Prostate brachytherapy seed migration to the right ventricle found at autopsy following acute cardiac dysrhythmia.
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Biomedical subjects
Publications and source records attributed to B F King.
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The pharmacological activity of diadenosine polyphosphates was investigated at three recombinant P2X receptors (rat P2X1, rat P2X3, rat P2X4) expressed in Xenopus oocytes and studied under voltage-clamp conditions. For the rat P2X1 receptor, only P1,P6-diadenosine hexaphosphate (Ap6A) was a full agonist yet 2-3 folds less potent than ATP. At rat P2X3, P1,p4-diadenosine tetraphosphate (Ap4A), P1,P5-diadenosine pentaphosphate (Ap5A) and Ap6A were full agonists and more potent than ATP. Ap4A alone was equipotent with ATP at rat P2X4, but only as a partial agonist. Compared to known data for rat P2X2 and human P2X1 receptors, our findings contrast with rat P2X2 where only Ap4A is a full agonist although four folds less potent than ATP. At rat and human orthologues of P2X1, Ap5A was a partial agonist with similar potency. These data provide a useful basis for selective agonists of P2X receptor subunits.
We have cloned and functionally characterised the mouse orthologue of the P2X4 receptor, mP2X4, and a splice variant of this receptor, mP2X4a. mP2X4 is 388 amino acids in length and shares 94% and 87% identity with the rat and human P2X4 receptors, respectively, while mP2X4a is 361 amino acids in length and lacks a 27-amino acid region in the extracellular domain corresponding to exon 6 of the known P2X receptor gene structures. When expressed in Xenopus laevis oocytes, mP2X4 produces a rapid inward current in response to ATP with an EC50 of 1.68+/-0.2 microM, consistent with the affinity of the rat and human P2X4 receptors for ATP. This agonist response is potentiated by the P2X receptor antagonists suramin, Reactive blue 2 and, over a limited concentration range, by PPADS. Although mP2X4a forms a poorly functional homomeric receptor, it appears able to interact with the full-length mP2X4 subunit to result in a functional channel with a reduced affinity for ATP. These results suggest a possible role for splice variants of P2X receptors in the formation of functional heteromeric ion channels.
Electronic imaging clinical implementation strategies and principles need to be developed as we move toward replacement of film-based radiology practices. During an 8-month period (1998 to 1999), an Electronic Imaging Clinical Implementation Work Group (EICIWG) was formed from sections of our department: Informatics Lab, Finance Committee, Management Section, Regional Practice Group, as well as several organ and image modality sections of the Department of Diagnostic Radiology. This group was formed to study and implement policies and strategies regarding implementation of electronic imaging into our practice. The following clinical practice issues were identified as key focus areas: (1) optimal electronic worklist organization; (2) how and when to link images with reports; (3) how to redistribute technical and professional relative value units (RVU); (4) how to facilitate future practice changes within our department regarding physical location and work redistribution; and (5) how to integrate off-campus imaging into on-campus workflow. The EICIWG divided their efforts into two phases. Phase I consisted of Fact finding and review of current practice patterns and current economic models, as well as radiology consulting needs. Phase II involved the development of recommendations, policies, and strategies for reengineering the radiology department to maintain current practice goals and use electronic imaging to improve practice patterns. The EICIWG concluded that electronic images should only be released with a formal report, except in emergent situations. Electronic worklists should support and maintain the physical presence of radiologists in critical areas and direct imaging to targeted subspecialists when possible. Case tools should be developed and used in radiology and hospital information systems (RIS/HIS) to monitor a number of parameters, including professional and technical RVU data. As communication standards improve, proper staffing models must be developed to facilitate electronic on-campus and off-campus consultation.
