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Biomedical subjects

W Harris

Publications and source records attributed to W Harris.

At least 19 recordsLinked to original sources
PubMed2001

Does the preoperative radiographic degree of osteoarthritis correlate to results in primary total knee arthroplasty?

The preoperative radiographs of 1,888 patients undergoing 2,759 consecutive total knee arthroplasties (TKAs) performed for osteoarthritis (OA) were retrospectively reviewed to test the hypothesis that patients with less severe OA changes experience less pain relief and lower function scores after TKA. Age at surgery averaged 70.6 years, and follow-up averaged 2.5 years. Preoperatively, pain was independent of an arthritis grade, and this likewise was noted at > or =3 years after surgery. Patients with severe and mild radiographic changes of OA eventually experienced the same degree of function and pain relief after TKA.

Adult↗
PubMed2001

Comparison of two commercial microimmunofluorescence kits and an enzyme immunoassay kit for detection of serum immunoglobulin G antibodies to Chlamydia pneumoniae.

We compared the MRL and the Labsystems Chlamydia pneumoniae microimmunofluorescence (MIF) immunoglobulin G (IgG) kits and the Labsystems enzyme immunoassay (EIA) kit in a blinded study of 83 serum samples in which we evaluated titers, cross-reactivity to other species, and reproducibility. There was no statistically significant difference between the MRL and the Labsystems MIF kits in the endpoint titers of IgG antibody to C. pneumoniae. The correlation between the results obtained with these two MIF kits was excellent (r = 0.95; P = 0.001). The cross-reactivity of the C. pneumoniae-positive sera with C. trachomatis- and C. psittaci-positive sera was assessed for each MIF kit. For C. pneumoniae-positive sera with titers of > or =32, the Labsystems MIF kit exhibited more cross-reactivity to C. psittaci than the MRL kit did. The values obtained with the Labsystems EIA kit represented single dilutions of serum specimens expressed as enzymeimmuno units on a continuous scale. The results obtained with the Labsystems EIA kit correlated moderately well with those obtained with each MIF kit when they were compared for their abilities to detect IgG antibodies to C. pneumoniae (for the MRL MIF kit, r = 0.79 [P = 0.001]; for the Labsystems MIF kit, r = 0.78 [P = 0.001]). The results obtained with the commercial MRL and Labsystems MIF kits and the Labsystems EIA kit tested were reproducible; and the kits were standardized, had quality control reagents, and are suitable for detection of C. pneumoniae antibodies in serum and for use in interlaboratory studies. Validation of the use of these kits for clinical diagnosis still needs further evaluation.

Antigens, Bacterial↗
PubMed2001

Threshold-level repetitive transcranial electrical stimulation for intraoperative monitoring of central motor conduction.

OBJECT: The authors conducted a study to evaluate repetitive transcranial electrical stimulation (TES) to assess spinal cord motor tract function in individuals undergoing spine surgery, with emphasis on safety and efficacy. METHODS: Somatosensory evoked potentials (SSEPs) were elicited using standard technique. Muscle electromyographic values were measured in response to a three- or four-pulse train of stimulation delivered to the motor cortex via subdermal electrodes. They also evaluated whether changes in the minimum stimulus intensity (that is, threshold level) needed to elicit a response from a given muscle predict motor status immediately postoperatively, as well as whether changes in SSEP response amplitude and latency predict sensory status immediately postoperatively. Anesthesia was routinely induced with intravenous propofol and remifentanil, supplemented with inhaled nitrous oxide. Use of neuromuscular block was avoided after intubation. Satisfactory monitoring of muscle response to threshold-level repetitive TES was achieved in all but nine of the 194 patients studied. In contrast, cortical SSEP responses could not be elicited in 42 of 194 individuals. In cases in which responses were present, TES-based evoked responses proved to be extremely accurate for predicting postoperative motor status. Somatosensory evoked potential monitoring was nearly as accurate for predicting postoperative sensory status. There were frequent instances of postoperative motor or sensory deficit that were not predicted by SSEP- and TES-based monitoring, respectively. There were no adverse events attributable to TES-based monitoring, although since this study ended we have had a single adverse event attributable to threshold-level repetitive TES. CONCLUSIONS: Intraoperative threshold-level repetitive TES-based monitoring of central motor conduction has proven to be a simple, safe, and highly accurate technique for the prevention or minimization of inadvertent motor deficit during surgery involving the spine or spinal cord.

