dissecans lesions of 3 patients were evaluated using micro-CT (0.125 mA, 40 keV, 60 m slice thickness, 60 m isotropic resolution, entire sample) and light microscopy (toluidine blue, 3-5 m slice thickness). The methods were compared regarding preparation time, detectability of tissue types and morphologic features of bone and cartilage. Results: Non-destructive micro-CT imaging of the entire sample was faster than histologic preparation of a single slice for light microscopy. Morphologic features of bone and cartilage could be imaged in a comparable way to histology. It was not possible to image cells or different tissue types of bone and cartilage with micro-CT. Conclusion: Micro-CT is a fast, non-destructive tool that may be a supplement or, if ...
The survival time of golden hamsters (Mesocricetus auratus) after whole-body "6"0Co-#gamma#-irradiation in the range of 600 to 200 000 rad was investigated. The two plateaus of the dose-survival curve which correspond to bone marrow and gastrointestinal death are similar to those of other species such as mice, rats and mongolian gerbils. A new plateau occurring 40-57 hours after doses of 30 000-60 000 rad, where there is a little reduction in survival time, has been found. It is in addition to the well recognized central nervous system (CNS) syndrome. This plateau is observed only in golden hamsters, presumably because of their relatively high resistance to CNS syndrome. Experiments involving partial body irradiation of the animals indicate that the target is in the cephalic one-third of abdomen. The new segment may well indicate a new type of acute somatic radiation injury different from the well known bone marrow, gastrointestinal and CNS ...
The synovial and bone uptake of tracer in the knees of patients with rheumatoid arthritis (RA) was quantified using {sup 99}Tc{sup m}-hexamethyl propylene amine oxime-labelled leucocytes and {sup 99}Tc{sup m}-methylene diphosphonate (MDP), respectively. Significant neutrophil migration and MDP uptake occurred in the knees of patients with RA irrespective of the disease duration. In all but one patient neutrophil migration was reduced after intra-articular steroid injection. The change in MDP uptake after steroid injection was variable. There was a significant correlation between the percentage reduction in neutrophil migration and pain score, while the latter correlated poorly with the change in MDP uptake. The quantification of the neutrophil component of the inflammatory process is a sensitive index for monitoring RA activity and response to pharmacological interventions, while quantitative bone scintigraphy should not be employed to monitor changes in joint ...
The synovial and bone uptake of tracer in the knees of patients with rheumatoid arthritis (RA) was quantified using "9"9Tc"m-hexamethyl propylene amine oxime-labelled leucocytes and "9"9Tc"m-methylene diphosphonate (MDP), respectively. Significant neutrophil migration and MDP uptake occurred in the knees of patients with RA irrespective of the disease duration. In all but one patient neutrophil migration was reduced after intra-articular steroid injection. The change in MDP uptake after steroid injection was variable. There was a significant correlation between the percentage reduction in neutrophil migration and pain score, while the latter correlated poorly with the change in MDP uptake. The quantification of the neutrophil component of the inflammatory process is a sensitive index for monitoring RA activity and response to pharmacological interventions, while quantitative bone scintigraphy should not be employed to monitor changes in joint inflammation in ...
The authors present a comparison between the diagnostic accuracy of Computed Tomography (CT) and Magnetic Resonance Imaging (MRI) in the articular pathologies of the knee. CT and MRI were performed in 30 patients who subsequently underwent surgery. CT and MRI results were compared on three bases: technique, definition of normal anatomy, and diagnostic accuracy. CT allowed a standardization of the technique, while MRI was superior in defining normal anatomy - especially in the study of capsulo-ligamentous structures. In all cases the use of both CT and MRI allowed a correct diagnosis, showing the site, extent and gravity of the lesion. CT proved to be superior to MRI in 4 cases, while MRI corrected CT diagnosis in 11 cases; in 15 patients both techniques had the same diagnostic accuracy. Meniscal tears were better identified by CT, while MRI was superior in the detection of ligamentous lesions and in the characterization of PVNS and tendinitis of the patellar ...
The chemopreventive efficacy of turmeric has been established in experimental systems. However, its mechanism(s) of action are not fully elucidated in vivo. The present study investigates the mechanism of turmeric-mediated chemoprevention in 7,12-dimethylbenz(a)anthracene (DMBA)-induced hamster buccal pouch (HBP) carcinogenesis at 2, 4, 6, 10 and 12?weeks. Dietary turmeric (1%) led to decrease in DMBA-induced tumor burden and multiplicity, and enhanced the latency period in parallel, to its modulatory effects on oncogene products and various cellular responses during HBP tumorigenesis. DMBA-induced expression of ras oncogene product, p21 and downstream target, the mitogen-activated protein kinases were significantly decreased by turmeric during HBP carcinogenesis. Turmeric also diminished ...
We have developed a cell-mediated mutagenesis assay in which cells with the appropriate markers for mutagenesis are co-cultivated with either lethally irradiated rodent embryonic cells that can metabolize carcinogenic hydrocarbons or with primary rat liver cells that can metabolize chemicals carcinogenic to the liver. During co-cultivation, the reactive metabolites of the procarcinogen appear to be transmitted to the mutable cells and induce mutations in them. Assays of this type make it possible to demonstrate a relationship between carcinogenic potency of the chemicals and their ability to induce mutations in mammalian cells. In addition, by simultaneously comparing the frequencies of transformation and mutation induced in normal diploid hamster cells by benzo(a)pyrene (BP) and one of its metabolites, it is possible to estimate the genetic target size for cell transformation in vitro.
Abstract This study reports the effects of varying concentrations of copper sulfate on the metabolic and gene transcriptional profile of a recombinant Chinese hamster ovary (CHO) cell line producing an immunoglobulin G (IgG)-fusion protein (B0). Addition of 50 M copper sulfate significantly decreased lactate accumulation in the cultures while increasing viable cell density and protein titer. These changes could be seen from day 6 and became increasingly evident with culture duration. Reducing the copper sulfate concentration to 5 M retained all the above beneficial effects, but with the added benefit of reduced levels of the aggregated form of the B0 protein. To profile the cellular changes due to copper sulfate addition at the transcriptional level, Affymetrix CHO microarrays were used to...
Large scale production of monoclonal antibodies has been accomplished using bioreactors with different length to diameter ratios, and diverse impeller and sparger designs. The differences in these physical attributes often result in dissimilar mass transfer, mechanical stresses due to turbulence and mixing inside the bioreactor that may lead to disparities in cell growth and antibody production. A rational analysis of impeller design parameters on cell growth, protein expression levels and subsequent antibody production is needed to understand such differences. The purpose of this study was to examine the impact of Rushton turbine and marine impeller designs on Chinese hamster ovary (CHO) cell growth and metabolism, and antibody production and quality. Experiments to evaluate mass transfer...
