Biomedical subjects
Publications and source records attributed to A Fine.
There's danger in direct contracting with employers, but big advantages if you know how to steer clear. In this excerpt from Dealing Direct, a recent book from American Hospital Publishing, the authors point out common obstacles--from retaliation by insurers to managing high-risk enrollees to shifts in the market.
Green fluorescent protein (hGFP-S65T) was expressed in transgenic mice under the control of the astrocyte-specific glial fibrillary acidic protein (GFAP) promoter. Tissues from two independent transgenic lines were characterized by Northern blot analysis and by confocal microscopy. The expression pattern in these two lines was identical in all tissues examined, and similar to that found previously with a lacZ transgene driven by the same promoter. Bright fluorescence was observed in the cell bodies and processes of unfixed or fixed astrocytes, using both whole mount and brain slice preparations, from multiple areas of the central nervous system. However, in contrast to GFAP-lacZ transgenics, retinal Müller cells expressed the GFP transgene in response to degeneration of neighboring photoreceptors. These data indicate that the 2.2-kb hGFAP promoter contains sufficient regulatory elements to direct expression in Müller cells, and that GFP is a suitable reporter gene for use in living preparations of the mammalian nervous system. Such mice should prove useful for studies of dynamic changes in astrocyte morphology during development, and in response to physiological and pathological conditions.
Two cases of severe hyperphosphatemia following phosphate bowel preparation are described and a review of the literature is presented. Impairment of renal function appears to be a risk factor in those patients without primary bowel pathology. One of our patients died, and the mortality rate combining our cases with all other reported cases is 33%. Repeated doses of phosphate bowel preparations/purgatives can be dangerous in patients with renal impairment.
Demand for dialysis for patients with end-stage renal disease is growing, as is the comorbidity of dialysis patients. Accurate prediction of those destined to die quickly despite dialysis could be useful to patients, providers, and society in making decisions about starting dialysis. To determine whether age and comorbidity accurately predict death within 6 months of first dialysis for end-stage renal disease, a prospective cohort study of 822 patients starting dialysis at one of 11 Canadian centers was performed. Patient characteristics were recorded at first dialysis. Follow-up continued until death or study end (at least 6 months after enrollment). One hundred thirteen of 822 (13.7%) patients died within 6 months. Although an existing scoring system predicted prognosis, adverse scores greater than 9 were found in only 9.7% of those who died; only 52% of those who scored higher than 9 died within 6 months. No score cutoff point combined high true-positive and low false-positive rates for predicting early death. Age, severity of heart failure or peripheral vascular disease, arrhythmias, malnutrition, malignancy, or myeloma were independent prognostic factors identified in multivariate models. However, the best fit discriminant and logistic models were also unable to accurately predict death within 6 months. Clinicians were very accurate in assigning patients to prognostic groups up to a 50% risk of death by 6 months, above which they tended to overestimate risk. However, clinicians were only marginally better than the predictive models in determining whether a given high-risk patient would die. The inability of a scoring system or clinical intuition to accurately predict death soon after starting dialysis for end-stage renal disease suggests that limiting access to dialysis on the basis of likely short survival may be inappropriate in Canada.
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Central nervous system neurons and glia arise from undifferentiated embryonic neuroepithelial cells. Such progenitor cells from the human fetal forebrain can be propagated in vitro for extended periods, when grown on non-adhesive substrates in medium containing epidermal growth factor and insulin-like growth factor-1. These actively-dividing cells can be induced to differentiate into a variety of histochemically-characterized neurons and glia consistent with their forebrain origin. Electrophysiological recording indicates that differentiated neurons derived from these progenitors mature slowly, and display a range of glutamate- and GABA-mediated conductances characteristic of normal mammalian forebrain neurons. Our observations support a role for these trophic factors in normal development of the human brain. The methods described here may provide abundant normal, untransformed human forebrain neurons and glia for research and therapeutic applications.