Adolescent↗
PubMed2000

Electromyography during stereotactic pallidotomy for Parkinson's disease.

In stereotactic pallidotomy for Parkinson's disease, care must be taken to avoid internal capsule injury while maximizing improvement of rigidity and tremor. In 21 patients, intraoperative electromyography (EMG) was used to assess stimulation thresholds required for capsular responses and to monitor muscle tone and tremor. Surface EMG electrodes were placed on the face and multiple muscle groups of the extremities. The stimulation and lesion electrode was introduced via MRI-guided stereotaxis toward a point 2-3 mm anterior to the midcommissural point, 5-6 mm inferior to the AC-PC plane, and 21-22 mm lateral to the midline. Exact targets were modified according to MRI-visualized anatomy. With stimulation at 5 and 50 Hz, thresholds for detection of EMG responses were usually seen at 4-5 mA. EMG responses were consistently seen prior to visual observation of muscle activity. Timing of EMG response relative to stimulus aided in differentiating stimulus-related movement from spontaneous tremor. Resting spontaneous EMG activity was seen to decrease as rigidity was improved by incremental lesion production. EMG activity related to tremor was recorded; tremor decrease by lesion production was documented by EMG recording. Patient cooperation with physiologic testing during stimulation and lesion production may become limited. Intraoperative EMG monitoring provides an adjunct to improve reliability of assessment of capsular stimulation and rigidity while providing documentation of lesion impact on rigidity and tremor.

Aged↗
PubMed1999

Comparative genomic hybridization of breast tumors stratified by histological grade reveals new insights into the biological progression of breast cancer.

How does breast cancer progress? There is evidence both to support (S. W. Duffy et al., Br. J. Cancer, 64: 1133-1138, 1991; R. Rajakariar et al., Br. J. Cancer, 71: 150-154, 1995) and refute (M. Hakama et al., Lancet, 345: 221-224, 1995; R. R. Millis et al., Eur. J. Cancer, 34: 548-553, 1998) the hypothesis of dedifferentiation; the theory that as breast cancers grow they evolve from well differentiated (grade I) to poorly differentiated (grade III) tumors. We provide evidence to support the view that the majority of grade I tumors do not progress to grade III tumors. Comparative genomic hybridization was used to screen entire genomes of a large sample (40 grade I and 50 grade III) of invasive ductal breast carcinomas, stratified by grade. We found distinct genetic differences between grade I and grade III tumors. Significantly, we found that 65% of grade I tumors lost the long arm of chromosome 16 compared with only 16% of grade III tumors. This pattern of loss leads us to conclude that the majority of grade I tumors do not progress to grade III tumors. These findings have important implications because they suggest that different breast tumor grades may have distinct molecular origins, pathogenesis, and behavior and, therefore, potentially present distinct molecular targets for research and treatment.

Breast Neoplasms↗
PubMed1999

Imaging protein kinase Calpha activation in cells.

Spatially resolved fluorescence resonance energy transfer (FRET) measured by fluorescence lifetime imaging microscopy (FLIM), provides a method for tracing the catalytic activity of fluorescently tagged proteins inside live cell cultures and enables determination of the functional state of proteins in fixed cells and tissues. Here, a dynamic marker of protein kinase Calpha (PKCalpha) activation is identified and exploited. Activation of PKCalpha is detected through the binding of fluorescently tagged phosphorylation site-specific antibodies; the consequent FRET is measured through the donor fluorophore on PKCalpha by FLIM. This approach enabled the imaging of PKCalpha activation in live and fixed cultured cells and was also applied to pathological samples.

