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At least 343 records · Page 19Linked to original sources
PubMed1988

Isolation, characterization, and long-term culture of fetal bovine tracheal epithelial cells.

Epithelial cells were isolated from fetal bovine trachea by exposing and stripping the mucosal epithelium from the adjacent connective tissue. The tissue was minced and enzymically dissociated in Ca-Mg-free medium containing dispase and dithiothreitol. The stripping procedure and selective trypsinization produced epithelial cell cultures free of fibroblasts. Seeded on plastic, the plating efficiency was 21.5% with a doubling time of 24 h. Dome formation, evidence of occluding junctions and active ion transport characteristic of epithelial cells, was common. Growth of the cells on glass, collagen, and Engelbreth-Holm-Swarm (EHS) substrate demonstrated a striking difference in morphology. Cells grown on EHS presented a more distinctly three-dimensional growth pattern and many more microvilli when compared to cells grown on glass or collagen. The cells retained their epithelioid characteristics through more than 30 passages as shown by the presence of distinct apical and basolateral membranes, tight junctions, and positive keratin staining.

Animals↗
PubMed1990

Microbiological cultivation of Mycoplasma hyorhinis from cell cultures.

The failure of many cell culture isolates of Mycoplasma hyorhinis to grow on microbiological media has stressed the need for alternate assays to detect these organisms. The use of freshly prepared yeast extract in mycoplasmal media together with incubation in 5% CO2/air successfully detected M. hyorhinis in 12 of 12 infected cultures. These were not detected by the use of conventional mycoplasmal media using aerobic or anaerobic incubation. This assay may also be helpful in detection of other mycoplasmal species commonly isolated from cell cultures.

Cells, Cultured↗
PubMedDate not supplied

Phenotypic differences between Syrian hamster embryo cells cultured at pH 6.7 or 7.3.

We have investigated the molecular phenotypic differences between Syrian hamster embryo (SHE) cells cultured at pH 6.7 or 7.3. Multiple pH-sensitive phenotypic differences were noted including changes in cellular morphology, a unique charge differential in a major cellular protein, nine uniquely expressed proteins, two unique phosphoserine/threonine phosphoproteins, one unique phosphotyrosine phosphoprotein, and the pH dependent mRNA level of a gap junctional gene (connexin 43). These differences, combined with previously described pH-specific differences (differential transformation rates and gap junctional communication), illustrate that culturing SHE cells in media that differ by 0.6 pH units (0.3 units on either side of pH 7.0) can have a profound influence on the cellular phenotype.

Animals↗
PubMedDate not supplied

Establishment of immortalized alveolar type II epithelial cell lines from adult rats.

We developed methodology to isolate and culture rat alveolar Type II cells under conditions that preserved their proliferative capacity, and applied lipofection to introduce an immortalizing gene into the cells. Briefly, the alveolar Type II cells were isolated from male F344 rats using airway perfusion with a pronase solution followed by incubation for 30 min at 37 degrees C. Cells obtained by pronase digestion were predominantly epithelial in morphology and were positive for Papanicolaou and alkaline phosphatase staining. These cells could be maintained on an extracellular matrix of fibronectin and Type IV collagen in a low serum, insulin-supplemented Ham's F12 growth medium for four to five passages. Rat alveolar epithelial cells obtained by this method were transformed with the SV40-T antigen gene and two immortalized cell lines (RLE-6T and RLE-6TN) were obtained. The RLE-6T line exhibits positive nuclear immunostaining for the SV40-T antigen and the RLE-6TN line does not. PCR analysis of genomic DNA from the RLE-6T and RLE-6TN cells demonstrated the T-antigen gene was present only in the RLE-6T line indicating the RLE-6TN line is likely derived from a spontaneous transformant. After more than 50 population doublings, the RLE-6T cells stained positive for cytokeratin, possessed alkaline phosphatase activity, and contained lipid-containing inclusion bodies (phosphine 3R staining); all characteristics of alveolar Type II cells. The RLE-6TN cells exhibited similar characteristics except they did not express alkaline phosphatase activity. Early passage RLE-6T and 6TN cells showed a near diploid chromosome number.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed2000

Impact of sentinel lymph node mapping on relative charges in patients with early-stage breast cancer.

