Biomedical subjects
Publications and source records attributed to A Peterson.
A protocol to recover Bacillus anthracis spores from a steel surface using macrofoam swabs was evaluated for its accuracy, precision, reproducibility, and limit of detection. Macrofoam swabs recovered 31.7 to 49.1% of spores from 10-cm2 steel surfaces with a < or =32.7% coefficient of variation in sampling precision and reproducibility for inocula of > or =38 spores.
Traditional models of hematopoiesis have been hierarchical in nature. Over the past 10 years, we have developed data indicating that hematopoiesis is regulated in a continuum with deterministic and stochastic components. We have shown that the most primitive stem cells, as represented by lineage negative rhodamine(low) Hoechst(low) murine marrow cells are continuously or intermittently cycling as determined by in vivo BrdU labeling. When marrow stem cells are induced to transit cell cycle by in vitro exposure to cytokines, either IL-3, IL-6, IL-11, and steel factor or thrombopoietin, FLT3 ligand, and steel factor, they progress through cycle in a highly synchronized fashion. We have determined that when the stem cells progress through a cytokine stimulated cell cycle the homing, engraftment, adhesion protein, global gene expression, and hematopoietic differentiation phenotypes all change in a reversible fashion. This has led to the continuum model, in which, with cycle transit, chromatin is continually changing altering open transcription areas and providing a continually changing landscape of transcriptional opportunity. More recently, we have extended the changing differentiation profiles to differentiation into lung cells and found that non-hematopoietic differentiation also shows cycle related reversibly modulation. These observations all together support a continuum model of stem cell regulation in which the phenotype of the marrow stem cells is continually and reversibly changing over time.
Neurofilaments are synthesised and assembled in neuronal cell bodies, transported along axons and degraded at the synapse. However, in several pathological situations they aggregate in cell bodies or axons. To investigate their turnover when separated from their normal site of degradation, we used a previously described transgenic model characterised by perikaryal retention of neurofilaments, and compared the basic features of both neurofilament synthesis and degradation with that observed in normal mice. Despite the massive perikaryal aggregates, neurofilament transcript levels were found to be unchanged, whereas the total accumulation of neurofilament proteins was markedly reduced. Neurofilaments isolated from transgenic samples are more sensitive to both trypsin and alpha-chymotrypsin mediated proteolysis. Consistent with their greater in vitro sensitivity, trypsin immunolabeling of cell bodies was stronger in transgenic mice. These results show a novel mechanism to regulate the amount of neurofilaments when they abnormally aggregate.
Although fish embryos have been used in a number of slow-freezing cryopreservation experiments, they have never been successfully cryopreserved. In part this is because little is known about whether ice forms within the embryo during the slow-freezing dehydration process. Therefore, we examined the temperature of intraembryonic ice formation (T(IIF)) and the temperature of extraembryonic ice formation (T(EIF)), using a cryomicroscope. We used both unmodified zebrafish embryos and those with water channels (aquaporin-3 or AQP3) inserted into their membranes to increase permeability to water and cryoprotectants, examined at 100% epiboly to the 6-somite stage. In these experiments we examined: (1) the spontaneous freezing of (external) solutions; (2) the spontaneous freezing of solutions containing embryos; (3) the effect of preloading the embryos with cryoprotectants on T(IIF); (4) whether preloading the embryos with cryoprotectant helps in survival after nucleating events in the solution; and (5) the damaging effects of extracellular nucleation events versus solution toxicity on the embryos. The solutes alone (embryo medium--EM, sucrose culture medium, 1 M propylene glycol in EM, and 1 M propylene glycol in a sucrose culture medium) froze at -14.9 +/- 1.1, -17.0 +/- 0.3, -17.8 +/- 1.0, and -17.7 +/- 1.4, respectively. There was no difference amongst these means (P > 0.05), thus adding cryoprotectant did not significantly lower the nucleation point. Adding embryos (preloaded with cryoprotectant or not) did not change the basic freezing characteristics of these solutes. In all these experiments, (T(EIF)) equaled (T(IIF)), and there was no difference in the freezing point of the solutions with or without the embryos (P > 0.05). Additionally, there was no difference in the freezing characteristics of embryos with and without aquaporins (P > 0.05). The formation of intraembryonic ice was lethal to the zebrafish embryos in all cases. But this lethal outcome was not related to solution injury effects, because 88-98% of embryos survived when exposed to a higher solute concentration with no ice present. Taken together, these data suggest that slow-freezing is not a suitable option for zebrafish embryos. The mechanism of this high temperature nucleation event in zebrafish embryos is still unknown.
