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Biomedical subjects

C Keller

Publications and source records attributed to C Keller.

At least 217 records · Page 12Linked to original sources
PubMed1997

Construction of cDNA bank from biopsy specimens for multiple gene analysis of cancer.

We constructed a "cDNA bank" of human colorectal cancer and surrounding normal tissues with our unique mRNA assay system. Total nucleic acids extracted from patients' tissues were applied to 96-well plates, where poly(dT) sequences of oligonucleotides were immobilized. After hybridization, the cDNA was reverse-transcribed on the plate with the captured mRNA as a template, followed by synthesis of double-stranded (ds) cDNA. The resulting sense cDNA was removed from the plate, then used in PCR for analysis of various genes. The sense strand of the cDNA was repeatedly synthesized by using the immobilized antisense cDNA as a template even from plates used once and stored at 4 degrees C for as long as 6 months. Furthermore, the results of PCR could be easily compared among different specimens if the same amount of total mRNA were applied to the plate for the ds cDNA synthesis. This demonstrated that the cDNA bank constructed from clinical materials provides almost unlimited supplies of cDNA for multiple gene analysis of cancer.

Actins↗
PubMed1997

Physical activity in the elderly: benefits and intervention strategies.

In spite of the well-documented evidence that physical activity is beneficial, only 30% of individuals over the age of 65 report exercising regularly. Regular physical activity can minimize and prevent chronic problems and increase functional ability in the elderly. Designing physical activity for the elderly requires initial assessment of functional ability, exercise tolerance, physical limitations, and psychologic and social support of the individual. The exercise prescription should address the intensity, frequency, and duration of the exercise. Several factors affect the elderly's initiation and adherence to a physical activity program. These include the elder's perception of factors preventing physical activity, individual goal setting, and personal and therapist support in the effort.

Aged↗
PubMed1997

Assessment of overweight and obese patients.

The increasing prevalence of overweight and obesity in the United States has significant impact on the assessment, diagnosis, and treatment skill requirements of clinicians. There are a number of parameters that are used in the appropriate assessment of overweight and obesity. These include knowledge of basic body composition, measurement techniques for the assessment of total and regional body fat, and awareness of cultural influences on overweight and obesity. The focus of this article is to define overweight and obesity and delineate appropriate assessment parameters for those conditions.

Anthropometry↗
PubMed1997

The usefulness of three biallelic restriction fragment length polymorphisms versus a polymorphic dinucleotide tandem repeat polymorphism at the low-density-lipoprotein receptor gene locus for diagnosis of familial hypercholesterolemia.

Indirect molecular diagnosis of familial hypercholesterolemia is possible based on genetic linkage analysis of DNA polymorphisms at the low-density-lipoprotein receptor gene locus in family studies. The use of biallelic restriction fragment length polymorphisms, however, is restricted by their low degree of heterogeneity, and therefore several of these markers have to be combined. To overcome these restrictions we examined the value and applicability of an (AT)n tandem repeat polymorphism at the 3' untranslated region of the gene, alone and in combination with three biallelic restriction fragment length polymorphisms in 35 independent healthy subjects and in familial hypercholesterolemia families with 23 parents and 52 children. For each family one of the parents had the clinical diagnosis of familial hypercholesterolemia. The probands were genotyped using the TaqI, HincII and NcoI restriction fragment length polymorphism and the (AT)n tandem repeat polymorphism of the gene. The heterozygosity index (0.60) and the polymorphism information content (PIC value) (0.53) of the ATn repeat polymorphism were higher compared to each single biallelic restriction fragment length polymorphism (heterozygosity index 0.26-0.54; PIC value 0.24-0.36). The combined PIC value of all three biallelic restriction fragment length polymorphisms (0.79) was comparable to the combination of the HincII and the ATn polymorphism (0.74). Using these two markers, a definitive molecular diagnosis could be made in 36 children from 15 parents compared to just 12 parents and their children using the three biallelic restriction fragment length polymorphisms. We conclude that the ATn tandem repeat polymorphisms is useful for indirect molecular diagnosis of familial hypercholesterolemia in affected families.

Chromosome Mapping↗
PubMed1997

Pragmatic assessment of physical activity.

Nurse practitioners are confronted with the need to measure the physical activity levels of clients. There is no "gold standard" for measurement in the field. Nurses increasingly use physical activity for assessment, intervention, and evaluation. Selection of the appropriate tool depends on the purpose of the assessment, the client population, practicality, and psychometric properties of the instrument. Nine physical activity instruments with good psychometric properties, economy, and acceptability are presented to assist with instrument selection.

Activities of Daily Living↗
PubMed1996

Thoracoscopic laser bullectomy: a prospective study with three-month results.