Although it is intuitively valuable that more expedient delivery of radiographic images and reports to clinicians would improve patient care, it is important to document these outcomes to validate further advances in these areas. We evaluated the care of 215 patients seen at a walk-in clinic to determine what benefit digital imaging is to the patient. Cohorts consisted of all patients for whom specified radiology examinations were ordered during a 7-day period. The first cohort was recruited when analog films were used. The second cohort received examinations performed with computed radiography (CR) acquisition and computer display, which had been in use for 2 years. Patients were categorized as to the type of study they received, as well as whether a staff radiologist was immediately available to read the study. Clinical behavior was characterized by outcome measures of time to final diagnosis, time to final treatment, and need for follow-up. Our analysis demonstrated a reduction in time to final diagnosis that was better appreciated during the times when a staff radiologist was not immediately available. It also suggested that greater time reductions were seen for patients who received extremity examinations than those who received chest, sinus, or rib films. These data suggest that digital imaging is a useful tool to improve clinical outcome of patients seen in the acute care setting.
We prospectively compared image and report delivery times in our Urgent Care Center (UCC) during a film-based practice (1995) and after complete implementation of an electronic imaging practice in 1997. Before switching to a totally electronic and filmless practice, multiple time periods were consistently measured during a 1-week period in May 1995 and then again in a similar week in May 1997 after implementation of electronic imaging. All practice patterns were the same except for a film-based practice in 1995 versus a filmless practice in 1997. The following times were measured: (1) waiting room time, (2) technologist's time of examination, (3) time to quality control, (4) radiology interpretation times, (5) radiology image and report delivery time, (6) total radiology turn-around time, (7) time to room the patient back in the UCC, and (8) time until the ordering physician views the film. Waiting room time was longer in 1997 (average time, 26:47) versus 1995 (average time, 15:54). The technologist's examination completion time was approximately the same (1995 average time, 06:12; 1997 average time, 05:41). There was also a slight increase in the time of the technologist's electronic verification or quality control in 1997 (average time, 7:17) versus the film-based practice in 1995 (average time, 2:35). However, radiology interpretation times dramatically improved (average time, 49:38 in 1995 versus average time 13:50 in 1997). There was also a decrease in image delivery times to the clinicians in 1997 (median, 53 minutes) versus the film based practice of 1995 (1 hour and 40 minutes). Reports were available with the images immediately upon completion by the radiologist in 1997, compared with a median time of 27 minutes in 1995. Importantly, patients were roomed back into the UCC examination rooms faster after the radiologic procedure in 1997 (average time, 13:36) than they were in 1995 (29:38). Finally, the ordering physicians viewed the diagnostic images and reports in dramatically less time in 1997 (median, 26 minutes) versus 1995 (median, 1 hour and 5 minutes). In conclusion, a filmless electronic imaging practice within our UCC greatly improved radiology image and report delivery times, as well as improved clinical efficiency.
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The modulatory effects of extracellular H+ and Zn2+ were tested against ATP-responses at rat P2X4 (rP2X4) receptors expressed in Xenopus oocytes under voltage-clamp conditions. ATP (0.1-100 microM, at pH 7.5), evoked inward currents via rP2X4 receptors (EC50 value, 4.1+/-0.98 microM; nH, 1.2+/-0.1). ATP potency was reduced 2 fold, at pH 6.5, without altering maximal activity. ATP potency was reduced by a further 4 fold, at pH 5.5, and the maximal activity of ATP was also reduced. Alkaline conditions (pH 8.0) had no effect on ATP-responses. Zn2+ (100 nM - 10 microM) potentiated ATP-responses at the rP2X4 receptor by 2 fold, whereas higher concentrations (30 microM - 1 mM) inhibited ATP-responses. Zn2+ potentiation was due to an increase in ATP potency, whereas its inhibitory action was due to a reduction in ATP efficacy. Zn2+ modulation of ATP-responses was pH-dependent. At pH 6.5, the bell-shaped curve for Zn2+ was shifted to the right by 1 log unit. At pH 5.5, Zn2+ potentiation was abolished and its inhibitory effect reduced considerably. Suramin (50 microM) also potentiated ATP-responses at rP2X4 receptors. Neither H+ (pH 6.5 and 5.5), Zn2+ (10-100 microM) or a combination of both failed to reveal an inhibitory action of suramin at rP2X4 receptors. In conclusion, H+ and Zn2+ exerted opposite effects on the rP2X4 receptor by lowering and raising agonist potency, respectively. H+ (> or = 3 microM) and Zn2+ (> or = 30 microM) also reduces agonist efficacy by lowering the number of rP2X4 receptors available for activation. The striking differences between the modulatory actions of H+ and Zn2+ at rP2X4 and rP2X2 receptors are discussed.