(1) To determine the accuracy of computed tomography (CT) in the evaluation of ligament tear and avulsion in patients with tibial plateau fracture. (2) To evaluate whether the presence or severity of fracture gap and articular depression can predict meniscal injury. A fellowship-trained musculoskeletal radiologist retrospectively reviewed knee CT and MRI examinations of 41 consecutive patients presenting to a level 1 trauma center with tibial plateau fractures. Fracture gap, articular depression, ligament tear and footprint avulsions were assessed on CT examinations. The MRI studies were examined for osseous and soft tissue injuries, including meniscal tear, meniscal displacement, ligament tear, and ligament avulsion. CT demonstrated torn ligaments with 80% sensitivity and 98% specificity. Only 2% of ligaments deemed intact on careful CT evaluation had partial or complete tears on MRI. Although the degree of fracture gap and ...
70 patients with squamouscell carcinoma of larynx and hypopharynx were examined by computed tomography; the TNM staging of tumors by CT, by endoscopy and by surgical operation was reviewed. CT proved to be reliable both to recognize the presence of neoplasms, with the exception of those very superficial, and their deep spreading to preepiglottic and paralaryngeal spaces, to the soft tissues of the neck and to the cartilages. Therefore CT is the examination of choice in laryngeal neoplasms staging, because it precisely completes the clinical and endoscopical informations.
Magnetic resonance imaging can be used in the trauma setting to detect fractures and associated soft tissue injuries of the musculoskeletal system. Magnetic resonance imaging complements plain radiography and CT for evaluating cancellous bone, cartilage and growth-plate injuries, and intra- and extraarticular supporting soft tissue structures. This review outlines typical applications and imaging consideration for injuries of the pelvis and lower extremity. (orig.) With 12 figs., 78 refs.
Abstract Unrefined and refined black raspberry seed oils (RSOs) were examined for their lipid-modulating effects in male Syrian hamsters fed high-cholesterol (0.12% g/g), high-fat (9% g/g) diets. Hamsters fed the refined and the unrefined RSO diets had equivalently lower plasma total cholesterol and high-density lipoprotein (HDL) cholesterol in comparison with the atherogenic coconut oil diet. The unrefined RSO treatment group did not differ in liver total and esterified cholesterol from the coconut oil-fed control animals, but the refined RSO resulted in significantly elevated liver total and esterified cholesterol concentrations. The unrefined RSO diets significantly lowered plasma triglycerides (46%; P=.0126) in comparison with the coconut oil diet, whereas the refined RSO only tended to lower plasma triglyceride (29%; P=.1630). Liver triglyceride concentrations were lower in the unrefined (46%; P=.0002) and refined (36%; P=.0005) RSO-fed ...
The ERA strain of rabies virus was propagated in a baby hamster kidney cell line (BHK-21/C13). The viral titer was 10(1.8) tissue culture infective doses (TCID) higher than that of commercial ERA vaccine....Full Text Available
PuO_2 particles behaved similarly in the lungs of rat and hamster. The relative concentrations of PuO_2 up to 1-yr postexposure were 1.0 for lung parenchyma, 1.3 for subpleural areas, 0.5 for peribronchiolar regions and 0.2 for perivascular regions in the lungs.
Chinese hamster lung fibroblasts V79 cells were treated with heat stress for 4 weeks with short duration (15 min) heat shock every alternate day in culture. It was observed that Hsp...Full Text Available
The state of chromatin during the cell cycle was examined using synchronized cultures of CHO hamster cells. Results support Mazia's dynamic chromosome cycle model and indicate that DNA-interactive chemotherapeutic agents elicit different types of kinetic responses in treated cells, suggesting a degree of specificity of interaction between various alkylating and intercalating agents and the genome. Effects of sarkosyl crystals, heparin, and chemotherapeutic agents, neocarzinostatin and adriamycin, on chromation are discussed. (HLW)
Twenty-seven patients with symptoms or clinical findings suggesting joint or bone disorders of the anterior chest wall (ACW) were evaluated by spiral CT with 3-dimensional (3-D) and multiplanar reconstructions. Preceding conventional ACW tomography was performed in 10 patients. ACW joint and bone changes were visualized more adequately by coronal 2-D reconstructions based on spiral CT than by conventional tomography. In addition, nonossified costal cartilages and soft tissue lesions were demonstrated. 3-D reconstructions sometimes added information, especially in patients with fracture and dislocation. (orig.).
Roentgenological analysis of the anterior chest wall was performed in twenty six patients with sternocostoclavicular hyperostosis. Initial hyperostotic change was seen at the first costosternal junction with ventral protrusion. Hyperostosis gradually developed around the first ribs, and irregular hyperostotic changes were also seen along the costoclavicular ligaments. Even in the final stage, however, sternoclavicular joint spaces were well preserved. These findings suggest that sternocostoclavicular hyperostosis is a disorder initiated around the costal cartilage including the periosteum and perichondrium, and that arthritis is not a condition stemming from that disorder.
Thermal neutron beam from the TNRF of JRR-3M was utilized for detection of mice lung cancer in vivo. Both the static and real-time neutron imaging were tried. Very strong thermal neutron beam is necessary to detect lung cancers because small changes are hidden by the very large attenuation of thermal neutron beam by the hydrogens in the tissues. The detection was successfully performed. Especially, the Super Eye of Hamamatsu Photonics contributed very effectively. Bone marrows and cartilages were also shown clearly. If an efficient and compact neutron source will become available in future, this method will grow up as a powerful tool for biomedical application. (author).
The human articular cartilagineous proteoglycans (PG) R.I.A. is highly specific. The PG used as the standard and the /sup 125/I labelled molecule appear to be pure. Under these conditions, all the potential interfering substances which have been tested show no cross reaction. For instance, the Ag-Ab equilibrium is not affected by adding human IgG, human albumin, hyaluronic acid, chondroitin sulfate, rat type II collagen or total human serum proteins. This R.I.A. also exhibits a species spcificity since there is no cross reaction with rat PG and negligible cross section with dog PG. The results obtained after addition of enzymes to the antigen demonstrate that the antigenic sites are localized on the protein region and not on the glycosaminoglycan region of the molecule.