In peripheral nerves, large caliber axons are ensheathed by myelin-elaborating Schwann cells. Multiple lines of evidence demonstrate that expression of the genes encoding myelin structural proteins occurs in Schwann cells in response to axonal instructions. To gain further insight into the mechanisms controlling myelin gene expression, we used reporter constructs in transgenic mice to search for the DNA elements that regulate the myelin basic protein (MBP) gene. Through this in vivo investigation, we provide evidence for the participation of multiple, widely distributed, positive and negative elements in the overall control of MBP expression. Notably, all constructs bearing a 0.6 kb far-upstream sequence, designated Schwann cell enhancer 1 (SCE1), expressed at high levels in myelin-forming Schwann cells. In addition, robust targeting activity conferred by SCE1 was shown to be independent of other MBP 5' flanking sequence. These observations suggest that SCE1 will make available a powerful tool to drive transgene expression in myelinating Schwann cells and that a focused analysis of the SCE1 sequence will lead to the identification of transcription factor binding sites that positively regulate MBP expression.
BACKGROUND: This study was designed to test the hypothesis that smokeless tobacco (SLT) serves as a gateway drug for smoking among young adult males. Methods. A cohort (n = 7,865) of U.S. Air Force recruits who claimed to have never smoked cigarettes was followed prospectively for 1 year. The participants were male, 32.9% were ethnic minorities, and their average age was 19.84 years (SD = 2.29). Among recruits entering basic military training, 403 (5.1%) reported current SLT use and 198 (2.5%) reported a past history of SLT use. RESULTS: At the 1-year follow-up current SLT users were 233% more likely to have initiated smoking than nonusers (odds ratio = 2.33, 95% CI = 1.84-2.94). Similarly, recruits who reported past SLT use were 227% more likely to begin smoking than participants who had never used SLT (odds ratio = 2.27, 95% CI = 1.64-3.15). SLT use remained a potent predictor of smoking initiation in a multivariate logistic model that included demographic factors and other risk factors for initiation. CONCLUSIONS: SLT use appears to be an important predictor of smoking initiation among young adult males. This study suggests that smoking prevention and cessation programs should also include strategies related to SLT use.
OBJECTIVES: To review our experiences with some of the preoperative and postoperative findings in five children who were diagnosed with auditory neuropathy and were provided with cochlear implants. We describe changes in auditory function, which enabled these children to have significant improvement in their hearing and communication skills. STUDY DESIGN: Pre- and postoperatively, these children received complete medical examinations at Mayo Clinic, including related consultations in audiology, pediatrics, neurology, medical genetics, otolaryngology, psychology, speech pathology, and radiology. METHODS: These children typically had additional medical and audiological examinations at more than one medical center. The hearing assessments of these children included appropriate behavioral audiometric techniques, objective measures of middle ear function, acoustic reflex studies, transient (TOAE) or distortion product (DPOAE) otoacoustic emissions, auditory brainstem responses (ABR), and, in some cases, transtympanic electrocochleography (ECoG). After placement of the internal cochlear implant devices (Nucleus CI24), intraoperatively we measured electrode impedances, visually detected electrical stapedius reflexes (VESR) and neural response telemetry (NRT). These intraoperative objective measures were used to help program the speech processor for each child. Postoperatively, each child has had regular follow-up to assure complete healing of the surgical incision, to assess their general medical conditions, and for speech processor programming. Their hearing and communication skills have been assessed on a regular basis. Postoperatively, we have also repeated electrode impedance measurements, NRT measurements, otoacoustic emissions, and electrical auditory brainstem responses (EABR). We now have 1 year or more follow-up information on the five children. RESULTS: The five children implanted at Mayo Clinic Rochester have not had any postoperative medical or cochlear implant device complications. All of the children have shown significant improvements in their sound detection, speech perception abilities and communication skills. All of the children have shown evidence of good NRT results. All but case D (who was not tested) showed evidence of good postoperative EABR results. Otoacoustic emissions typically remained in the non-operated ear but, as expected, they are now absent in the operated ear. CONCLUSION: Our experiences with cochlear implantation for children diagnosed with auditory neuropathy have been very positive. The five children we have implanted have not had any complications postoperatively, and each child has shown improved listening and communication skills that have enabled each child to take advantage of different communication and educational options.