1. Whole-cell patch-clamp recording was used to determine the functional expression and pharmacological properties of P2X receptors in chromaffin cells dissociated from adrenal medullae of rats and guinea-pigs. 2. In rat chromaffin cells maintained in culture for 1 - 7 days, ATP and UTP failed to evoke any detectable response. 3. Guinea-pig chromaffin cells responded to ATP (100 microM) with a rapidly activating inward current. The amplitude of the response to ATP increased over the period cells were maintained in culture and so did the number of cells giving a detectable response, with 69% of cells responding after >/=4 days of culture. 4. The response to ATP desensitized slowly, and had a reversal potential of 2.5 mV. The EC50 for ATP was 43 microM. The potency order for ATP analogues was 2-MeSATP>ATP>ADP. Adenosine, UTP and alpha,beta-meATP were inactive. 5. Suramin (100 microM) and Cibacron blue (50 microM) inhibited the ATP (100 microM)-activated current by 51 and 47%, respectively. PPADS antagonized the response to ATP (100 microM) with an IC50 of 3.2 microM. 6. The ATP concentration-response curve shifted to the left at pH 6.8 (EC50, 19 microM) and right at pH 8.0 (EC50, 96 microM), without changing the maximal response. Zn2+ inhibited the response to ATP (100 microM) with an IC50 of 48 microM. 7. This study indicates that expression of ATP-gated cation channels in chromaffin cells is species dependent. The P2X receptors in guinea-pig chromaffin cells show many characteristics of the P2X2 receptor subtype.
1 The modulatory activity of extracellular H+ and Zn2+ was examined on ATP-responses at rat P2X1 (rP2X1) and rat P2X3 (rP2X3) receptors expressed in Xenopus oocytes and studied under voltage-clamp conditions. 2 Superfused ATP (0.03-30 microM, at pH 7.5) evoked inward currents at rP2X1 receptors (EC50 value, 300+/-7 nM). ATP potency was reduced 2 fold at pH 6.5, and 6 fold at pH 5.5, without altering the maximum ATP effect. Alkaline conditions (pH 8.0) did not alter ATP activity. 3 Superfused ATP (0.01 - 300 microM, at pH 7. 5) evoked inward currents at rP2X3 receptors (EC50 value, 1.8+/-0.3 microM). ATP activity was affected only at pH 5.5, reducing agonist potency 15 fold without altering the maximum ATP effect. 4 Extracellular Zn2+ inhibited ATP-responses at rP2X1 receptors in a time-dependent manner, a 20 min pre-incubation being optimal (IC50 value, 1.0+/-0.2 microM). However, the Zn2+ effect was pH-independent, suggesting Zn2+- and H+-inhibition of ATP-responses occur through independent processes. 5 Extracellular Zn2+ weakly potentiated ATP-responses at rP2X3 receptors (EC50 value, 11+/-1 microM). The Zn2+ effect was dependent on pre-incubation time and, with 20 min pre-incubation periods, Zn2+ potentiated then inhibited ATP-responses in a concentration-dependent, but pH-independent, manner. 6 In summary, ATP activity at rP2X1 receptors was decreased by both extracellular H+ and Zn2+ and their effects were additive. ATP activity at rP2X3 receptors was less sensitive to H+-inhibition and, in contrast, was potentiated by Zn2+ in a pH-independent manner. These differential effects may help distinguish P2X1 and P2X3 receptors in whole tissues.