Magnetic resonance imaging has proven an invaluable method in the diagnosis of joint diseases associated with osteonecrotic, inflammatory, traumatic and degenerative processes. At the clinical level, it has an important role in decisions about the method of treatment and evaluations of the therapeutic success. When the merits of MRT are balanced against those of conventional radiography including tomography and CT, which both ensure better spatial resolution in the visualisation of cortical and spongy bone structures, it becomes quite evident that MRT must not be regarded as an alternative method of imaging but as one that can be used additionally to obtain the most information for the diagnosis of arthropathy. The question as to whether new pulse sequences (snap shots) or invasive techniques like intra-articular injection of paramagnetic substances (MR arthrography) are likely to become routine procedures in the detection of joint diseases using MRT remains to be ...
(Materials and Methods) (I) VX/sub 2/ carcinoma was transplanted to the rabbit tibia, sup(99m)Tc-MDP was intravenously injected when bone destruction was radiologically seen. Preoperative scintigraphs, postoperative one of the resected tibia and fibra and scintigraphs of sliced tumor were compared with each other. Tissue fragments were collected from the sliced tumor, and RI was measured to compare concentration rates at different parts together with histological investigations. (II) Tetracycline was given to the rabbit of Experiment (I) to compare with the concentration of sup(99m)Tc-MDP. (III) Microautoradiography method was done on the experimental animals for investigation of the concentration of sup(99m)Tc-MDP. (Results) (I) Localization of sup(99m)Tc-MDP was increased at the sites of tumorous bone formation, of reactive bone formation at tumor edges, and of cartilage calcification, with little concentration at tumor cells or necrotic tissue. (II) Deposition ...
Tc-MDP 10mCi was intravenously given preoperatively to 20 patients with primary bone tumors and 1 with a metastatic bone tumor. Preoperative and postoperative scintigraphs of resected tumor and scintigraphs of sliced tumor were compared. Tissue fragments were collected from the sliced tumor, and RI measured to compare concentration rates at different areas. Histological investigations were also done. Localization of Tc-MDP was increased at sites of tumorous bone formation, of reactive bone formation at tumor edges, and of cartilage calcification, with little concentration at tumor cells or necrotic tissue. VX/sub 2/ carcinoma was transplanted into the rabbit tibia, Tc-MDP was intravenously injected when bone destruction was radiologically evident, and investigations were done as in the clinical cases. In experimental bone tumor, increased uptake localization of Tc-MDP was much the same as in clinical cases. Tetracycline was given to the rabbit of Experiment to ...
Abstract MicroRNAs (miRNAs) play important roles in global gene regulation. Researchers in recombinant protein production have proposed miRNAs as biomarkers and cell engineering targets. However, miRNA expression remains understudied in Chinese Hamster Ovary cells, one of the most commonly used host cell systems for therapeutic protein production. To profile highly conserved miRNA expression, we used the miRCURY- miRNA array for screening miRNAs in CHO cells. The selection criteria for further miRNA profiling included positive hybridization signals and experimentally validated predicted regulatory targets. On the basis of screening, we selected 16 miRNAs for quantitative RT-PCR profiling. We profiled miR expression in parental CHO DG44 and CHO K1 cell lines as well as four recombinant DG44...
Marmesin was isolated from the medicinal plant, Afraegle paniculata. Its cytotoxicity and mutagenicity in Chinese hamster V79 cells when sensitized to near ultraviolet (NUV) and long wavelength ultraviolet light or black light (BL) were assayed. Marmesin was extremely cytotoxic in the dark. This cytotoxicity was photoenhanced in NUV and BL; the photoenhanced lethality being higher in NUV than in BL. The LD/sub 50/ of marmesin under NUV and BL photosensitization were 0.002 ..mu..M and (0.012 ..mu..M), respectively. In the absence of NUV and BL, marmesin's LD/sub 50/ was 0.013 ..mu..M.NUV and BL without marmesin were not significantly cytotoxic at the fluence rates of 0.29 W/m/sup 2/ and 4.2 W/m/sup 2/, respectively, for up to 20 min. In contrast to the observed high cytotoxicity of marmesin, its mutagenicity at the HGPRT locus (Asub(z)Gsup(r)) was weak. The implication of this result in the high incidence of skin cancer in Nigeria in which A. paniculata is ...
The usefulness of the mouse, rat, and Syrian golden hamster as experimental models for studies of the development of respiratory tract cancer is considered. Three different application methods for examining the effects of environmental chemicals on the respiratory tract in these species are discussed: implantation, intratracheal instillation, and inhalation. The results of various investigations are presented to demonstrate that each particular method has both advantages and disadvantages. The techniques are also considered in comparison with the human situation and in the light of obtaining the most valid data for extrapolation to man. Brief attention is paid to the relevance to respiratory tract carcinogenesis of more recently developed organ and cell culture systems.
A comparison of the concentration-response effects of inhaled ozone (O/sub 3/) and phosgene (COCl/sub 2/) in different species of laboratory animals was made in order to better understand the influence of the choice of species in inhalation toxicity studies. The effect of 4-h exposures to ozone at concentrations of 0.2, 0.5, 1.0, and 2.0 ppm, and to COCl/sub 2/ and 0.1, 0.2, 0.5, and 1.0 ppm was determined in rabbits, guinea pigs, rats, hamsters, and mice. Lavage fluid protein (LFP) accumulation 18-20 h after exposure was used as the indicator of O3- and COCl/sub 2/-induced pulmonary edema. All species had similar basal levels of LFP (250-350 mg/ml) when a volume of saline that approximated the total lung capacity was used to lavage the collapsed lungs. Ozone effects were most marked in guinea pigs, which showed significant effects at 0.2 ppm and above. Mice, hamsters, and rats showed effects at 1.0 ppm O3 and above, while rabbits responded ...
This paper summarizes results of preliminary experiments to develop cytological and biochemical indicators for estimating damage to respiratory cells in test animals exposed by inhalation to toxic agents associated with nonnuclear energy production, the specific goal being the application of advanced multiparameter flow-systems technologies to the detection of early atypical cellular changes in lung epithelium. Normal Syrian hamster lung cell samples composed of histiocytes, leukocytes, macrophages, ciliated columnar cells, and epithelial cells were stained with fluorescent dyes specific for different biochemical parameters and were analyzed in liquid suspension as they flowed through a chamber intersecting a laser beam of exciting light. Multiple sensors measured the total or two-color fluorescence and light scatter on a cell-by-cell basis. Cellular parameters proportional to optical measurements (i.e., cell size, DNA content, total protein, nonspecific esterase ...
