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Biomedical subjects

M L King

Publications and source records attributed to M L King.

At least 37 records · Page 2Linked to original sources

Xenopus VegT RNA is localized to the vegetal cortex during oogenesis and encodes a novel T-box transcription factor involved in mesodermal patterning.

An RNA localized to the vegetal cortex of Xenopus oocytes encodes a novel T-box protein (VegT) capable of inducing either dorsal or posterior ventral mesoderm at different times in development. VegT is a nuclear protein and its C-terminal domain can activate transcription in a yeast reporter assay, observations consistent with VegT functioning as a transcription factor. Zygotic expression is dynamic along the dorsoventral axis, with transcripts first expressed in the dorsal marginal zone. By the end of gastrulation, VegT is expressed exclusively in posterior ventral and lateral mesoderm and is excluded from the notochord. Later expression is confined to a subset of Rohon-Beard cells, a type of primary sensory neuron. In animal cap assays, VegT is capable of converting prospective ectoderm into ventral lateral mesoderm. Such ectopic expression of VegT induces its own expression as well as that of Xwnt-8 in caps, suggesting that a Wnt pathway may be involved. Mis-expression of VegT in dorsal animal blastomeres fated to contribute to brain suppresses head formation. Our results suggest that VegT is a localized transcription factor, which operates sequentially in several developmental pathways during embryogenesis, including dorsoventral and posterior patterning of mesoderm.

Amino Acid Sequence↗

Localization of Xcat-2 RNA, a putative germ plasm component, to the mitochondrial cloud in Xenopus stage I oocytes.

The mitochondrial cloud is a unique cell structure found in stage I Xenopus oocytes that plays a role in mitochondriogenesis and in the distribution of germ plasm to the vegetal pole. Xcat-2 RNA specifically localizes to the mitochondrial cloud and moves with it to the vegetal subcortex in stage II oocytes. Later, in the 4-cell embryo, it is found in a pattern identical to the germ plasm. Following microinjection into stage I oocytes, synthetic Xcat-2 RNAs localize to the mitochondrial cloud within 22 hours. Transcripts are stable over this time period with very little evidence of degradation. The Xcat-2 3'untranslated region was found to be both required and sufficient for mitochondrial cloud localization. Further deletion analysis narrowed this localization signal to a 250 nucleotide region at the proximal end of the 3'untranslated region. This region is different from, but overlaps with, a domain previously shown to be sufficient to direct Xcat-2 to the vegetal cortex in stage IV oocytes. Examination of early stage I oocytes reveals a time when Xcat-2 is uniformly distributed, arguing against vectorial nuclear export into the mitochondrial cloud. Analysis of localization at different time points does not suggest active transport to the mitochondrial cloud. We postulate that localization occurs by selective entrapment of Xcat-2 within the cloud by localized binding sites.

Animals↗

Adjuvant radiation therapy for rectal carcinoma: predictors of outcome.

PURPOSE: To review predictors of outcome, including sequencing of modalities and pretreatment findings for adjuvantly treated rectal cancer. METHODS AND MATERIALS: From 1975 through 1990, 307 patients with adenocarcinoma of the rectum underwent adjuvant radiation therapy. In 251 cases the radiation therapy was administered preoperatively, either 40-50 Gy (median dose 45 Gy) followed in 6-7 weeks by surgery (210 cases), or 20 Gy in five fractions immediately prior to surgery (41 cases). In 56 cases, patients were referred postoperatively for radiation (median dose 50 Gy). Adjuvant chemotherapy was never given concurrently with the preoperative radiation (RT), although 43 of the cases (including 14 of the preoperative RT cases) received postoperative chemotherapy. RESULTS: Multivariate analysis (Cox model) indicated that significant predictors of better overall freedom from disease were preoperative rather than postoperative RT (p < 0.001), low surgical stage (p < 0.0001), specialist surgeon (p = 0.007), low or moderate histologic grade (p = 0.026), and proximal lesion (p = 0.033). The significant predictors for better local control included use of preoperative RT (p < 0.001), low or moderate grade (p = 0.001), and low surgical stage (p = 0.015). The 5-year local control and freedom from disease for the preoperative RT patients were 90% +/- 2% and 73% +/- 3%, respectively. The selected cases that received the short course of 20 Gy preoperatively did well. Although 24 out of 41 patients proved to have Astler Coller B2 or C disease, local control at last follow-up was 39 out of 41 (95%). A second multivariate analysis of pretreatment factors was performed on the preoperative RT cases. The significant factors for both local control and overall freedom from disease were noncircumferential vs. circumferential tumor, proximal vs. distal lesion, and background of the surgeon. Additional negative factors on univariate analysis (although not achieving independent significance on multivariate analysis) included the finding of near-obstructing lesions and elevated carcinoembryonic antigen (CEA). Grade > or = 3 sequelae occurred in 8% of cases (including 3% bowel obstruction). The only significant factor for complications was background of the surgeon (4% for colorectal specialists vs. 12% for nonspecialists, p = 0.015). CONCLUSIONS: Significant factors for better tumor control included preoperative as opposed to postoperative RT and the experience of the surgeon. In selected cases, excellent results can be obtained with a short course of preoperative radiation. Concurrent chemotherapy need not be given routinely with preoperative radiation. Subgroups of preoperative RT cases at risk for distant metastases (who might benefit from postoperative chemotherapy), and at high risk for local failure (for whom concurrent preoperative chemotherapy and radiation might be considered), are identified.