1. The antagonist activity of a series of diinosine polyphosphates (IpnI, where n=3, 4, 5) was assessed against ATP-activated inward currents at rat P2X(1-4) receptors expressed in Xenopus oocytes and studied under voltage-clamp conditions. 2. Diinosine polyphosphates were prepared by the enzymatic degradation of their corresponding diadenosine polyphosphates (e.g., Ap5A into Ip5I) using 5'-adenylic deaminase, and purified using reverse-phase chromatography. 3. Against ATP-responses at rP2X1 receptors, the potency order for antagonism was (pIC50): Ip5I (8.5)>Ip4I (6.3)>Ip3I (>4.5). Ip5I (10-100 nM) caused a concentration-dependent rightwards displacement of the ATP concentration-response curve without reducing the maximum ATP effect. However, the Schild plot was non-linear which indicated Ip5I is not a competitive antagonist. Blockade by micromolar concentrations of Ip5I was not surmountable. Ip4I also behaved as a non-surmountable antagonist. 4. Against ATP-responses at rP2X3 receptors, the potency order for antagonism was (pIC50): Ip4I (6. 0)>Ip5I (5.6)>Ip3I (>4.5). Blockade by Ip4I (pA2, 6.75) and Ip5I (pA2, 6.27) was surmountable at micromolar concentrations. 5. Diinosine polyphosphates failed to inhibit ATP-responses at rP2X2 receptors, whereas agonist responses at rP2X4 were reversibly potentiated by Ip4I and Ip5I. None of the parent diadenosine polyphosphates behave as antagonists at rP2X1 - 4 receptors. 6. Thus, Ip5I acted as a potent and relatively-selective antagonist at the rP2X1 receptor. This dinucleotide pentaphosphate represents a high-affinity antagonist for the P2X1 receptor, at which it acts in a competitive manner at low (</=100 nM) concentrations but has more complex actions at higher (>100 nM) concentrations.
Ultrasonography (US) is the modality of choice for prenatal screening, but occasionally additional imaging information is needed. Magnetic resonance (MR) imaging is an attractive alternative but until recently has been limited by motion artifact. Single-shot fast spin-echo MR imaging was used to depict normal and abnormal anatomy in 26 fetuses. Thirteen studies were performed for maternal indications and 13 were performed to evaluate fetal abnormalities identified or suspected at US. Three of the fetal abnormalities involved the central nervous system (CNS) and 10 involved other anatomic sites. Results were correlated with findings at postnatal clinical examination, imaging, and pathologic analysis. MR imaging demonstrated normal fetal anatomy without substantial motion artifact. CNS structures were well visualized as early as 18-20 weeks gestation, as were most other normal anatomic structures except the heart. MR imaging also allowed characterization of a variety of abnormalities of the CNS (Arnold-Chiari malformation, Walker-Warburg syndrome, amniotic band syndrome) as well as of other structures (renal agenesis, multicystic dysplastic kidney, abdominal masses, severe limb-body wall defect, clubfoot with arthrogryposis, diaphragmatic hernia). US findings were confirmed in most cases, and additional information about the precise diagnosis or the severity or location of the anomaly often helped guide clinical management. Single-shot fast spin-echo MR imaging of the fetus is a useful adjunct to US in difficult diagnostic situations.