The distribution of neurones that could be stained immunohistochemically with antibody to the p75 neurotrophin protein was studied in the forebrain of the common marmoset. The p75-immunoreactive forebrain cells appear to correspond to choline acetyltransferase-immunoreactive (i.e., cholinergic) neurones. Two populations of cells could be distinguished on the basis of the intensity of p75 immunostaining. Moderately stained cells correspond to cholinergic interneurones of the caudate and putamen, while intensely stained cells correspond to the cholinergic neurones projecting to the cortex, amygdala, and hippocampus, located in the septum, diagonal band, and basal nucleus of Meynert. The distribution of cells of the diagonal band/basal nucleus complex is more extensive in the marmoset than in other primate species, extending into parts of the postcommissural fornix via the posterior septum, and by small projections dorsal to the anterior commissure and via the thalamic fasciculus from the basal nucleus; the posterior extent of the basal nucleus continues extensively into the lamina between the globus pallidus and the putamen.
Apoptotic cells in tissue sections can be localized by in situ labelling of partly degraded DNA. In a heterogeneous population of cells, however, the specific identity of cell types undergoing apoptosis often cannot be reliably achieved at the light microscope level because of the marked alterations in cellular morphology that characterize apoptosis. In order to clearly specify cell types undergoing apoptosis, in situ end labelling has been coupled to immunohistochemistry. This method is limited by the availability of antibodies that bind to cell-specific protein markers in tissue sections. In contrast, we describe a method that combines in situ end labellin with in situ hybridization, a technique that specifies cell types based on mRNA expression. Taking advantage of the specific expression of surfactant protein C mRNA in type II alveolar epithelial cells, we demonstrate that this technique has the ability to localize alveolar type II cells undergoing apoptosis in vivo after the intratracheal instillation of an antibody that activates the cell surface Fas protein. The wide availability of cell-specific gene markers suggests that this method can be adapted to define cell types that undergo apoptosis during various physiological and pathological states in vivo.
Fas, a type I membrane receptor protein, transduces a signal culminating in apoptosis after binding to the Fas ligand. Information regarding the expression of Fas in nonlymphoid tissues, although limited, suggests a role for Fas in epithelial progenitor cell populations. In this paper, we provide several lines of evidence indicating that the progenitor cell of the alveolus, the type II cell, displays restricted expression of Fas. We found 1) Fas gene expression in RNA derived from fresh isolates of primary rat type II cells; 2) restriction of Fas expression to a subset of alveolar type II cells by in situ hybridization and immunohistochemistry of the normal mouse lung; 3) induction of apoptosis in a mouse lung type II epithelial cell line (MLE) after activation of Fas; and 4) induction of apoptosis in a subpopulation of type II cells after the intratracheal instillation of an activating anti-Fas antibody in mice. These findings suggest that Fas-dependent apoptosis is involved in regulating turnover of the alveolar epithelium.
Salt restriction in continuous ambulatory peritoneal dialysis (CAPD) patients is widely prescribed and thereby may reduce quality of life. It is presumed that this has a beneficial effect on BP and reduces the need for hypertonic dialysate. However, this has never been formally evaluated. A double-blind crossover study of placebo versus sodium chloride pills (60 mEq of sodium per day) is presented in 20 stable CAPD patients, 10 of whom were hypertensive. Dietary sodium was quantified throughout the study by 3-d dietary histories and remained unaltered throughout. There was a clinically unimportant but statistically significant rise in BP with added salt: 135/77 to 144/82 (P < 0.05). No rise in BP occurred in the hypertensive patients. Weights, use of hypertonic dialysate, and BP medications remained unaltered throughout the study. In conclusion, 200 mEq of sodium per day, i.e., a normal sodium intake, is easily tolerated in stable CAPD patients, and the recommended sodium intake commonly prescribed is too restrictive.
During development, most neurons become polarized when one neurite, generally the longest, becomes the axon and the other neurites become dendrites. The physical mechanism responsible for such length-related differentiation has not been established. Here, we present a model of neuronal polarization based upon the existence of a "determinant chemical' whose concentration at the neurite tips influences the growth rate of the neurite. Over an extended parameter range the equations describing the formation, transport, and consumption of this chemical and the resulting neurite growth undergo a winner-take-all instability, yielding rapid growth of one neurite (the axon) and diminished growth of all others. The behaviour of this model agrees well with the results of axotomy experiments and experiments in which growth-modulating substances are applied to individual growth cones. Possible candidates for the determinant chemical are discussed, and further experiments are proposed to test the model.