Adenocarcinoma↗

Celahinine A, a new sesquiterpene pyridine alkaloid from Celastrus hindsii.

A new sesquiterpene pyridine alkaloid, celahinine A [1], and the related known polyester celahin A, as well as the known cytotoxic sesquiterpene pyridine alkaloid emarginatine A [2], were isolated from Celastrus hindsii. The structure of 1 was determined by 2D nmr techniques and was also confirmed by spectral comparison with the related 2.

Antineoplastic Agents, Phytogenic↗

Effect of a tracheostomy speaking valve on secretions, arterial oxygenation, and olfaction: a quantitative evaluation.

Tracheostomy speaking valves consist of a one-way valve that closes upon exhalation, causing a redirection of exhaled gas into the upper airway, thus allowing for the primary benefit of speech. The present study was undertaken to test various hypotheses concerning the secondary benefits of speaking valves. We hypothesized that use of a speaking valve will result in a decrease in accumulated secretions, an increase in arterial oxygenation and an improvement in olfactory function. A total of 8 tracheotomized patients met the following inclusion criteria: age > 18; ability to tolerate wearing a speaking valve for at least 3 hours; no unstable medical conditions; no use of thrombolytic agents. While using the speaking valve patients accumulated fewer secretions (74.3 +/- 63.6 vs. 122.8 +/- 44.6 ml/day, p = 0.004, n = 7) and had improved olfactory function (accuracy = 28.4 +/- 5.2 vs 8.1 +/- 2.9%, p = 0.02; and percent correct = 64.2 +/- 2.6 vs 50.0 +/- 3.9%, p = 0.03, n = 6) than when off the speaking valve. No significant differences were found in 24-hour arterial oxygen saturation (pulse oximetry and ABG analysis respectively, n = 7), arterial PO2, pH, PCO2, HCO3, or 24-hour heart rate (n = 7). Thus, the present study found a significant decrease in secretions and improvements in olfaction when tracheotomized patients wore a speaking valve, but no difference in arterial oxygenation.

Adult↗

Patterns of localization and cytoskeletal association of two vegetally localized RNAs, Vg1 and Xcat-2.

In Xenopus, localization of a rare class of mRNAs during oogenesis is believed to initiate pattern formation in the early embryo. We have determined the pattern of RNA localization for one of these RNAs, Xcat-2, which encodes a putative RNA-binding protein related to Drosophila nanos (Mosquera, L., Forristall, C., Zhou, Y. and King, M. L. (1993) Development 117, 377-386). Xcat-2 is exclusively localized to the mitochondrial cloud in stage I oocytes, moves with this body into the vegetal cortex during stage II and, later, partitions into islands consistent with it being a component of the germ plasm. As previously shown, Vg1 is not localized to the vegetal cortex until stage IV and distributes to all vegetal blastomeres during development. We found a direct correlation between the localized condition of these RNAs and their recovery in a detergent-insoluble fraction. We present evidence suggesting that differential RNA binding to a cytoskeletal component(s) in the vegetal cortex determines the pattern of inheritance for that RNA in the embryo.

Animals↗

A double-blind randomized controlled trial of colchicine in patients with hepatitis B virus-related postnecrotic cirrhosis.

The preliminary results of a prospective double-blind controlled trial of colchicine in 100 patients with hepatitis B virus-related cirrhosis are reported. The patients, 94 males and 6 females, aged 32-80, were assigned to receive either 1 mg of colchicine or an identical placebo orally on a daily basis. The duration of the follow up ranged from 15 to 51 months (median 26 months). Seventy percent had histological proof of cirrhosis. On entry, 80 patients were in Child-Pugh class A, 19 were in class B, and one was in class C. Compared to the placebo group, there was no improvement in the colchicine group after a 24-month follow up in any of the biochemistry data, for example, serum albumin, alkaline phosphatase, alanine and aspartate aminotransferase, bilirubin, and prothrombin time. The difference in the cumulative survival rates at 51 months did not reach statistical significance (p = 0.8) in either group. There was no histological improvement in 30 patients who were willing to undergo repeated liver biopsies. No trend toward improvement of the hepatic pressure gradient was observed in these patients. The serum levels of aminopropeptide of type III procollagen increased significantly in patients in both groups after 24 months of therapy (1.07 +/- 0.06 vs. 1.36 +/- 0.06 U/ml in the colchicine group, 0.93 +/- 0.09 vs. 1.25 +/- 0.07 U/ml in the placebo group; p < 0.05). In addition, neither the clinical deterioration of cirrhosis nor death was prevented in patients receiving colchicine therapy. This report indicates that colchicine has no effect in the treatment of HBV-related postnecrotic cirrhosis.