One hundred forty-one patients were prospectively enrolled in a study of contact-tip laser bullectomy at four institutions. Ninety-one have had both preoperative and postoperative testing at 3 months. Nonsmoking patients with disabling dyspnea at less than 50 yards and with a forced expiratory volume in 1 second of 35% or less were enrolled. Testing included formal pulmonary function tests, arterial blood gasses, computed tomographic scans, ventilation/perfusion scans, echocardiograms, electrocardiograms, 6-minute walk testing, transdiaphragmatic pressures, and quality of life and dyspnea index questionnaires. A modest 16% improvement was noted in forced expiratory volume in 1 second (0.69 to 0.80 L), and there was a 29% improvement in 6-minute walk distances (655.2 to 846.3 feet). Oxygen use was completely discontinued in 16%. Risk factors for mortality included age, 6-minute walk distances, low diffusing capacity for carbon monoxide, high carbon dioxide tension, and high base excess. Minor improvement was judged from the dyspnea index and the Medical Outcome Study Short Form-36. Preoperative predictors of good outcome included heterogeneous disease, lack of carbon dioxide retention, and no emaciation (weight < 40 kg). Comparison of our results with those in the literature suggests that the improvement seen with the contact neodymium:yttrium-aluminum-garnet laser is not as good as that provided by the stapled techniques for volume reduction.

Acid-Base Imbalance↗
PubMed1996

Nephropathy of type II diabetes mellitus.

In recent years there has been a dramatic increase of almost epidemic proportions in the incidence of patients with type II diabetes mellitus who reach end-stage renal failure and enter renal replacement programmes. This is mainly due to the greater prevalence and better survival of patients with type II diabetes and diabetic nephropathy. Against this background measures to prevent the appearance and progression of diabetic nephropathy are of immense interest. Apart from the undoubted role of hyperglycaemia, the importance of genetic determinants of nephropathy has recently been recognized. Factors of proven or suspected efficacy in attenuating progression include: hypertension, hyperglycaemia, smoking and proteinuria. The role of dietary protein intake is less well documented. Nephropathy in type II diabetes has become the single most common cause of end-stage renal failure in Germany and is today a major challenge to clinical nephrology.

Blood Pressure↗
PubMed1996

Multiplex-PCR-based single-strand conformation polymorphism protocol for simultaneous analysis of up to five fragments of the low-density-lipoprotein receptor gene.

Single-strand conformation polymorphism has become a screening method for the detection of mutations in different genes. For analysis of the promotor region and the coding sequence of the low-density-lipoprotein receptor gene by standard protocols, 21 radiolabeled PCRs and electrophoreses have to be performed. To accelerate this procedure, we developed a nonradioactive multiplex approach of the single-strand conformation polymorphism analysis. Multiplex PCRs were established, each resulting in the amplification of 4 or 5 fragments of this gene. The heat-denatured, single-stranded multiplex-PCR products were electrophoresed, blotted on a nylon membrane and visualized using a chemiluminescence detection system. The simultaneously amplified fragments were clearly resolved by their different mobility on the gel. Comparing the pattern of bands of each separately amplified PCR product and the multiplex-PCR products allowed identification of each band as one exon, part of an exon or the promotor region of the gene. To determine the sensitivity of this method, the low-density-lipoprotein receptor gene of 11 patients with 11 different mutations was analyzed. All mutations could be identified in the multiplex reactions. We conclude that a multiplex-PCR-based, single-strand conformation polymorphism protocol is much faster but equally sensitive compared to standard protocols.

Codon↗
PubMed1996

Plasma thrombomodulin: a marker for microvascular complications in diabetes mellitus.

Thrombomodulin (TM), a marker of endothelial cell damage was studied in 183 patients with diabetes mellitus and different stages of complications. Thrombomodulin plasma levels correlated with duration of diabetes in patients with type I and type II diabetes. Thrombomodulin levels were higher in patients with increasing numbers of complications (nephropathy, retinopathy, arterio-occlusive disease, neuropathy). Neither the presence of retinopathy, nor neuropathy alone significantly increased plasma thrombomodulin in patients with similar urinary albumin concentration. The plasma level of thrombomodulin was more prominent in hypertensive than normotensive patients. Multivariate analysis showed that albuminuria is the factor which influences the most the increase of thrombomodulin in serum of diabetic patients.

Adult↗
PubMed1996

Effect of fluvastatin or bezafibrate on the distribution of high density lipoprotein subpopulations in patients with familial hypercholesterolemia.