Purpose: to compare several morphometric parameters in MRI with the functional status of the articular disc in a large patient group suffering from internal derangement (ID) of the temporomandibular joint (TMJ). Materials and methods: in a retrospective study, 320 analyzable high resolution MRI examinations of the TMJs obtained in a 1.5 T unit were evaluated in 184 patients with clinically suspected ID. The analysis included the anatomical structures and a number of morphometric parameters previously described in the literature. The parameters were compared with the position of the articular disc. Results: the disc position was categorized as ''normal'' (NDP, 21.9%, n = 70), ''anterior displacement with reduction on opening'' (AMR, 51.6%, n = 165) and ''anterior displacement without reduction on opening'' (AOR, 26.6%, n = 85). With ...
A prospective study was performed on 50 patients suffering from osteochondritis dissecans of the knee and ankle to define criteria for stability and fixation of osteochondral lesions. Morphological parameters in MRI (size, fragmentation, cartilage, interface) and conventional radiology (separation, fragmentation) were registered and compared with arthroscopical staging. MRI staging based on different types of interfaces was demonstrated on T_1- and T_2-weighted images. MRI could correctly predict a Grade 1 lesion in 50%, a Grade 2 lesion in 90%, a Grade 3 lesion on 0%, and a Grade 4 lesion in 79%. Stable lesions were differentiated from unstable lesions in 90%. Radiographic findings corresponded with arthroscopic staging in only 56% of the cases because fibrotic connection may guarantee stability in case of bony separation. (orig./MG).
Background Photopolymerizable poly(ethylene glycol) (PEG) hydrogels offer a platform to deliver cells in vivo and support three-dimensional cell culture but should be designed to degrade in sync with neotissue development and endure the physiologic environment. Questions/purposes We asked whether (1) incorporation of degradation into PEG hydrogels facilitates tissue development comprised of essential cartilage macromolecules; (2) with early loading before pericellular matrix formation, the duration of load affects matrix production; and (3) dynamic loading in general influences macroscopic tissue development. Methods Primary bovine chondrocytes were encapsulated in hydrogels (n?=?3 for each condition). The independent variables were hydrogel degradation (nondegrading PEG and degrading olig...
Fuji Computed Radiography (FCR) is a new system of digital radiography that is based on the latest computer technologies. We applied this FCR system to the X-ray diagnosis of the laryngeal disease. Twenty patients with various laryngeal diseases were evaluated by both FCR frontal tomography and conventional screen-film frontal tomography under the same exposure condition. Both tomograms were then compared on the subject of the clarity of delineation of laryngeal structures. As the result, FCR tomograms revealed the feature of the laryngeal lumen and cartilages more clearly than conventional tomograms. In addition, energy subtraction method, which is one of FCR image processing systems, was presented as a new technical procedure to extinguish cervical spine shadows in plain anteroposterior laryngeal X-photograph. We stress that FCR system is the suitable procedure for X-ray analysis of laryngeal diseases, especially in the region related with the phono-surgery. ...
The aim of this study was to evaluate typical magnetic resonance imaging (MRI) findings in early rheumatic diseases manifesting at the soft tissues of the hand using a retrospective analysis. A total of 186 MRI examinations of patients with clinical suspicion of a rheumatic disease were evaluated in a consensus reading by two experienced radiologists. All imaging patterns were assessed with respect to their type and localization. Under blinded and non-blinded conditions diagnoses were correlated with final clinical diagnosis. The most frequent diagnoses were rheumatoid arthritis (RA, 45.7%) and psoriatic arthritis (PsA, 15.6%). The mean correlation between clinical and MRI diagnosis (r) was 0.75 in blinded and 0.853 in non-blinded reading (p <0.001). The following extra-articular imaging patterns were found: synovitis (59.1%), tendovaginitis (91.4%), dactylitis (14.5%), and bone marrow edema (18.3%). Only dactylitis was specific for a particular rheumatic ...
Correct ortopedic therapy for traumas of the cervical rachis requires perfect knowledge of the spatial balance of the fracture focus. The authors belive Computed Tomography (CT) to be the most suitable, and often indispensable, method for this purpose. Twenty-four patients were examined for traumatic pathology of the cervical rachis. In 7 cases with clinically minor traumas, the negative outcome of the traditional traumas, the negative outcome of the traditional exam was considered reliable and sufficient for therapeutic purposes. The extant 17 patients were examined also by means of CT, either to better dermine the characteristics of skeletal lesions already ascertained with traditional techniques or to asses the presence of clinically-suspected osteo-articular lesions, even with negative conventional X-rays. For 9 of these patients orthopedic treatment was considered sufficient, whereas 8 patients underwent surgery and were subsequently examined with CT, which ...
The purpose of this study was to evaluate the usefulness of dynamic MRI for femoral head perfusion. Objects were 64 femoral heads of 44 normal cases (20-95 years old), 10 cases (33-90 y) of femoral neck fracture and 8 cases (61-86 y) using steroid due to articular rheumatism, nephrosis and other diseases. Siemens 1.0 T Magneton Impact with body array coil was used for dynamic MRI by the fast low-angle shot 2D method for every 17 sec interval. Gd-DTPA was injected intravenously. ROI was defined in the center of femur head and of trochanter to monitor their values for the dynamic curve. Evaluation was done on the ratio of enhancement ratio (ER) in the head/trochanter and on the curve in both heads. In normal cases, the head ER was significantly lower in >60 years old cases. There was neither significant correlation between their head/trochanter ER ratios nor significant difference between their dynamic curves. In fracture cases, there were dynamic curves showing ...
Abstract The objective of this study was to determine the functional properties of cartilaginous tissues generated by porcine MSCs isolated from different tissue sources, and to compare these properties to those derived from chondrocytes (CCs). MSCs were isolated from bone marrow (BM) and infrapatellar fat pad (FP), while CCs were harvested from the articular surface of the femoro-patellar joint. Culture-expanded CCs and MSCs were encapsulated in agarose hydrogels and cultured in the presence of TGF3. Samples were analysed biomechanically, biochemically and histologically at days 0, 21 and 42. After 42 days in free swelling culture, mean GAG content was 1.50% w/w in CC-seeded constructs, compared to 0.95% w/w in FP- and 0.43% w/w in BM-seeded constructs. Total collagen accumulation was hig...