Adult↗

Two new macrolide sesquiterpene pyridine alkaloids from Maytenus emarginata: emarginatine G and the cytotoxic emarginatine F.

Two new macrolide sesquiterpene pyridine alkaloids, emarginatine F [1] and emarginatine G [2], were isolated from Maytenus emarginata. The structural determinations of 1 and 2 by 2D nmr techniques and spectral comparison with a related compound, emarginatine A [3], are discussed. Biological evaluation showed that emarginatine F [1] demonstrated strong cytotoxicity against human epidermoid carcinoma of the nasopharynx (KB), ileocecal adenocarcinoma (HCT-8), melanoma (RPMI-7951) and medulloblastoma (TE-671) tumor cells, and against murine leukemia (P-388).

Animals↗

Exertional hypotension in spinal cord injury.

STUDY OBJECTIVE: To assess blood pressure (BP) response to continuous maximal arm ergometry in patients with spinal cord injury (SCI). DESIGN: Cross-sectional analysis of data collected for a prospective study of functional electrical stimulation in patients with SCI. SETTING: Short-term rehabilitation hospital. PARTICIPANTS: Twenty individuals with SCI; 4 cervical (C6 to C8), 10 high thoracic (T1 to T6), and 6 low thoracic (T7 to T12). MEASUREMENTS AND RESULTS: Each subject performed continuous maximal arm ergometry with expired gas analysis. Blood pressure was measured using a technician-assisted protocol. The BP at maximal exercise was compared with the highest submaximal BP reached during the test (delta BP = final BP minus highest submaximal BP). All 20 subjects had a negative delta BP (mean +/- SD; -22.8 +/- 12.1 mm Hg) for mean BP and 19 of 20 had a negative delta BP (-25.8 +/- 14.4 mm Hg) for systolic BP. The delta BP was not significantly related to maximum exercise parameters, resting BP, or level of lesion. Four able-bodied subjects and six wheelchair-bound individuals without SCI showed no exertional hypotension. Repeated testing on the four able-bodied subjects showed excellent reproducibility for mean BP (coefficient of variation [CV] = 3.6 percent; r = 0.98; p < 0.01) and systolic BP (CV = 2.2 percent; r = 0.99; p < 0.01) using this protocol. CONCLUSIONS: These data describe, for the first time to our knowledge, that exertional hypotension is present in all individuals with SCI during continuous arm ergometry. Further studies are needed to clarify the mechanisms responsible for this phenomenon and to evaluate the long-term consequences for individuals with SCI.

Adult↗

Application of physiotherapy outcome measures to the managed care model.

Managed Care (MC) is a multidisciplinary model for health care delivery that organizes and sequences the caregiving process. Its objectives include: 1) to reduce length of stay and resource consumption, and 2) to measure, maintain or improve patient outcomes related to care received. Our tertiary care facility is the first Canadian hospital to implement MC. Patient care is directed through the use of a Care Map. Each map is specific to a pathological state and its treatment, i.e. Total Knee Replacement (TKR), and consists of a Patient Problem List, with related patient-centred outcomes, and a Critical Path. The Critical Path outlines the temporal sequence of the provision of care. Most key events on a Care Map are determined anecdotally. The purpose of this project was to collect outcome information in patients assigned to the Total Knee Replacement Care Map in an attempt to validate the existing Care Map or make recommendations for revisions. Inter-rater and intra-rater reliability of knee range of motion-was calculated using the Intra Class Correlation Coefficient (ICC). ICC values ranged from .64-.97. Seventeen patients were assessed. All patients were measured on Day 6 and 8 of the Care Map. This process has resulted in validation of certain range estimates and recommendations for revision of others.

Aged↗

Isolated vegetal cortex from Xenopus oocytes selectively retains localized mRNAs.

The process of converting an egg to a multicellular organism often begins with a polarization of information within the oocyte. The cytoskeleton plays an important role in maintaining such asymmetries. Here we describe the isolation and characterization of cortices, rich in cytoskeletal elements, from the animal and vegetal poles of stage VI Xenopus oocytes. Ultrastructural and immunocytochemical analyses revealed that the 10-microns-thick isolates included a cytokeratin network, mitochondria, and other elements, found in the oocyte cortex. Furthermore, three vegetally localized mRNAs, Vg1, Xcat-2, and Xcat-3, were all found to be specifically retained within the isolated vegetal cortices, in sharp contrast to histone RNA. Our findings support the view that the vegetal cortex represents a unique cytoskeletal domain in which a rare class of maternal mRNAs becomes localized.