PURPOSE: Previous studies have suggested that central zone prostatic volume may be more strongly correlated with lower urinary tract symptom severity and peak urinary flow rates than total prostatic volume. We determine whether prostatic central zone volume and central zone index volume correlate better with these measures than total prostate volume in an age stratified, community based random sample of healthy white men. MATERIALS AND METHODS: A cohort of 474 men were randomly selected from the 2,115 community dwelling men, 40 to 79 years old, who participated in the Olmsted County study of urinary symptoms and health status among men. All men had undergone transrectal ultrasound of the prostate. The total prostate and hypoechoic central zone volumes were caliper measured by 1 operator on static ultrasounds from baseline. Volumes were calculated with the prolate ellipsoid formula. The operator was blinded to clinical information and outcome. The associations between total prostate volume and central zone index (central zone volume/total volume), and American Urological Association (AUA) symptom index and peak urinary flow rates, respectively, were quantified with the Spearman rank correlation coefficient and least squares regression models. RESULTS: There was a moderately strong correlation between patient age and central zone volume (rs 0.54, p <0.001), total prostate volume (rs 0.45, p <0.001) and central zone index (rs 0.38, p <0.001). The AUA symptom index and peak flow rates correlated less strongly with central zone volume (rs 0.17, p = 0.001 and rs -0.20, p <0.001, respectively) and total volume (rs 0.16, p <0.001 and rs -0.16, p <0.001, respectively). Central zone index weakly correlated with AUA symptom index (rs 0.08, p = 0.103) and peak urinary flow rate (rs -0.08, p = 0.0823). In regression models predicting AUA symptom index and peak flow rates central zone volume added little information after accounting for age and total prostatic volume in predicting AUA symptom index (p = 0.55) and peak flow rate (p = 0.84). CONCLUSIONS: Central zone volume measured from static images optimized for total prostate volume no more closely correlated with lower urinary tract symptom severity or peak urinary flow rates than total prostate volume. Thus, the potentially greater imprecision in measuring central zone volume may not be offset by gains in strength of association with lower urinary tract symptom severity or peak urinary flow rates.
1. There is increasing evidence that extracellular ATP acts directly on bone cells via P2 receptors. In normal rat osteoclasts, ATP activates both non-selective cation channels and Ca2+-dependent K+ channels. In this study we investigated the action of ATP on the formation of osteoclasts and on the ultimate function of these cells, namely resorption pit formation. 2. We found that ATP stimulated resorption pit formation up to 5.6-fold when osteoclast-containing bone cell populations from neonatal rats were cultured for 26 h on ivory discs, with a maximum effect occurring at relatively low concentrations (0.2-2 microM). The stimulatory effect of ATP was amplified greatly when osteoclasts were activated by culture in acidified media (pH 6.9-7.0). Pit formation by acid-activated osteoclasts in the absence of ATP was inhibited by apyrase, an ecto-ATPase and by suramin, an antagonist of P2 receptors. 3. Over the same concentration range at which rat osteoclast activation occurred (0.2-2 microM), ATP also enhanced osteoclast formation in 10 day mouse marrow cultures, by up to 3.3-fold, with corresponding increases in resorption pit formation. Higher concentrations of ATP (20-200 microM) reduced or blocked osteoclast formation. Adenosine, a P1 receptor agonist, was without effect on either osteoclast activation or formation. 4. These results suggest that low levels of extracellular ATP may play a fundamental role in modulating both the resorptive function and formation of mammalian osteoclasts.
Analogues of the P2 receptor antagonists pyridoxal-5'-phosphate and the 6-azophenyl-2',4'-disulfonate derivative (PPADS), in which the phosphate group was cyclized by esterification to a CH2OH group at the 4-position, were synthesized. The cyclic pyridoxine-alpha4, 5-monophosphate, compound 2 (MRS 2219), was found to be a selective potentiator of ATP-evoked responses at rat P2X1 receptors with an EC50 value of 5.9 +/- 1.8 microM, while the corresponding 6-azophenyl-2',5'-disulfonate derivative, compound 3 (MRS 2220), was a selective antagonist. The potency of compound 3 at the recombinant P2X1 receptor (IC50 10.2 +/- 2.6 microM) was lower than PPADS (IC50 98.5 +/- 5.5 nM) or iso-PPADS (IC50 42.5 +/- 17.5 nM), although unlike PPADS its effect was reversible with washout and surmountable. Compound 3 showed weak antagonistic activity at the rat P2X3 receptor (IC50 58.3 +/- 0.1 microM), while at recombinant rat P2X2 and P2X4 receptors no enhancing or antagonistic properties were evident. Compounds 2 and 3 were found to be inactive as either agonists or antagonists at the phospholipase C-coupled P2Y1 receptor of turkey erythrocytes, at recombinant human P2Y2 and P2Y4 receptors, and at recombinant rat P2Y6 receptors. Similarly, compounds 2 and 3 did not have measurable affinity at adenosine A1, A2A, or A3 receptors. The lack of an aldehyde group in these derivatives indicates that Schiff's base formation with the P2X1 receptor is not necessarily required for recognition of pyridoxal phosphate derivatives. Thus, compounds 2 and 3 are relatively selective pharmacological probes of P2X1 receptors, filling a long-standing need in the P2 receptor field, and are also important lead compounds for future studies.