3T3 Cells↗
PubMed1999

'Loop' domain deletional mutant of Bcl-xL is as effective as p29Bcl-xL in inhibiting radiation-induced cytosolic accumulation of cytochrome c (cyt c), caspase-3 activity, and apoptosis.

PURPOSE/OBJECTIVE: To investigate the effect of the enforced expression of p29Bcl-xL or its loop deletional mutant, p18Bcl-xLdelta, on irradiation-induced apoptosis and cell-cycle distribution of HL-60 cells. MATERIALS & METHODS: We compared the irradiation-induced molecular cascade of apoptosis in control human AML HL-60/neo versus Bcl-xL overexpressing (approximately 8-fold) (HL-60/Bcl-xL) and HL-60/Bcl-XLdelta cells that express the loop domain deletional mutant construct (delta26-83 AA) of Bcl-xL. The three cell lines were irradiated with 6MV photons to varying doses up to 20 Gy. Following this, cytosolic cyt c levels, caspase-3 activity, and the Bcl-2 family of proteins were evaluated utilizing Western blot analysis (whole cell lysate or cytosolic S-100 fraction). Apoptosis was assessed by internucleosomal DNA fragmentation, Annexin-V staining and FACS analysis, as well as by morphologic criteria. The cell-cycle effects of radiation were analyzed by flow cytometry. RESULTS: Eight hours following irradiation (12 Gy) of HL-60/neo cells, a marked increase (approximately 8-fold) in the cytosolic accumulation of cyt c in the S-100 fraction was observed. This was associated with the cleavage of caspase-3, as well as the generation of its poly (ADP-ribose) polymerase (PARP) and DFF (DNA fragmentation factor)-45 cleavage activity. Twenty-four to forty-eight hours after irradiation, internucleosomal DNA fragmentation and positive Annexin-V staining (32.3+/-3.3%) was detected in HL-60/neo cells. In contrast, in both HL-60/Bcl-xL and HL-60/Bcl-xLdelta cells, a significantly lower percentage of apoptotic cells (p<0.05) were detected and internucleosomal DNA fragmentation was not induced. Following irradiation, Western analysis neither demonstrated any significant alteration in Bcl-2, p29Bcl-xL, p18Bcl-xLdelta, or Bax; nor induced CD95 (Fas receptor) or Fas ligand expression in any cell type. However, in all cell types, irradiation produced approximately a 2-fold increase in the percentage of cells in the G2/M phase of the cell cycle. CONCLUSION: These results demonstrate that an intact loop domain is not necessary for the full antiapoptotic function of Bcl-xL against irradiation-induced cytosolic accumulation of cyt c, caspase activation, and apoptosis of HL-60 cells. Additionally, the cell-cycle effects of ionizing radiation in HL-60 cells are not affected by enforced expression of Bcl-xL or Bcl-xLdelta.

Apoptosis↗
PubMed1999

The bisindolylmaleimide protein kinase C inhibitor, Ro 32-2241, reverses multidrug resistance in KB tumour cells.