Abstract The glycosylation profile of therapeutic antibodies is routinely analyzed throughout development to monitor the impact of process parameters and to ensure consistency, efficacy, and safety for clinical and commercial batches of therapeutic products. In this study, unusually high levels of the mannose-5 (Man5) glycoform were observed during the early development of a therapeutic antibody produced from a Chinese hamster ovary (CHO) cell line, model cell line A. Follow up studies indicated that the antibody Man5 level was increased throughout the course of cell culture production as a result of increasing cell culture medium osmolality levels and extending culture duration. With model cell line A, Man5 glycosylation increased more than twofold from 12% to 28% in the fed-batch process...
We determined the chromosomal localization and structure of the gene encoding human type II inosine 5{prime}-monophosphate dehydrogenase (IMPDH, EC 1.1.1.205), an enzyme associated with cellular proliferation, malignant transformation, and differentiation. Using polymerase chain reaction (PCR) primers specific for type II IMPDH, we screened a panel of human-Chinese hamster cell somatic hybrids and a separate deletion panel of chromosome 3 hybrids and localized the gene to 3p21.1{yields}p24.2. Two overlapping yeast artificial chromosome clones containing the full gene for type II IMPDH were isolated and a physical map of 117 kb of human genomic DNA in this region of chromosome 3 was constructed. The gene for type II IMPDH was localized and oriented on this map and found to span no more than 12.5 kb.
In many species social behaviors are dependent on integration of chemosensory and hormonal cues. Many chemosensory stimuli are detected by the vomeronasal organ, which projects to many regions that contain steroid receptors, including the medial amygdala. In male hamsters, testosterone is known to acutely increase in response to chemosensory stimulation, and can facilitate sexual behavior by direct action within the medial amygdala. Conspecific stimuli activate the anterior (MeA) and posterior (MeP) medial amygdala, while heterospecific stimuli activate only MeA. Chemosensory stimuli with different social significance differentially activate the dorsal and ventral subdivisions of MeA and MeP. Therefore, it is likely that steroids differentially facilitate stimulation of the medial amygdala...
Ionizing radiations that differ in number, size, and distribution of energy deposition events might be expected to cause different effects for the same absorbed dose. Furthermore, microdosimetry calculations suggest that large variations in biological effectiveness might be expected for internally deposited alpha-emitting radionuclides for the same absorbed dose, depending upon the specific activity and spatial distribution of the sources in tissue. The study described in this article was designed to demonstrate these phenomena in vitro. Cultured Chinese hamster ovary (CHO-K1 BH4) cells were exposed in vitro to inert, insoluble ceramic microspheres in zirconium dioxide labeled with "2"3"9Pu. The average exit energy of the 5.15 MeV alpha particle was calculated to be 4.3 MeV. Exposure times varied from four to seven hours to achieve the desired dose level.
Abstract One of the most significant problems in industrial bioprocessing of recombinant proteins using engineered mammalian cells is the phenomenon of cell line instability, where a production cell line suffers a loss of specific productivity (qP). This phenomenon occurs with unpredictable kinetics and has been widely observed in Chinese hamster ovary (CHO) cell lines and with all commonly used gene expression systems. The underlying causes (both genetic and physiological) and the precise molecular mechanisms underpinning cell line instability have yet to be fully elucidated, although recombinant gene silencing and loss of recombinant gene copies have been shown to cause qP loss. In this work we have investigated the molecular mechanisms underpinning qP instability over long-term sub-cult...
In order to demonstrate possible specific effects of microwaves at the cellular level V-79 Chinese hamster cells were exposed to 2.45-GHz radiation at power levels of 20-200 mW/cm2 and at specific absorption rates of 10-100 mW/g. Intracellular cytoplasmic changes were observed by fluorescence polarization using a method based on the intracellular enzymatic hydrolysis of nonfluorescent fluorescein diacetate (FDA). At levels of absorbed energy below 90 J/g, modifications of microviscosity and mitochondrial state were absent, but a slight stimulation of enzymatic hydrolysis of FDA was observed which may be explained by microwave-induced alterations of cellular membranes possibly due to differences in heating pattern of microwaves compared to water-bath heating. At levels of absorbed energy above 90 J/g, the decrease of enzymatic hydrolysis of FDA, increase in degree of polarization, and increase of permeation of the fluorescent marker correlated well with the decrease ...
The bed nucleus of the stria terminalis (BST) and the medial amygdala (MeA) are anatomically connected sites necessary for chemosensory regulation of social behaviors in rodents. Prairie voles (Microtus ochrogaster) are a valuable model for studying the neural regulation of social behaviors because, unlike many other rodents, they are gregarious, pair bond after copulating, and are biparental. We herein describe sex and species differences in immunoreactivity for tyrosine hydroxylase (TH), the rate-limiting enzyme for catecholamine synthesis, in the BST and MeA. Virgin male prairie voles had a large number of TH-immunoreactive cells in areas analogous to the rat principal nucleus of the BST (pBST) and the posterodorsal medial amygdala (MeAPd). Virgin female prairie voles had far fewer TH-immunoreactive cells in these sites ( approximately 17% of the number of cells as males in the pBST, approximately 35% of the number of cells in the MeAPd). A few TH-immunoreactive cells were found in ...
Alcaptonuria is a rare autosomal recessive metabolic disease, due to the lack of homogentisic acid oxidase. The following accumulation of homogentisic acid brings about a black discoloration of both the urine alcaptonuria) and connective tissue (ochronosis). The ochronotic alterations into joint cartilages cause degenerative arthropathy and osteopenia. The radiological features of three unrelated cases of alcaptonuria are reported. Radiographic abnormalities of ochconotic arthropathy are found in both the spine and the extraspinal joints. In the spine, the progressive calcification and ''vacuum'' phenomenon of disc spaces are the most characteristic findings. Disc space narrowing is associated with calcification and marginal sclerosis of vertebral bodies and is accentuated by osteopenia. Osteophytes are usually absent or of small size; neverthless progressive formation of marginal intervertebral bridges and obliteration of disc spaces at multiple levels ...
Patients with haemarthros of the knee were examined by MRI and arthroscopy to identify the bleeding source and to determine surgical or conservative therapy. A prospective study was performed on 120 patients who underwent MRI and arthroscopy, point lesions and bleeding sources were documented separately on a standardised form. Arthroscopy demonstrated a ruptured anterior cruciate ligament in 86%, meniscal tears in 55%, disruption of collateral ligaments in 17%, osteochondral lesions in 5% and tears of capsule or plicae in 13%. Disruption of cruciate ligaments caused haemorrhage in all but three cases. In contrast, only meniscal tears located very adjacent to the meniscal basement had bled. Those types of osteochondral lesions caused haemorrhage characterised on MRI by destruction of the subchondral plate and overlying cartilage. Severe lesions of the knee were diagnosed by MRI in 71% to 100%, slight lesions in 13% to 40% of the cases. Single, combined and bland ...