BACKGROUND: The introduction of SLNB has allowed accurate staging in early-stage breast carcinomas and has minimized the number of unnecessary ALNDs. Intraoperative frozen-section analysis is a fundamental component of the sentinel lymph node biopsy (SLNB) procedure. Some patients have positive nodes on frozen-section analysis and thus undergo a conventional axillary lymph node dissection (ALND) at the time of the SLNB. A few patients have negative nodes on frozen-section analysis but have subsequent evidence of metastases on final pathologic examination. The purpose of our study was 2-fold: to compare the hospital-related charges of patients undergoing staging by SLNB with those of patients undergoing conventional ALND and to assess whether the different outcomes associated with SLNB adversely affect the charges incurred with this procedure. METHODS: Our study group consisted of 100 patients with T1 breast cancer and breast conservation therapy who underwent either SLNB or ALND from July 1, 1997, to June 30, 1998. We identified the first 50 consecutive patients to undergo SLNB during this period. We chose a similar cohort of 50 patients for ALND. Mean hospital-related charges for the SLNB patients were categorized and compared with those for the ALND patients. RESULTS: Results for the two groups were analyzed using a two-sample Wilcoxon rank-sum test. Charges for the OR and hospital stay were less for the SLNB group (P < .05). Frozen-section analysis in the SLNB group contributed to the significant difference in charges for pathologic evaluation. Overall, the two groups showed no significant difference in total hospital-related charges. CONCLUSIONS: When SLNB is used for T1 tumors, a small percentage of patients (10% in our study) will return to the operating room for an ALND. This small percentage does not increase the charges related to SLNB, however, as the reduced stay for most patients offsets this subgroup's contribution to the total hospital-related charges. Thus, in patients with clinical stage I breast cancer, SLNB does not cause significantly higher hospital-related charges compared with conventional ALND.

Axilla↗
PubMed2002

Tactile inhibition of return: non-ocular response inhibition and mode of response.

Inhibition of return (IOR) refers to the slowing of responses to stimuli presented at the same location as a preceding cue. However, the cue-target paradigm used in most previous studies may have overestimated the contribution of eye movement programming to IOR, due to the existence of manual response inhibition effects. This confound can be circumvented by using a target-target paradigm in which participants respond to all stimuli. Here, we compared IOR magnitude from tactile cue-target and target-target tasks involving identical interstimulus intervals of 1,400 and 1,800 ms. Reaction times were measured using a foot pedal toe-lift response and a vocal response. Tactile IOR was observed using both modes of response, demonstrating IOR for the first time using a non-spatial, vocal response. Moreover, IOR effects were significantly smaller in target-target compared to cue-target conditions, thereby confirming the existence of the response inhibition confound.

Adolescent↗
PubMed2005

Moderate sedation for MRI in young children with autism.

UNLABELLED: Autism is a pervasive neurodevelopmental disorder. Because of the deficits associated with the condition, sedation of children with autism has been considered more challenging than sedation of other children. OBJECTIVE: To test this hypothesis, we compared children with autism against clinical controls to determine differences in requirements for moderate sedation for MRI. MATERIALS AND METHODS: Children ages 18-36 months with autism (group 1, n = 41) and children with no autistic behavior (group 2, n = 42) were sedated with a combination of pentobarbital and fentanyl per sedation service protocol. The sedation nurse was consistent for all patients, and all were sedated to achieve a Modified Ramsay Score of 4. Demographics and doses of sedatives were recorded and compared. RESULTS: There were no sedation failures in either group. Children in group 1 (autism) were significantly older than group 2 (32.02+/-3.6 months vs 28.16+/-6.7 months) and weighed significantly more (14.87+/-2.1 kg vs 13.42+/-2.2 kg). When compared on a per-kilogram basis, however, group 1 had a significantly lower fentanyl requirement than group 2 (1.25+/-0.55 mcg/kg vs 1.57+/-0.81 mcg/kg), but no significant difference was found in pentobarbital dosing between groups 1 and 2, respectively (4.92+/-0.92 mg/kg vs 5.21+/-1.6 mg/kg). CONCLUSION: Autistic children in this age range are not more difficult to sedate and do not require higher doses of sedative agents for noninvasive imaging studies.

Anesthetics, Intravenous↗
PubMed2004

Dual-channel photobleaching FRET microscopy for improved resolution of protein association states in living cells.

Fluorescence resonance energy transfer (FRET) from a donor-labelled molecule to an acceptor-labelled molecule is a useful, proximity-based fluorescence tool to discriminate molecular states on the surface and in the interior of cells. Most microscope-based determinations of FRET yield only a single value, the interpretation of which is necessarily model-dependent. In this paper we demonstrate two new measurements of FRET heterogeneity using selective donor photobleaching in combination with synchronous donor/acceptor detection based on either (1) full kinetic analysis of donor-detected and acceptor-detected donor photobleaching or (2) a simple time-based ratiometric approach. We apply the new methods to study the cell surface distribution of concanavalin A yielding estimates of FRET and non-FRET population distributions, as well as FRET efficiencies within the FRET populations.