Ro 32-2241 is a bisindolylmaleimide that selectively inhibits protein kinase C (PKC) as compared with other protein kinases. Experiments were carried out to examine its potential as a multidrug resistance-reversing agent. Ro 32-2241 inhibited efflux, and increased accumulation, of [3H]-daunomycin in multidrug-resistant (MDR) KB-8-5 and KB-8-5-11 cells and had no effect on drug-sensitive KB-3-1 cells. Ro 32-2241 completely reversed the doxorubicin resistance of KB-8-5 and KB-8-5-11 cells, showing no effect on the sensitivity of drug-sensitive KB-3-1 cells. The potency of Ro 32-2241 was comparable with that of cyclosporin A and better than that of verapamil, known modulators of multidrug resistance. Ro 32-2241 also completely reversed the taxol resistance of KB-8-5 cells and partially reversed the resistance of KB-8-5-11 cells. Vinblastine resistance was also partially reversed. Mechanistic experiments were carried out to determine whether Ro 32-2241 interacted with P-glycoprotein (Pgp) directly. Increased efflux of [14C]-Ro 32-2241 was seen with the more resistant KB-8-5-11 cells (although the percentage effluxed was very low as compared with [3H]-daunomycin), suggesting that Ro 32-2241 can act as a substrate for Pgp. Direct interaction of Ro 32-2241 with Pgp was confirmed by demonstration that it inhibited binding of [3H]-azidopine to Pgp in KB-8-5-11 membranes. In conclusion, Ro 32-2241, acting directly on Pgp (rather than, or in addition to, an effect on PKC), is effective in reducing or reversing resistance to doxorubicin, taxol and vinblastine in human tumour cells with a clinically relevant degree of MDR. However, results of in vivo experiments conducted to investigate the effects of Ro 32-2241 on resistance to doxorubicin suggest that it may not be possible to achieve sufficiently high levels of Ro 32-2241 in vivo to modulate MDR.

ATP Binding Cassette Transporter, Subfamily B, Mem↗
PubMed1998

The role of GSK3beta in regulating neuronal differentiation in Xenopus laevis.

The serine threonine protein kinase encoded by the shaggy locus has been implicated in neurogenesis in Drosophila. In vertebrates, the shaggy homolog, GSK3beta, is involved in early pattern formation, specifically in setting up the dorsal ventral axis. In the present study we have cloned the Xenopus homolog of the shaggy kinase and show (1) that GSK3beta is expressed in the right time and place to play a role in primary neurogenesis in Xenopus; (2) that overexpression of wild-type GSK3beta leads to a decrease in the number of primary neurons; (3) that inhibition of endogenous GSK3beta activity with overexpression of a dominant negative GSK3beta construct leads to an increase in the number of primary neurons; and (4) that GSK3beta inhibits the ability of neurogenin and NeuroD to produce ectopic tubulin expression, but does not inhibit the ability of neurogenin to produce ectopic NeuroD. On the basis of these data we propose that GSK3beta inhibits the function of NeuroD and therefore prevents neuronal differentiation at a relatively late stage in the developmental pathway.

Amino Acid Substitution↗
PubMed1998

Recurrent mixed germ cell-sex cord-stromal tumor of the ovary in an adult.

A 32-year-old woman underwent laparotomy and right salpingo-oophorectomy for a right adnexal mass. Examination of the specimen revealed a 77 g well-encapsulated tumor that was diagnosed as a mixed germ cell-sex cord-stromal tumor with annular tubules. Sixty-seven months after an uneventful recovery, a large recurrence was treated by total abdominal hysterectomy, left salpingo-oophorectomy, removal of peritoneal implants, lymphadenectomy, omentectomy, and chemotherapy. Histologically, the neoplasm was identical to the one found in the right ovary. To our knowledge, no similar case has been previously reported.

Adult↗
PubMed1998

"Threshold-level" multipulse transcranial electrical stimulation of motor cortex for intraoperative monitoring of spinal motor tracts: description of method and comparison to somatosensory evoked potential monitoring.