In eight patients with familial hypercholesterolemia the effects of two lipid reducing drugs on subpopulations of high density lipoproteins (HDL) were examined. After a 14-week period of diet and diet/placebo a 12-week therapy followed with either bezafibrate (CAS 41859-67-0) or fluvastatin (CAS 93957-55-2). Throughout both treatments a significant decrease of total and low density lipoprotein (LDL)-cholesterol, apolipoprotein B and triglycerides during both therapies were noted as well as an insignificant increase of HDL-cholesterol during bezafibrate. The nondenaturating gradient gel electrophoresis is a valid method for the investigation of the behaviour of HDL and was therefore chosen for this investigation. The individual HDL pattern and HDL diameters did not change in these subjects. The effect on the amounts of HDL 3b and HDL 3c was significantly more extensive during fluvastatin (+2.4% resp. + 2.9%) as compared to during bezafibrate therapy (+1.7% resp. + 2.9%). The changes noted in the HDL subclasses are probably due to a variable lipoprotein metabolism, for example increased activity of lipoprotein lipase, hepatic triglyceride lipase, lecithin-cholesterol acyltransferase and cholesterol ester transfer protein.

Adult↗
PubMed1996

Differential regulation of transcription by the NURR1/NUR77 subfamily of nuclear transcription factors.

NURR1 is an orphan member of the nuclear receptor superfamily of transcription factors that shares close sequence homology to the orphan nuclear receptor and immediate early gene product NUR77(NGF1 beta). The physiological role of NURR1 has not been established in mammalian cells. However, the observation that NURR1 and NUR77 interact with at least one common enhancer element (AAAAGGTCA), together with their partly overlapping but differential expression patterns in mammalian tissues, suggests that these proteins may have both shared and independent transcription regulatory functions. To identify potential target genes that may be regulated by NURR1, we analyzed its DNA binding properties to potential cis-acting enhancer elements. Using point mutagenesis of the AAAAGGTCA motif, we have identified three additional sequences that bind specifically to both NURR1 and NUR77, one of which serves as a functional enhancer element. Comparative analysis of the transcription regulatory properties of NURR1 and NUR77 indicates that the proteins can display opposing transregulatory activities that are influenced by the specific cis-acting sequences to which they bind. Our results indicate that the transcriptional responses of specific target genes to the NURR1/NUR77 subfamily may be differentially regulated by the relative cellular levels of NURR1 and NUR77 and influenced by the specific enhancer sequences that mediate their activity. Finally, we have identified several potential target genes of neuronal and neuroendocrine origin whose promoters contain this element.

Base Sequence↗
PubMed1996

Assessment, etiology, and intervention in obesity in children.

Obesity is a common nutritional disturbance of children and affects 25% to 30% of children and adolescents. This paper examines obesity in childhood, the measurement of obesity in children, and the relationship of obesity to coronary heart disease risks and discusses weight reduction issues in children. Clinically useful definitions of obesity in children have not been established, although the body mass index, together with anthropometric measurements, may provide the practitioner with useful assessment parameters. Discussions of weight in children must acknowledge the nutritional requirements of the normally occurring growth process. Because growth (and subsequently weight) varies widely among children even of the same age, measurement, classification, and control of weight must take into account the growth process and growth requirements. Interventions in childhood obesity should be directed toward family involvement in the chosen strategy and nutritional prudence coupled with typical activity.

Adolescent↗
PubMed1995

Donor-specific transfusions have long-term beneficial effects for human renal allografts.

From April, 1980 to November, 1988, 163 one or two haplotype-mismatch living related (LR) or living unrelated (LUR) potential renal transplant recipients received three 200 ml aliquots of donor specific transfusion (DST) at biweekly intervals with concomitant azathioprine (2 mg/kg/day). Following transplantation, only prednisone and azathioprine were given for immunosuppression. The results for the DST group are compared with those for HLA identical living recipients (57 patients) transplanted during this same interval (1980-1988). Comparison is also made with a group of one or two haplotype-mismatched living donor recipients (54 patients) not treated with DST but with triple drug therapy (prednisone, azathioprine, cyclosporine) and antithymocyte globulin (ATG) or OKT-3 induction. Permanent T cell crossmatch sensitization occurred in 11 of 163 patients (7%). Successful DST donor transplants were performed between 121 one HLA haplotype-mismatched, 14 two HLA haplotype-mismatched LR, and 7 two haplotype-mismatched LUR pairs. Actual one- and five-year graft survivals were 94%, 100%, 100%, and 72%, 85%, and 71%, respectively, for these three subgroups of DST treated patients. The graft survival for all DST pretreated recipients at one, five, and ten years was comparable to the HLA-identical group (94%, 79%, 64% vs. 91%, 80% and 77%). At a mean follow-up of 10 1/2 years, 54% (80 patients) of the entire group of 147 patients transplanted after DST have functioning transplants with a mean serum creatinine of 1.7 mg/dl. Fifteen percent of DST patients (21 patients) died with a functioning graft 2 to 132 months after transplantation, 26% (37 patients) rejected the DST graft after 1 to 128 months, and 6% (9 patients) were lost for nonimmunological reasons. No lymphoproliferative disease developed in the DST group and the incidence of cytomegalovirus sepsis was only 2% (3 patients). The long-term beneficial effects of DST on renal allograft survival and function and the lower incidence of the complications of nonspecific immunosuppression should encourage increased utilization of DST in renal transplantation.

Azathioprine↗