We have investigated the binding and internalization of alpha2-macroglobulin and serum albumin by human placental syncytiotrophoblast cells in vitro. The time course (obtained at 4 degrees C) of alpha2-macroglobulin binding indicated that an equilibrium was reached after 4 h. The binding of 125I-labelled alpha2-macroglobulin to syncytiotrophoblast cells was competitively reduced in the presence of excess unlabelled alpha2-macroglobulin. When the concentration-dependence of binding was examined over a wide concentration range, non-linear regression analysis yielded a Kd of 6.4 nM. In the case of albumin, binding was weak and ligand dissociated from the cell surface during aqueous washing making it impractical to analyze the binding reaction. In other experiments, syncytiotrophoblast cells were incubated with 125I-labelled alpha2-macroglobulin at 37 degrees C. Under these conditions, trypsin-resistant cell-associated radioactivity increased with time consistent with ligand internalization. 125I-labelled-ligand was internalized with a t1/2 of about 5 min. After a lag period some radioactivity was released back into the incubation medium. When measured at times up to 210 min, this was found to consist of mostly TCA-precipitable material that had been lost from the cell surface. However, when the incubation was extended to 24 h, almost 15% of the initial cell-associated radioactivity was released to the extracellular medium as TCA-soluble material, consistent with a slow rate of ligand degradation. The specific binding of 65Zn-labelled alpha2M was similar to that of the 125I-labelled ligand and trypsin-resistance measurements provided evidence of alpha2M-mediated 65Zn uptake. These results support a role for syncytiotrophoblast in the metabolism of alpha2-macroglobulin during pregnancy and are also consistent with a role for alpha2-macroglobulin in the maternal-fetal transport of zinc.
PURPOSE: We examined and defined anatomical structures relevant to radical prostatectomy using magnetic resonance imaging. MATERIALS AND METHODS: Before radical prostatectomy, 15 men underwent high-resolution magnetic resonance imaging studies of their pelvic floors (fast spin echo, T2 weighting of 3- to 4-mm. contiguous or overlapping slices) in axial, coronal, and sagittal planes. RESULTS: Pubovesical ligaments, rather than the commonly reported puboprostatic ligaments, were observed attaching the bladder-prostate unit to the pubis. We suggest that the part of the urethra that extends from the apex of the prostate to the bulb of the penis, which is surrounded by the striated sphincter, should be termed the sphincteric urethra rather than the membranous urethra. Further, we found no evidence that supports the traditional concept of a urogenital diaphragm. The lower part of the striated urethral sphincter was flanked on its sides by the anterior recesses of the ischioanal fossae. The portion of the levator ani, which we have termed the puboanalis sling, flanked the apex of the prostate. The most anteromedial portion of this sling inserts into the perineal body and should be termed the puboperinealis. The terminal part of the gastrointestinal tract (the part continued beyond the levator ani) should be termed the anal canal, not the rectum, as used frequently in the urologic literature. Therefore, the initial plane of dissection in radical perineal prostatectomy passes along the anterior portion of the anal canal, not the rectum. CONCLUSION: We used magnetic resonance imaging to study male pelvic floor and perineal anatomy without the artifact of dissection. This study allowed us to devise a more precise nomenclature with respect to radical prostatectomy and, in so doing, to provide a better understanding of both the retropubic and the perineal operations.