Differential phase-contrast X-ray imaging has been performed in a Talbot-Lau configuration, where a multiline X-ray source was used instead of a combination of a hard-X-ray multiple slit and a normal focus X-ray generator. When the multiple slit is used, a high aspect ratio structure is needed and slit width should be below 10 ?m for its function. The fabrication and use of such a multislit can be omitted using the presented configuration. The multiline X-ray source was developed by making grooves on a tungsten rotating anode, which was irradiated by an electron beam to generate X-rays. An array of 10?m line sources with a pitch of 30?m was formed and combined with a 4.5 ?m pitch phase grating and a 5.3 ?m pitch amplitude grating to generate differential phase contrast. With a total exposure time of 40 s, a differential phase image depicting cartilages was obtained. (author)
Four species of nanocomposites organized by hydroxyapatite (HAp), hyaluronic acid (HyA), chondroitin sulfate (ChS) and type II collagen (Col), i.e. HAp/HyA, HAp/ChS, HAp/HyA/Col and HAp/ChS/Col composites, were synthesized by coprecipitation methods. The composites could retain lots of water: 40%, 32%, 42% and 58% for the HAp/HyA, HAp/ChS, HAp/HyA/Col and HAp/ChS/Col composites, respectively. Transmission electron microscopy observations revealed that the HAp/HyA and HAp/ChS composites consisted of island-like aggregations whose sizes were 300 nm in length and 30 nm in width, and 150 nm in length and 30 nm in width, respectively. In the aggregations, there were many HAp nanocrystals of 20 nm in length, and their c-axes were aligned along the respective polymer molecules through a self-organization process. Animal tests showed that chondrocyte-like cells penetrated into the HAp/ChS/Col composites 4 weeks after implantation. It was shown that the HAp/ChS/Col composite has a potential for ...
The human zinc finger protein genes (ZFX/Y) were identified as a result of a systematic search for the testis-determining factor gene on the human Y chromosome. Although they play no direct role in sex determination, they are of particular interest because they are highly conserved among mammals, birds, and amphibians and because, in eutherian mammals at least, they have active alleles on both the X and the Y chromosomes outside the pseudoautosomal region. We used in situ hybridization to localize the homologues of the zinc finger protein gene to chromosome 1 of the Australian echidna and to an equivalent position on chromosomes 1 and 2 of the playtpus. The localization to platypus chromosome 1 was confirmed by Southern analysis of a Chinese hamster [times] platypus cell hybrid retaining most of platypus chromosome 1. This localization is consistent with the cytological homology of chromosome 1 between the two species. The zinc finger protein gene homologues were ...
Radixin is a cytoskeletal protein that may be important in linking actin to the plasma membrane. Recent cloning of the murine and porcine radixin cDNAs revealed a protein highly homologous to ezrin and moesin. The authors have cloned and sequenced the human radixin cDNA and found the predicted amino acid sequence for the human protein to be nearly identical to those predicted for radixin in the two other species. By Southern analyses of Chinese hamster x human somatic cell hybrid DNA and of PCR products derived from hybrids, the coding gene (RDX) was mapped to 11q. Fluorescence chromosomal in situ hybridization with a cDNA plasmid further localized this gene to band 11q23. However, PCR amplification with [open quotes]radixin-specific[close quotes] primers on the hybrid DNA panel yielded an additional, very similar DNA sequence that was further characterized by direct sequencing of PCR products. This sequence represents a truncated version and the respective locus ...
The product of the human GRO gene is a cytokine with inflammatory and growth-regulatory properties; GRO is also called MGSA for melanoma growth-stimulatory activity. The authors have identified two additional genes, GRO#beta# and GRO#gamma#, that share 90% and 86% identity at the deduced amino acid level with the original GRO#alpha# isolate. One amino acid substitution of proline in GRO#alpha# by leucine in GRO#beta# and GRO#gamma# leads to a large predicted change in protein conformation. Significant differences also exist in the 3' untranslated region, including different numbers of ATTTA repeats associated with mRNA instability. A 122-base-pair region in the 3' region is conserved among the three GRO genes, and a part of it is also conserved in the Chinese hamster genome, suggesting a role in regulation. DNA hybridization with oligonucleotide probes and partial sequence analysis of the genomic clones confirm that the three forms are derived from related but ...
RNA interference has become a powerful biological tool over the last decade. In this study, a tetracycline-inducible shRNA vector system was designed for silencing CFP expression and delivered alongside the yfp marker gene into Chinese hamster ovary cells using impalefection on spatially indexed vertically aligned carbon nanofiber arrays (VACNFs). The VACNF architecture provided simultaneous delivery of multiple genes, subsequent adherence and proliferation of interfaced cells, and repeated monitoring of single cells over time. Following impalefection and tetracycline induction, 53.1% 10.4% of impalefected cells were fully silenced by the inducible CFP-silencing shRNA vector. Additionally, efficient CFP-silencing was observed in single cells among a population of cells that remained CFP-expressing. This effective transient expression system enables rapid analysis of gene silencing effects using RNAi in single cells and cell populations.
Macrophage differentiation antigen associated with complement three receptor function (Mac-1) belongs to #beta#_2 subfamily of integrins that mediate important cell-cell and cell-extracellular matrix interactions. Biochemical studies have indicated that Mac-1 is a constitutive heterodimer in vitro. Here, we detected the heterodimerization of Mac-1 subunits in living cells by means of two fluorescence resonance energy transfer (FRET) techniques (fluorescence microscopy and fluorescence spectroscopy) and our results demonstrated that there is constitutive heterodimerization of the Mac-1 subunits and this constitutive heterodimerization of the Mac-1 subunits is cell-type independent. Through FRET imaging, we found that heterodimers of Mac-1 mainly localized in plasma membrane, perinuclear, and Golgi area in living cells. Furthermore, through analysis of the estimated physical distances between cyan fluorescent protein (CFP) and yellow fluorescent protein (YFP) fused to Mac-1 subunits, we ...
The human CDC25 tyrosine phosphatases trigger activation of CDC2 by removing inhibitory phosphates; thus the genes encoding these phosphatases may be suspected as potential oncogenes due to their role in promoting cell division. To date, three human CDC25 genes have been identified: CDC25A, B, and C. This communication describes the mapping of CDC25A to chromosome 3p21 and CDC25B to chromosome 20p13 by fluorescence in situ hybridization with confirmation by the polymerase chain reaction of hamster-human somatic cell hybrid DNA. 3p21 is near an area frequently involved in karyotypic abnormalities in renal carcinomas, small cell carcinomas of the lung, and benign tumors of the salivary gland. 20p13 does not seem to be a common area for karyotypic alteration in tumors. Mapping of these genes to their chromosomal loci may help identify tumors with abnormal regulation of CDC25 genes due to genomic alterations. 15 refs., 3 figs.