UNLABELLED: Numerous methods have been pursued to evaluate function in central motor pathways during surgery in the anesthetized patient. At this time, no standard has emerged, possibly because each of the methods described to date requires some degree of compromise and/or lacks sensitivity. OBJECT: The goal of this study was to develop and evaluate a protocol for intraoperative monitoring of spinal motor conduction that: 1) is safe; 2) is sensitive and specific to motor pathways; 3) provides immediate feedback; 4) is compatible with anesthesia requirements; 5) allows monitoring of spontaneous and/or nerve root stimulus-evoked electromyography; 6) requires little or no involvement of the surgical team; and 7) requires limited equipment beyond that routinely used for somatosensory evoked potential (SSEP) monitoring. Using a multipulse electrical stimulator designed for transcranial applications, the authors have developed a protocol that they term "threshold-level" multipulse transcranial electrical stimulation (TES). METHODS: Patients considered at high risk for postoperative deficit were studied. After anesthesia had been induced and the patient positioned, but prior to incision, "baseline" measures of SSEPs were obtained as well as the minimum (that is, threshold-level) TES voltage needed to evoke a motor response from each of the muscles being monitored. A brief, high-frequency pulse train (three pulses; 2-msec interpulse interval) was used for TES in all cases. Data (latency and amplitude for SSEP; threshold voltage for TES) were collected at different times throughout the surgical procedure. Postoperative neurological status, as judged by evaluation of sensory and motor status, was compared with intraoperative SSEP and TES findings for determination of the sensitivity and specificity of each electrophysiological monitoring technique. Of the 34 patients enrolled, 32 demonstrated TES-evoked responses in muscles innervated at levels caudal to the lesion when examined after anesthesia induction and positioning but prior to incision (that is, baseline). In contrast, baseline SSEPs could be resolved in only 25 of the 34 patients. During surgery, significant changes in SSEP waveforms were noted in 12 of these 25 patients, and 10 patients demonstrated changes in TES thresholds. Fifteen patients experienced varying degrees and durations of postoperative neurological deficit. Intraoperative changes in TES thresholds accurately predicted each instance of postoperative motor weakness without error, but failed to predict four instances of postoperative sensory deficit. Intraoperative SSEP monitoring was not 100% accurate in predicting postoperative sensory status and failed to predict five instances of postoperative motor deficit. As a result of intraoperative TES findings, the surgical plan was altered or otherwise influenced in six patients (roughly 15% of the sample population), possibly limiting the extent of postoperative motor deficit experienced by these patients. CONCLUSIONS: This novel method for intraoperative monitoring of spinal motor conduction appears to meet all of the goals outlined above. Although the risk of postoperative motor deficit is relatively low for the majority of spine surgeries (for example, a simple disc), high-risk procedures, such as tumor resection, correction of vascular abnormalities, and correction of major deformities, should benefit from the virtually immediate and accurate knowledge of spinal motor conduction provided by this new monitoring approach.

Adolescent↗
PubMed1997

Humanization of a mouse monoclonal antibody that blocks the epidermal growth factor receptor: recovery of antagonistic activity.

BACKGROUND: Antibody humanization by transplanting the complementarity determining regions (CDRs) of a murine antibody to a human framework aims to reduce the response of the human immune system against a foreign molecule. Frequently, however, some murine amino acids from the framework have to be retained to recover binding affinity. OBJECTIVES: To redesign R3, a mouse monoclonal antibody (mAb) that binds the human epidermal growth factor (EGF)-receptor and inhibits the binding of EGF, to be a human IgG1. STUDY DESIGN: The light and heavy chains of REI and Eu, respectively, were selected as human immunoglobulin (Ig) frameworks for CDR-grafting based on their high homology with the corresponding sequences of murine R3. Molecular modeling was used to analyze the possible effects of mutating murine residues that underlie the CDRs. RESULTS: CDR-grafting dramatically reduced the binding capability of the antibody. Molecular modeling suggested that two amino acids (Thr 76 and Thr 93), among five immunoglobulin heavy chain variable region (VH) residues underlying the CDRs, were critical for antigen binding. The five residues were mutated back to the original murine amino acids in different combinations contained in six variants of humanized antibodies. In agreement with molecular modeling analysis. The variant in which three murine residues were retained (Ser 75, Thr 76 and Thr 93) exhibited a similar capacity to inhibit the binding of 125I-labeled EGF to its receptor as compared with the original antibody. This humanized antibody was at least 2-fold less immunogenic in African Green monkeys than the chimeric antibody. CONCLUSIONS: Only very few mutations in the frameworks may be necessary to recover the binding capability of a humanized antibody. Molecular modeling can serve as a powerful tool to identify residues critical for binding.