Group randomized trials (GRTs) in public health research typically use a small number of randomized groups with a relatively large number of participants per group. Two fundamental features characterize GRTs: a positive correlation of outcomes within a group, and the small number of groups. Appropriate consideration of these fundamental features is essential for design and analysis. This paper presents the fundamental features of GRTs and the importance of considering these features in design and analysis. It also reviews and contrasts the main analytic methods proposed for GRTs, emphasizing the assumptions required to make these methods valid and efficient. Also discussed are various design issues, along with guidelines for choosing among them. A real data example illustrates these issues and methods.
The biologically active form of vitamin D, 1,25-dihydroxyvitamin D3 (1,25-(OH)2D3) has been shown to regulate the proliferation of human prostate epithelial cell lines. Since the insulin-like growth factor (IGF) system is involved in the transformation process of epithelial cells, the following study was undertaken to determine if the IGF system, in particular IGF binding protein-3 (IGFBP-3), is altered by 1,25-(OH)2D3 in normal prostate epithelial cells as part of a mechanism for inhibition of transformation. Two cell systems were used in this study: (1) primary cultures of benign human prostate epithelial cells (PECs) and (2) an SV40-T immortalized prostate epithelial cell line (P153) that is non-tumorigenic. 1,25-(OH)2D3 was added to parallel sets of PECs and P153 cells in addition to the presence or absence of IGF-I or des(1-3)IGF-I. Treatment with 1,25-(OH)2D3 resulted in significant growth inhibition of both PECs and P153 cells. Furthermore, 1,25-(OH)2D3 inhibited IGF-induced proliferation, but this was partially reversed by high concentrations of IGF-I. Western ligand blots of condition media demonstrated a significant increase in IGFBP-3; likewise Northern blots demonstrated an increase in mRNA for IGFBP-3. Proliferation assays using an antibody designed to block the IGF-independent effects of IGFBP-3 failed to reverse the inhibitory effect of 1,25-(OH)2D3. Thus, IGFBP-3 acts in an IGF-dependent manner to inhibit cell growth of benign prostate epithelial cells.
The present investigation examined neuropsychological functioning in 50 male batterers court-ordered into treatment and 23 nonpatient controls. Subjects were administered a neuropsychological screening battery consisting of the Screening Test for the Luria-Nebraska Neuropsychological Battery, the Stroop Color and Word Test, two memory subtests from the Luria-Nebraska Neuropsychological Battery-III (Figural Memory and Delayed Figural Memory), and two subtests from the Halstead-Reitan Neuropsychological Battery (Trails A & B). Subjects were categorized as having neuropsychological dysfunction if their scores exceeded the statistical cut offs on two or more subtests. Results indicated that 24 (48%) of the male batterers exhibited cognitive dysfunction, as compared to only 1 (4.3%) of the nonpatient controls. Inspection of individual neuropsychological measures indicated poorer performance across all subtests for impaired male batterers as compared to both nonimpaired batterers and normal controls. In contrast, no significant differences on any of these measures emerged between nonimpaired male batterers and normal controls. Implications for the appropriate screening and treatment of male batterers are discussed.
This patient displayed a pattern of neuropsychological test results that indicated early Alzheimer's dementia. Using neuropsychological testing to make the diagnosis provided an answer for the patient, family, and psychiatrist, who all recognized a problem, and allowed for future planning. Recommendations for the patient included: psychoeducation and support for the patient and family assistance with planning for the future, including discussions of advance directives and financial management re-evaluation of the patient's ability to drive a motor vehicle a trial of a cholinesterase inhibitor such as donepezil or rivastigmine ongoing health maintenance, including periodic screening for depression and other psychiatric symptoms repeat neuropsychological assessment in 6 months to 1 year to assess rate of decline and confirm the diagnosis.