The type I transforming growth factor-beta (TGF-beta) receptors are serine/threonine kinases that are essential for the action of TGF-beta. In this paper, we describe the molecular cloning and expression of the R4/ALK5 human type I TGF-beta receptor promoter. DNA sequence analysis indicates that the promoter lacks a TATA and CAAT box but is highly GC-rich and contains putative Sp1 binding sites. The transcriptional start site is approx. 232 base pairs upstream of the AUG start codon. In human lung fibroblasts, TGF-beta induced a 3-fold increase in steady-state level for type I receptor mRNA. Exposure of cells transfected with a 618 bp promoter fragment to TGF-beta 1 up-regulated transcriptional activity indicating that a TGF-beta response element is contained within this region.
Highly branched dendritic shapes are distinguishing characteristics of neurons and certain other cell types, but the physical mechanisms responsible for their formation are not well understood. Here, we model the growth of cells under the control of diffusible growth-regulating factors (morphogens such as calcium ion) whose local internal concentration results from influx and active extrusion across the cell membrane. Nonlinearities in voltage-dependent ionic permeabilities enhance unstable growth, so that branching dendritic outgrowths results from self-sustaining internal morphogen gradients. Simulations display complex patterns of branching growth, influenced by membrane conductance, galvanotropism and chemotropism. This self-organizing pattern formation is in agreement with the development of real neurons under corresponding conditions.
Ultrasound examination of the liver and right lung was performed in 260 sheep and 320 goats from the Turkana district of Kenya. Hydatid cysts were visualized in 9.2% of the sheep and 2.5% of the goats. Of the animals positive on ultrasound, 87.5% received post-mortem examinations. Eighteen (6.9%) sheep and 5 (1.5%) goats were positive for hydatid cysts on ultrasound and post-mortem examination. False-positives were a result of Taenia hydatigena cysticerci present in the liver in all but 1 case. Positive predictive value of ultrasound for diagnosis of hydatidosis in sheep and goats was 82.1%.
A significant proportion of hemodialysis patients have subnormal body temperature. Dialysis against cool dialysate has been frequently shown to reduce the incidence of symptomatic hypotension (SH), although only one of these reports included patient temperatures. Our hypothesis was that the response to cool or normal temperature dialysis could depend on a patient's baseline temperature. Of 128 patients in two hemodialysis units, 28 had a (mean of 5) baseline temperature less than 36 degrees C and 48 patients had a temperature higher than 36.5 degrees C. A crossover study was performed by dialyzing patients for 10 consecutive treatments with the same dialysate temperature, either 37 degrees C or 35 degrees C. All patients combined had a significant reduction in SH with 35 degrees C dialysate, 11.2% versus 5.5% with 37 degrees C dialysate (P = 0.001). The incidence of SH in euthermic patients was not affected by dialysate temperature. Hypothermic patients dialyzed against 37 degrees C dialysate had the highest incidence of SH, which decreased markedly with 35 degrees C dialysate (15.9% v 3.4%; P = 0.0001). There were no differences in age, duration of dialysis, gender, hemoglobin, urea, creatinine, or volume removed per dialysis between the two groups. In conclusion, subnormal temperature is common in dialysis patients but the etiology is unclear. The hemodynamic protective effect of cool dialysate only occurs in patients with subnormal temperatures. Only the subpopulation of patients with SH and low body temperature should be dialyzed against cool dialysate.