Animals↗

A mRNA localized to the vegetal cortex of Xenopus oocytes encodes a protein with a nanos-like zinc finger domain.

mRNAs concentrated in specific regions of the oocyte have been found to encode determinants that specify cell fate. We show that an intermediate filament fraction isolated from Xenopus stage VI oocytes specifically contains, in addition to Vg1 RNA, a new localized mRNA, Xcat-2. Like Vg1, Xcat-2 is found in the vegetal cortical region, is inherited by the vegetal blasomeres during development, and is degraded very early in development. Sequence analysis suggests that Xcat-2 encodes a protein that belongs to the CCHC RNA-binding family of zinc finger proteins. Interestingly, the closest known relative to Xcat-2 in this family is nanos, an RNA localized to the posterior pole of the Drosophila oocyte whose protein product suppresses the translation of the transcription factor hunchback. The localized and maternally restricted expression of Xcat-2 RNA suggests a role for its protein in setting up regional differences in gene expression that occur early in development.

Amino Acid Sequence↗

Determination of fenoverine, a modulator of smooth muscle motility, in capsules and in human plasma: application to dosage form stability and a pilot study in humans.

Fenoverine is a novel, potent, musculotropic, spasmolytic agent that affects primarily the gastrointestinal tract, bile duct, and female genital organs. A simple, specific, and accurate HPLC method was developed for the determination of fenoverine in capsules and plasma. This method has been successfully applied to stability studies of fenoverine capsules and to a pilot study in a normal, healthy volunteer following oral administration of fenoverine. For the determination of fenoverine in capsules, a Nucleosil 5-micron CN column, with acetonitrile:0.1 M ammonium acetate (60:40) as mobile phase and detection at 254 nm, was employed. The mean correlation coefficient of the calibration curve (n = 6) for the assay was 0.9999 over a concentration range of 24.6 to 147.6 micrograms/mL of fenoverine standard solutions. Fenoverine did not decompose significantly at 4, 45, 55, and 65 degrees C for 3 months. The mean correlation coefficients of within-day and between-day calibration curves were 0.9995 and 0.9999, respectively, over a range of 10 to 1000 ng/mL of fenoverine in plasma. The limit of detection was 10 ng in plasma.

Administration, Oral↗

Exertional hypotension in thoracic spinal cord injury: case report.

Exertional hypotension is well described in quadraplegics, but there are few descriptions of this hemodynamic response in paraplegics or of treatment modalities to correct this condition. We describe a patient with a complete T3-4 spinal cord lesion who repeatedly demonstrated symptomatic hypotension with wheelchair sports and arm ergometry. We used gas exchange analysis and exercise echocardiography to delineate the mechanism for hypotension. These results enabled us to develop a simple treatment plan consisting of abdominal binding and elastic stockings to avoid recurrent symptoms.

Adult↗

Case management.

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Humans↗

Potential liability in utilization review: the risk grows.

The use of cost containment measures in utilization review is a continuing topic of concern to physicians and payors. Focusing on a recent California case that dealt with this issue, this article discusses the potential liability of physicians and payors when such measures are used.

Cost Control↗

Developmental expression of regionally specific cell surface antigens in the Xenopus gastrula.

Molecular markers for specific cell lineages would be useful in studies of cellular differentiation. To isolate such markers monoclonal antibodies (MoABs) were raised against plasma membranes isolated from gastrulating Xenopus embryos. Those antibodies that recognized subsets of cells within the embryo were selected by indirect immunofluorescence. The analysis of eight such MoAbs is presented. Western blot analysis showed that all but one MoAb recognized a complex pattern of glycoconjugates associated with glycoproteins. All the antigens recognized by the MoAbs were maternal in origin and displayed similar spatial patterns of pregastrular expression. This pattern of immunoreactivity at the apical surface was inherited passively during cleavage by the resulting superficial blastomeres suggesting that ectodermal specific markers of maternal origin are pre-localized to the cortical ooplasm in mature oocytes. We suggest that these maternal components may be specific glycosyl transferases. Three different patterns of expression were observed during gastrulation as exemplified by MoAbs 1F10C1, 3A4D1, and 6F10B6. MoAb 6F10B6 was specific for both neural and non-neural epithelium. MoAb 3A4D1 was specific for non-neural epidermis. MoAb 1F10C1 appeared to recognize a protein epitope on an extracellular component expressed by the superficial and involuting epithelial cells. The pattern of expression for the 1F10C1 antigen suggests that it may play a role in facilitating the movement of the involuting cells during gastrulation.

Animals↗