BACKGROUND AND PURPOSE Human K2P3.1 (TASK1) channels represent potential targets for pharmacological management of atrial fibrillation. K2P channels control excitability by stabilizing membrane potential and by expediting repolarization. In the heart, inhibition of K2P currents by class III antiarrhythmic drugs results in action potential prolongation and suppression of electrical automaticity. Carvedilol exerts antiarrhythmic activity and suppresses atrial fibrillation following cardiac surgery or cardioversion. The objective of this study was to investigate acute effects of carvedilol on human K2P3.1 (hK2P3.1) channels. EXPERIMENTAL APPROACH Two-electrode voltage clamp and whole-cell patch clamp electrophysiology was used to record hK2P3.1 currents from Xenopus oocytes, Chinese hamster o...
Aim: Rheumatoid arthritis is a limiting disease having, among its pathological features, the inflammation of synovial tissue with progressive and later destruction of the articulation. This leads to joint deformation and loss of its function, generating pain and reducing the mobility of the affected articulation. The aim was to evaluate "1"6"6Ho-Hydroxyapatite ("1"6"6 Ho-HA) as potential radiopharmaceutical for the symptomatic treatment of chronic and acute arthritis. Materials and Methods: Holmiun-166 was produced by irradiation of Ho_2O_3 at La Reina Research Reactor, Nuclear Chilean Energy Commission. Hydroxyapatite was in-house synthesized. Its labelling and quality controls follows the internationally accepted procedures. An antigen's arthritis was induced to eight New Zealand rabbits with the "1"6"6 Ho-HA radiochemical being administered thereafter in two dosage modalities (single and double). The compound therapeutic efficiency was evaluated based upon clinical improvement and ...
Cytochrome c, labeled with /sup 125/I, has been used by us for staining glycosaminoglycans (GAG) separated by electrophoresis on cellulose acetate strips. As GAG between 1-10 ng could be quantified by autoradiography and densitometry, the reagent is approximately 100-fold more sensitive than currently used non-radiolabeled stains. The authors extend now the use of radiolabeled cytochrome c to the quantification of PG transblotted to solid supports subsequent to separation on polyacrylamine slab gels. Dot blotting used for exploring optimal conditions for detecting PG indicated that because of low background positively charged Nylon 66 was superior to nitrocellulose. Increasing the positive charge of the staining reagent by cationization decreased background radiation even further so that 1 ng PG could be seen readily after 5 hrs of autoradiography. Use of cationized /sup 125/I-cytochrome c has been made in detecting PG of bovine fetal epiphyseal cartilage (> 1 x ...
A hyaluronic acid binding fraction was purified from the supernatant media of both 3T3 and murine sarcoma virus (MSV) transformed 3T3 cultures by hyaluronate and immunoaffinity chromatography. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis resolved the hyaluronate affinity-purified fraction into three major protein bands of estimated molecular weight (M/sub r,e/) 70K, 66K, and 56K which contained hyaluronate binding activity and which were termed hyaluronate binding proteins (HABP). Hyaluronate affinity chromatography combined with immunoaffinity chromatography, using antibody directed against the larger HABP, allowed a 20-fold purification of HABP. Fractions isolated from 3T3 supernatant medium also contained additional binding molecules in the molecular weight range of 20K. This material was present in vanishingly small amounts and was not detected with a silver stain or with (/sup 35/S)methionine label. The three protein species isolated by hyaluronate affinity ...
A comparison of the concentration-response effects of inhaled ozone (O/sub 3/) in different species of laboratory animals was made in order to better understand the influence of the choice of species in inhalation studies of this gas. The effect of 4-hour exposure to ozone (O/sub 3/) at concentrations of 0.0, 0.2, 0.5, 1.0, and 2.0 ppm was determined in rabbits, guinea pigs, rats, hamsters and mice. Lavage fluid protein (LFP) accumulation 18 hr after exposure was used as the indicator of O/sub 3/-induced pulmonary edema. All species had similar basal levels of LFP (250-350 ug/ml) when a volume of saline which approximated the total lung capacity was used for lavage of the collapsed lungs. Exponential dose-response curves were seen in all species except guinea pigs, which showed significant increases in LFP at low O/sub 3/ concentrations (0.2 ppm) and a leveling off of response at the higher O/sub 3/ levels. Other species usually showed significant elevations in LFP ...
Propionibacterium freudenreichii ET-3 culture, a cell-free product of whey fermentation using P. freudenreichii ET-3 (7025), has been shown to promote the growth of Bifidobacteria through the action of 1,4-dihydroxy-2-naphthoic acid (DHNA), and therefore, has potential use in the food and supplement industries. Although currently used as a food ingredient in Japan, the safety of this novel ingredient has not been previously evaluated through traditional toxicity testing. Therefore, here we report the results of standard toxicological testing performed on P. freudenreichii ET-3 culture. In a 4-week oral toxicity study, administration of 6000mg/kg body weight/day P. freudenreichii ET-3 culture was without compound-related adverse effects on clinical signs, body weights, food consumption, ophthalmology, hematology, clinical chemistry, urinalysis, organ weights, and gross and microscopic findings in male and female Sprague-Dawley rats. Furthermore, in vitro mutagenicity testing ...
The nuclear lamina proteins, prelamin A, lamin B, and a 70-kD lamina-associated protein, are posttranslationally modified by a metabolite derived from mevalonate. This modification can be inhibited by treatment with (3-R,S)-3-fluoromevalonate, demonstrating that it is isoprenoid in nature. We have examined the association between isoprenoid metabolism and processing of the lamin A precursor in human and hamster cells. Inhibition of 3-hydroxy-3-methylglutaryl coenzyme A reductase by mevinolin (lovastatin) specifically depletes endogenous isoprenoid pools and inhibits the conversion of prelamin A to lamin A. Prelamin A processing is also blocked by mevalonate starvation of Mev-1, a CHO cell line auxotrophic for mevalonate. Moreover, inhibition of prelamin A processing by mevinolin treatment is rapidly reversed by the addition of exogenous mevalonate. Processing of prelamin A is, therefore, dependent on isoprenoid metabolism. Analysis of the conversion of prelamin A ...