The effects and interaction of transforming growth factor-beta (TGF-beta) and insulin on collagen production in human fetal lung fibroblasts was examined. Fibroblasts were labeled with [3H]proline and collagen production was analyzed by polyacrylamide gel electrophoresis. The addition of insulin (2 micrograms/ml) increased collagen production 5 fold and TGF-beta (5 ng/ml) increased collagen production 6-fold. The combination of TGF-beta and insulin further increased type I collagen production (12 fold). We found that TGF-beta increased pro-alpha 1 (I) collagen mRNA levels 2-3 fold, insulin increased mRNA levels by less than 2 fold, and the combination stimulated a 3-4 fold increase. In a nuclear run-on assay, we found a 1.7 fold increase in the rate of transcription for the pro-alpha 1 (I) collagen gene in insulin-treated cultures and a 2-fold increase in TGF-treated cultures. In fibroblasts transfected with a plasmid containing 2.4 kb of the 5' flanking sequences of the human pro-alpha 1 (I) collagen gene, TGF-beta stimulated a 2.8 fold increase in promoter activity. In contrast, the addition of insulin stimulated a small increase (less than 2 fold) in the pro-alpha 1 (I) collagen promoter activity when administered alone or in combination with TGF-beta. Insulin prolonged the half-life of pro-alpha 1 (I) collagen mRNA from 9.1 h to 14.3 h as assessed by treatment with actinomycin D. The insulin-induced increase in pro-alpha 1 (I) collagen mRNA was blocked by the presence of cycloheximide indicating a requirement for new protein synthesis. These results show that the combination of TGF-beta and insulin stimulate large increases in type I collagen formation by acting at different sites in the collagen biosynthetic pathway.
Restless legs syndrome (RLS) and periodic limb movements during sleep (PLMS) are sleep disorders that are common and distressing to uremic patients. There are few data regarding effective treatment in this population. Five chronic hemodialysis patients completed a double-blind, placebo-controlled, crossover study using a single bedtime dose of controlled release L-DOPA/carbidopa (100/25 mg) for treatment of RLS and sleep disruption. Leg movements per hour of sleep and percentage of sleep time accompanied by leg movements were decreased with treatment (101.0 +/- 29.1 events/hour on placebo vs. 61.0 +/- 28.3 events per hour on drug, p = 0.006; and 15.1 +/- 4.9% of sleep time with leg movements on placebo vs. 8.6 +/- 4.0% on drug, p = 0.014). In addition, arousals associated with leg movements (mean 209 +/- 49 events on placebo, mean 108 +/- 46 events on drug) and the leg movement arousal index (mean 59 +/- 23 events/hour on placebo, mean 23 +/- 9 events/hour on drug) were decreased by active medication (p = 0.03 and 0.04, respectively). Patients, however, continued to have very disrupted sleep and we could not document consistent subjective or objective improvement in overall sleep except for an increase in slow-wave sleep (SWS) from 9.0% to 22.8% (p = 0.01). The patterns of movements during sleep were not uniform in different patients, and the movements, although often periodic, were much longer than defined for PLMS. Because of this, finding suitable objective parameters to analyze was problematic. Measuring the percentage of sleep time during which there were leg movements was probably the most efficient and reproducible means of quantitating this disorder. Thus, although controlled-release L-DOPA/carbidopa at a dose of 100/25 mg given once nightly reduced leg movements and increased SWS, sleep continued to be disrupted. Whether higher doses or more frequent dosing is effective requires further investigation.
The effect of PGE2 on TGF-beta receptor binding was assessed for both signaling (type I and type II) and non-signaling (type III) TGF-beta receptors. We found in cross-linking studies that PGE2 treatment (24 h) decreased binding of TGF-beta to the type I and type II receptors by approx. 50% and markedly increased binding of TGF-beta to type III receptors nearly 10-fold. Northern analyses indicated that PGE2 decreased type I receptor mRNA levels by approximately 30-50%, decreased type II receptor mRNA levels by approximately 60%, and markedly increased type III receptor mRNA levels. Coincubation with cycloheximide partially blocked the PGE2-induced inhibition of type II receptor mRNA. In contrast, cycloheximide minimally affected PGE2-induced type III receptor mRNA levels. Activation of protein kinase C by phorbol esters had no apparent effect on TGF-beta receptor mRNA levels. Our data suggest that alterations in TGF-beta receptor expression could modify the response of tissues to TGF-beta during injury.