Transducin-like Enhancer of split (TLE) 1 is a mammalian transcriptional corepressor homologous to Drosophila Groucho. In Drosophila, Groucho acts together with bHLH proteins of the Hairy/Enhancer of split (HES) family to negatively regulate neuronal differentiation. Loss of the functions of Groucho or HES proteins results in supernumerary central and peripheral neurons. This suggests that mammalian TLE/Groucho family members may also be involved in the regulation of neuronal differentiation. Consistent with this possibility, TLE1 is expressed in proliferating neural progenitor cells of the central nervous system, but its expression is transiently down-regulated in newly generated postmitotic neurons. Based on these observations, we investigated whether persistent TLE1 expression in postmitotic neurons would perturb the normal course of neuronal development. Transgenic mice were derived in which the human TLE1 gene is regulated by the promoter of the Talpha1 alpha-tubulin gene, which is exclusively expressed in postmitotic neurons. In these mice, constitutive expression of TLE1 inhibits neuronal development in the embryonic forebrain leading to increased apoptosis and neuronal loss in the ventral and dorsal telencephalon. These results provide the first direct evidence that TLE1 is an important negative regulator of postmitotic neuronal differentiation in the mammalian central nervous system.
A clone of cardenolide 16'-O-glucohydrolase cDNA (CGH I) was obtained from Digitalis lanata which encodes a protein of 642 amino acids (calculated molecular mass 73.2 kDa). The amino acid sequence derived from CGH I showed high homology to a widely distributed family of beta-glucohydrolases (glycosyl hydrolases family 1). The recombinant CGH I protein produced in Escherichia coli had CGH I activity. CGH I mRNA was detected in leaves, flowers, stems and fruits of D. lanata.
BACKGROUND: Short-term clinical studies have indicated the possibility of one-stage surgery and early loading of machined titanium implants. However, long-term data comparing the outcome to the conventional two-stage technique are missing. PURPOSE: A clinical and radiographic study was performed to compare the outcome of oral rehabilitation of the edentulous mandible by fixed suprastructures connected to implants installed according to either (1) a one-stage surgical procedure and early loading (experimental group--EG) or (2) the original two-stage concept (reference group--RG). The EG and RG comprised 16 and 11 subjects, respectively. MATERIALS AND METHODS: The following specific inclusion criterion were adopted: (1) all patients had to consider themselves to be in good general health, (2) the amount of bone had to enable the installation of five to six, at least 10-mm long fixtures (Mk II fixtures; Nobel Biocare AB, Göteborg, Sweden) between the mental foramina, and (3) the patients had to be available for the follow-up and maintenance program. A total of 88 implants were placed in the EG compared to 30 in the RG. In the EG, fixed appliances were connected to the implants within 20 days following implant installation while the fixed appliances in the RG were connected about 4 months following fixture installation. At delivery of the suprastructures, all patients were radiographically examined, an examination that was repeated at the 18- and 60-month follow-ups. RESULTS: The analysis of the radiographs from the EG disclosed that during the observation period, between 18 and 60 months, the mean loss of bone support amounted to 0.2 mm (SD = 0.4). The corresponding value observed in the RG was 0.0 mm (SD = 0.5). During the 60-month observation period, no fixture was lost in any of the two groups examined. The implants under study as well as those in the reference material were at all observation intervals found to be clinically stable. CONCLUSIONS: This clinical study demonstrated that it is, at least based on a 5-year observation period, possible to successfully load via a permanent fixed rigid cross-arch suprastructure titanium dental implants soon after installation. However, such a treatment approach has to be strictly limited to the interforamina area of the edentulous mandible. Furthermore, the bone resorption was found to be within the same range around such implants as around implants installed and loaded according to the original two-stage protocol.