Animals↗
PubMed2004

The distal femoral and proximal tibial growth plates: MR imaging, three-dimensional modeling and estimation of area and volume.

OBJECTIVE: To explore how the size of the growth plate changes with age using three-dimensional (3D) models of the distal femoral and proximal tibial growth plates in pediatric patients. DESIGN AND PATIENTS: We retrospectively created 3D models of the normal unaffected distal femoral (n=20) and proximal tibial (n=10) growth plates in 14 patients (9 males, 5 females) age range 3.8-15.6 years who were referred for evaluation of premature partial closure of the growth plate or hyaline cartilage abnormality. All patients had one or more 3D fat-suppressed spoiled GRASS sequence from which models were made of normal growth plates. Total projected area was estimated from standardized maximum intensity projection (MIP) views, and volume was computed from the entire model. We also included the total projected area of the distal femur (n=7) or proximal tibia (n=8) in 11 patients (8 males, 3 females, 5-13 years) who had previously been evaluated for bone bridging. RESULTS: The 3D femoral and tibial growth plate anatomy was displayed. Femoral growth plate area varied from 804 mm2 to 3,463 mm2. Femoral physeal cartilage volume varied from 2.1 cm3 to 12.6 cm3. Tibial growth plate area varied from 736 mm2 to 3,026 mm2. Tibial physeal cartilage volume varied from 1.9 cm3 to 13.2 cm3. The growth plate area values appear to increase linearly with increasing age. CONCLUSIONS: The distal femoral and proximal tibial physeal plates have complex anatomy. Both area and volume of the growth plates appeared to follow a linear increase with age and reached a plateau in adolescence, although there was some scatter. Area appears to have less measurement variability than volume, and may be a more reliable predictor of growth plate tissue quantity.

Adolescent↗
PubMed2000

Bone density distribution and gender dominate femoral neck fracture risk predictors.

OBJECTIVE: To determine whether regional characteristics of the proximal femur could discriminate between a group of patients who had just sustained a first low-trauma femoral neck fracture (n=50) from a group of healthy volunteers (n=123). DESIGN: The application of an integral bone measurement (dual-energy X-ray absorptiometry) in conjunction with a volumetric cancellous bone density measurement (quantitative computed tomography) to the proximal femur in vivo provided an estimate of the contribution of the spatial distribution of bone density to hip fracture risk prediction. RESULTS: The primary finding of this study was a significant difference between male and female hip fracture risk predictor variables. In men with femoral neck fracture, a significant decrease in bone density throughout the proximal femur was observed. In women with femoral neck fracture, a combination of local bone deficits (significant decrease in cancellous bone at the site of fracture, and a decrease in cortical bone at the site of impact) and significantly larger proximal femur dimensions (femoral neck and head widths) was evident. CONCLUSIONS: These results imply that effective hip fracture prevention strategies may require separate approaches for men and women. Screening programs for diminished bone density at the proximal femur have proved effective in previous studies. An approach which includes examining these local bone characteristics may further improve our ability to accurately determine hip fracture risk in vivo.

Absorptiometry, Photon↗
PubMed2005

Orthopaedic applications of gene therapy.

Gene therapy presents a novel approach to biological treatment. Several orthopaedic diseases can cause changes in biological signalling at the tissue level that potentially can be repaired or modified by inserting genes into the cells or tissues to modulate gene expression. Impaired bone healing, need for extensive bone formation, cartilage repair and metabolic bone diseases are all conditions where alterations of the signalling peptides involved may provide cure or improvement. In orthopaedic oncology, gene therapy may achieve induction of tumour necrosis and increased tumour sensitivity to chemotherapy. In the last decade, extensive improvements have been made to optimise gene therapy and have been tested on several orthopaedic conditions. How far this development has come in orthopaedics is highlighted in this paper.

Bone Diseases↗
PubMed2003

Natural mechanisms for evading graft rejection: the fetus as an allograft.

Most primitive multicellular animals mount allogeneic immune responses to protect themselves from invasion by foreign organisms. The reproductive success of eutherian mammals, in which the maternal immune system is in direct contact with the semi-allogeneic fetus, depends on the ability to control allogeneic immune responses. Multiple, overlapping mechanisms exist to prevent maternal allogeneic immune responses towards the fetus while maintaining the capacity to mount a defense against infectious organisms. The formation of an anatomical barrier between mother and fetus, lack of maternal immune responsiveness, and a lack of expression of allogeneic molecules by the fetus have been proposed as mechanisms to account for the lack of fetal rejection during pregnancy. These mechanisms have helped us begin to understand how rejection of the fetus is avoided; however, these mechanisms do not completely explain how the fetus evades the maternal immune system. Site-specific suppression, in which maternal immune responses are controlled locally at the maternal- fetal interface, plays a fundamental role in controlling maternal allogeneic immune responses. Stem cells, both adult and embryonic, might use mechanisms similar to those of the fetus to avoid rejection. Future discoveries in the field of reproductive immunology will help us understand not only immune regulation during pregnancy, but also how immune responses towards organ and cellular transplants might be controlled.