An analogue of hGH isolated from plerocercoids of the tapeworm Spirometra mansonoides displaces ["1"2"5I]hGH from its receptors in rabbit, rat, and hamster liver membranes. Biologically, plerocercoid growth factor (PGF) is more similar to hGH than to other mammalian GH's but has not been shown to bond human cells. Receptors specific for hGH have been described on cultured human lymphocytes (IM-9). In this study, the authors compared the binding of PGF and hGH in IM-9 cells and in rabbit hepatic membranes. IM-9 lymphocytes (12 x 10"6 cells/tube) were incubated with ["1"2"5I]hGH and increasing concentrations of hGH (ng/ml) or PGF (serial dilutions) for 90 min at 30"0 C. Specific binding (B_0 - NSB) was determined for each dose of hGH or PGF and the binding curves were analyzed by logit-log regression. The results show that PGF displaced ["1"2"5I]hGH from human cells in a dose dependent manner (r = 0.98). Based on the IM-9 assay, 1 ml of the PGF had an activity ...
Asynchronous Chinese hamster ovary (CHO) cells treated with conditioning hyperthermia (10 min at 45/sup 0/C) followed immediately by x irradiation with 4 Gy at room temperature display a decreased radiation response when either irradiated, or heated and irradiated again 24 h later. In addition, conditioning by 4 Gy of x radiation alone followed at 24 h by the combined heat and radiation treatment als resulted in a larger radiation D/sub 0/. Since the differential cell-cycle responses to heat- and x-ray-induced lethality are well established for CHO cells, the potential role of cell-cycle redistribution in the observed radiation desensitization was investigated by flow cytometry. At 24 h after the combined treatment, the fraction of the cells found in the G/sub 2/ + M portion of the DNA histograms was about three times that of the controls and there was a concomitant reduction in both the G/sub 1/ and S fractions. Similar observations were made after the ...
Under certain growth conditions, the human colon adenocarcinoma cell line HT-29 exhibits intestinal enterocyte-like properties. The differentiated cells possess a brush border with the enzyme markers (aminopeptidase and sucrase) normally associated with the intestine. To aid in the characterization of the transport properties of these cells, the uptake of a non-metabolizable amino acid analog, "1"4C-#alpha#-methylaminoisobutyric acid (MeAIB) as examined in the HT-29-Al subclone which possesses a brush border. The cells exhibited a time-dependent uptake of MeAIB which was concentrative and sodium-dependent. The pH optimum for uptake was about 7.8. Uptake was inhibited by low temperature, 1 mM ouabain, or 0.5 mM dinitrophenol. A 1 hr-preincubation of the cells in an isotonic KCl solution resulted in a decreased uptake rate, suggesting that a negative membrane potential is important for MeAIB uptake. The rate of 0.5 mM MeABIB uptake was inhibited by 40 to 90% by 5 mM of certain small ...
Individuals with male factor infertility resulting from idiopathic oligo-, astheno- or teratozoospermia are frequently offered IVF in an attempt to increase their chances of having a child. A concern remains whether these infertile males have an elevated risk of transmitting chromosomal abnormalities to their offspring. Sperm chromosomal complements from these men were assayed using the human sperm/hamster oocyte fusion system and fluorescence in situ hybridization (FISH) on sperm nuclei. For each of 5 infertile patients, 100 sperm karyotypes were analyzed and multicolour FISH analysis was performed on a minimum of 10,000 sperm nuclei for each chromosome-specific DNA probe for chromosomes 1 (pUC1.77), 12 (D12Z3), X (XC) and Y (DYZ3). As a group, the infertile patients showed increased frequencies of both numerical ({chi}{sup 2}=17.26, {proportional_to} <0.001) and total abnormalities ({chi}{sup 2}=7.78, {proportional_to} <0.01) relative to control donors when ...
A prospective study was performed on 50 patients suffering from osteochondritis dissecans of the knee and ankle to define criteria for stability and fixation of osteochondral lesions. Morphological parameters in MRI (size, fragmentation, cartilage, interface) and conventional radiology (separation, fragmentation) were registered and compared with arthroscopical staging. MRI staging based on different types of interfaces was demonstrated on T{sub 1}- and T{sub 2}-weighted images. MRI could correctly predict a Grade 1 lesion in 50%, a Grade 2 lesion in 90%, a Grade 3 lesion on 0%, and a Grade 4 lesion in 79%. Stable lesions were differentiated from unstable lesions in 90%. Radiographic findings corresponded with arthroscopic staging in only 56% of the cases because fibrotic connection may guarantee stability in case of bony separation. (orig./MG) [Deutsch] In einer prospektiven Studie an 50 Patienten mit einer Osteochondrosis dissecans (OD) des Knie- und ...
The aim of this study was to evaluate typical magnetic resonance imaging (MRI) findings in early rheumatic diseases manifesting at the soft tissues of the hand using a retrospective analysis. A total of 186 MRI examinations of patients with clinical suspicion of a rheumatic disease were evaluated in a consensus reading by two experienced radiologists. All imaging patterns were assessed with respect to their type and localization. Under blinded and non-blinded conditions diagnoses were correlated with final clinical diagnosis. The most frequent diagnoses were rheumatoid arthritis (RA, 45.7%) and psoriatic arthritis (PsA, 15.6%). The mean correlation between clinical and MRI diagnosis (r) was 0.75 in blinded and 0.853 in non-blinded reading (p <0.001). The following extra-articular imaging patterns were found: synovitis (59.1%), tendovaginitis (91.4%), dactylitis (14.5%), and bone marrow edema (18.3%). Only dactylitis was specific for a particular rheumatic ...
Patients with haemarthros of the knee were examined by MRI and arthroscopy to identify the bleeding source and to determine surgical or conservative therapy. A prospective study was performed on 120 patients who underwent MRI and arthroscopy, point lesions and bleeding sources were documented separately on a standardised form. Arthroscopy demonstrated a ruptured anterior cruciate ligament in 86%, meniscal tears in 55%, disruption of collateral ligaments in 17%, osteochondral lesions in 5% and tears of capsule or plicae in 13%. Disruption of cruciate ligaments caused haemorrhage in all but three cases. In contrast, only meniscal tears located very adjacent to the meniscal basement had bled. Those types of osteochondral lesions caused haemorrhage characterised on MRI by destruction of the subchondral plate and overlying cartilage. Severe lesions of the knee were diagnosed by MRI in 71% to 100%, slight lesions in 13% to 40% of the cases. Single, combined and bland ...