Myelination of peripheral nerve fibres is performed by Schwann cells and is associated with the coordinate upregulation of lipid synthesis and multiple genes encoding myelin-specific proteins. Both the decision to enter into a myelinating phenotype and subsequently, the quantity of myelin that each Schwann cell elaborates appear to be controlled by axonal signals. Understanding of the relevant signaling pathways and the downstream transcription factors and cis elements that confer myelin gene expression is notably limited. In large part, this has resulted directly from a lack of methods for obtaining nuclear extracts from myelinating Schwann cells thus precluding the application of numerous powerful molecular techniques. In this report, we describe a method that overcomes this limitation for the myelinating Schwann cells in the sciatic nerves of the mouse. During the evolution of the method, its effectiveness was monitored using an oligonucleotide containing the binding site for KROX-20, a transcription factor known to be present in myelinating Schwann cells. Following technical development, the optimized protocol has proven to be entirely reliable and thus novel experimental strategies now can be applied to the investigation of the molecular mechanisms controlling gene expression in peripheral nerves.
The development of tissue engineered cartilage is emerging as a potential treatment for the repair of cartilage defects. By seeding chondrocytes onto poly-glycolic acid (PGA) biodegradable scaffolds within a convective-flow bioreactor, the synthesis of tissue-engineered articular cartilage has been recently demonstrated. The ability to cultivate and manipulate this cell-polymer construct to possess specific dimensions, as well as biochemical and biomechanical properties is critical for potential application as an in vivo therapy of damaged articular surfaces. Bioreactor design requirements for stages from research to development to commercialization are discussed. Advantages and limitations to various bioreactor designs are critiqued. These studies illustrate the ability to synthesize tissue-engineered cartilage under convective-flow conditions for potential human tissue repair.
OBJECTIVES: To review our intraoperative and postoperative testing protocol for cochlear implant patients. This study describes the methodologies and applications of a new technique called neural response telemetry (NRT) for the Nucleus CI24M cochlear implant system. NRT uses radiofrequency telemetry technology to measure the action potentials of the auditory nerve. STUDY DESIGN: We have developed a specific protocol for intraoperative testing of the implant device before, during, and after implantation. This includes device integrity tests, visual detection of electrical stapedius muscle reflexes (VESR), and NRT. METHODS: Our methodologies use the commercial software (Windows-based Diagnostic and Programming System [WIN-DPS] and NRT) for the Nucleus CI24M. We describe the details of our protocol used on all of the patients (14 adults and 14 children) who received CI24M implants at Mayo Clinic (Rochester, MN). Our protocol correlates the NRT threshold with the behavioral responses for each patient on at least four electrodes. RESULTS: From August 1, 1998, to December 31,1998, we completed electrode integrity tests, NRT, and VESR testing intraoperatively on 12 patients with the Nucleus CI24M. We have measured normal implant function on all 28 of our CI24M patients with one exception. One of our children had a device failure after approximately 4 months as a result of head trauma. We have also obtained NRT results from an additional 10 patients postoperatively. CONCLUSIONS: The measurement of device and electrode array function is quite simple with the CI24M software. These measurements can be obtained intraoperatively as well as postoperatively. We conclude that VESR and NRT measurements can be very helpful in programming for patients with cochlear implants, especially children, because they provides us with target settings for the speech processor.
The telencephalic vesicles form in the mouse embryo by the expansion of precursor regions in the anterior neural tube. Once the vesicles have formed, discrete dorsal and ventral territories can be recognized that later give rise to cortical and subcortical structures, respectively. To investigate the mechanisms that regulate the expansion and regionalization of the telencephalon, we have carried out a screen to identify recessive mutations that disrupt these events. We isolated a mouse mutant in which an early and critical step in development of the telencephalic vesicles is disrupted. Telencephalic primordia are present in flat-top embryos but they fail to progress to form the telencephalic vesicles. An increased rate of proliferation in the forebrain neurectoderm that accompanies telencephalic expansion in wild-type embryos fails to occur in flat-top embryos. Regionalization events that would normally take place during expansion of the primordia also fail to occur. Thus the phenotype of the flat-top mouse reveals that outgrowth of the telencephalic vesicles and their regionalization are coupled processes.