Animals↗
PubMed2002

Concepts in developing health-based indicators for ozone.

OBJECTIVES: The traditional manner to evaluate whether regulatory controls meet their public health goals of reducing adverse health effects associated with exposure to environmental pollutants is to compare measured concentrations of the target pollutant in the environment with a standard. A complementary approach is also to measure health-based indicators, e.g., changes in the prevalence of adverse health outcomes attributed to the pollutant. This manuscript presents the concepts of using asthma emergency room (ER) visits and hospital admission as potential health-based indicators for ozone. METHODS: The frequency of ER visits and hospital admissions for asthma in New Jersey in 1995 was compared with daily ozone concentrations, to establish the consistency of the relationship and the presence of potential confounders, and to establish whether routinely documented adverse outcomes in asthmatics could serve as health-based indicators. RESULTS: A mathematical model relating ER visits and hospital admissions of asthmatics to ozone concentration was developed for 1995, which was to be used as a baseline year within a health-based indicator program. A coherent relationship was found between same-day ambient air ozone concentrations and ER visits and 2-day time-lagged ambient ozone and hospital admissions during 1995; pollen was identified as a confounder and the association between ER visits and ozone concentration was similar to that determined for 1986 to 1990. CONCLUSIONS: Sufficient databases exist for ER visits by asthmatics in Northern and Central New Jersey, and throughout the state for hospital admissions, for these health outcomes to be used as health-based indicators, complementing air-monitoring data in assessing whether improvements in public health are occurring because of reduction in emissions of precursors of ozone.

Air Pollutants↗
PubMed1999

Gene expression patterns in cell lines from patients with 18q- syndrome.

Some studies have suggested that for trisomies, some genes are expressed far in excess of the expected 150% level and that this "dysregulation" is one of the mechanisms for the pathogenesis of trisomies. In an attempt to generalize this result to a monosomy, we examined mRNA isolated from lymphoblastoid cell lines derived from patients with 18q- syndrome, a deletion syndrome involving loss of the distal long arm of chromosome 18. Expression levels of ten chromosome 18 genes were compared between cell lines from eight patients with 18q- syndrome and four diploid controls. Gene expression was investigated by a quantitative reverse-transcription polymerase chain reaction (RT-PCR) method. With the exception of the transcription factor NFATC1, which shows a tendency towards gene dosage compensation (the expression pattern correlates with IgA deficiency), all of the other genes were expressed at a level proportional to their gene copy number. This was true regardless of mRNA abundance or different patterns of gene expression (ubiquitous versus tissue-specific gene expression). These results indicate that, unlike dysregulated gene expression apparent in some trisomies, this monosomic syndrome is largely due to consequences of reduced gene expression.

Blotting, Northern↗
PubMed1999

Haplosufficiency of the melancortin-4 receptor gene in individuals with deletions of 18q.

The melanocortin-4 receptor (MC4R) is a seven, transmembrane G-protein-coupled receptor whose ligand, alpha-melanocyte-stimulating hormone (alpha-MSH), is a post-translational derivative of pro-opiomelanocortin (POMC). The regulatory pathway, of which MC4R is a part, has become an area of intense interest because of its potential role in obesity. Three studies have identified individuals with dominantly inherited obesity segregating with mutations in the MC4R gene. It has been hypothesized that the mutation found in these subjects resulted in a loss of gene function resulting in obesity due to haploinsufficiency of the MC4R gene. We have been studying the molecular basis of the phenotype of individuals with large deletions of chromosome 18q. Due to its location at 18q21.3, the MC4R gene is hemizygous in approximately one-third of the individuals in our study. If hemizygosity of the MC4R gene results in haploinsufficiency-induced obesity, then individuals with deletions of 18q whose deletions include the MC4R gene should be obese in comparison with those individuals whose deletion does not include the gene. Our data indicate no difference in obesity among those deleted and not deleted for the gene. This supports the hypothesis that the MC4R gene product is haplosufficient and the involvement of MC4R in obesity may reflect a dominant negative effect.

Base Sequence↗