Amino Acid Sequence↗
PubMed1996

Management of limb length inequality during total hip replacement.

Significant limb length inequality is not an uncommon problem after total hip replacement. Preoperative measurement of limb length inequality, preoperative planning with radiographic templates, and intraoperative correction with measurements of limb lengths before and after the insertion of the trial components using special calipers can reduce the incidence and magnitude of this problem. A review of 85 consecutive patients who had primary total hip arthroplasty in which these techniques were used by a single surgeon, showed that 43 had limb inequality preoperatively ranging from 0.5 to 7.25 cm, but only 14 (16%) had limb length inequality after surgery. Eleven limbs (13%) had been lengthened 0.5 to 1 cm compared with the contralateral limb. Of the 42 patients with equal limb lengths preoperatively, 3 had a lengthened limb postoperatively compared with their contralateral limb. Four patients were using lifts on the same side because the limb was too short, and 2 were using lifts on the other side because the limb was too long. None of the other patients complained about limb length inequality. The techniques described above are helpful in minimizing limb length inequality during total hip replacements.

Hip Prosthesis↗
PubMed1995

Anti-CD3-induced changes in protein kinase C isozymes expression in human CD4+ and CD8+ T lymphocytes.

In order to determine whether there is a differential expression and activation of PKC isozymes between CD4+ and CD8+ T cells, peripheral blood mononuclear cells were stimulated with anti-CD3 monoclonal antibody (moAb) for various time intervals and the expression of calcium-dependent PKC isozymes (alpha, beta, gamma) and calcium-independent PKC isozymes (delta, epsilon, zeta) was analyzed with dual color flow cytometry, using anti-PKC isozyme antibodies and anti-CD4 or anti-CD8 antibodies. The basal fluorescence intensity of all PKC isozymes was comparable between CD4+ T cells and CD8+ T cells. Following activation with anti-CD3 moAb a marked increase in the fluorescence intensity of all PKC isozymes in both CD4+ and CD8+ T cells, albeit to a different extent and with different kinetics was observed. Among all PKC isozymes studied, the least striking changes were observed in PKC zeta isozyme and the most striking changes were observed in PKC-epsilon isozyme. Laser-based confocal microscopic studies confirmed that the increase in fluorescence intensity of PKC isozymes following anti-CD3 moAb stimulation, as measured by flow cytometry was accompanied by the translocation of PKC isozymes from cytosol to the plasma membrane. This study demonstrates a differential effect of anti-CD3 moAb on the expression of PKC isozymes between CD4+ and CD8+ T cells and suggests that flow cytometry can be used to study the translocation of PKC isozymes from cytosol to the plasma membrane.

Antibodies, Monoclonal↗
PubMed1995

Tissue response to particulate polymethylmethacrylate in mice with various immune deficiencies.

We examined the tissue response to subcutaneous injections of particulate polymethylmethacrylate powder in fully immunocompetent C3Hf/Sed mice as well as three strains of mice with different levels of lymphocyte dysfunction. Five weeks after the injection, we found clearly demarcated granulomas. Histological and immunohistochemical studies showed that these granulomas were similar among all strains, with either paucity or absence of lymphoid cells. In situ hybridization with use of complementary RNA probes indicated that macrophages were synthesizing interleukin-1 beta messenger RNA (mRNA), a marker of macrophage activation, and a cytokine implicated in pathological bone resorption. We concluded that, in mice, there is a lymphocyte-independent pathway of macrophage activation in response to particulate polymethylmethacrylate. This suggests that the foreign-body response to particulate orthopaedic biomaterials is macrophage-initiated and maintained and that lymphocytes are not essential to this response, although they may modulate it.

Analysis of Variance↗